| United States Patent Application |
20050071901
|
| Kind Code
|
A1
|
|
Sebastian, Scott
|
March 31, 2005
|
Soybean plants having superior agronomic performance and methods for their
production
Abstract
This invention provides compositions including favorable alleles of marker
loci associated with genetic elements contributing to superior agronomic
performance. Also provided are markers for identifying favorable alleles
of marker loci associated with genetic elements involved in superior
agronomic performance, as well as methods employing the markers.
| Inventors: |
Sebastian, Scott; (Polk City, IA)
|
| Correspondence Address:
|
LAW OFFICES OF JONATHAN ALAN QUINE
P.O. BOX 458
ALAMEDA
CA
94501
US
|
| Assignee: |
PIONEER HI-BRED INTERNATIONAL, INC.
Des Moines
IA
|
| Family ID:
|
34119823
|
| Appl. No.:
|
10/901425
|
| Filed:
|
July 27, 2004 |
Related U.S. Patent Documents
| | | | |
|
| Application Number | Filing Date | Patent Number | |
|---|
| | 60492074 | Aug 1, 2003 | | |
| | 60547811 | Feb 25, 2004 | | |
| | 60582241 | Jun 22, 2004 | | |
|
|
| Current U.S. Class: |
800/312 ; 435/6.12; 536/24.3; 800/267 |
| Current CPC Class: |
A01H 5/10 20130101; C12Q 1/6895 20130101; C12Q 2600/156 20130101; A01H 1/04 20130101; C12Q 2600/13 20130101; A01H 1/02 20130101 |
| Class at Publication: |
800/312 ; 435/006; 800/267; 536/024.3 |
| International Class: |
A01H 001/00; C12N 015/11; A01H 005/00; C12Q 001/68 |
Claims
What is claimed is:
1. A method of identifying a soybean subline with increased yield, the
method comprising: determining in a genome of each of a plurality of
progeny of a progenitor soybean, at least one allelic form of a plurality
of chromosome segments each comprising a genetic element contributing to
increased yield and each comprising or proximal to a marker locus
selected from the set of marker loci consisting of: set of loci
consisting of: Satt684, Satt165, Satt042, Satt364, Satt300, Satt591,
Satt155, Satt385, Satt385, Satt225, Satt236, Satt511, P12390B-1, Satt480,
Satt632-TB, Satt233, Satt327, Satt329, Satt508, P10635A-1, Satt409,
Satt228, Satt429, Satt426, Satt509, SAT.sub.--261, Satt197, Satt519,
Satt597, SCT.sub.--026, Satt415, Satt583, Satt430, P12198A-1, P8584A-1,
Satt359, P10648A-1, P12105A-1, P10641A-1, Satt168, Satt556, Satt272,
Satt020, Satt066, Satt534, P10638B-2, Satt399, Satt361, P10639A-1,
Satt661-TB, Satt190, SAT.sub.--311-DB, Satt338, Satt227, Satt640-TB,
Satt422, Satt457, Satt457, Satt557, Satt319, SAT.sub.--142-DB, Satt460,
P13073A-1, Satt307, SCT.sub.--028, Satt433, Satt357, Satt321, Satt267,
Satt383, Satt295, Satt203, Satt507, SAT.sub.--110, P10620A-1, Satt129,
Satt147, Satt216, SAT.sub.--351, P10621B-2, Satt701, Satt634, Satt558,
Satt266, Satt282, Satt537, Satt506, Satt546, P13072A-1, Satt582, Satt389,
Satt461, Satt311, Satt514, Satt464, Satt662, Satt543, Satt186, Satt413,
Satt672, P13074A-1, P10624A-1, Satt573, Satt598, Satt204, Satt263,
Satt491, Satt602, Satt151, Satt355, Satt452, SAT.sub.--273-DB, Satt146,
Satt193, Satt569, Satt343, Satt586, Satt040, Satt423, Satt348, Satt595,
P10782A-1, P3436A-1, P10598A-1, Satt334, Satt510, Satt510, Satt144,
Satt522, Satt522, P9026A-1, P10646A-1, P5219A-1, P7659A-2, Satt570,
Satt356, Satt130, Satt115, Satt594, Satt533, Satt303, Satt352, Satt566,
Satt199, Satt503, Satt517, Satt191, SAT.sub.--117, Satt353, Satt442,
Satt279, Satt314, Satt142, Satt181, Satt367, Satt127, SCTT012, Satt270,
Satt292, Satt440, P10640A-1, Satt249, SAG1223, SAC1699, SCT.sub.--065,
Satt596, Satt280, Satt406, Satt380, Satt183, Satt529, Satt431, Satt242,
Satt102, Satt441, Satt544, Satt617, Satt240, P10618A-1, Satt475, Satt196,
SAT.sub.--301, Satt523, Satt418, Satt418, Satt398, Satt497, Satt284,
Satt166, Satt448, Satt373, Satt513, P12394A-1, Satt590, Satt567, Satt220,
SAG1048, Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt250,
Satt346, Satt336, SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2,
Satt584, SAT.sub.--084, P3050A-2, SAT.sub.--275-DB, Satt387, Satt549,
Satt660, Satt339, Satt255, Satt257, Satt358, P12396A-1, Satt487, Satt259,
Satt259, Satt347, Satt420, Satt576, Satt550, Satt633, Satt262, Satt473,
Satt477, Satt581, P11070A-1, Satt153, Satt243, P8230A-1, P10623A-1,
P10632A-1, P10793A-1, P12391A-1, P13560A-1, P13561A-1, P13561A-1,
P2481A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB S60149-TB,
S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB, S60361-TB,
S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB, S60519-TB,
S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB, S60812-TB,
SAC1677, SAC1724, SAG1055, Satt040, Satt108, Satt109, Satt111, Satt176,
Satt176, Satt219, Satt299, and Satt512; thereby identifying a subline
with increased yield.
2. The method of claim 1, comprising determining the allelic form of
between about 10% and about 100% of the chromosome segments comprising or
proximal to markers from the set: Satt684, Satt165, Satt042, Satt364,
Satt300, Satt591, Satt155, Satt385, Satt385, Satt225, Satt236, Satt511,
P12390B-1, Satt480, Satt632-TB, Satt233, Satt327, Satt329, Satt508,
P10635A-1, Satt409, Satt228, Satt429, Satt426, Satt509, SAT.sub.--261,
Satt197, Satt519, Satt597, SCT.sub.--026, Satt415, Satt583, Satt430,
P12198A-1, P8584A-1, Satt359, P10648A-1, P12105A-1, P10641A-1, Satt168,
Satt556, Satt272, Satt020, Satt066, Satt534, P10638B-2, Satt399, Satt361,
P10639A-1, Satt661-TB, Satt190, SAT.sub.--311-DB, Satt338, Satt227,
Satt640-TB, Satt422, Satt457, Satt457, Satt557, Satt319,
SAT.sub.--142-DB, Satt460, P13073A-1, Satt307, SCT.sub.--028, Satt433,
Satt357, Satt321, Satt267, Satt383, Satt295, Satt203, Satt507,
SAT.sub.--110, P10620A-1, Satt129, Satt147, Satt216, SAT.sub.--351,
P10621B-2, Satt701, Satt634, Satt558, Satt266, Satt282, Satt537, Satt506,
Satt546, P13072A-1, Satt582, Satt389, Satt461, Satt311, Satt514, Satt464,
Satt662, Satt543, Satt186, Satt413, Satt672, P13074A-1, P10624A-1,
Satt573, Satt598, Satt204, Satt263, Satt491, Satt602, Satt151, Satt355,
Satt452, SAT.sub.--273-DB, Satt146, Satt193, Satt569, Satt343, Satt586,
Satt040, Satt423, Satt348, Satt595, P10782A-1, P3436A-1, P10598A-1,
Satt334, Satt510, Satt510, Satt144, Satt522, Satt522, P9026A-1,
P10646A-1, P5219A-1, P7659A-2, Satt570, Satt356, Satt130, Satt115,
Satt594, Satt533, Satt303, Satt352, Satt566, Satt199, Satt503, Satt517,
Satt191, SAT.sub.--117, Satt353, Satt442, Satt279, Satt314, Satt142,
Satt181, Satt367, Satt127, SCTT012, Satt270, Satt292, Satt440, P10640A-1,
Satt249, SAG1223, SAC1699, SCT.sub.--065, Satt596, Satt280, Satt406,
Satt380, Satt183, Satt529, Satt431, Satt242, Satt102, Satt441, Satt544,
Satt617, Satt240, P10618A-1, Satt475, Satt196, SAT.sub.--301, Satt523,
Satt418, Satt418, Satt398, Satt497, Satt284, Satt166, Satt448, Satt373,
Satt513, P12394A-1, Satt590, Satt567, Satt220, SAG1048, Satt536, Satt175,
Satt677, Satt680, P10615A-1, Satt551, Satt250, Satt346, Satt336,
SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2, Satt584, SAT.sub.--084,
P3050A-2, SAT.sub.--275-DB, Satt387, Satt549, Satt660, Satt339, Satt255,
Satt257, Satt358, P12396A-1, Satt487, Satt259, Satt259, Satt347, Satt420,
Satt576, Satt550, Satt633, Satt262, Satt473, Satt477, Satt581, P11070A-1,
Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1, P10793A-1, P12391A-1,
P13560A-1, P13561A-1, P13561A-1, P2481A-1, S60021-TB, S60048-TB,
S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB,
S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB,
S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,
S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt040,
Satt108, Satt109, Satt111, Satt176, Satt176, Satt219, Satt299, and
Satt512.
3. The method of claim 1, comprising determining the allelic form of each
of a majority of chromosome segments comprising or proximal to markers
from the set: Satt684, Satt165, Satt042, Satt364, Satt300, Satt591,
Satt155, Satt385, Satt385, Satt225, Satt236, Satt511, P12390B-1, Satt480,
Satt632-TB, Satt233, Satt327, Satt329, Satt508, P10635A-1, Satt409,
Satt228, Satt429, Satt426, Satt509, SAT.sub.--261, Satt197, Satt519,
Satt597, SCT.sub.--026, Satt415, Satt583, Satt430, P12198A-1, P8584A-1,
Satt359, P10648A-1, P12105A-1, P10641A-1, Satt168, Satt556, Satt272,
Satt020, Satt066, Satt534, P10638B-2, Satt399, Satt361, P10639A-1,
Satt661-TB, Satt190, SAT.sub.--311-DB, Satt338, Satt227, Satt640-TB,
Satt422, Satt457, Satt457, Satt557, Satt319, SAT.sub.--142-DB, Satt460,
P13073A-1, Satt307, SCT.sub.--028, Satt433, Satt357, Satt321, Satt267,
Satt383, Satt295, Satt203, Satt507, SAT.sub.--110, P10620A-1, Satt129,
Satt147, Satt216, SAT.sub.--351, P10621B-2, Satt701, Satt634, Satt558,
Satt266, Satt282, Satt537, Satt506, Satt546, P13072A-1, Satt582, Satt389,
Satt461, Satt311, Satt514, Satt464, Satt662, Satt543, Satt186, Satt413,
Satt672, P13074A-1, P10624A-1, Satt573, Satt598, Satt204, Satt263,
Satt491, Satt602, Satt151, Satt355, Satt452, SAT.sub.--273-DB, Satt146,
Satt193, Satt569, Satt343, Satt586, Satt040, Satt423, Satt348, Satt595,
P10782A-1, P3436A-1, P10598A-1, Satt334, Satt510, Satt510, Satt144,
Satt522, Satt522, P9026A-1, P10646A-1, P5219A-1, P7659A-2, Satt570,
Satt356, Satt130, Satt115, Satt594, Satt533, Satt303, Satt352, Satt566,
Satt199, Satt503, Satt517, Satt191, SAT.sub.--117, Satt353, Satt442,
Satt279, Satt314, Satt142, Satt181, Satt367, Satt127, SCTT012, Satt270,
Satt292, Satt440, P10640A-1, Satt249, SAG1223, SAC1699, SCT.sub.--065,
Satt596, Satt280, Satt406, Satt380, Satt183, Satt529, Satt431, Satt242,
Satt102, Satt441, Satt544, Satt617, Satt240, P10618A-1, Satt475, Satt196,
SAT.sub.--301, Satt523, Satt418, Satt418, Satt398, Satt497, Satt284,
Satt166, Satt448, Satt373, Satt513, P12394A-1, Satt590, Satt567, Satt220,
SAG1048, Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt250,
Satt346, Satt336, SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2,
Satt584, SAT.sub.--084, P3050A-2, SAT.sub.--275-DB, Satt387, Satt549,
Satt660, Satt339, Satt255, Satt257, Satt358, P12396A-1, Satt487, Satt259,
Satt259, Satt347, Satt420, Satt576, Satt550, Satt633, Satt262, Satt473,
Satt477, Satt581, P11070A-1, Satt153, Satt243, P8230A-1, P10623A-1,
P10632A-1, P10793A-1, P12391A-1, P13560A-1, P13561A-1, P13561A-1,
P2481A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB, S60149-TB,
S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB, S60361-TB,
S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB, S60519-TB,
S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB, S60812-TB,
SAC1677, SAC1724, SAG1055, Satt040, Satt108, Satt109, Satt111, Satt176,
Satt176, Satt219, Satt299, and Satt512.
4. The method of claim 1, comprising determining the allelic form of the
chromosome segments comprising or proximal to the set of markers
consisting essentially of: Satt684, Satt165, Satt042, Satt364, Satt300,
Satt591, Satt155, Satt385, Satt385, Satt225, Satt236, Satt511, P12390B-1,
Satt480, Satt632-TB, Satt233, Satt327, Satt329, Satt508, P10635A-1,
Satt409, Satt228, Satt429, Satt426, Satt509, SAT.sub.--261, Satt197,
Satt519, Satt597, SCT.sub.--026, Satt415, Satt583, Satt430, P12198A-1,
P8584A-1, Satt359, P10648A-1, P12105A-1, P10641A-1, Satt168, Satt556,
Satt272, Satt020, Satt066, Satt534, P10638B-2, Satt399, Satt361,
P10639A-1, Satt661-TB, Satt190, SAT.sub.--311-DB, Satt338, Satt227,
Satt640-TB, Satt422, Satt457, Satt457, Satt557, Satt319,
SAT.sub.--142-DB, Satt460, P13073A-1, Satt307, SCT.sub.--028, Satt433,
Satt357, Satt321, Satt267, Satt383, Satt295, Satt203, Satt507,
SAT.sub.--110, P10620A-1, Satt129, Satt147, Satt216, SAT.sub.--351,
P10621B-2, Satt701, Satt634, Satt558, Satt266, Satt282, Satt537, Satt506,
Satt546, P13072A-1, Satt582, Satt389, Satt461, Satt311, Satt514, Satt464,
Satt662, Satt543, Satt186, Satt413, Satt672, P13074A-1, P10624A-1,
Satt573, Satt598, Satt204, Satt263, Satt491, Satt602, Satt151, Satt355,
Satt452, SAT.sub.--273-DB, Satt146, Satt193, Satt569, Satt343, Satt586,
Satt040, Satt423, Satt348, Satt595, P10782A-1, P3436A-1, P10598A-1,
Satt334, Satt510, Satt510, Satt144, Satt522, Satt522, P9026A-1,
P10646A-1, P5219A-1, P7659A-2, Satt570, Satt356, Satt130, Satt115,
Satt594, Satt533, Satt303, Satt352, Satt566, Satt199, Satt503, Satt517,
Satt191, SAT.sub.--117, Satt353, Satt442, Satt279, Satt314, Satt142,
Satt181, Satt367, Satt127, SCTT012, Satt270, Satt292, Satt440, P10640A-1,
Satt249, SAG1223, SAC1699, SCT.sub.--065, Satt596, Satt280, Satt406,
Satt380, Satt183, Satt529, Satt431, Satt242, Satt102, Satt441, Satt544,
Satt617, Satt240, P10618A-1, Satt475, Satt196, SAT.sub.--301, Satt523,
Satt418, Satt418, Satt398, Satt497, Satt284, Satt166, Satt448, Satt373,
Satt513, P12394A-1, Satt590, Satt567, Satt220, SAG1048, Satt536, Satt175,
Satt677, Satt680, P10615A-1, Satt551, Satt250, Satt346, Satt336,
SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2, Satt584, SAT.sub.--084,
P3050A-2, SAT.sub.--275-DB, Satt387, Satt549, Satt660, Satt339, Satt255,
Satt257, Satt358, P12396A-1, Satt487, Satt259, Satt259, Satt347, Satt420,
Satt576, Satt550, Satt633, Satt262, Satt473, Satt477, Satt581, P11070A-1,
Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1, P10793A-1, P12391A-1,
P13560A-1, P13561A-1, P13561A-1, P2481A-1, S60021-TB, S60048-TB,
S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB,
S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB,
S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,
S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt040,
Satt108, Satt109, Satt111, Satt176, Satt176, Satt219, Satt299, and
Satt512.
5. The method of claim 1, comprising determining the favorable allelic
form of at least one chromosome segment, wherein the favorable allelic
form of at least one chromosome segment is determined by: a) identifying
at least one polymorphic marker locus selected from the set in a
plurality of sublines of progeny of a progenitor soybean; b) assessing
yield in at least two sublines of progeny, which sublines of progeny have
different allelic forms of at least one marker locus; and, c) identifying
the subline of progeny with increased yield relative to the progenitor
soybean.
6. A method of identifying a soybean subline with increased yield, the
method comprising: determining in a genome of a progeny of a progenitor
soybean, at least one allele of one or more marker loci segregating in a
plurality of progeny of the progenitor soybean, each of which segregating
marker loci comprises a first allele and a second allele, which first
allele correlates with increased yield and which second allele does not
correlate with increased yield relative to the mean yield of the
plurality of progeny of the progenitor soybean, wherein the one or more
marker loci are selected from the group consisting of: Satt684, Satt165,
Satt042, Satt364, Satt300, Satt591, Satt155, Satt385, Satt385, Satt225,
Satt236, Satt511, P12390B-1, Satt480, Satt632-TB, Satt233, Satt327,
Satt329, Satt508, P10635A-1, Satt409, Satt228, Satt429, Satt426, Satt509,
SAT.sub.--261, Satt197, Satt519, Satt597, SCT.sub.--026, Satt415,
Satt583, Satt430, P12198A-1, P8584A-1, Satt359, P10648A-1, P12105A-1,
P10641A-1, Satt168, Satt556, Satt272, Satt020, Satt066, Satt534,
P10638B-2, Satt399, Satt361, P10639A-1, Satt661-TB, Satt190,
SAT.sub.--311-DB, Satt338, Satt227, Satt640-TB, Satt422, Satt457,
Satt457, Satt557, Satt319, SAT.sub.--142-DB, Satt460, P13073A-1, Satt307,
SCT.sub.--028, Satt433, Satt357, Satt321, Satt267, Satt383, Satt295,
Satt203, Satt507, SAT.sub.--110, P10620A-1, Satt129, Satt147, Satt216,
SAT.sub.--351, P10621B-2, Satt701, Satt634, Satt558, Satt266, Satt282,
Satt537, Satt506, Satt546, P13072A-1, Satt582, Satt389, Satt461, Satt311,
Satt514, Satt464, Satt662, Satt543, Satt186, Satt413, Satt672, P13074A-1,
P10624A-1, Satt573, Satt598, Satt204, Satt263, Satt491, Satt602, Satt151,
Satt355, Satt452, SAT.sub.--273-DB, Satt146, Satt193, Satt569, Satt343,
Satt586, Satt040, Satt423, Satt348, Satt595, P10782A-1, P3436A-1,
P10598A-1, Satt334, Satt510, Satt510, Satt144, Satt522, Satt522,
P9026A-1, P10646A-1, P5219A-1, P7659A-2, Satt570, Satt356, Satt130,
Satt115, Satt594, Satt533, Satt303, Satt352, Satt566, Satt199, Satt503,
Satt517, Satt191, SAT.sub.--117, Satt353, Satt442, Satt279, Satt314,
Satt142, Satt181, Satt367, Satt127, SCTT012, Satt270, Satt292, Satt440,
P10640A-1, Satt249, SAG1223, SAC1699, SCT.sub.--065, Satt596, Satt280,
Satt406, Satt380, Satt183, Satt529, Satt431, Satt242, Satt102, Satt441,
Satt544, Satt617, Satt240, P10618A-1, Satt475, Satt196, SAT.sub.--301,
Satt523, Satt418, Satt418, Satt398, Satt497, Satt284, Satt166, Satt448,
Satt373, Satt513, P12394A-1, Satt590, Satt567, Satt220, SAG1048, Satt536,
Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt250, Satt346, Satt336,
SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2, Satt584, SAT.sub.--084,
P3050A-2, SAT.sub.--275-DB, Satt387, Satt549, Satt660, Satt339, Satt255,
Satt257, Satt358, P12396A-1, Satt487, Satt259, Satt259, Satt347, Satt420,
Satt576, Satt550, Satt633, Satt262, Satt473, Satt477, Satt581, P11070A-1,
Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1, P10793A-1, P12391A-1,
P13560A-1, P13561A-1, P13561A-1, P2481A-1, S60021-TB, S60048-TB,
S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB,
S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB,
S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,
S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt040,
Satt108, Satt109, Satt111, Satt176, Satt176, Satt219, Satt299, and
Satt512.
7. The method of claim 6, wherein the first allele that correlates with
increased yield and the second allele that does not correlate with
increased yield are determined by: a) identifying at least one
polymorphic marker locus selected from the group, which polymorphic
marker locus comprises at least two alleles segregating in a plurality of
sublines of progeny of a progenitor soybean; b) assessing yield in at
least two sublines of progeny, which sublines of progeny have different
alleles of the at least one polymorphic marker locus; and, c) identifying
the subline of progeny with increased yield relative to the mean yield of
the plurality of sublines.
8. The method of claim 1 or 6, wherein the at least two sublines of
progeny are near isogenic sublines.
9. The method of claim 1 or 6, wherein the at least two sublines of
progeny are random sublines.
10. The method of claim 1 or 6, wherein the plurality of progeny are
obtained by selfing a progenitor soybean.
11. The method of claim 1 or 6, wherein the plurality of progeny are
obtained by crossing a first progenitor soybean and a second progenitor
soybean.
12. The method of claim 11, wherein a first progenitor soybean comprising
a strain of elite germplasm is crossed with a second progenitor soybean
comprising a different strain of elite germplasm to generate a population
of soybean plants from which the progeny is selected.
13. The method of claim 11, wherein a first progenitor soybean comprising
a strain of elite germplasm is crossed with a second progenitor soybean
comprising a strain of exotic germplasm to generate a population of
soybean plants from which the progeny is selected.
14. The method of claim 12 or 13, wherein the elite strain of germplasms
are selected from the group consisting of: 90A07, 90B11, 90B31, 90B43,
90B72, 90B73, 91B01, 91B12, 91B33, 91B52, 91B53, 91B64, 91B91, 91B92,
92B05, 92B12, 92B23, 92B38, 92B52, 92B63, 92B74, 92B75, 92B84, 92B95,
92M30, 92M31, 92M70, 92M71, 92M72, 92M80, 92M91, 93B01, 93B09, 93B11,
93B15, 93B25, 93B26, 93B36, 93B41, 93B45, 93B46, 93B66, 93B67, 93B68,
93B72, 93B82, 93B84, 93B85, 93B86, 93B87, 93M10, 93M30, 93M40, 93M50,
93M60, 93M80, 93M90, 93M92, 93M93, 94B01, 94B23, 94B24, 94B53, 94B54,
94B73, 95B32, 95B33, 95B34, 95B53, 95B95, 95B96, 95B97, 96B21, 96B51,
97B52, 97B61, A1395, A2722, A2835, A2943, A3127, A3237, A3242, A3322,
A3431, A4009, A4138, A4415, A4595, A4715, A5403, A5560, A5843, A5885,
A5979, A5980, A6297, BEDFORD, CM428, CX105, CX232, CX253, CX289, CX394C,
CX469C, D00566D362, ESSEX, EX04C00, EX06A00, EX10F01, EX13P01, EX13Q01,
EX15N01, EX16N00, EX16P01, EX22Y01, EX22Z01, EX23B03, EX34T03, EX35F03,
EX36Y01, EX39E00, EX40T03, EX44V03, FORREST, G3362, HS93-4118, HUTCHESON,
JIM, KORADA, MO15733, MO400644-02, MO413735-11-52, MO501577-27-23,
MO505469-61-89, MP39009, P1677, P9007, P9008, P9041, P9042, P9061, P9062,
P9063, P9071, P9092, P9132, P9141, P9151, P9163, P9182, P9203, P9233,
P9244, P9273, P9281, P9035, P9306, P9321, P9341, P9392, P9395, P9481,
P9482, P9492, P9521, P9552, P9561, P9584, P9591, P9592, P9594, P9631,
P9641, PHARAOH, RA451, R01154R002, S0066, S03W4, S0880, S1550, S1990,
S19T9, S20F8, S22C3, S24L2, S25J5, S32Z3, S33N1, S38T8, S3911, S4260,
S42H1, S43B5, S5960, S6189, S6262, ST0653, ST1073, ST1090, ST1570,
ST1690, ST1970, ST2250, ST2488, ST2660, ST2686, ST2688, ST2788, ST2870,
ST3171, ST3380, ST3630, ST3660, ST3870, ST3883, TRACY, TRAILL, X9916,
YB03E00, XB03F01, XB07E01, XB10D01, XB15M01, XB19U04, XB20M01, XB22C04,
XB22R01, XB23W03, XB23Y02, XB25E02, XB25L04, XB25X04, XB25W01, XB26L04,
XB27P04, XB29A04, XB29D01, XB29K04, XB29L04, XB30E04, XB31C01, XB31R04,
XB33B, XB34D04, XB34F01, XB35D, XB35L04, XB35W00, XB38A01, XB41M01,
XB42J00, XB42M01, XB48H01, XB54K01, XB55J01, XB58P99, XB63D00, XB67A00,
YB03G01, YB08D01, YB09F01, YB09G01, YB10E01, YB11D01, YB14H01, YB15K99,
YB21F01, YB21G01, YB22S00, YB22V01, YB22W01, YB22X01, YB24Z01, YB25R03,
YB25R99, YB25X00, YB25Y01, YB25Z01, YB27L03, YB27S00, YB27X01, YB27Y01,
YB28A03, YB28N01, YB29H01, YB29J01, YB29T04, YB30J01, YB30N01, YB30P01,
YB31E01, YB32K01, YB33K01, YB34H01, YB34R03, YB34S03, YB35C01, YB36E03,
YB36V00, YB38E03, YB38G03, YB39M01, YB39V03, YB40M01, YB40N01, YB41Q01,
YB48L01, YB52J00, YB53E00, YB54H00, YB54J00, YB54L00, YB55H00, YB56E00,
YB60N01, and YOUNG.
15. The method of claim 1 or 6, the set of marker loci comprising about
10% and about 100% of the set of markers consisting of: Satt642, Satt042,
Satt364, Satt454, Satt526, Satt300, Satt591, Satt155, Satt385, Satt511,
P12390B-1, Satt632-TB, Satt429, SAT.sub.--261, Satt197, P10641A-1,
Satt556, Satt534, P10638B-2, Satt399. Satt361, P10639A-1, Satt661-TB,
Satt190, SAT.sub.--311-DB, Satt338, Satt640-TB, Satt557, Satt319,
SAT.sub.--142-DB, Satt460, Satt433, Satt357, Satt321, Satt295, Satt203,
Satt507, Satt129, Satt147, SAT.sub.--351, P10621B-2, Satt558, Satt701,
Satt634, Satt582, Satt389, Satt464, Satt662, Satt672, Satt573, Satt598,
Satt263, Satt602, Satt151, SAT.sub.--273-DB, Satt146, Satt193, Satt569,
Satt176, Satt343, Satt586, Satt040, Satt595, P10782A-1, Satt334, Satt144,
Satt522, Satt570, Satt356, Satt533, Satt199, Satt517, Satt191,
SAT.sub.--117, Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223,
SAC1699, SAT.sub.--065, Satt596, Satt406, Satt380, Satt183, Satt529,
Satt242, Satt617, Satt240, SAT.sub.--301, Satt418, Satt398, Satt497,
Satt166, Satt448, Satt373, Satt513, P12394A-1, SAG1048, Satt536, Satt175,
Satt677, Satt680, P10615A-1, Satt551, Satt346, Satt336, SAT.sub.--330-DB,
P13069A-1, P5467A-1, P5467A-2, SAT.sub.--084, SAT.sub.--275-DB, Satt660,
Satt339, P12396A-1, Satt358, Satt487, Satt259, Satt420, Satt576, Satt633,
Satt477, Satt581, Satt153, Satt243, P10793A-1, P12391A-1, P12392A-1,
P13560A-1, P13561A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB,
S60149-TB, S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB,
S60361-TB, S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB,
S60519-TB, S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB,
S60812-TB, SAC1677, SAC1724, SAG1055, Satt040, Satt111, Satt176, Satt219
and Satt299.
16. The method of claim 1 or 6, the set of marker loci comprising between
about 10% and about 100% of the set of markers consisting of: Satt684,
Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155, Satt385,
Satt632-TB, Satt429, SAT-.sub.--261, P10641A-1, Satt556, P10638B-2,
Satt399, Satt361, Satt661-TB, Satt190, SAT.sub.--311-DB, Satt338,
Satt640-TB, Satt557, Satt319, SAT.sub.--142-DB, Satt321, Satt203,
Satt129, Satt147, SAT351, P10621B-2, Satt701, Satt634, Satt582, Satt389,
Satt464, Satt662, Satt672, Satt573, Satt598, Satt263, Satt151,
SAT.sub.--273-DB, Satt146, Satt193, Satt569, Satt343, Satt586, Satt040,
Satt595, Satt334, Satt144, Satt522, Satt570, Satt356, Satt199, Satt517,
Satt191, Sat.sub.--117, Satt279, Satt181, Satt127, Satt270, Satt292,
SAG1223, SAC1699, Sat.sub.--065, Satt596, Satt406, Satt380, Satt183,
Satt529, Satt242, Satt617, Satt240, SAT.sub.--301, Satt418, Satt398,
Satt497, Satt166, Satt448, Satt373, Satt513, P12394A-1, SAG1048, Satt536,
Satt175, Satt677, Satt680, P10615A-1, Satt551, SAT.sub.--330-DB, Satt358,
Satt487, Satt487, Satt420, Satt576, Satt633, Satt581, Satt153, Satt243,
P10793A-1, P13560A-1, P13561A-1, S60021-TB, S60048-TB, S60076-TB,
S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB, S60338-TB,
S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB, S60505-TB,
S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB, S60630-TB,
S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt111, Satt219 and
Satt299.
17. The method of claim 1 or 6, the set of marker loci comprising between
about 10% and about 100% of the set of markers consisting of: Satt684,
Satt526, Satt591, Satt385, Satt632-TB, Satt429, SAT-.sub.--261,
P10641A-1, Satt556, P10638B-2, Satt190, SAT.sub.--311-DB, Satt338,
Satt640-TB, Satt557, SAT.sub.--142-DB, Satt321, Satt203, Satt129,
SAT.sub.--351, Satt701, Satt582, Satt389, Satt464, Satt672, Satt598,
Satt343, Satt595, Satt334, Satt144, Satt522, Satt570, Satt356, Satt199,
Sat.sub.--117, Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223,
Sat.sub.--065, Satt529, Satt242, Satt617, SAT.sub.--301, Satt398,
Satt497, Satt166, Satt373, SAG1048, Satt680, P10615A-1, SAT.sub.--330-DB,
P13069A-1, SAT.sub.--275-DB, Satt339, Satt487, Satt420, Satt581 and
Satt153.
18. The method of claim 1 or 6, the set of marker loci consisting
essentially of: Satt684, Satt526, Satt591, Satt385, Satt632-TB, Satt429,
SAT-.sub.--261, P10641A-1, Satt556, P10638B-2, Satt190, SAT.sub.--311-DB,
Satt338, Satt640-TB, Satt557, SAT.sub.--142-DB, Satt321, Satt203,
Satt129, SAT.sub.--351, Satt701, Satt582, Satt389, Satt464, Satt672,
Satt598, S Satt595, Satt334, Satt144, Satt522, Satt570, Satt356, Satt199,
Sat.sub.--117, Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223,
Sat.sub.b 065, Satt529, Satt242, Satt617, SAT.sub.--301, Satt398,
Satt497, Satt166, Satt373, SAG1048, Satt680, P10615A-1, SAT.sub.--330-DB,
P13069A-1, SAT.sub.--275-DB, Satt339, Satt487, Satt420, Satt581 and
Satt153.
19. The method of claim 1 or 6, the set of marker loci comprising between
about 10% and about 100% of the set of markers consisting of: Satt526,
Satt591, Satt429, SAT-.sub.--261, P10641A-1, Satt190, Satt557,
SAT.sub.--142-DB, Satt129, SAT.sub.--351, Satt464, Satt343, Satt595,
Satt570, Satt181, Satt127, Satt270, SAG1223, Sat.sub.--065, Satt529,
Satt242, Satt398, Satt166, Satt373, SAG1048, Satt680, SAT.sub.--330-DB,
P13069A-1, Satt339, Satt487, Satt581 and Satt153.
20. The method of claim 1 or 6, the set of marker loci consisting
essentially of: Satt526, Satt591, Satt429, SAT-.sub.--261, P10641A-1,
Satt190, Satt557, SAT.sub.--142-DB, Satt129, SAT.sub.--351, Satt464,
Satt343, Satt595, Satt570, Satt181, Satt127, Satt270, SAG1223,
Sat.sub.--065, Satt529, Satt242, Satt398, Satt166, Satt373, SAG1048,
Satt680, SAT.sub.--330-DB, P13069A-1, Satt339, Satt487, Satt581 and
Satt153.
21. The method of claim 1 or 6, further comprising electronically
transmitting or electronically storing data representing the determined
allelic forms in a computer readable medium.
22. The method of claim 1 or 6, further comprising selecting at least one
plant of the identified soybean subline.
23. The method of claim 22, wherein the selected soybean plant comprises a
whole plant, a plant organ, a plant seed, a plant cell or a plant tissue
culture.
24. The method of claim 22, wherein the selected soybean plant or a
progeny thereof is crossed with a second soybean plant, which second
soybean plant lacks the determined alleles of marker loci or the
determined allelic forms of the plurality of chromosome segments.
25. The method of claim 24, the second soybean plant comprising an elite
strain of germplasm.
26. The method of claim 24, the second soybean plant comprising exotic
germplasm.
27. The method of claim 1 or 6, wherein the allelic form of each of a
plurality of marker loci are determined in a first soybean plant genome
and at least a second soybean plant genome, wherein the first soybean
plant comprises a parent soybean plant and the at least second soybean
plant comprises at least one progeny of the first soybean plant.
28. A soybean subline with increased yield produced by the method of claim
1 or 6.
29. A plant organ, a plant seed, a plant cell or a plant tissue culture
from a plant of the soybean subline of claim 28.
30. A set of markers useful for identifying a soybean plant with increased
yield, the set of markers comprising between about 10% and about 100% of
the set of marker loci consisting of: Satt684, Satt165, Satt042, Satt364,
Satt454, Satt526, Satt300, Satt591, Satt155, Satt385, Satt385, Satt225,
Satt236, Satt511, P12390B-1, Satt480, Satt632-TB, Satt233, Satt327,
Satt329, Satt508, P10635A-1, Satt409, Satt228, Satt429, Satt426, Satt509,
SAT.sub.--261, Satt197, Satt519, Satt597, SCT.sub.--026, Satt415,
Satt583, Satt430, P12198A-1, P8584A-1, Satt359, P10648A-1, P12105A-1,
P10641A-1, Satt168, Satt556, Satt272, Satt020, Satt066, Satt534,
P10638B-2, Satt399, Satt361, P10639A-1, Satt661-TB, Satt190,
SAT.sub.--311-DB, Satt338, Satt227, Satt640-TB, Satt422, Satt457,
Satt457, Satt557, Satt319, SAT.sub.--142-DB, Satt460, P13073A-1, Satt307,
SCT.sub.--028, Satt433, Satt357, Satt321, Satt267, Satt383, Satt295,
Satt203, Satt507, SAT.sub.--110, P10620A-1, Satt129, Satt147, Satt216,
SAT.sub.--351, P10621B-2, Satt701, Satt634, Satt558, Satt266, Satt282,
Satt537, Satt506, Satt546, P13072A-1, Satt582, Satt389, Satt461, Satt311,
Satt514, Satt464, Satt662, Satt543, Satt186, Satt413, Satt672, P13074A-1,
P10624A-1, Satt573, Satt598, Satt204, Satt263, Satt491, Satt602, Satt151,
Satt355, Satt452, SAT.sub.--273-DB, Satt146, Satt193, Satt569, Satt343,
Satt586, Satt040, Satt423, Satt348, Satt595, P10782A-1, P3436A-1,
P10598A-1, Satt334, Satt510, Satt510, Satt144, Satt522, Satt522,
P9026A-1, P10646A-1, P5219A-1, P7659A-2, Satt570, Satt356, Satt130,
Satt115, Satt594, Satt533, Satt303, Satt352, Satt566, Satt199, Satt503,
Satt517, Satt191, SAT.sub.--117, Satt353, Satt442, Satt279, Satt314,
Satt142, Satt181, Satt367, Satt127, SCTT012, Satt270, Satt292, Satt440,
P10640A-1, Satt249, SAG1223, SAC1699, SCT.sub.--065, Satt596, Satt280,
Satt406, Satt380, Satt183, Satt529, Satt431, Satt242, Satt102, Satt441,
Satt544, Satt617, Satt240, P10618A-1, Satt475, Satt196, SAT.sub.--301,
Satt523, Satt418, Satt418, Satt398, Satt497, Satt284, Satt166, Satt448,
Satt373, Satt513, P12394A-1, Satt590, Satt567, Satt220, SAG1048, Satt536,
Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt250, Satt346, Satt336,
SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2, Satt584, SAT.sub.--084,
P3050A-2, SAT.sub.--275-DB, Satt387, Satt549, Satt660, Satt339, Satt255,
Satt257, Satt358, P12396A-1, Satt487, Satt259, Satt259, Satt347, Satt420,
Satt576, Satt550, Satt633, Satt262, Satt473, Satt477, Satt581, P11070A-1,
Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1, P10793A-1, P12391A-1,
P13560A-1, P13561A-1, P13561A-1, P2481A-1, S60021-TB, S60048-TB,
S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB,
S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB,
S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,
S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt040,
Satt108, Satt109, Satt111, Satt176, Satt176, Satt219, Satt299, and
Satt512.
31. A set of markers useful for identifying a soybean plant with increased
yield, the set of markers comprising a majority of the markers: Satt684,
Satt165, Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155,
Satt385, Satt385, Satt225, Satt236, Satt511, P12390B-1, Satt480,
Satt632-TB, Satt233, Satt327, Satt329, Satt508, P10635A-1, Satt409,
Satt228, Satt429, Satt426, Satt509, SAT.sub.--261, Satt197, Satt519,
Satt597, SCT.sub.--026, Satt415, Satt583, Satt430, P12198A-1, P8584A-1,
Satt359, P10648A-1, P12105A-1, P10641A-1, Satt168, Satt556, Satt272,
Satt020, Satt066, Satt534, P10638B-2, Satt399, Satt361, P10639A-1,
Satt661-TB, Satt190, SAT.sub.--311-DB, Satt338, Satt227, Satt640-TB,
Satt422, Satt457, Satt457, Satt557, Satt319, SAT.sub.--142-DB, Satt460,
P13073A-1, Satt307, SCT.sub.--028, Satt433, Satt357, Satt321, Satt267,
Satt383, Satt295, Satt203, Satt507, SAT.sub.--110, P10620A-1, Satt129,
Satt147, Satt216, SAT.sub.--351, P10621B-2, Satt701, Satt634, Satt558,
Satt266, Satt282, Satt537, Satt506, Satt546, P13072A-1, Satt582, Satt389,
Satt461, Satt311, Satt514, Satt464, Satt662, Satt543, Satt186, Satt413,
Satt672, P13074A-1, P10624A-1, Satt573, Satt598, Satt204, Satt263,
Satt491, Satt602, Satt151, Satt355, Satt452, SAT.sub.--273-DB, Satt146,
Satt193, Satt569, Satt343, Satt586, Satt040, Satt423, Satt348, Satt595,
P10782A-1, P3436A-1, P10598A-1, Satt334, Satt510, Satt510, Satt144,
Satt522, Satt522, P9026A-1, P10646A-1, P5219A-1, P7659A-2, Satt570,
Satt356, Satt130, Satt115, Satt594, Satt533, Satt303, Satt352, Satt566,
Satt199, Satt503, Satt517, Satt191, SAT.sub.--117, Satt353, Satt442,
Satt279, Satt314, Satt142, Satt181, Satt367, Satt127, SCTT012, Satt270,
Satt292, Satt440, P10640A-1, Satt249, SAG1223, SAC1699, SCT.sub.--065,
Satt596, Satt280, Satt406, Satt380, Satt183, Satt529, Satt431, Satt242,
Satt102, Satt441, Satt544, Satt617, Satt240, P10618A-1, Satt475, Satt196,
SAT.sub.--301, Satt523, Satt418, Satt418, Satt398, Satt497, Satt284,
Satt166, Satt448, Satt373, Satt513, P12394A-1, Satt590, Satt567, Satt220,
SAG1048, Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt250,
Satt346, Satt336, SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2,
Satt584, SAT.sub.--084, P3050A-2, SAT.sub.--275-DB, Satt387, Satt549,
Satt660, Satt339, Satt255, Satt257, Satt358, P12396A-1, Satt487, Satt259,
Satt259, Satt347, Satt420, Satt576, Satt550, Satt633, Satt262, Satt473,
Satt477, Satt581, P11070A-1, Satt153, Satt243, P8230A-1, P10623A-1,
P10632A-1, P10793A-1, P12391A-1, P13560A-1, P13561A-1, P13561A-1,
P2481A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB, S60149-TB,
S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB, S60361-TB,
S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB, S60519-TB,
S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB, S60812-TB,
SAC1677, SAC1724, SAG1055, Satt040, Satt108, Satt109, Satt111, Satt176,
Satt176, Satt219, Satt299, and Satt512.
32. A set of markers useful for identifying a soybean plant with increased
yield, the set of markers consisting essentially of: Satt684, Satt165,
Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155, Satt385,
Satt385, Satt225, Satt236, Satt511, P12390B-1, Satt480, Satt632-TB,
Satt233, Satt327, Satt329, Satt508, P10635A-1, Satt409, Satt228, Satt429,
Satt426, Satt509, SAT.sub.--261, Satt197, Satt519, Satt597,
SCT.sub.--026, Satt415, Satt583, Satt430, P12198A-1, P8584A-1, Satt359,
P10648A-1, P12105A-1, P10641A-1, Satt168, Satt556, Satt272, Satt020,
Satt066, Satt534, P10638B-2, Satt399, Satt361, P10639A-1, Satt661-TB,
Satt190, SAT.sub.--311-DB, Satt338, Satt227, Satt640-TB, Satt422,
Satt457, Satt457, Satt557, Satt319, SAT.sub.--142-DB, Satt460, P13073A-1,
Satt307, SCT.sub.--028, Satt433, Satt357, Satt321, Satt267, Satt383,
Satt295, Satt203, Satt507, SAT.sub.--110, P10620A-1, Satt129, Satt147,
Satt216, SAT.sub.--351, P10621B-2, Satt701, Satt634, Satt558, Satt266,
Satt282, Satt537, Satt506, Satt546, P13072A-1, Satt582, Satt389, Satt461,
Satt311, Satt514, Satt464, Satt662, Satt543, Satt186, Satt413, Satt672,
P13074A-1, P10624A-1, Satt573, Satt598, Satt204, Satt263, Satt491,
Satt602, Satt151, Satt355, Satt452, SAT.sub.--273-DB, Satt146, Satt193,
Satt569, Satt343, Satt586, Satt040, Satt423, Satt348, Satt595, P10782A-1,
P3436A-1, P10598A-1, Satt334, Satt510, Satt510, Satt144, Satt522,
Satt522, P9026A-1, P10646A-1, P5219A-1, P7659A-2, Satt570, Satt356,
Satt130, Satt115, Satt594, Satt533, Satt303, Satt352, Satt566, Satt199,
Satt503, Satt517, Satt191, SAT.sub.--117, Satt353, Satt442, Satt279,
Satt314, Satt142, Satt181, Satt367, Satt127, SCTT012, Satt270, Satt292,
Satt440, P10640A-1, Satt249, SAG1223, SAC1699, SCT.sub.--065, Satt596,
Satt280, Satt406, Satt380, Satt183, Satt529, Satt431, Satt242, Satt102,
Satt441, Satt544, Satt617, Satt240, P10618A-1, Satt475, Satt196,
SAT.sub.--301, Satt523, Satt418, Satt418, Satt398, Satt497, Satt284,
Satt166, Satt448, Satt373, Satt513, P12394A-1, Satt590, Satt567, Satt220,
SAG1048, Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt250,
Satt346, Satt336, SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2,
Satt584, SAT.sub.--084, P3050A-2, SAT.sub.--275-DB, Satt387, Satt549,
Satt660, Satt339, Satt255, Satt257, Satt358, P12396A-1, Satt487, Satt259,
Satt259, Satt347, Satt420, Satt576, Satt550, Satt633, Satt262, Satt473,
Satt477, Satt581, P11070A-1, Satt153, Satt243, P8230A-1, P10623A-1,
P10632A-1, P10793A-1, P12391A-1, P13560A-1, P13561A-1, P13561A-1,
P2481A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB, S60149-TB,
S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB, S60361-TB,
S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB, S60519-TB,
S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB, S60812-TB,
SAC1677, SAC1724, SAG1055, Satt040, Satt108, Satt109, Satt111, Satt176,
Satt176, Satt219, Satt299, and Satt512.
33. A set of markers useful for identifying a soybean plant with increased
yield, the set of markers comprising between about 10% and about 100% of
the set of marker loci consisting of: Satt642, Satt042, Satt364, Satt454,
Satt526, Satt300, Satt591, Satt155, Satt385, Satt511, P12390B-1,
Satt632-TB, Satt429, SAT.sub.--261, Satt197, P10641A-1, Satt556, Satt534,
P10638B-2, Satt399. Satt361, P10639A-1, Satt661-TB, Satt190,
SAT.sub.--311-DB, Satt338, Satt640-TB, Satt557, Satt319,
SAT.sub.--142-DB, Satt460, Satt433, Satt357, Satt321, Satt295, Satt203,
Satt507, Satt129, Satt147, SAT.sub.--351, P10621B-2, Satt558, Satt701,
Satt634, Satt582, Satt389, Satt464, Satt662, Satt672, Satt573, Satt598,
Satt263, Satt602, Satt151, SAT.sub.--273-DB, Satt146, Satt193, Satt569,
Satt176, Satt343, Satt586, Satt040, Satt595, P10782A-1, Satt334, Satt144,
Satt522, Satt570, Satt356, Satt533, Satt199, Satt517, Satt191,
SAT.sub.--117, Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223,
SAC1699, SAT.sub.--065, Satt596, Satt406, Satt380, Satt183, Satt529,
Satt242, Satt617, Satt240, SAT.sub.--301, Satt418, Satt398, Satt497,
Satt166, Satt448, Satt373, Satt513, P12394A-1, SAG1048, Satt536, Satt175,
Satt677, Satt680, P10615A-1, Satt551, Satt346, Satt336, SAT.sub.--330-DB,
P13069A-1, P5467A-1, P5467A-2, SAT.sub.--084, SAT.sub.--275-DB, Satt660,
Satt339, P12396A-1, Satt358, Satt487, Satt259, Satt420, Satt576, Satt633,
Satt477, Satt581, Satt153, Satt243, P10793A-1, P12391A-1, P12392A-1,
P13560A-1, P13561A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB,
S60149-TB, S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB,
S60361-TB, S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB,
S60519-TB, S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB,
S60812-TB, SAC1677, SAC1724, SAG1055, Satt040, Satt111, Satt176, Satt219
and Satt299.
34. A set of markers useful for identifying a soybean plant with increased
yield, the set of markers comprising between about 10% and about 100% of
the set of marker loci consisting of: Satt684, Satt042, Satt364, Satt454,
Satt526, Satt300, Satt591, Satt155, Satt385, Satt632-TB, Satt429,
SAT-261, P10641A-1, Satt556, P10638B-2, Satt399, Satt361, Satt661-TB,
Satt190, SAT.sub.--311-DB, Satt338, Satt640-TB, Satt557, Satt319,
SAT.sub.--142-DB, Satt321, Satt203, Satt129, Satt147, SAT.sub.--351,
P10621B-2, Satt701, Satt634, Satt582, Satt389, Satt464, Satt662, Satt672,
Satt573, Satt598, Satt263, Satt151, SAT.sub.--273-DB Satt146, Satt193,
Satt569, Satt343, Satt586, Satt040, Satt595, Satt334, Satt144, Satt522,
Satt570, Satt356, Satt199, Satt157, Satt191, Sat.sub.--117, Satt279,
Satt181, Satt127, Satt270, Satt292, SAG1223, SAC1699, Sat.sub.--065,
Satt596, Satt406, Satt380, Satt183, Satt529, Satt242, Satt617, Satt240,
SAT.sub.--301, Satt418, Satt398, Satt497, Satt166, Satt448, Satt373,
Satt513, P12394A-1, SAG1048, Satt536, Satt175, Satt677, Satt680,
P10615A-1, Satt551, SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2,
SAT.sub.--084, SAT.sub.--275-DB, Satt660, Satt339, Satt358, Satt487,
Satt487, Satt420, Satt576, Satt633, Satt581, Satt 153, Satt243,
P10793A-1, P13560A-1, P13561A-1, S60021A-TB, S60048-TB, S60076-TB,
S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB, S60338-TB,
S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB, S60505-TB,
S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB, S60630-TB,
S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt111, Satt219 and
Satt299.
35. A subset of the set of markers of claim 34, the subset of marker loci
comprising between about 10% and about 100% of the set of markers
consisting of: Satt684, Satt526, Satt591, Satt385, Satt632-TB, Satt429,
SAT-.sub.--261, P10641A-1, Satt556, P10638B-2, Satt190, SAT.sub.--311-DB,
Satt338, Satt640-TB, Satt557, SAT.sub.--142-DB, Satt321, Satt203,
Satt129, SAT.sub.--351, Satt701, Satt582, Satt389, Satt464, Satt672,
Satt598, S Satt595, Satt334, Satt144, Satt522, Satt570, Satt356, Satt199,
Sat.sub.--117, Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223,
Sat.sub.b 065, Satt529, Satt242, Satt617, SAT.sub.--301, Satt398,
Satt497, Satt166, Satt373, SAG1048, Satt680, P10615A-1, SAT.sub.--330-DB,
P13069A-1, SAT.sub.--275-DB, Satt339, Satt487, Satt420, Satt581 and
Satt153.
36. A subset of the set of markers of claim 34, the subset of marker loci
consisting essentially of: Satt684, Satt526, Satt591, Satt385,
Satt632-TB, Satt429, SAT-.sub.--261, P10641A-1, Satt556, P10638B-2,
Satt190, SAT.sub.--311-DB, Satt338, Satt640-TB, Satt557,
SAT.sub.--142-DB, Satt321, Satt203, Satt129, SAT.sub.--351, Satt701,
Satt582, Satt389, Satt464, Satt672, Satt598, S Satt595, Satt334, Satt144,
Satt522, Satt570, Satt356, Satt199, Sat.sub.--117, Satt279, Satt181,
Satt127, Satt270, Satt292, SAG1223, Sat.sub.b 065, Satt529, Satt242,
Satt617, SAT.sub.--301, Satt398, Satt497, Satt166, Satt373, SAG1048,
Satt680, P10615A-1, SAT.sub.--330-DB, P13069A-1, SAT.sub.--275-DB,
Satt339, Satt487, Satt420, Satt581 and Satt153.
37. A subset of the set of markers of claim 34, the subset of marker loci
comprising between about 10% and about 100% of the set of markers
consisting of: Satt526, Satt591, Satt429, SAT-.sub.--261, P10641A-1,
Satt190, Satt557, SAT.sub.--142-DB, Satt129, SAT.sub.--351, Satt464,
Satt343, Satt595, Satt570, Satt181, Satt127, Satt270, SAG1223,
Sat.sub.--065, Satt529, Satt242, Satt398, Satt166, Satt373, SAG1048,
Satt680, SAT.sub.--330-DB, P13069A-1, Satt339, Satt487, Satt581 and
Satt153.
38. A subset of the set of markers of claim 34, the subset of marker loci
consisting essentially of: Satt526, Satt591, Satt429, SAT-.sub.--261,
P10641A-1, Satt190, Satt557, SAT.sub.--142-DB, Satt129, SAT.sub.--351,
Satt464, Satt343, Satt595, Satt570, Satt181, Satt127, Satt270, SAG1223,
Sat.sub.--065, Satt529, Satt242, Satt398, Satt166, Satt373, SAG1048,
Satt680, SAT.sub.--330-DB, P13069A-1, Satt339, Satt487, Satt581 and
Satt153.
39. A system comprising a data input device for inputting genotyping data
corresponding to soybean germplasm; a computer readable medium comprising
genotyping data corresponding to the allelic forms of a plurality of
markers selected from the set consisting of: Satt684, Satt165, Satt042,
Satt364, Satt454, Satt526, Satt300, Satt591, Satt155, Satt385, Satt385,
Satt225, Satt236, Satt511, P12390B-1, Satt480, Satt632-TB, Satt233,
Satt327, Satt329, Satt508, P10635A-1, Satt409, Satt228, Satt429, Satt426,
Satt509, SAT.sub.--261, Satt197, Satt519, Satt597, SCT.sub.--026,
Satt415, Satt583, Satt430, P12198A-1, P8584A-1, Satt359, P10648A-1,
P12105A-1, P10641A-1, Satt168, Satt556, Satt272, Satt020, Satt066,
Satt534, P10638B-2, Satt399, Satt361, P10639A-1, Satt661-TB, Satt190,
SAT.sub.--311-DB, Satt338, Satt227, Satt640-TB, Satt422, Satt457,
Satt457, Satt557, Satt319, SAT.sub.--142-DB, Satt460, P13073A-1, Satt307,
SCT.sub.--028, Satt433, Satt357, Satt321, Satt267, Satt383, Satt295,
Satt203, Satt507, SAT.sub.--110, P10620A-1, Satt129, Satt147, Satt216,
SAT.sub.--351, P10621B-2, Satt701, Satt634, Satt558, Satt266, Satt282,
Satt537, Satt506, Satt546, P13072A-1, Satt582, Satt389, Satt461, Satt311,
Satt514, Satt464, Satt662, Satt543, Satt186, Satt413, Satt672, P13074A-1,
P10624A-1, Satt573, Satt598, Satt204, Satt263, Satt491, Satt602, Satt151,
Satt355, Satt452, SAT.sub.--273-DB, Satt146, Satt193, Satt569, Satt343,
Satt586, Satt040, Satt423, Satt348, Satt595, P10782A-1, P3436A-1,
P10598A-1, Satt334, Satt510, Satt510, Satt144, Satt522, Satt522,
P9026A-1, P10646A-1, P5219A-1, P7659A-2, Satt570, Satt356, Satt130,
Satt115, Satt594, Satt533, Satt303, Satt352, Satt566, Satt199, Satt503,
Satt517, Satt191, SAT.sub.--117, Satt353, Satt442, Satt279, Satt314,
Satt142, Satt181, Satt367, Satt127, SCTT012, Satt270, Satt292, Satt440,
P10640A-1, Satt249, SAG1223, SAC1699, SCT.sub.--065, Satt596, Satt280,
Satt406, Satt380, Satt183, Satt529, Satt431, Satt242, Satt102, Satt441,
Satt544, Satt617, Satt240, P10618A-1, Satt475, Satt196, SAT.sub.--301,
Satt523, Satt418, Satt418, Satt398, Satt497, Satt284, Satt166, Satt448,
Satt373, Satt513, P12394A-1, Satt590, Satt567, Satt220, SAG1048, Satt536,
Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt250, Satt346, Satt336,
SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2, Satt584, SAT.sub.--084,
P3050A-2, SAT.sub.--275-DB, Satt387, Satt549, Satt660, Satt339, Satt255,
Satt257, Satt358, P12396A-1, Satt487, Satt259, Satt259, Satt347, Satt420,
Satt576, Satt550, Satt633, Satt262, Satt473, Satt477, Satt581, P11070A-1,
Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1, P10793A-1, P12391A-1,
P13560A-1, P13561A-1, P13561A-1, P2481A-1, S60021-TB, S60048-TB,
S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB,
S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB,
S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,
S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt040,
Satt108, Satt109, Satt111, Satt176, Satt176, Satt219, Satt299, and
Satt512. an instruction set for selecting soybean germplasm comprising a
plurality of favorable alleles allelic forms of chromosome segments
comprising or proximal to a plurality of the markers; and, a data output
device providing results of the selecting.
40. A computer readable medium comprising genotyping data corresponding to
the allelic forms of a plurality of markers selected from the set
consisting of: Satt684, Satt165, Satt042, Satt364, Satt454, Satt526,
Satt300, Satt591, Satt155, Satt385, Satt385, Satt225, Satt236, Satt511,
P12390B-1, Satt480, Satt632-TB, Satt233, Satt327, Satt329, Satt508,
P10635A-1, Satt409, Satt228, Satt429, Satt426, Satt509, SAT.sub.--261,
Satt197, Satt519, Satt597, SCT.sub.--026, Satt415, Satt583, Satt430,
P12198A-1, P8584A-1, Satt359, P10648A-1, P12105A-1, P10641A-1, Satt168,
Satt556, Satt272, Satt020, Satt066, Satt534, P10638B-2, Satt399, Satt361,
P10639A-1, Satt661-TB, Satt190, SAT.sub.--311-DB, Satt338, Satt227,
Satt640-TB, Satt422, Satt457, Satt457, Satt557, Satt319,
SAT.sub.--142-DB, Satt460, P13073A-1, Satt307, SCT.sub.--028, Satt433,
Satt357, Satt321, Satt267, Satt383, Satt295, Satt203, Satt507,
SAT.sub.--110, P10620A-1, Satt129, Satt147, Satt216, SAT.sub.--351,
P10621B-2, Satt701, Satt634, Satt558, Satt266, Satt282, Satt537, Satt506,
Satt546, P13072A-1, Satt582, Satt389, Satt461, Satt311, Satt514, Satt464,
Satt662, Satt543, Satt186, Satt413, Satt672, P13074A-1, P10624A-1,
Satt573, Satt598, Satt204, Satt263, Satt491, Satt602, Satt151, Satt355,
Satt452, SAT.sub.--273-DB, Satt146, Satt193, Satt569, Satt343, Satt586,
Satt040, Satt423, Satt348, Satt595, P10782A-1, P3436A-1, P10598A-1,
Satt334, Satt510, Satt510, Satt144, Satt522, Satt522, P9026A-1,
P10646A-1, P5219A-1, P7659A-2, Satt570, Satt356, Satt130, Satt115,
Satt594, Satt533, Satt303, Satt352, Satt566, Satt199, Satt503, Satt517,
Satt191, SAT.sub.--117, Satt353, Satt442, Satt279, Satt314, Satt142,
Satt181, Satt367, Satt127, SCTT012, Satt270, Satt292, Satt440, P10640A-1,
Satt249, SAG1223, SAC1699, SCT.sub.--065, Satt596, Satt280, Satt406,
Satt380, Satt183, Satt529, Satt431, Satt242, Satt102, Satt441, Satt544,
Satt617, Satt240, P10618A-1, Satt475, Satt196, SAT.sub.--301, Satt523,
Satt418, Satt418, Satt398, Satt497, Satt284, Satt166, Satt448, Satt373,
Satt513, P12394A-1, Satt590, Satt567, Satt220, SAG1048, Satt536, Satt175,
Satt677, Satt680, P10615A-1, Satt551, Satt250, Satt346, Satt336,
SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2, Satt584, SAT.sub.--084,
P3050A-2, SAT.sub.--275-DB, Satt387, Satt549, Satt660, Satt339, Satt255,
Satt257, Satt358, P12396A-1, Satt487, Satt259, Satt259, Satt347, Satt420,
Satt576, Satt550, Satt633, Satt262, Satt473, Satt477, Satt581, P11070A-1,
Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1, P10793A-1, P12391A-1,
P13560A-1, P13561A-1, P13561A-1, P2481A-1, S60021-TB, S60048-TB,
S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB,
S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB,
S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,
S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt040,
Satt108, Satt109, Satt111, Satt176, Satt176, Satt219, Satt299, and
Satt512; which genotyping data is obtained from one or more progeny of at
least one soybean comprising an elite germplasm selected from the group
consisting of: 90A07, 90B11, 90B31, 90B43, 90B72, 90B73, 91B01, 91B12,
91B33, 91B52, 91B53, 91B64, 91B91, 91B92, 92B05, 92B12, 92B23, 92B38,
92B52, 92B63, 92B74, 92B75, 92B84, 92B95, 92M30, 92M31, 92M70, 92M71,
92M72, 92M80, 92M91, 93B01, 93B09, 93B11, 93B15, 93B25, 93B26, 93B36,
93B41, 93B45, 93B46, 93B66, 93B67, 93B68, 93B72, 93B82, 93B84, 93B85,
93B86, 93B87, 93M10, 93M30, 93M40, 93M50, 93M60, 93M80, 93M90, 93M92,
93M93, 94B01, 94B23, 94B24, 94B53, 94B54, 94B73, 95B32, 95B33, 95B34,
95B53, 95B95, 95B96, 95B97, 96B21, 96B51, 97B52, 97B61, A1395, A2722,
A2835, A2943, A3127, A3237, A3242, A3322, A3431, A4009, A4138, A4415,
A4595, A4715, A5403, A5560, A5843, A5885, A5979, A5980, A6297, BEDFORD,
CM428, CX105, CX232, CX253, CX289, CX394C, CX469C, D00566D362, ESSEX,
EX04C00, EX06A00, EX10F01, EX13P01, EX13Q01, EX15N01, EX16N00, EX16P01,
EX22Y01, EX22Z01, EX23B03, EX34T03, EX35F03, EX36Y01, EX39E00, EX40T03,
EX44V03, FORREST, G3362, HS93-4118, HUTCHESON, JIM, KORADA, MO15733,
MO400644-02, MO413735-11-52, MO501577-27-23, MO505469-61-89, MP39009,
P1677, P9007, P9008, P9041, P9042, P9061, P9062, P9063, P9071, P9092,
P9132, P9141, P9151, P9163, P9182, P9203, P9233, P9244, P9273, P9281,
P9035, P9306, P9321, P9341, P9392, P9395, P9481, P9482, P9492, P9521,
P9552, P9561, P9584, P9591, P9592, P9594, P9631, P9641, PHARAOH, RA451,
R01154R002, S0066, S03W4, S0880, S1550, S1990, S19T9, S20F8, S22C3,
S24L2, S25J5, S32Z3, S33N1, S38T8, S3911, S4260, S42H1, S43B5, S5960,
S6189, S6262, ST0653, ST1073, ST1090, ST1570, ST1690, ST1970, ST2250,
ST2488, ST2660, ST2686, ST2688, ST2788, ST2870, ST3171, ST3380, ST3630,
ST3660, ST3870, ST3883, TRACY, TRAILL, X9916, YB03E00, XB03F01, XB07E01,
XB10D01, XB15M01, XB19U04, XB20M01, XB22C04, XB22R01, XB23W03, XB23Y02,
XB25E02, XB25L04, XB25X04, XB25W01, XB26L04, XB27P04, XB29A04, XB29D01,
XB29K04, XB29L04, XB30E04, XB31C01, XB31R04, XB33B, XB34D04, XB34F01,
XB35D, XB35L04, XB35W00, XB38A01, XB41M01, XB42J00, XB42M01, XB48H01,
XB54K01, XB55J01, XB58P99, XB63D00, XB67A00, YB03G01, YB08D01, YB09F01,
YB09G01, YB10E01, YB11D01, YB14H01, YB15K99, YB21F01, YB21G01, YB22S00,
YB22V01, YB22W01, YB22X01, YB24Z01, YB25R03, YB25R99, YB25X00, YB25Y01,
YB25Z01, YB27L03, YB27S00, YB27X01, YB27Y01, YB28A03, YB28N01, YB29H01,
YB29J01, YB29T04, YB30J01, YB30N01, YB30P01, YB31E01, YB32K01, YB33K01,
YB34H01, YB34R03, YB34S03, YB35C01, YB36E03, YB36V00, YB38E03, YB38G03,
YB39M01, YB39V03, YB40M01, YB40N01, YB41Q01, YB48L01, YB52J00, YB53E00,
YB54H00, YB54J00, YB54L00, YB55H00, YB56E00, YB60N01, and YOUNG.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to and benefit of U.S. Provisional
Application Nos. 60/492,074, filed Aug. 1, 2003; 60/547,811, filed Feb.
25, 2004; and, 60/582,241, filed Jun. 22, 2004, the disclosures of each
of which are incorporated herein for all purposes.
FIELD OF THE INVENTION
[0002] The invention relates to the field of agricultural biotechnology
and, more specifically, to molecular marker assisted selection and
breeding of soybean plants.
BACKGROUND OF THE INVENTION
[0003] One of the most challenging aspects of plant breeding is to
identify plant varieties that are superior to the currently available
varieties used in commerce. Herein, the term "variety" and "genotype"
will be used interchangeably since genetic differences are what make each
variety unique and what make one variety superior to another in terms of
commercial value.
[0004] For commodity crops like soybeans and corn, the most universal
measure of commercial value is grain productivity per unit area or
"yield". Since a farmer is paid according to the quantity (weight) of
grain he delivers to an elevator, a farmer typically wants to plant a
variety that produces the most grain per acre.
[0005] Although yield is arguably the most important trait that a plant
breeder is concerned with, it is also the least understood genetically.
There are many different plant traits that control the efficiency of
converting nutrients and light into grain. Yield is therefore the final
culmination of many different traits that contribute to productivity over
the growing season. These would include seedling emergence vigor,
photosynthetic ability, disease resistance, ability to mine nutrients
from the soil, ability to produce flowers, and ability to shuttle
photosynthate into grain, etc. The genetic bases of these individual
traits that contribute to yield are largely unknown. Each trait that
contributes to "yield" could be controlled by several or many genetic
loci. Therefore, the overall genetic basis for yield is undoubtedly very
complex. This is just one reason why traditional methods of determining
the genetic basis of yield have not been very successful. To make
incremental improvements in yield potential, for the most part, plant
breeders are still using the same resource intensive methods that have
been in use for the last 80 or more years. Existing varieties are crossed
to produce an array of new genotypes which are then exhaustively tested
over many locations and replications in order to get enough yield data to
differentiate the few consistently superior genotypes. This is one of the
most expensive and time-consuming aspects of plant breeding.
[0006] During the 1990's, genetic markers linked to genes that contribute
to yield emerged as a means to improve efficiency in certain aspects of
the breeding process. These success stories have been limited to traits
that are controlled by relatively few genes that are highly heritable. In
this case, it is fairly routine to make a reliable association between a
DNA sequence and a phenotype that can be confirmed with a greenhouse or
field assay. However, until very recently, it has been extremely
difficult to make reliable associations between specific DNA sequences
and a very complex quantitative trait such as yield.
[0007] "Breeding bias," described in U.S. Pat. No. 5,437,697, which is
incorporated herein in its entirety for all purposes, is a unique way to
determine which genetic loci have been affected by extended periods of
recurrent selection for yield. By comparing the genetic marker profiles
of modern high yielding varieties to their most distant ancestors,
breeding bias can quickly leverage an entire century of yield data to
determine which specific alleles of which genetic markers have increased
in frequency over time due to selection. Since increased yield has been
the main criteria for selection, these markers are those most likely to
be associated with yield progress over time. The present invention
provides genetic markers that are associated with yield performance in a
variety of geographic regions, as well as methods for utilizing these
markers to efficiently identify soybean lines and sublines with increased
yield.
SUMMARY OF THE INVENTION
[0008] The present invention provides representative markers that
correspond to, and identify, chromosome segments important for superior
agronomic performance in a variety of geographic regions and growing
conditions. The markers described herein are shown to be associated with
genetic elements contributing to increased yield in soybean. The markers,
and methods for their use, described herein provide the means for
defining and identifying soybean plants with improved yield relative to
existing elite lines. Using the markers and methods described herein,
identification of residual allelic variation among segregating lines of
soybeans derived from elite strains can be used to increase the
efficiency of the breeding program to develop novel sublines of soybean
with increased yield relative to existing elite strains.
[0009] In a first aspect, the invention provides methods for identifying
soybean sublines with increased yield relative to existing elite lines of
soybeans. The methods of the invention involve detecting at least one
allelic form of a plurality of chromosome segments, each of which a)
includes a genetic element contributing to increased yield; and, b)
includes or is proximal to a marker locus shown to be associated with
increased yield in soybean.
[0010] For example, detecting an allelic form involves identifying at
least one favorable allelic form of a chromosome segment, where the
identified chromosome segment includes a genetic element contributing to
increased yield and either includes or is proximal to (linked to) a
marker selected from the specified set of markers which have been shown
to be associated with yield. The favorable allelic form of a chromosome
segment can be confirmed by identifying a polymorphic marker locus
selected from the set, which is segregating in various sublines of
progeny derived from a progenitor soybean. Yield is assessed in at least
two sublines of progeny with different allelic forms of the marker locus,
and a subline of progeny with increased yield relative to the progenitor
soybean is identified.
[0011] Exemplary marker loci shown by Breeding Bias analysis to be
associated with genetic elements that contribute to increased yield in
one or more geographic growing region are included in the following set
of markers: Satt684, Satt165, Satt042, Satt364, Satt454, Satt526,
Satt300, Satt591, Satt155, Satt385, Satt385, Satt225, Satt236, Satt511,
P12390B-1, Satt480, Satt632-TB, Satt233, Satt327, Satt329, Satt508,
P10635A-1, Satt409, Satt228, Satt429, Satt426, Satt509, SAT.sub.--261,
Satt197, Satt519, Satt597, SCT.sub.--026, Satt415, Satt583, Satt430,
P12198A-1, P8584A-1, Satt359, P10648A-1, P12105A-1, P10641A-1, Satt168,
Satt556, Satt272, Satt020, Satt066, Satt534, P10638B-2, Satt399, Satt361,
P10639A-1, Satt661-TB, Satt190, SAT.sub.--311-DB, Satt338, Satt227,
Satt640-TB, Satt422, Satt457, Satt457, Satt557, Satt319,
SAT.sub.--142-DB, Satt460, P13073A-1, Satt307, SCT.sub.--028, Satt433,
Satt357, Satt321, Satt267, Satt383, Satt295, Satt203, Satt507,
SAT.sub.--110, P10620A-1, Satt129, Satt147, Satt216, SAT.sub.--351,
P10621B-2, Satt701, Satt634, Satt558, Satt266, Satt282, Satt537, Satt506,
Satt546, P13072A-1, Satt582, Satt389, Satt461, Satt311, Satt514, Satt464,
Satt662, Satt543, Satt186, Satt413, Satt672, P13074A-1, P10624A-1,
Satt573, Satt598, Satt204, Satt263, Satt491, Satt602, Satt151, Satt355,
Satt452, SAT.sub.--273-DB, Satt146, Satt193, Satt569, Satt343, Satt586,
Satt040, Satt423, Satt348, Satt595, P10782A-1, P3436A-1, P10598A-1,
Satt334, Satt510, Satt510, Satt144, Satt522, Satt522, P9026A-1,
P10646A-1, P5219A-1, P7659A-2, Satt570, Satt356, Satt130, Satt115,
Satt594, Satt533, Satt303, Satt352, Satt566, Satt199, Satt503, Satt517,
Satt191, SAT.sub.--117, Satt353, Satt442, Satt279, Satt314, Satt142,
Satt181, Satt367, Satt127, SCTT012, Satt270, Satt292, Satt440, P10640A-1,
Satt249, SAG1223, SAC1699, SCT.sub.--065, Satt596, Satt280, Satt406,
Satt380, Satt183, Satt529, Satt431, Satt242, Satt102, Satt441, Satt544,
Satt617, Satt240, P10618A-1, Satt475, Satt196, SAT.sub.--301, Satt523,
Satt418, Satt418, Satt398, Satt497, Satt284, Satt116, Satt448, Satt373,
Satt513, P12394A-1, Satt590, Satt567, Satt220, SAG1048, Satt536, Satt175,
Satt677, Satt680, P10615A-1, Satt551, Satt250, Satt346, Satt336,
SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2, Satt584, SAT.sub.--084,
P3050A-2, SAT.sub.--275-DB, Satt387, Satt549, Satt660, Satt339, Satt255,
Satt257, Satt358, P12396A-1, Satt487, Satt259, Satt259, Satt347, Satt420,
Satt576, Satt550, Satt633, Satt262, Satt473, Satt477, Satt581, P11070A-1,
Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1, P10793A-1, P12391A-1,
P13560A-1, P13561A-1, P13561A-1, P2481A-1, S60021-TB, S60048-TB,
S60076-TB, S60148-TB S60149-TB, S60201-TB, S60243-TB, S60326-TB,
S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB,
S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,
S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt040,
Satt108, Satt109, Satt111, Satt176, Satt176, Satt219, Satt299, and
Satt512. As such, marker loci of the set identify chromosome segments
contributing to increased yield. Any number of additional marker loci
linked to a marker locus selected from the set can be identified and will
function as equivalents in the methods of the invention.
[0012] For example, in some embodiments the favorable allelic form of at
least one chromosome segment, i.e., the allelic form associated with
increased yield, is determined by a) identifying at least one polymorphic
marker locus selected from the set in a plurality of sublines of progeny
of a progenitor soybean, and b) assessing yield in at least two sublines
of progeny having different allelic forms of the marker locus.
[0013] In other embodiments, the methods of identifying a soybean subline
with increased yield involve detecting at least one allele of a marker
locus segregating among progeny of a progenitor soybean. The marker locus
includes at least two alleles, one of which correlates with increased
yield whereas the other allele(s) does not correlate with increased
yield. Increase in yield is measured relative to the mean yield of the
progeny. Optionally, more than one marker locus is evaluated. The marker
loci are selected from the set of loci consisting of: Satt684, Satt165,
Satt042, Satt364, Satt300, Satt591, Satt155, Satt385, Satt385, Satt225,
Satt236, Satt511, P12390B-1, Satt480, Satt632-TB, Satt233, Satt327,
Satt329, Satt508, P10635A-1, Satt409, Satt228, Satt429, Satt426, Satt509,
SAT.sub.--261, Satt197, Satt519, Satt597, SCT.sub.--026, Satt415,
Satt583, Satt430, P12198A-1, P8584A-1, Satt359, P10648A-1, P12105A-1,
P10641A-1, Satt168, Satt556, Satt272, Satt020, Satt066, Satt534,
P10638B-2, Satt399, Satt361, P10639A-1, Satt661-TB, Satt190,
SAT.sub.--311-DB, Satt338, Satt227, Satt640-TB, Satt422, Satt457,
Satt457, Satt557, Satt319, SAT.sub.--142-DB, Satt460, P13073A-1, Satt307,
SCT.sub.--028, Satt433, Satt357, Satt321, Satt267, Satt383, Satt295,
Satt203, Satt507, SAT.sub.--110, P10620A-1, Satt129, Satt147, Satt216,
SAT.sub.--351, P10621B-2, Satt701, Satt634, Satt558, Satt266, Satt282,
Satt537, Satt506, Satt546, P13072A-1, Satt582, Satt389, Satt461, Satt311,
Satt514, Satt464, Satt662, Satt543, Satt186, Satt413, Satt672, P13074A-1,
P10624A-1, Satt573, Satt598, Satt204, Satt263, Satt491, Satt602, Satt151,
Satt355, Satt452, SAT.sub.--273-DB, Satt146, Satt193, Satt569, Satt343,
Satt586, Satt040, Satt423, Satt348, Satt595, P10782A-1, P3436A-1,
P10598A-1, Satt334, Satt510, Satt510, Satt144, Satt522, Satt522,
P9026A-1, P10646A-1, P5219A-1, P7659A-2, Satt570, Satt356, Satt130,
Satt115, Satt594, Satt533, Satt303, Satt352, Satt566, Satt199, Satt503,
Satt517, Satt191, SAT.sub.--117, Satt353, Satt442, Satt279, Satt314,
Satt142, Satt181, Satt367, Satt127, SCTT012, Satt270, Satt292, Satt440,
P10640A-1, Satt249, SAG1223, SAC1699, SCT.sub.--065, Satt596, Satt280,
Satt406, Satt380, Satt183, Satt529, Satt431, Satt242, Satt102, Satt441,
Satt544, Satt617, Satt240, P10618A-1, Satt475, Satt196, SAT.sub.--301,
Satt523, Satt418, Satt418, Satt398, Satt497, Satt284, Satt166, Satt448,
Satt373, Satt513, P12394A-1, Satt590, Satt567, Satt220, SAG1048, Satt536,
Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt250, Satt346, Satt336,
SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2, Satt584, SAT.sub.--084,
P3050A-2, SAT.sub.--275-DB, Satt387, Satt549, Satt660, Satt339, Satt255,
Satt257, Satt358, P12396A-1, Satt487, Satt259, Satt259, Satt347, Satt420,
Satt576, Satt550, Satt633, Satt262, Satt473, Satt477, Satt581, P11070A-1,
Satt153, Satt243, P8230A-1, P10623A-1, P10632A-1, P10793A-1, P12391A-1,
P13560A-1, P13561A-1, P13561A-1, P2481A-1, S60021-TB, S60048-TB,
S60076-TB, S60148-TB S60149-TB, S60201-TB, S60243-TB, S60326-TB,
S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB,
S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,
S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt040,
Satt108, Satt109, Satt111, Satt176, Satt176, Satt219, Satt299, and
Satt512.
[0014] In some embodiments, the allelic form of between about 10% and
about 100% of chromosome segments in the set, or in a specified subset of
the markers relevant in a particular geographic region, as enumerated in
Tables 3 through 12, are detected. Usually the allelic form of between
about 10% and about 90% of the chromosome segments relevant in a
particular geographic region are determined. Commonly, a majority of the
allelic forms are determined. In an embodiment, the allelic forms of
essentially all of the chromosome segments are determined.
[0015] For example, in a breeding program aimed at developing soybeans
with increased yield in the central growing region (e.g., Iowa) allelic
forms of between about 10% and about 100% of the chromosome segments
including or proximal to the markers from the set including Satt642,
Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155, Satt385,
Satt511, P12390B-1, Satt632-TB, Satt429, SAT.sub.--261, Satt197,
P10641A-1, Satt556, Satt534, P10638B-2, Satt399. Satt361, P10639A-1,
Satt661-TB, Satt190, SAT.sub.--311-DB, Satt338, Satt640-TB, Satt557,
Satt319, SAT.sub.--142-DB, Satt460, Satt433, Satt357, Satt321, Satt295,
Satt203, Satt507, Satt129, Satt147, SAT.sub.--351, P10621B-2, Satt558,
Satt701, Satt634, Satt582, Satt389, Satt464, Satt662, Satt672, Satt573,
Satt598, Satt263, Satt602, Satt151, SAT.sub.--273-DB, Satt146, Satt193,
Satt569, Satt176, Satt343, Satt586, Satt040, Satt595, P10782A-1, Satt334,
Satt144, Satt522, Satt570, Satt356, Satt533, Satt199, Satt517, Satt191,
SAT.sub.--117, Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223,
SAC1699, SAT.sub.--065, Satt596, Satt406, Satt380, Satt183, Satt529,
Satt242, Satt617, Satt240, SAT.sub.--301, Satt418, Satt398, Satt497,
Satt166, Satt448, Satt373, Satt513, P12394A-1, SAG1048, Satt536, Satt175,
Satt677, Satt680, P10615A-1, Satt551, Satt346, Satt336, SAT.sub.--330-DB,
P13069A-1, P5467A-1, P5467A-2, SAT.sub.--084, SAT.sub.--275-DB, Satt660,
Satt339, P12396A-1, Satt358, Satt487, Satt259, Satt420, Satt576, Satt633,
Satt477, Satt581, Satt153, Satt243, P10793A-1, P12391A-1, P12392A-1,
P13560A-1, P13561A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB,
S60149-TB, S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB,
S60361-TB, S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB,
S60519-TB, S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB,
S60812-TB, SAC1677, SAC1724, SAG1055, Satt040, Satt111, Satt176, Satt219
and Satt299 are determined. In an embodiment, the set of markers includes
Satt684, Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155,
Satt385, Satt632-TB, Satt429, SAT-.sub.--261, P10641A-1, Satt556,
P10638B-2, Satt399, Satt361, Satt661-TB, Satt190, SAT.sub.--311-DB,
Satt338, Satt640-TB, Satt557, Satt319, SAT.sub.--142-DB, Satt321,
Satt203, Satt129, Satt147, SAT.sub.--351, P10621B-2, Satt701, Satt634,
Satt582, Satt389, Satt464, Satt662, Satt672, Satt573, Satt598, Satt263,
Satt151, SAT.sub.--273-DB, Satt146, Satt193, Satt569, Satt343, Satt586,
Satt040, Satt595, Satt334, Satt144, Satt522, Satt570, Satt356, Satt199,
Satt517, Satt191, Sat.sub.--117, Satt279, Satt181, Satt127, Satt270,
Satt292, SAG1223, SAC1699, Sat.sub.--065, Satt596, Satt406, Satt380,
Satt183, Satt529, Satt242, Satt617, Satt240, SAT.sub.--301, Satt418,
Satt398, Satt497, Satt166, Satt448, Satt373, Satt513, P12394A-1, SAG1048,
Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551, SAT.sub.--330-DB,
P13069A-1, P5467A-1, P5467A-2, SAT.sub.--084, SAT.sub.--275-DB, Satt660,
Satt339, Satt358, Satt487, Satt487, Satt420, Satt576, Satt633, Satt581,
Satt153, Satt243, P10793A-1, P13560A-1, P13561A-1, S60021-TB, S60048-TB,
S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB,
S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB,
S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,
S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt111,
Satt219 and Satt299 are determined. In an embodiment, the set of markers
includes Satt684, Satt526, Satt591, Satt385, Satt632-TB, Satt429,
SAT-.sub.--261, P10641A-1, Satt556, P10638B-2, Satt190, SAT.sub.--311-DB,
Satt338, Satt640-TB, Satt557, SAT.sub.--142-DB, Satt321, Satt203,
Satt129, SAT.sub.--351, Satt701, Satt582, Satt389, Satt464, Satt672,
Satt598, Satt343, Satt595, Satt334, Satt144, Satt522, Satt570, Satt356,
Satt199, Sat.sub.--117, Satt279, Satt181, Satt127, Satt270, Satt292,
SAG1223, Sat.sub.--065, Satt529, Satt242, Satt617, SAT.sub.--301,
Satt398, Satt497, Satt166, Satt373, SAG1048, Satt680, P10615A-1,
SAT.sub.--330-DB, P13069A-1, SAT.sub.--275-DB, Satt339, Satt487, Satt420,
Satt581 and Satt153.
[0016] In other embodiments, the methods of identifying a soybean subline
with increased yield involve detecting at least one allele of a marker
locus segregating among progeny of a progenitor soybean. The marker locus
includes at least two alleles, one of which correlates with increased
yield whereas the other allele(s) does not correlate with increased
yield. Increase in yield is measured relative to the mean yield of the
progeny. Optionally, more than one marker locus is evaluated. The marker
loci are selected from the set of loci consisting of: Satt684, Satt165,
Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155, Satt385,
Satt385, Satt225, Satt236, Satt511, P12390B-1, Satt480, Satt632-TB,
Satt233, Satt327, Satt329, Satt508, P10635A-1, Satt409, Satt228, Satt429,
Satt426, Satt509, SAT.sub.--261, Satt197, Satt519, Satt597,
SCT.sub.--026, Satt415, Satt583, Satt430, P12198A-1, P8584A-1, Satt359,
P10648A-1, P12105A-1, P10641A-1, Satt168, Satt556, Satt272, Satt020,
Satt066, Satt534, P10638B-2, Satt399, Satt361, P10639A-1, Satt661-TB,
Satt190, SAT.sub.--311-DB, Satt338, Satt227, Satt640-TB, Satt422,
Satt457, Satt457, Satt557, Satt319, SAT.sub.--142-DB, Satt460, P13073A-1,
Satt307, SCT.sub.--028, Satt433, Satt357, Satt321, Satt267, Satt383,
Satt295, Satt203, Satt507, SAT.sub.--110, P10620A-1, Satt129, Satt147,
Satt216, SAT.sub.--351, P10621B-2, Satt701, Satt634, Satt558, Satt266,
Satt282, Satt537, Satt506, Satt546, P13072A-1, Satt582, Satt389, Satt461,
Satt311, Satt514, Satt464, Satt662, Satt543, Satt186, Satt413, Satt672,
P13074A-1, P10624A-1, Satt573, Satt598, Satt204, Satt263, Satt491,
Satt602, Satt151, Satt355, Satt452, SAT.sub.--273-DB, Satt146, Satt193,
Satt569, Satt343, Satt586, Satt040, Satt423, Satt348, Satt595, P10782A-1,
P3436A-1, P10598A-1, Satt334, Satt510, Satt510, Satt144, Satt522,
Satt522, P9026A-1, P10646A-1, P5219A-1, P7659A-2, Satt570, Satt356,
Satt130, Satt115, Satt594, Satt533, Satt303, Satt352, Satt566, Satt199,
Satt503, Satt517, Satt191, SAT.sub.--117, Satt353, Satt442, Satt279,
Satt314, Satt142, Satt181, Satt367, Satt127, SCTT012, Satt270, Satt292,
Satt440, P10640A-1, Satt249, SAG1223, SAC1699, SCT.sub.--065, Satt596,
Satt280, Satt406, Satt380, Satt183, Satt529, Satt431, Satt242, Satt102,
Satt441, Satt544, Satt617, Satt240, P10618A-1, Satt475, Satt196,
SAT.sub.--301, Satt523, Satt418, Satt418, Satt398, Satt497, Satt284,
Satt166, Satt448, Satt373, Satt513, P12394A-1, Satt590, Satt567, Satt220,
SAG1048, Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt250,
Satt346, Satt336, SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2,
Satt584, SAT.sub.--084, P3050A-2, SAT.sub.--275-DB, Satt387, Satt549,
Satt660, Satt339, Satt255, Satt257, Satt358, P12396A-1, Satt487, Satt259,
Satt259, Satt347, Satt420, Satt576, Satt550, Satt633, Satt262, Satt473,
Satt477, Satt581, P11070A-1, Satt153, Satt243, P8230A-1, P10623A-1,
P10632A-1, P10793A-1, P12391A-1, P13560A-1, P13561A-1, P13561A-1,
P2481A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB, S60149-TB,
S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB, S60361-TB,
S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB, S60519-TB,
S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB, S60812-TB,
SAC1677, SAC1724, SAG1055, Satt040, Satt108, Satt109, Satt111, Satt176,
Satt176, Satt219, Satt299, and Satt512.
[0017] In some embodiments, at least one allele of between about 10% and
about 100% of the markers in the set, or in a specified subset of the
markers relevant in a particular geographic region, as enumerated in
Tables 3 through 12, are detected. Usually at least one allele of between
about 10% and about 90% of the marker loci relevant in a particular
geographic region are determined. Commonly, a majority of the marker loci
are evaluated. In an embodiment, essentially all of the marker loci are
evaluated.
[0018] For example, in a breeding program aimed at developing soybeans
with increased yield in the central growing region (e.g., Iowa) at least
one allele of between about 10% and about 100% of the marker loci from
the set including Satt642, Satt042, Satt364, Satt454, Satt526, Satt300,
Satt591, Satt155, Satt385, Satt511, P12390B-1, Satt632-TB, Satt429,
SAT.sub.--261, Satt197, P10641A-1. Satt556, Satt534, P10638B-2, Satt399,
Satt361, P10639A-1, Satt661-TB, Satt190, SAT.sub.--311-DB, Satt338,
Satt640-TB, Satt557, Satt319, SAT.sub.--142-DB, Satt460, Satt433,
Satt357, Satt321, Satt295, Satt203, Satt507, Satt129, Satt147,
SAT.sub.--351, P10621B-2, Satt558, Satt701, Satt634, Satt582, Satt389,
Satt464, Satt662, Satt672, Satt573, Satt598, Satt263, Satt602, Satt151,
SAT.sub.--273-DB, Satt146, Satt193, Satt569, Satt176, Satt343, Satt586,
Satt040, Satt595, P10782A-1, Satt334, Satt144, Satt522, Satt570, Satt356,
Satt533, Satt199, Satt517, Satt191, SAT.sub.--117, Satt279, Satt181,
Satt127, Satt270, Satt292, SAG1223, SAC1699, SAT.sub.--065, Satt596,
Satt406, Satt380, Satt183, Satt529, Satt242, Satt617, Satt240,
SAT.sub.--301, Satt418, Satt398, Satt497, Satt166, Satt448, Satt373,
Satt513, P12394A-1, SAG1048, Satt536, Satt175, Satt677, Satt680,
P10615A-1, Satt551, Satt346, Satt336, SAT.sub.--330-DB, P13069A-1,
P5467A-1, P5467A-2, SAT.sub.--084, SAT.sub.--275-DB, Satt660, Satt339,
P12396A-1, Satt358, Satt487, Satt259, Satt420, Satt576, Satt633, Satt477,
Satt581, Satt153, Satt243, P10793A-1, P12391A-1, P12392A-1, P13560A-1,
P13561A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB, S60149-TB,
S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB, S60361-TB,
S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB, S60519-TB,
S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB, S60812-TB,
SAC1677, SAC1724, SAG1055, Satt040, Satt111, Satt176, Satt219 and Satt299
are determined. In an embodiment, the set of markers includes Satt684,
Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155, Satt385,
Satt632-TB, Satt429, SAT-.sub.--261, P10641A-1, Satt556, P10638B-2,
Satt399, Satt361, Satt661-TB, Satt190, SAT.sub.--311-DB, Satt338,
Satt640-TB, Satt557, Satt319, SAT.sub.--142-DB, Satt321, Satt203,
Satt129, Satt147, SAT.sub.--351, P10621B-2, Satt701, Satt634, Satt582,
Satt389, Satt464, Satt662, Satt672, Satt573, Satt598, Satt263, Satt151,
SAT.sub.--273-DB, Satt146, Satt193, Satt569, Satt343, Satt586, Satt040,
Satt595, Satt334, Satt144, Satt522, Satt570, Satt356, Satt199, Satt517,
Satt191, Sat.sub.--117, Satt279, Satt181, Satt127, Satt270, Satt292,
SAG1223, SAC1699, Sat.sub.--065, Satt596, Satt406, Satt380, Satt183,
Satt529, Satt242, Satt617, Satt240, SAT.sub.--301, Satt418, Satt398,
Satt497, Satt166, Satt448, Satt373, Satt513, P12394A-1, SAG1048, Satt536,
Satt175, Satt677, Satt680, P10615A-1, Satt551, SAT.sub.--330-DB,
P13069A-1, P5467A-1, P5467A-2, SAT.sub.--084, SAT.sub.--275-DB, Satt660,
Satt339, Satt358, Satt487, Satt487, Satt420, Satt576, Satt633, Satt581,
Satt153, Satt243, P10793A-1, P13560A-1, P13561A-1, S60021-TB, S60048-TB,
S60076-TB, S60148-TB, S60149-TB, S60201-TB, S60243-TB, S60326-TB,
S60338-TB, S60350-TB, S60361-TB, S60422-TB, S60440-TB, S60446-TB,
S60505-TB, S60513-TB, S60519-TB, S60536-TB, S60552-TB, S60585-TB,
S60630-TB, S60728-TB, S60812-TB, SAC1677, SAC1724, SAG1055, Satt111,
Satt219 and Satt299 are determined. In an embodiment, the set of markers
includes Satt684, Satt526, Satt591, Satt385, Satt632-TB, Satt429,
SAT-.sub.--261, P10641A-1, Satt556, P10638B-2, Satt190, SAT.sub.--311-DB,
Satt338, Satt640-TB, Satt557, SAT.sub.--142-DB, Satt321, Satt203,
Satt129, SAT.sub.--351, Satt701, Satt582, Satt389, Satt464, Satt672,
Satt598, Satt343, Satt595, Satt334, Satt144, Satt522, Satt570, Satt356,
Satt199, Sat.sub.--117, Satt279, Satt181, Satt127, Satt270, Satt292,
SAG1223, Sat.sub.--065, Satt529, Satt242, Satt617, SAT.sub.--301,
Satt398, Satt497, Satt166, Satt373, SAG1048, Satt680, P10615A-1,
SAT.sub.--330-DB, DB, P13069A-1, SAT.sub.--275-DB, Satt339, Satt487,
Satt420, Satt581 and Satt153.
[0019] In some embodiments, the allele correlated with increased yield,
and conversely, the allele not correlated with increased yield, are
determined as follows. At least one polymorphic marker locus having at
least two segregating alleles in a plurality of sublines of progeny
soybean plants is selected. The yield is assessed in at least two
sublines of progeny with different alleles of the marker. A subline with
increased yield relative to the mean yield of the sublines is then
identified confirming a correlation between one of the segregating
alleles and increased yield.
[0020] The methods for identifying soybean sublines with increased yield
involve detecting at least one allelic form of multiple marker loci.
Typically, the number of marker loci is greater than two, and typically
is between 10% and 100% of the set of marker loci including: Satt684,
Satt165, Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155,
Satt385, Satt385, Satt225, Satt236, Satt511, P12390B-1, Satt480,
Satt632-TB, Satt233, Satt327, Satt329, Satt508, P10635A-1, Satt409,
Satt228, Satt429, Satt426, Satt509, SAT.sub.--261, Satt197, Satt519,
Satt597, SCT.sub.--026, Satt415, Satt583, Satt430, P12198A-1, P8584A-1,
Satt359, P10648A-1, P12105A-1, P10641A-1, Satt168, Satt556, Satt272,
Satt020, Satt066, Satt534, P10638B-2, Satt399, Satt361, P10639A-1,
Satt661-TB, Satt190, SAT.sub.--311-DB, Satt338, Satt227, Satt640-TB,
Satt422, Satt457, Satt457, Satt557, Satt319, SAT.sub.--142-DB, Satt460,
P13073A-1, Satt307, SCT.sub.--028, Satt433, Satt357, Satt321, Satt267,
Satt383, Satt295, Satt203, Satt507, SAT.sub.--110, P10620A-1, Satt129,
Satt147, Satt216, SAT.sub.--351, P10621B-2, Satt701, Satt634, Satt558,
Satt266, Satt282, Satt537, Satt506, Satt546, P13072A-1, Satt582, Satt389,
Satt461, Satt311, Satt514, Satt464, Satt662, Satt543, Satt186, Satt413,
Satt672, P13074A-1, P10624A-1, Satt573, Satt598, Satt204, Satt263,
Satt491, Satt602, Satt151, Satt355, Satt452, SAT.sub.--273-DB, Satt146,
Satt193, Satt569, Satt343, Satt586, Satt040, Satt423, Satt348, Satt595,
P10782A-1, P3436A-1, P10598A-1, Satt334, Satt510, Satt510, Satt144,
Satt522, Satt522, P9026A-1, P10646A-1, P5219A-1, P7659A-2, Satt570,
Satt356, Satt130, Satt115, Satt594, Satt533, Satt303, Satt352, Satt566,
Satt199, Satt503, Satt517, Satt191, SAT.sub.--117, Satt353, Satt442,
Satt279, Satt314, Satt142, Satt181, Satt367, Satt127, SCTT012, Satt270,
Satt292, Satt440, P10640A-1, Satt249, SAG1223, SAC1699, SCT.sub.--065,
Satt596, Satt280, Satt406, Satt380, Satt183, Satt529, Satt431, Satt242,
Satt102, Satt441, Satt544, Satt617, Satt240, P10618A-1, Satt475, Satt196,
SAT.sub.--301, Satt523, Satt418, Satt418, Satt398, Satt497, Satt284,
Satt166, Satt448, Satt373, Satt513, P12394A-1, Satt590, Satt567, Satt220,
SAG1048, Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt250,
Satt346, Satt336, SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2,
Satt584, SAT.sub.--084, P3050A-2, SAT.sub.--275-DB, Satt387, Satt549,
Satt660, Satt339, Satt255, Satt257, Satt358, P12396A-1, Satt487, Satt259,
Satt259, Satt347, Satt420, Satt576, Satt550, Satt633, Satt262, Satt473,
Satt477, Satt581, P11070A-1, Satt153, Satt243, P8230A-1, P10623A-1,
P10632A-1, P10793A-1, P12391A-1, P13560A-1, P13561A-1, P13561A-1,
P2481A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB, S60149-TB,
S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB, S60361-TB,
S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB, S60519-TB,
S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB, S60812-TB,
SAC1677, SAC1724, SAG1055, Satt040, Satt108, Satt109, Satt111, Satt176,
Satt176, Satt219, Satt299, and Satt512, or a geographically relevant
subset thereof as indicated in Tables 3 through 12. Usually, the methods
involve detecting between about 10% and about 90% of the markers in the
set. Frequently, at least 50% of the markers are detected, i.e., a
majority of the markers in the set. In some instances essentially all of
the markers are detected. Each of the detected marker loci identifies a
chromosome segment shown to include a genetic element which contributes
to increased yield in at least one geographic growing region. Thus, by
identifying alleles of the markers associated with increased yield,
sublines of soybeans with increased yield are identified.
[0021] The population of progeny soybeans utilized can be obtained by
crossing a first progenitor soybean with a second progenitor soybean.
Alternatively, the population of progeny can be obtained by selfing a
single progenitor soybean. In some embodiments, the sublines of progeny
evaluated include random sublines. In some embodiments, the sublines
include near isogenic sublines.
[0022] Typically, the progenitor soybean is selected from an elite strain
of germplasm. Such a progenitor can be self-fertilized to generate
progeny. More commonly, in the methods of the invention, the progenitor
soybean is crossed to a second soybean selected from a different elite
strain of germplasm. Alternatively, the progenitor soybean can be crossed
to a soybean with an exotic strain of germplasm.
[0023] The elite strain of germplasm is typically selected from among the
following strains of germplasm: 90A07, 90B11, 90B31, 90B43, 90B72, 90B73,
91B01, 91B12, 91B33, 91B52, 91B53, 91B64, 91B91, 91B92, 92B05, 92B12,
92B23, 92B38, 92B52, 92B63, 92B74, 92B75, 92B84, 92B95, 92M30, 92M31,
92M70, 92M71, 92M72, 92M80, 92M91, 93B01, 93B09, 93B11, 93B15, 93B25,
93B26, 93B36, 93B41, 93B45, 93B46, 93B66, 93B67, 93B68, 93B72, 93B82,
93B84, 93B85, 93B86, 93B87, 93M10, 93M30 93M40, 93M50, 93M60, 93M80,
93M90, 93M92, 93M93, 94B01, 94B23, 94B24, 94B53, 94B54, 94B73, 95B32,
95B33, 95B34, 95B53, 95B95, 95B96, 95B97, 96B21, 96B51, 97B52, 97B61,
A1395, A2722, A2835, A2943, A3127, A3237, A3242, A3322, A3431, A4009,
A4138, A4415, A4595, A4715, A5403, A5560, A5843, A5885, A5979, A5980,
A6297, BEDFORD, CM428, CX105, CX232, CX253, CX289, CX394C, CX469C,
D00566D362, ESSEX, EX04C00, EX06A00, EX10F01, EX13P01, EX13Q01, EX15N01,
EX16N00, EX16P01, EX22Y01, EX22Z01, EX23B03, EX34T03, EX35F03, EX36Y01,
EX39E00, EX40T03, EX44V03, FORREST, G3362, HS93-4118, HUTCHESON, JIM,
KORADA, MO15733, MO400644-02, MO413735-11-52, MO501577-27-23,
MO505469-61-89, MP39009, P1677, P9007, P9008, P9041, P9042, P9061, P9062,
P9063, P9071, P9092, P9132, P9141, P9151, P9163, P9182, P9203, P9233,
P9244, P9273, P9281, P9305, P9306, P9321, P9341, P9392, P9395, P9481,
P9482, P9492, P9521, P9552, P9561, P9584, P9591, P9592, P9594, P9631,
P9641, PHARAOH, RA451, R01154R002, S0066, S03W4, S0880, S1550, S1990,
S19T9, S20F8, S22C3, S24L2, S25J5, S32Z3, S33N1, S38T8, S3911, S4260,
S42H1, S43B5, S5960, S6189, S6262, ST0653, ST1073, ST1090, ST1570,
ST1690, ST1970, ST2250, ST2488, ST2660, ST2686, ST2688, ST2788, ST2870,
ST3171, ST3380, ST3630, ST3660, ST3870, ST3883, TRACY, TRAILL, X9916,
YB03E00, XB03F01, XB07E01, XB10D01, XB15M01, XB19U04, XB20M01, XB22C04,
XB22P04, XB23W03, XB23Y02, XB25E02, XB25L04, XB25X04, XB25W01, XB26L04,
XB27L04, XB29A04, XB29D01, XB29K04, XB29L04, XB30E04, XB31C01, XB31R04,
XB33B, XB34D04, XB34F01, XB35D, XB35L04, XB35W00, XB38A01, XB41M01,
XB42J00, XB42M01, XB48H01, XB54K01, XB55J01, XB58P99, XB63D00, XB67A00,
YB03G01, YB08D01, YB09F01, YB09G01, YB10E01, YB11D01, YB14H01, YB15K99,
YB21R99, YB21G01, YB22S00, YB22V01, YB22W01, YB22X01, YB24Z01, YB25R03,
YB25R99, YB25X00, YB25Y01, YB25Z01, YB27L03, YB27S00, YB27X01, YB27Y01,
YB28A03, YB28N01, YB29H01, YB29J01, YB29T04, YB30J01, YB30N01, YB30P01,
YB31E01, YB32K01, YB33K01, YB34H01, YB34R03, YB34S03, YB35C01, YB36E03,
YB36V00, YB38E03, YB38G03, YB39M01, YB39V03, YB40M01, YB40N01, YB41Q01,
YB48L01, YB52J00, YB53E00, YB54H00, YB54J00, YB54L00, YB55H00, YB56E00,
YB60N01, and YOUNG.
[0024] In some embodiments, the methods include electronically
transmitting or electronically storing data representing the determined
marker alleles or allelic forms or chromosome segments in a computer
readable medium. Accordingly, another aspect of the invention includes
computer systems including a data input device for inputting genotyping
data, and a computer readable medium incorporating the genotyping data
corresponding to the markers of the invention. Computer readable medium
including the genotyping data are also a feature of the invention.
[0025] In some embodiments, the methods further include selecting at least
one plant of the identified soybean subline. The selected plant can be a
whole plant, a plant organ, a plant seed, a plant cell, a plant tissue
culture, or the like. Optionally, the selected soybean plant, or a
progeny thereof is crossed with a second soybean plant. Typically the
second soybean plant lacks the determined allele of the marker locus (or
allelic form of the chromosome segment). In some embodiments, the second
soybean plant is from an elite strain of germplasm. In other embodiments,
the second soybean plant is from an exotic strain of germplasm.
[0026] Soybean plants with increased yield produced according to the
methods of the invention are also a feature of the invention.
[0027] In another aspect, the invention includes sets of markers useful
for identifying soybean plants with increased yield. The marker sets
include markers selected from the set of markers including Satt684,
Satt165, Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155,
Satt385, Satt385, Satt225, Satt236, Satt511, P12390B-1, Satt480,
Satt632-TB, Satt233, Satt327, Satt329, Satt508, P10635A-1, Satt409,
Satt228, Satt429, Satt426, Satt509, SAT.sub.--261, Satt197, Satt519,
Satt597, SCT.sub.--026, Satt415, Satt583, Satt430, P12198A-1, P8584A-1,
Satt359, P10648A-1, P12105A-1, P10641A-1, Satt168, Satt556, Satt272,
Satt020, Satt066, Satt534, P10638B-2, Satt399, Satt361, P10639A-1,
Satt661-TB, Satt190, SAT.sub.--311-DB, Satt338, Satt227, Satt640-TB,
Satt422, Satt457, Satt457, Satt557, Satt319, SAT.sub.--142-DB, Satt460,
P13073A-1, Satt307, SCT.sub.--028, Satt433, Satt357, Satt321, Satt267,
Satt383, Satt295, Satt203, Satt507, SAT.sub.--110, P10620A-1, Satt129,
Satt147, Satt216, SAT.sub.--351, P10621B-2, Satt701, Satt634, Satt558,
Satt266, Satt282, Satt537, Satt506, Satt546, P13072A-1, Satt582, Satt389,
Satt461, Satt311, Satt514, Satt464, Satt662, Satt543, Satt186, Satt413,
Satt672, P13074A-1, P10624A-1, Satt573, Satt598, Satt204, Satt263,
Satt491, Satt602, Satt151, Satt355, Satt452, SAT.sub.--273-DB, Satt146,
Satt193, Satt569, Satt343, Satt586, Satt040, Satt423, Satt348, Satt595,
P10782A-1, P3436A-1, P10598A-1, Satt334, Satt510, Satt510, Satt144,
Satt522, Satt522, P9026A-1, P10646A-1, P5219A-1, P7659A-2, Satt570,
Satt356, Satt130, Satt115, Satt594, Satt533, Satt303, Satt352, Satt566,
Satt199, Satt503, Satt517, Satt191, SAT.sub.--117, Satt353, Satt442,
Satt279, Satt314, Satt142, Satt181, Satt367, Satt127, SCTT012, Satt270,
Satt292, Satt440, P10640A-1, Satt249, SAG1223, SAC1699, SCT.sub.--065,
Satt596, Satt280, Satt406, Satt380, Satt183, Satt529, Satt431, Satt242,
Satt102, Satt441, Satt544, Satt617, Satt240, P10618A-1, Satt475, Satt196,
SAT.sub.--301, Satt523, Satt418, Satt418, Satt398, Satt497, Satt284,
Satt166, Satt448, Satt373, Satt513, P12394A-1, Satt590, Satt567, Satt220,
SAG1048, Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt250,
Satt346, Satt336, SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2,
Satt584, SAT.sub.--084, P3050A-2, SAT.sub.--275-DB, Satt387, Satt549,
Satt660, Satt339, Satt255, Satt257, Satt358, P12396A-1, Satt487, Satt259,
Satt259, Satt347, Satt420, Satt576, Satt550, Satt633, Satt262, Satt473,
Satt477, Satt581, P11070A-1, Satt153, Satt243, P8230A-1, P10623A-1,
P10632A-1, P10793A-1, P12391A-1, P13560A-1, P13561A-1, P13561A-1,
P2481A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB, S60149-TB,
S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB, S60361-TB,
S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB, S60519-TB,
S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB, S60812-TB,
SAC1677, SAC1724, SAG1055, Satt040, Satt108, Satt109, Satt111, Satt176,
Satt176, Satt219, Satt299, and Satt512. Typically a subset of markers
shown to be relevant in a geographic growing region are selected, as
indicated in Tables 3 through 12. For example, in a breeding program
designed to develop strains of soybean with increased yield in the
Central (e.g., Iowa) region, a set of markers selected from among the
following marker set is preferred: Satt642, Satt042, Satt364, Satt454,
Satt526, Satt300, Satt591, Satt155, Satt385, Satt511, P12390B-1,
Satt632-TB, Satt429, SAT.sub.--261, Satt197, P10641A-1, Satt556, Satt534,
P10638B-2, Satt399. Satt361, P10639A-1, Satt661-TB, Satt190,
SAT.sub.--311-DB, Satt338, Satt640-TB, Satt557, Satt319,
SAT.sub.--142-DB, Satt460, Satt433, Satt357, Satt321, Satt295, Satt203,
Satt507, Satt129, Satt147, SAT.sub.--351, P10621B-2, Satt558, Satt701,
Satt634, Satt582, Satt389, Satt464, Satt662, Satt672, Satt573, Satt598,
Satt263, Satt602, Satt151, SAT.sub.--273-DB, Satt146, Satt193, Satt569,
Satt176, Satt343, Satt586, Satt040, Satt595, P10782A-1, Satt334, Satt144,
Satt522, Satt570, Satt356, Satt533, Satt199, Satt517, Satt191,
SAT.sub.--117, Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223,
SAC1699, SAT.sub.--065, Satt596, Satt406, Satt380, Satt183, Satt529,
Satt242, Satt617, Satt240, SAT.sub.--301, Satt418, Satt398, Satt497,
Satt166, Satt448, Satt373, Satt513, P12394A-1, SAG1048, Satt536, Satt175,
Satt677, Satt680, P10615A-1, Satt551, Satt346, Satt336, SAT.sub.--330-DB,
P13069A-1, P5467A-1, P5467A-2, SAT.sub.--084, SAT.sub.--275-DB, Satt660,
Satt339, P12396A-1, Satt358, Satt487, Satt259, Satt420, Satt576, Satt633,
Satt477, Satt581, Satt153, Satt243, P10793A-1, P12391A-1, P12392A-1,
P13560A-1, P13561A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB,
S60149-TB, S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB,
S60361-TB, S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB,
S60519-TB, S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB,
S60812-TB, SAC1677, SAC1724, SAG1055, Satt040, Satt111, Satt176, Satt219
and Satt299. In some embodiment, the markers of the set art selected from
among: Satt684, Satt042, Satt364, Satt454, Satt526, Satt300, Satt591,
Satt155, Satt385, Satt632-TB, Satt429, SAT-.sub.--261, P10641A-1,
Satt556, P10638B-2, Satt399, Satt361, Satt661-TB, Satt190,
SAT.sub.--311-DB, Satt338, Satt640-TB, Satt557, Satt319,
SAT.sub.--142-DB, Satt321, Satt203, Satt129, Satt147, SAT.sub.--351,
P10621B-2, Satt701, Satt634, Satt582, Satt389, Satt464, Satt662, Satt672,
Satt573, Satt598, Satt263, Satt151, SAT.sub.--273-DB, Satt146, Satt193,
Satt569, Satt343, Satt586, Satt040, Satt595, Satt334, Satt144, Satt522,
Satt570, Satt356, Satt199, Satt517, Satt191, Sat.sub.--117, Satt279,
Satt181, Satt127, Satt270, Satt292, SAG1223, SAC1699, Sat.sub.--065,
Satt596, Satt406, Satt380, Satt183, Satt529, Satt242, Satt617, Satt240,
SAT.sub.--301, Satt418, Satt398, Satt497, Satt166, Satt448, Satt373,
Satt513, P12394A-1, SAG1048, Satt536, Satt175, Satt677, Satt680,
P10615A-1, Satt551, SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2,
SAT.sub.--084, SAT.sub.--275-DB, Satt660, Satt339, Satt358, Satt487,
Satt487, Satt420, Satt576, Satt633, Satt581, Satt153, Satt243, P10793A-1,
P13560A-1, P13561A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB,
S60149-TB, S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB,
S60361-TB, S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB,
S60519-TB, S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB,
S60812-TB, SAC1677, SAC1724, SAG1055, Satt111, Satt219 and Satt299. In
certain embodiments the markers are selected from the set including:
Satt684, Satt526, Satt591, Satt385, Satt632-TB, Satt429, SAT-.sub.--261,
P10641A-1, Satt556, P10638B-2, Satt190, SAT.sub.--311-DB, Satt338,
Satt640-TB, Satt557, SAT.sub.--142-DB, Satt321, Satt203, Satt129,
SAT.sub.--351, Satt701, Satt582, Satt389, Satt464, Satt672, Satt598,
Satt343, Satt595, Satt334, Satt144, Satt522, Satt570, Satt356, Satt199,
Sat.sub.--117, Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223,
Sat.sub.--065, Satt529, Satt242, Satt617, SAT.sub.--301, Satt398,
Satt497, Satt166, Satt373, SAG1048, Satt680, P10615A-1, SAT.sub.--330-DB,
P13069A-1, SAT.sub.--275-DB, Satt339, Satt487, Satt420, Satt581 and
Satt153.
[0028] Typically the set of markers includes between about 10% and about
100% of the markers shown to be relevant in a selected geographic region.
Usually, the set includes between about 10% and about 90% of the relevant
markers. Frequently, the set includes a majority of the relevant markers.
In some embodiments, the set includes essentially all of the markers
shown to be relevant in a particular geographic region.
BRIEF DESCRIPTION OF THE DRAWINGS
[0029] FIGS. 1A-E. Schematic illustration of genetic map indicating
positions of markers.
[0030] FIG. 2. Genetic map indicating position of marker loci associated
with yield in Iowa.
DETAILED DESCRIPTION
[0031] The present invention provides soybean markers associated with loci
important for soybean yield. Using methods described in U.S. Pat. No.
5,437,697, which is incorporated in its entirety for all purposes, a
series of breeding bias analyses were conducted to identify genetic
markers that define regions of the soybean genome that are important for
yield. Each analysis identified loci that were affected by selection in
one of seven different soybean growing regions in North America. For each
geographic region, an "elite population" of between 38 and 86
representative elite lines was chosen by an experienced soybean breeder.
Each elite line was evaluated for up to 309 molecular markers to
determine its allelic genotype at each of 309 different genetic loci
spanning the soybean genome.
[0032] In addition to determining the marker genotypes for these elite
lines, the most relevant leaf ancestors for each representative elite
line were genotyped with the same 309 genetic markers. A "leaf ancestor"
is an ancestor for whom the previous parents are unknown, representing an
endpoint in the pedigree of each elite line. The breeding bias analysis
uses computer simulation and the known pedigree structure between the
elite lines and the leaf ancestors to determine the "expected" frequency
of each marker allele within the elite population assuming no bias due to
selection. The expected frequency is merely the average frequency with
which a given allele would be expected in the elite population due to the
rules of random Mendelian segregation. The simulation uses the actual
pedigrees of each elite line to determine the path that each allele must
take during the simulated inheritance process. For example, for any
diploid biparental cross within a pedigree, the simulation assumes a
50-50 chance that a given parent allele will be passed on to a given
progeny from that cross. By knowing the genotypes of the ancestors and
the pedigree of a given elite line, one can simulate the inheritance of
each allele through a pedigree structure to determine how often that
allele would be expected in the elite line according to a purely random
process.
[0033] However, plant breeding is not a random process; breeders purposely
select for characteristics that provide adaptation and high grain yield
in specific geographic regions. By practicing many cycles of genetic
recombination and selection of the best genotypes for a given region,
breeders indirectly "bias" the gene pool towards the alleles that provide
the best grain yield in that locale. Using this logic, any marker allele
that was inherited significantly more frequently than expected by random
simulation, must reside in a genomic region (chromosome segment) that
contributes either directly or indirectly to high grain yield.
[0034] Following identification of regions of the soybean genome important
for yield using the Breeding Bias analysis, specific marker alleles
associated with increased yield can be identified in lines and sublines
of soybeans within a breeding program. Since favorable allelic forms of
chromosome segments are linked to and defined by genetic markers,
accurate selection based on genotype can then replace inefficient
selection based solely on phenotype. The end result is more efficient
progress towards a genotype with the favorable allelic forms with respect
to yield being fixed within the elite gene pool.
[0035] Definitions
[0036] Before describing the present invention in detail, it is to be
understood that this invention is not limited to particular biological
systems, e.g., soybean lines, or reagents, such as particular markers,
which can, of course, vary. It is also to be understood that the
terminology used herein is for the purpose of describing particular
embodiments only, and is not intended to be limiting. As used in this
specification and the appended claims, the singular forms "a," "an" and
"the" include plural referents unless the content clearly dictates
otherwise. The term "plurality" is used to mean "two or more." Thus, for
example, reference to "a marker" includes a single marker as well as a
plurality of markers, such as two or more markers; and the like.
[0037] Unless defined otherwise, all technical and scientific terms used
herein have the same meaning as commonly understood by one of ordinary
skill in the art to which the invention pertains. Exemplary methods and
materials are described herein, however, any methods and materials
similar or equivalent to those described herein, such as additional or
alternative markers physically and genetically linked to the markers (and
alleles) described herein, can be used in the practice of the present
invention. In describing and claiming the present invention, the
following terminology will be used in accordance with the definitions set
out below.
[0038] The term "germplasm" refers to an individual, a group of
individuals, or a clone representing a genotype, variety, species or
culture, or the genetic material thereof.
[0039] In the context of this disclosure, the term "yield" refers to the
productivity per unit area of a particular plant product of commercial
significance. For example, yield of soybean is commonly measured in
bushels of seed per acre or metric tons of seed per hectare per season.
Yield is affected by both genetic and environmental factors.
"Agronomics," "agronomic traits," and "agronomic performance" refer to
the traits (and underlying genetic elements) of a given plant variety
that contribute to yield over the course of growing season. Individual
agronomic traits include emergence vigor, vegetative vigor, stress
tolerance, disease resistance, herbicide resistance, branching,
flowering, seed set, seed size, seed density, standability, threshability
and the like. Yield is, therefore, the final culmination of all agronomic
traits.
[0040] The term "genetic element" or "gene" refers to a heritable sequence
of DNA, i.e., a genomic sequence, with functional significance. The term
"gene" can be used to refer to, e.g., a cDNA and/or an mRNA encoded by a
genomic sequence, as well as to that genomic sequence.
[0041] "Locus" refers to a specific chromosome location in the genome of a
species where a specific gene can be found. The term "quantitative trait
locus" or "QTL" refers to a genetic locus with at least two alleles that
differentially affect the expression of a phenotypic trait on at least
one genetic background, e.g., in at least one breeding population or
sample of progeny.
[0042] The term "chromosome segment" designates a contiguous linear span
of genomic DNA that resides in planta on a single chromosome. The genetic
elements or genes located on a single chromosome segment are physically
linked. In the context of the present invention the genetic elements
located within a chromosome segment are also genetically linked,
typically within a genetic recombination distance of less than or equal
to 10 centimorgan (CM). That is, two genetic elements within a single
chromosome segment undergo recombination during meiosis with each other
at a frequency of less than or equal to about 10%.
[0043] "Allele" refers to one of two or more different DNA sequences at a
specific locus. In the example of a specific locus where a gene for
growth habit is located, one allele is a specific DNA sequence that,
e.g., codes for determinate growth habit while another allele is a
different DNA sequence that codes for indeterminate growth habit. A
"favorable allele" is the allele at a particular locus that confers, or
contributes to, an agronomically desirable phenotype, e.g., increased
yield. A favorable allele of a marker is an allele associated with a
favorable allele at a linked locus which confers or contributes to an
agronomically desirable phenotype, e.g., increased yield. A favorable
allelic form of a chromosome segment is a chromosome segment including a
DNA sequence that contributes to superior agronomic performance at one or
more genetic loci physically located on the chromosome segment.
[0044] "Allele frequency" refers to the frequency (proportion or
percentage) at which an allele is present at a locus within an
individual, within a line, or within a population of lines. For example,
regarding the allele "A," diploid individuals of genotype "AA," "Aa," or
"aa" have allele frequencies of 1.0, 0.5, or 0.0, respectively. One can
estimate the allele frequency within a line by averaging the allele
frequencies of a sample of individuals from that line. Similarly, one can
calculate the allele frequency within a population of lines by averaging
the allele frequencies of lines that make up the population. For a
population with a finite number of individuals or lines, an allele
frequency can be expressed as a count of individuals or lines containing
the allele.
[0045] A "genetic marker" is any qualitatively (discretely) inherited
phenotype that can be used to monitor the segregation of alleles at loci
that are genetically linked to the marker. Genetic markers include
visible traits such as flower color; enzyme variants such as isozymes and
molecular markers such as simple sequence repeats (SSRs), single
nucleotide polymorphisms (SNPs), e.g., allele specific hybridization
(ASH) markers, restriction fragment length polymorphisms (RFLPs) or
randomly amplified polymorphic DNA (RAPDs), etc. Thus, a "marker allele,"
alternatively an "allele of a marker locus" is one of a plurality of
polymorphic nucleotide sequences at a marker locus.
[0046] "Codominant markers" reveal the presence of each allele (two per
diploid individual) at a locus, e.g., SSR, SNP (e.g., ASH), RFLP, AFLP
markers. "Dominant markers" reveal the presence of only a single allele
per locus, e.g., RAPD markers. The presence of the dominant marker
phenotype (e.g., a band of DNA) is an indication that one allele is
present in either the homozygous or heterozygous condition. The absence
of the dominant marker phenotype (e.g., absence of a DNA band) is merely
evidence that some other, undefined, allele is present. In the case of
populations where individuals are predominantly homozygous and loci are
predominantly dimorphic, dominant and codominant markers are equally
valuable. As individuals within populations become more heterozygous and
multi-allelic, codominant markers become more informative of genotype
than dominant markers.
[0047] A "set" of markers refers to a collection or group of markers, or
the data derived therefrom, used for a common purpose, e.g., identifying
soybean plants with increased yield. Frequently, the data is stored in an
electronic medium. While each of the members of the set has been shown to
possess utility with respect to the specified purpose: individual markers
selected from the set as well as subsets including some, but not all of
the markers, are also effective in achieving the specified purpose.
[0048] A "genetic map" is a description of the genetic linkage
relationships among loci on one or more chromosomes (or linkage groups)
within a given species, generally depicted in a diagrammatic or tabular
form. "Mapping" is the process of defining the linkage relationships of
loci through the use of genetic markers, populations segregating for the
markers, and standard genetic principles of recombination frequency. A
"map location" is an assigned location on a genetic map relative to
linked genetic markers where a specified marker can be found within a
given species.
[0049] A "genotype" is the genetic constitution of an individual (or group
of individuals) at one or more genetic loci. Genotype is defined by the
allele(s) of one or more known loci that the individual has inherited
from its parents. A "haplotype" is the genotype of an individual at a
plurality of genetic loci. Typically, the genetic loci described by a
haplotype are physically and genetically linked, i.e., on the same
chromosome segment.
[0050] An individual is "homozygous" if the individual has only one type
of allele at a given locus (e.g., a diploid individual with two copies of
the same allele at a locus). An individual is "Heterozygous" if more than
one allele type is present at a given locus (e.g., a diploid individual
with one copy each of two different alleles). The term "homogeneity"
indicates that members of a group have the same genotype at one or more
specific loci. In contrast, the term "heterogeneity" is used to indicate
that individuals within the group differ in genotype at one or more
specific loci.
[0051] A "line" or "strain" is a group of individuals of identical
parentage that are generally inbred to some degree and are generally
homozygous and homogeneous at most loci.
[0052] An "elite line" or "elite strain" is a genetically superior line
that has resulted from many cycles of breeding and selection for superior
agronomic performance. Numerous elite lines are available and known to
those of skill in the art of soybean breeding. An "elite population" is
an assortment of elite lines that can be used to represent the state of
the art in terms of agronomically superior genotypes of a given crop
species, such as soybean. Similarly, an "elite germplasm" or elite strain
of germplasm is a genetically superior germplasm, typically derived from
and/or capable of giving rise to a plant with superior agronomic
performance, such as an existing or newly developed elite line of
soybean.
[0053] In contrast, an "exotic germplasm" is a germplasm derived from a
soybean not belonging to an available elite soybean line or strain of
germplasm. In the context of a cross between two soybean plants or
strains of germplasm, an exotic germplasm is unrelated by descent to the
elite germplasm with which it is crossed. Most commonly, the exotic
germplasm is not derived from any known elite line of soybean, but rather
is selected to introduce novel genetic elements (novel alleles) into a
breeding program.
[0054] An "ancestral line" is a parent used as a source of genes for the
development of elite lines. An "ancestral population" is a group of
ancestors that have contributed the bulk of the genetic variation that
was used to develop elite lines. "Descendants" are the progeny of
ancestors, and may be separated from their ancestors by many generations
of breeding. For example, elite lines are the descendants of their
ancestors. A "pedigree structure" defines the relationship between a
descendant and each ancestor that gave rise to that descendant. A
pedigree structure can span one or more generations, describing
relationships between the descendant and it's parents, grand parents,
great-grand parents, etc.
[0055] The term "subline" refers to a an inbred subset of descendents that
genetically distinct from other similarly inbred subsets descended from
the same progenitor. Traditionally, a "subline" has been derived by
inbreeding the seed from an individual soybean plant selected and at the
F3 to F5 generation until the residual segregating loci are "fixed" or
homozygous across most or all loci. Commercial soybean varieties (or
lines) are typically produced by aggregating ("bulking") the
self-pollinated progeny of a single F3 to F5 plant from a controlled
cross between 2 genetically different parents. While the variety
typically appears uniform, the self-pollinating variety derived from the
selected plant eventually (e.g., F8) becomes a mixture of homozygous
plants that can vary in genotype at any locus that was heterozygous in
the originally selected F3 to F5 plant. In the context of the invention,
marker-based sublines, that differ from each other based on qualitative
polymorphism at the DNA level at one or more specific marker loci, are
derived by genotyping a sample of seed derived from individual
self-pollinated progeny derived from a selected F3-F5 plant. The seed
sample can be genotyped directly as seed, or as plant tissue grown from
such a seed sample. Optionally, seed sharing a common genotype at the
specified locus (or loci) are bulked providing a subline that is
genetically homogenous at identified loci important for increased yield.
[0056] The term "near-isogenic" lines refers to lines that are genetically
similar to each other except at one or a small number of genetic loci
(e.g., at 1, 2, or about 5 to about 10 specified genetic loci). These can
be created as described for marker-based sublines or based on differences
for any qualitative trait that can serve as an effective genetic marker.
Percent similarity between near-isogenic lines is a function of the
similarity of the parents of the original cross and the generation at
which self-pollination is performed. On average, the relatedness between
members of a given inbred line increases 50% with each cycle of
inbreeding, due to a 50% increase in homozygosity at each cycle of
inbreeding. Percent similarity can be more accurately determined with
genetic markers that span the genome. In some cases, near-isogenic lines
differ from each other at one defined genetic locus.
[0057] "Transgressive segregation" is an inheritance pattern that results
in a phenotype (e.g., agronomic performance) of an individual that is
more extreme than either parent. For example, with respect to agronomic
performance, transgressive segregation results in a progeny with yield
that is greater than a best parent or less than a worst parent. Desirable
transgressive segregation is the case where the progeny are better than
either parent. Transgressive segregation can also be measured in terms of
the number of favorable alleles that an individual inherits in relation
to the number of favorable alleles of each of its parents. A "target
segregant" is a progeny from a specific cross that includes only
favorable alleles at each defined locus segregating in the cross. The
target segregant therefore, has the best possible genotype that can
result from a cross between parents that differ in genotype at known
loci. "Target genotype" refers to an individual containing the favorable
allelic forms at all chromosome segments or loci known to affect a
particular trait or phenotype, such as agronomic performance. With
respect to agronomic performance, the target genotype is that of the
target segregant from a cross between parents that complement in terms of
favorable alleles at all defined loci affecting agronomic performance.
[0058] A "survey" or "genetic survey" or "genetic marker survey" is the
process of determining and recording the genotype of individuals or lines
(e.g., ancestral and elite lines), at any number of defined loci, with
the use of genetic markers.
[0059] The term "associated with" or "associated," when referring to a
nucleic acid (e.g., a genetic marker) and a phenotype in the context of
the present invention, refers to a nucleic acid and a phenotypic trait
that are in linkage phase disequilibrium. The term "linkage phase
disequilibrium" or "linkage disequilibrium" refers to a non-random
segregation of genetic loci. This implies that such loci are in
sufficient physical proximity along a length of a chromosome that they
tend to segregate together with greater than random frequency.
[0060] The term "genetically linked" refers to genetic loci (including
genetic marker loci) that are physically close enough to each other on
the same chromosome such that they have a recombination frequency of less
than 0.5. When referring to the relationship between two genetic
elements, such as a genetic element contributing to yield and a proximal
marker, "Coupling" phase linkage indicates the state where the
"favorable" allele at the yield locus is physically associated on the
same chromosome strand as the "favorable" allele of the respective linked
marker locus. In coupling phase, both favorable alleles are inherited
together by progeny that inherit that chromosome strand. In "repuslsion"
phase linkage, the "favorable" allele at the yield locus is physically
linked with an "unfavorable" allele at the proximal marker locus, and the
two "favorable" alleles are not inherited together (i.e., the two loci
are "out of phase" with each other).
[0061] The term "physically linked" is used to indicate that two genetic
loci, e.g., two marker loci, a marker locus and a locus contributing to
variation in a phenotype, are physically present on the same chromosome.
Typically, the two loci are located in close proximity, such that
recombination between homologous chromosome pairs does not occur between
the two loci with high frequency. That is, recombination between two
physically linked loci typically occurs with a frequency of less than
about 10%, favorably with a frequency of less than 5%, more favorably
with a frequency of 2% or less or a frequency of 1% or less. Thus, two
loci that are localized to the same chromosome, and at such a distance
that recombination between the two loci occurs at a frequency of less
than 10% are said to be "proximal to" each other.
[0062] "Marker Assisted Selection" or "MAS" refers to the practice of
selecting for desired phenotypes among members of a breeding population
using genetic markers.
[0063] The phrase "hybrid plants" refers to plants which result from a
cross between genetically different individuals.
[0064] The term "crossed" or "cross" in the context of this invention
means the fusion of gametes, e.g., via pollination to produce progeny
(i.e., cells, seeds, or plants) in the case of plants. The term
encompasses both sexual crosses (the pollination of one plant by another)
and, in the case of plants, selfing (self-pollination, i.e., when the
pollen and,ovule are from the same plant).
[0065] "Random mating" is the mating of individuals within a population in
a way that insures the equal probability of any two individuals mating
regardless of genotype. "Non-random mating" is any deviation from random
mating in which specific crosses between individuals occur with greater
frequency than others.
[0066] The term "introgression" refers to the transmission of a desired
allele of a genetic locus from one genetic background to another. For
example, introgression of a desired allele at a specified locus can be
transmitted to at least one progeny via a sexual cross between two
parents of the same species, where at least one of the parents has the
desired allele in its genome. Alternatively, for example, transmission of
an allele can occur by recombination between two donor genomes, e.g., in
a fused protoplast, where at least one of the donor protoplasts has the
desired allele in its genome. The desired allele can be, e.g., a selected
allele of a marker or QTL or a transgene.
[0067] The terms "nucleic acid," "polynucleotide," "polynucleotide
sequence" and "nucleic acid sequence" refer to single-stranded or
double-stranded deoxyribonucleotide or ribonucleotide polymers, or
chimeras thereof. As used herein, the term can additionally or
alternatively include analogs of naturally occurring nucleotides having
the essential nature of natural nucleotides in that they hybridize to
single-stranded nucleic acids in a manner similar to naturally occurring
nucleotides (e.g., peptide nucleic acids). Unless otherwise indicated, a
particular nucleic acid sequence of this invention optionally encompasses
complementary sequences, in addition to the sequence explicitly
indicated.
[0068] The term "homologous" refers to nucleic acid sequences that are
derived from a common ancestral gene through natural or artificial
processes (e.g., are members of the same gene family), and thus,
typically, share sequence similarity. Typically, homologous nucleic acids
have sufficient sequence identity that one of the sequences or its
complement is able to selectively hybridize to the other under selective
hybridization conditions. The term "selectively hybridizes" includes
reference to hybridization, under stringent hybridization conditions, of
a nucleic acid sequence to a specified nucleic acid target sequence to a
detectably greater degree (e.g., at least 2-fold over background) than
its hybridization to non-target nucleic acid sequences and to the
substantial exclusion of non-target nucleic acids. Selectively
hybridizing sequences have about at least 80% sequence identity,
preferably at least 90% sequence identity, and most preferably 95%, 97%,
99%, or 100% sequence identity with each other. A nucleic acid that
exhibits at least some degree of homology to a reference nucleic acid can
be unique or identical to the reference nucleic acid or its complementary
sequence.
[0069] The term "isolated" refers to material, such as a nucleic acid or a
protein, which is partially or substantially free from components that
normally accompany or interact with it in its naturally occurring
environment. The isolated material optionally comprises material not
found with the material in its natural environment, e.g., a cell. In
addition, if the material is in its natural environment, such as a cell,
the material has been placed at a location in the cell (e.g., genome or
subcellular organelle) not native to a material found in that
environment. For example, a naturally occurring nucleic acid (e.g., a
promoter) is considered to be isolated if it is introduced by
non-naturally occurring means to a locus of the genome not native to that
nucleic acid. Nucleic acids which are "isolated" as defined herein, are
also referred to as "heterologous" nucleic acids.
[0070] The term "recombinant" when referring to a molecular species, such
as a nucleic acid or protein, indicates that the material (e.g., a
nucleic acid or protein) has been synthetically (non-naturally) altered
by human intervention. The alteration to yield the synthetic material can
be performed on the material within or removed from its natural
environment or state. For example, a naturally occurring nucleic acid is
considered a recombinant nucleic acid if it is altered, or if it is
transcribed from DNA which has been altered, by means of human
intervention, e.g., performed on the cell from which it originates. When
the term "recombinant" is used in the classical genetic sense, it refers
to an individual with one of the many possible rearrangments of genes due
to natural sexual recombination. For example, "recombinant inbred lines"
or "RILs" are merely the variety of inbred progeny from specific crosses
of divergent parents. The manner in which the term recombinant is
employed will be self evident from the context of its use.
[0071] The term "introduced" when referring to a heterologous or isolated
nucleic acid refers to the incorporation of a nucleic acid into a
eukaryotic or prokaryotic cell where the nucleic acid can be incorporated
into the genome of the cell (e.g., chromosome, plasmid, plastid or
mitochondrial DNA), converted into an autonomous replicon, or transiently
expressed (e.g., transfected mRNA). The term includes such nucleic acid
introduction means as "transfection," "transformation" and
"transduction."
[0072] The term "host cell" means a cell that contains a heterologous
nucleic acid, such as a vector, and supports the replication and/or
expression of the nucleic acid. Host cells may be prokaryotic cells such
as E. coli , or eukaryotic cells such as yeast, insect, amphibian, or
mammalian cells. In the context of the present invention, a eukaryotic
host cell is most commonly a soybean cell.
[0073] The term "transgenic" plant (or animal) refers to a plant (or
animal) which comprises within its genome a heterologous polynucleotide.
Generally, the heterologous polynucleotide is stably integrated within
the genome such that the polynucleotide is passed on to successive
generations. The heterologous polynucleotide may be integrated into the
genome alone or as part of a recombinant expression cassette.
"Transgenic" is used herein to refer to any cell, cell line, tissue, part
or organisms, the genotype of which has been altered by the presence of
heterologous nucleic acid including those transgenic organisms or cells
initially so altered, as well as those created by crosses or asexual
propagation from the initial transgenic organism or cell. The term
"transgenic" as used herein does not encompass the alteration of the
genome (chromosomal or extra-chromosomal) by conventional breeding
methods (i.e., crosses) or by naturally occurring events such as random
cross-fertilization, viral infection with non-recombinant nucleic acids,
bacterial transformation with non-recombinant nucleic acids,
transposition with non-recombinant nucleic acids, or spontaneous
mutation. Examples of processes by which a transgenic organism can be
produced are described below, and include electroporation,
microinjection, Agrobacterium-mediated transformation, biolistic methods,
in planta techniques, and the like.
[0074] The term "plant" includes any of: whole plants, plant organs (e.g.,
leaves, stems, roots, etc.), tissues, seeds, plant cells, and/or progeny
of the same. Similarly, "plant cell," as used herein includes, without
limitation, seeds, suspension cultures, embryos, meristematic regions,
callus tissue, leaves, roots, shoots, gametophytes, sporophytes, pollen,
and microspores. In addition, the term "plant" encompasses in silico
representations of part or all of a plant's genetic constitution.
[0075] Introduction
[0076] The present invention addresses the need in the field of
agriculture to more efficiently develop soybean plants having superior
agronomic performance. Superior agronomic performance, as measured by
increased yields of soybeans, is a function of such diverse factors as
seedling emergence vigor, resistance to environmental stress, pest
resistance, insect resistance, disease resistance, ability to mine
nutrients from the soil, ability to produce flowers, photosynthetic
ability, and ability to shuttle photosynthate into grain, etc. As
disclosed herein, genetic loci associated with superior agronomic
performance have been identified by analysis of a plurality of soybean
breeding programs (some of which extend back more than 70 years) in a
variety of geographic zones in the United States and Canada.
[0077] Soybean germplasm having superior agronomic performance (relative
to their parents and sibs developed in these same breeding programs) show
a statistically significant retention of particular allelic forms
(alternatively, "alleles") encompassed within the chromosome segments of
the present invention. This highly significant statistical association
demonstrates that loci within these chromosome segments are linked
(genetically and physically) to genetic elements associated with the
various traits involved in determining soybean yield in the context of
modem agricultural practices. Collectively, favorable attributes
corresponding to such traits are descriptively referred to as "superior
agronomic performance," and contribute to increased yield. Consequently,
molecular markers localized within these chromosome segments can be used
to define (and identify) soybean plants with superior agronomic
performance, and in marker assisted selection ("MAS") and marker assisted
breeding strategies (alternatively referred to as "molecular breeding")
to create soybean plants with superior agronomic performance.
Furthermore, chromosome segments encompassing the genetic elements
associated with the phenotype of superior agronomic performance can be
isolated and transformed into soybean or other monocot or dicot plant
species.
[0078] The methodology used to identify these chromosome segments,
referred to as "Breeding Bias" is disclosed in U.S. Pat. No. 5,437,697 to
Sebastian et al., incorporated herein in its entirety for all purposes.
[0079] Briefly, loci (e.g., markers and chromosome segments encompassing
genetic elements contributing to superior agronomic performance) that
have been affected by selection for agronomic performance, are identified
by comparing the genotype of modern elite lines with that of their
ancestors. Because domesticated soybeans are known to have a fairly
narrow gene pool and have been selected to fairly stringent standards, a
relatively small number of elite lines is adequate to identify relevant
chromosome regions associated with superior agronomic performance. The
identity of elite lines used for each geographical region is indicated in
Table 1.
[0080] Based on the selection of elite lines, the relevant ancestral
population, e.g., ancestors that were used most frequently by breeders to
develop the elite population is identified. A pedigree tracing the
relationship between each elite line and its earliest known ancestors is
obtained, and the genetic contribution of each ancestor is converted into
a proportional or percentage representation, assuming that on average 50%
of each parent's genome is passed on to each progeny as a result of a
two-way cross with another parent. By tracing the pedigree back until no
more branch points are found, the earliest known ancestors can be
identified and their contribution to each elite line calculated.
Calculations are performed, generally in a computerized format, for each
of the elite lines included in the genetic survey.
[0081] Once the appropriate elite and ancestral lines have been chosen,
the genotype of each line is determined through the use of genetic
markers. Genetic markers include any qualitative phenotype that can be
used as a direct measure of genotype at a specific locus. Markers include
visual traits such as flower color, enzyme variants such as isozymes, and
molecular markers, which can be detected by a variety of means such as
simple sequence repeats (SSR), single nucleotide polymorphism (SNP),
allele specific hybridization (ASH), restriction fragment length
polymorphism (RFLP), amplified variable sequences, single strand
conformation polymorphism (SSCP), amplified fragment length polymorphism
(AFLP) and randomly amplified polymorphic DNA (RAPD).
[0082] Regardless of which genetic markers are used to monitor genotype,
the end result is a marker genotype for each of the elite and ancestral
lines. The genotype of each line is merely an indication of which allele
the line possesses at any number of loci defined by the genetic markers.
[0083] Once one has determined the genotypes of ancestral and elite lines,
statistical analyses are used to determine whether selection for superior
agronomic performance has favored particular alleles at certain loci. The
first statistic to calculate is the probability of finding each allele
within the elite population with the assumption that selection had no
effect on allele frequency. This expected allele frequency within the
elite population serves as a basis for comparison to the observed allele
frequency.
[0084] Expected allele frequency within the elite population is a function
of the genotype of each ancestor and the pedigrees of elite lines
representing the elite population. In a random mating population, the
allele frequency among descendants should be similar to allele frequency
among ancestors unless breeding and selection has favored particular
alleles. However, since breeding of many crops (including soybeans) is
not done through random mating, one can use the pedigree of each
descendant (e.g., elite line) to calculate the probability of inheriting
a given allele from its ancestors. Within non-random mating populations,
expected allele frequency can be obtained by averaging the individual
probabilities of inheriting an allele over any number of descendants
(that may differ greatly in pedigree).
[0085] By comparing the observed frequency of a given allele in the elite
population to the average probability of inheriting that allele (i.e.,
comparing observed count to expected count), one can determine which loci
have been affected by historical selection for agronomic traits.
Favorable alleles are identified as the ones that have been inherited
more frequently than expected (i.e., have been favored by selection).
Unfavorable alleles are those inherited less frequently than expected
(i.e., selected against). A statistical test is then used, e.g., as
described in detail in U.S. Pat. No. 5,437,697, to determine the
significance of a difference between observed and expected allele
frequency.
[0086] According to these methods, loci and alleles with significant
deviations from expected allele frequency correlating with superior
agronomic performance in different growing environments have been
identified. These loci can be used to define and identify, as well as
select for soybeans with superior agronomic performance. For example, the
markers disclosed herein can be used to 1) identify soybean germplasm
with superior and/or improved agronomic performance, e.g., relative to
presently existing or parental soybean lines; 2) identify parents that
will produce superior transgressive segregants; 3) select superior lines
from crosses that are segregating at loci (e.g., QTL loci) which
contribute to superior agronomic performance; 4) select parents that will
produce the best hybrids; 5) purify heterogeneous lines, i.e., by
selecting only those individuals that include the favorable allele(s) at
loci that are still segregating within the line; 6) select for and
maintain desirable heterogeneity; 7) maintain favorable alleles at
multiple loci that have been assembled by many years of selection while
incorporating exotic alleles from new germplasm at other loci; 8) to test
the effects of exotic allele substitution at loci that have proven
important for domestication, e.g., in the event that an exotic allele
provides better agronomic performance at the loci identified by breeding
bias; and, 9) in any process where it is important to prioritize which
loci are more important for agronomic fitness than random loci within the
genome.
[0087] Markers and Alleles Correlating with Superior Agronomic Performance
in Soybeans
[0088] In one aspect, the present invention provides marker loci
correlated with superior agronomic performance in soybean. Each of the
identified markers is expected to be in close physical and genetic
proximity (i.e., physically and genetically linked) to a genetic element,
e.g., a quantitative trait locus or QTL, that contributes to superior
agronomic performance. If a particular marker were not in proximity to an
important QTL, the extended period of recurrent selection (70+ years for
soybean) would have provided many opportunities for crossing over between
the marker and QTL. This would result in disassociation between the
marker allele and the QTL allele resulting in "linkage equilibrium" and
statistically non-significant estimates of correlation between the marker
and yield, i.e., LOD or probability scores. Breeding bias does not detect
poorly-linked markers, that is, markers that are distant from important
loci, even if both the marker and the genetic element (e.g., QTL)
contributing to yield are found on the same chromosome. In addition,
since thousands of different environments have been sampled to test
performance during each cycle of selection, alleles that provide
adaptation to a wide range of environments will be most readily
identified. Alleles that are only favorable under rare environmental
conditions will not consistently increase in frequency due to selection
in different environments and will, therefore, not be detected by
breeding bias analysis. Furthermore, the narrow gene pool and high
relatedness among elite soybean germplasm shared by both public and
private institutions (Delanney et al, 1983, Crop Science 23:944-949) act
to homogenize the gene pool and make marker-QTL associations more
reliable. Taken together, these features in combination with the large
body of data analyzed by breeding bias, ensure that QTL with a
substantial contribution to yield are located in close proximity to the
markers disclosed herein.
[0089] Accordingly, each of the disclosed markers defines a chromosome
segment associated with a genetic element, e.g., a QTL contributing to
superior agronomic performance. One of skill in the art will recognize
that for each of the chromosome segments encompassing QTL related to
yield, identification of and/or selection for the QTL is optimized by
using a genetic marker that is as close as possible to the actual QTL
locus that is responsible for the phenotype in question. Thus, a
"perfect" marker would be one that is localized within the genetic
element or QTL itself, and corresponds to the DNA polymorphism that is
responsible for the superior phenotype. That is, the marker polymorphism
is the mutation underlying the improvement in yield. However, since most
of the genetic markers available for soybean consist of RFLPs, SSRs,
RAPDs and SNPs of unknown function, it would be highly unlikely that a
given marker from those currently available was already "perfect."
Nevertheless, the markers described herein constitute a set of tools for
detecting important chromosome segments comprising QTL associated with
yield. These markers are sufficiently close to their respective linked
QTL to detect a statistically significant shift in allele frequency (due
to selection) over 70 years of recurrent selection for yield, thus, are
useful for the purposes of identifying desirable germplasm and for MAS.
In addition, these markers are useful for identifying additional markers,
e.g., including a "perfect" marker within the QTL gene of interest.
[0090] Tables 3 through 12 disclose exemplary molecular, which define the
chromosome segments that are embodiments of the present invention. Each
of Tables 3 through 12 provides marker loci and favorable alleles,
identified by Breeding Bias, relevant in a specified growing environment
distinguished by geographic region.
[0091] The exemplary markers provided in Tables 3 through 12 identify
chromosome segments including genetic elements (genes) important for
yield in soybean. The chromosome segments of the present invention are
contiguous lengths of chromosome delimited by a specified crossover
frequency or map distance (centimorgans or CM) from a molecular marker of
the present invention. The chromosome segments of the present invention
are delimited by a crossover frequency of up to about 10%, i.e., 10 CM,
from a marker locus known to be associated with superior agronomic
performance, e.g., Satt165; Satt042; Satt364; Satt454; Satt526; Satt300;
Satt591; Satt155; Satt385; Satt511; P12390B-1; Satt327; Satt329; Satt508;
P10635A-1; Satt409; Satt228; Satt429; Satt509; Satt197; SCT.sub.--026;
Satt415; Satt583; Satt430; P12198A-1; P8584A-1; Satt359; P10648A-1;
P10641A-1; Satt168; Satt556; Satt272; Satt020; Satt534; P10638B-2;
Satt399; Satt361; P10639A-1; Satt190; Satt338; Satt227; Satt457; Satt557;
Satt319; Satt460; Satt307; SCT.sub.--028; Satt433; Satt357; Satt321;
Satt267; Satt295; Satt203; Satt507; SAT.sub.--110; P10620A-1; Satt129;
Satt147; Satt216; P10621B-2; Satt558; Satt266; Satt282; Satt537; Satt506;
P13072A-1; Satt582; Satt389; Satt461; Satt514; Satt464; Satt543;
P13074A-1; P10624A-1; Satt573; Satt598; Satt204; Satt263; Satt491;
Satt602; Satt151; Sat355; Satt452; Satt146; Satt193; Satt569; Satt343;
Satt586; Satt423; Satt348; Satt595; P10782A-1; P3436A-1; Satt334;
Satt510; Satt144; Satt522; P9026A-1; P10646A-1; P5219A-1; P7659A-2;
Satt570; Satt356; Satt130; Satt115; Satt594; Satt533; Satt303; Satt352;
Satt566; Satt199; Satt503; Satt517; Satt191; SAT.sub.--117; Satt353;
Satt442; Satt279; Satt314; Satt181; Satt367; Satt127; Satt270; Satt440;
P10640A-1; Satt249; SCT.sub.--065; Satt596; Satt280; Satt406; Satt380;
Satt183; Satt529; Satt431; Satt242; Satt102; Satt240; P10618A-1; Satt523;
Satt398; Satt497; Satt166; Satt448; Satt373; Satt513; P12394A-1; Satt590;
Satt220; Satt536; Satt175; P10615A-1; Satt250; Satt346; Satt336;
P13069A-1; P5467A-1; Satt584; SAT.sub.--084; Satt387; Satt339; P12396A-1;
Satt487; Satt259; Satt347; Satt420; Satt576; Satt550; Satt262; Satt473;
Satt477; Satt581; P11070A-1; Satt153; Satt243; P8230A-1; P10623A-1;
P10632A-1; P12391A-1; P12392A-1; P13560A-1; P13561A-1; p2481A-1; SAC1677;
Satt040; Satt109; Satt111; Satt176; Satt219; Satt299; and Satt512. The
genetic element contributing to yield can be localized to any portion of
the chromosome segment defined by these molecular markers. For example, a
gene encoding a function leading to improved yield can reside within the
chromosome segment at a distance of less than 1 CM (at a distance
resulting in recombination in fewer than 1% of mitotic events), or at any
distance between about 1 CM and 10 CM, e.g., values of approximately 2
CM, 3 CM, 4 CM, 5 CM, 6 CM, 7 CM, 8 CM, or 9 CM, or any value between
about 0 CM and about 10CM. Thus, a chromosome segment is defined as a
contiguous length of chromosome extending up to 10 CM in either direction
from a designated marker, and including the portion of the chromosome
that undergoes crossover with the marker locus at a frequency of no
greater than 10%. In many cases, the chromosome segment of interest, is
in fact a continuous length extending less than 10 CM, i.e., a length of
chromosome that undergoes crossover with the marker at a frequency of
less than 10%, e.g., 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less, or any
value therebetween. Alternatively, physical distances may be utilized to
determine the aforementioned chromosome segments using a conversion
factor of 1 (one) Mbp (one million base pairs) for 1 (one) CM of map
distance or 1% (one percent) crossover frequency. The actual conversion
factor in soybean ranges from about 100 Kbp to close to 1 Mbp depending
on the chromosome region. Accordingly, the arbitrary but reasonable
conversion factor of 1 Mbp is to be understood in the context of the
present invention.
[0092] The exemplary favorable alleles of marker loci enumerated herein
have been shown, e.g., by breeding bias analysis, to be associated with
genetic elements contributing to improved yield in various lines of
soybeans. One of skill will appreciate, that this is an empirical
association, and that a particular marker locus (and the designated
favorable alleles thereof) are used to identify a corresponding
chromosome segment having a genetic element that contributes to yield.
However, because a marker locus is not necessarily, or even typically,
identical to the genetic element which results in enhanced yield, in some
percentage of crosses, genetic recombination will occur between the
enumerated favorable allele and the genetic element contributing to
yield. This does not negate the importance of either the genetic element
or the identification of the marker locus and exemplary favorable
alleles. Any such recombination events can be detected and subsequent
marker assisted selection can be performed using the allele determined to
be in coupling linkage phase with the favorable genetic element as
discussed in more detail in EXAMPLE 4: SEGREGATION FOR YIELD IN NEAR
ISO-GENIC SUBLINES, in the context of detecting residual polymorphisms
associated with yield in near iso-genic sublines, and in EXAMPLE 5:
ALLELE CONFIRMATION USING RANDOM CROSSES.
[0093] In the context of the present invention, the allelic form of
multiple chromosome segments is determined, whether the purpose is to
define the genotype of a soybean or for marker assisted selection (MAS).
In most circumstances, it is desirable to ascertain the allelic form for
a large proportion, e.g., all, of the chromosome segments known to
contribute to yield in a particular geographic region. However, in some
circumstances, particularly when the parents are known to share
particular favorable alleles, a subset of the chromosome segments can be
employed, reducing time and cost, without losing efficiency. Thus, in the
context of the present invention it is common to assess at least 10%,
typically between at least 10% and about 90% of the relevant chromosome
segments, e.g., by determining the allelic form of the marker loci
specified in Tables 3 through 12. Commonly, at least about 25%,
frequently at least about 50%, often at least about 75% or more of the
allelic forms are determined. For example, at least about 80%, 85%, 90%,
95%, 97%, 98%, 99%. Accordingly, it is generally desirable to determine
the allele at a majority of the loci shown to be relevant to agronomic
performance in a particular geographic region or growing environment. In
one favorable embodiment, the allelic form of a set of markers defining a
set of chromosome segments consisting essentially of the markers shown to
be relevant to agronomic performance in a particular region are
determined. A set of markers is deemed to consist essentially of the
markers relevant in a particular region if it includes a sufficient
number of markers to prevent a diminishing yield phenotype or to prevent
a decrease in efficiency of selection when the set of markers is used for
marker assisted selection. Typically such a set will include at least
about 90%, 95%, 97%, 98%, 99% or more of the markers. That is, no more
than 10% of the markers shown to be relevant in a particular region will
be omitted, e.g., no more than 5%, 3%, 2%, or 1% of the allelic forms of
the relevant markers will be left undetermined.
[0094] Typically, the marker loci evaluated are shown to correlate with
superior agronomic performance with a significance of greater that 95%.
Often the marker loci are selected that exhibit a significance of greater
than 99% within a geographic region of interest. If desired, the allelic
form of multiple marker loci within the same chromosome segment can be
determined. Frequently, the allelic form of a single marker representing
the chromosome segment is determined.
[0095] Identification of Residual Polymorphism
[0096] The marker loci identified by Breeding Bias are associated with
genetic elements contributing to increased yield in soybean. Although
particular alleles of the marker loci have been found to be statistically
correlated with increased yield according to the Breeding Bias analysis,
alleles at one or more marker loci are typically not fixed within an
elite line or among progeny derived from a progenitor soybean selected
from an elite strain of germplasm. Indeed, such residual polymorphisms
provide valuable genetic variation from which improved sublines can be
selected. By identifying particular marker loci from among the set of
loci shown to be associated with genetic elements contributing to yield,
the efficiency of selection can be improved.
[0097] To this end, marker loci with one or more segregating alleles in a
population of progeny derived from an elite progenitor are evaluated to
identify the specific allele correlating with increased yield in the
population of progeny. Detailed descriptions of exemplary methods for
confirming the favorable allele in a population of progeny are provided
in EXAMPLES 4 and 5.
[0098] Definition and Identification of Target Germplasm
[0099] The marker loci of the present invention can be used as proxies for
loci contributing to improved yield to define the theoretically optimal
or "target" soybean genotype. Accordingly, alleles of marker loci
identified by Breeding Bias, and confirmed to correlate with increased
yield in a given environmental or geographic context can be used to
define or identify a soybean plant with superior agronomic performance.
While existing elite lines include favorable alleles at numerous marker
loci (and corresponding functional loci contributing to yield), none of
the existing elite lines yet incorporates all of the genetic elements
contributing to the ideal soybean genotype. Indeed, prior to the markers
of the present invention, it would not have been possible to predict the
ideal soybean genotype (or define a target genotype) with respect to
yield. Thus, a feature of the invention is the definition of a target
soybean genotype for superior agronomic performance. Any soybean plant
having an increased number of favorable allelic forms of chromosome
segments defined according to the markers of the invention, i.e., that
more closely approximates the target soybean genotype than existing elite
lines, constitutes a feature of the present invention.
[0100] The markers and methods described herein provide the means for
identifying and developing soybean plants having the target soybean
genotype for superior agronomic performance and genomes having increased
numbers of favorable allelic forms of the relevant chromosome segments
relative to existing elite lines, and or relative to either parent giving
rise to the soybean plant genome. Accordingly, the methods of the
invention can be used to produce compositions, including whole plants,
plant organs, seeds, and isolated genetic constituents, e.g., including
the entire chromosomal complement of the genome, or a subset thereof,
such as an individual chromosome or chromosome fragment (all of which are
collectively described by the term "soybean plant genome") with an
increased number of genetic elements contributing to yield. Methods for
separating and isolating genomes or individual chromosomes are well known
in the art and include, e.g., flow cytometry and pulse field gel
electrophoresis. Replicates of the soybean plant genome are also
encompassed within the meaning of the term soybean plant genome.
Replicates are identical or substantially identical (i.e., a mutant
arising from mitotic division from the same progenitor cell) to the
initial soybean plant genome. Replicates can be created, for example, by
yeast or bacterial artificial chromosomes or any of a variety of nucleic
acid vectors or replication methods such as PCR (polymerase chain
reaction).
[0101] A soybean plant genome(s) of the present invention can be from an
individual soybean plant. As used herein, the term "plant" includes whole
plants, plant organs (e.g., leaves, stems, roots, etc.), seeds,
suspension cultures, embryos, meristematic regions, callus tissue,
leaves, roots, shoots, gametophytes, sporophytes, pollen, and
microspores.
[0102] Marker Assisted Selection and Breeding
[0103] The ultimate goal of any breeding program is to combine as many
favorable alleles as possible into elite varieties of germplasm that are
genetically superior (with respect to one or more agronomic traits) to
their ancestors. The markers provided herein identify chromosome
segments, i.e., genomic regions, and alleles (allelic forms) that have
been favored by long-term selection for yield. Accordingly, these markers
can be used for marker assisted selection of soybean plants with superior
agronomic performance. For example, in a cross between parents that
complement favorable alleles at the target loci, progeny can be selected
that include more favorable alleles than either parent. Such progeny are
predicted to be phenotypically superior to either parent as illustrated
in Table 2.
[0104] Marker assisted selection (MAS), employing the markers of the
present invention, and the chromosome segments they identify are useful
in the context of a soybean breeding program to increase efficiency in
yield improvements. Phenotypic screening for a trait of interest, such as
yield, for large numbers of samples can be expensive, as well as time
consuming. In addition, phenotypic screening alone is often unreliable
due to the effects of epistasis and non-genetic (e.g., environmental)
contributions to the phenotype. MAS offers the advantage over field
evaluation that it can be performed at any time of year regardless of the
growing season or developmental stage. In addition, MAS facilitates
evaluation of organisms grown in disparate regions or under different
conditions.
[0105] A breeder of ordinary skill, desiring to breed soybean plants with
increased yield, can apply the methods for MAS described herein, using,
e.g., the exemplary markers provided herein or linked markers localized
to the chromosome segments identified by markers provided in Tables 3
through 12, to derive soybean lines with superior agronomic performance.
[0106] Genetic marker alleles, e.g., the exemplary markers provided Tables
3 through 12, linked markers, QTL, identifying the chromosome segments
encompassing genetic elements that are important for yield, are used to
identify plants that contain a desired genotype at one or more loci, and
that are expected to transfer the desired genotype, along with a desired
phenotype to their progeny. Marker alleles (or QTL alleles) can be used
to identify plants that contain a desired genotype at one locus, or at
several unlinked or linked loci (e.g., a haplotype), and that would be
expected to transfer the desired genotype, along with a desired phenotype
to their progeny. Similarly, by identifying plants lacking the desired
allele, plants with an undesirable phenotype, e.g., plants with poor
yield, can be identified, and, e.g., eliminated from subsequent crosses.
It will be appreciated that for the purposes of MAS, the term marker can
encompass both marker and QTL loci as both can be used to identify plants
with a desired phenotype.
[0107] For example, MAS can be used to develop lines or strains of soybean
and soybean germplasm with superior agronomic performance by identifying
favorable allelic forms of chromosome segments shown to be important,
e.g., that include a genetic element, for yield. Favorable alleles of
markers defining the chromosome segments of interest, e.g., Satt684,
Satt165, Satt042, Satt364, Satt454, Satt526, Satt300, Satt591, Satt155,
Satt385, Satt385, Satt225, Satt236, Satt511, P12390B-1, Satt480,
Satt632-TB, Satt233, Satt327, Satt329, Satt508, P10635A-1, Satt409,
Satt228, Satt429, Satt426, Satt509, SAT.sub.--261, Satt197, Satt519,
Satt597, SCT.sub.--026, Satt415, Satt583, Satt430, P12198A-1, P8584A-1,
Satt359, P10648A-1, P12105A-1, P10641A-1, Satt168, Satt556, Satt272,
Satt020, Satt066, Satt534, P10638B-2, Satt399, Satt361, P10639A-1,
Satt661-TB, Satt190, SAT.sub.--311-DB, Satt338, Satt227, Satt640-TB,
Satt422, Satt457, Satt457, Satt557, Satt319, SAT.sub.--142-DB, Satt460,
P13073A-1, Satt307, SCT.sub.--028, Satt433, Satt357, Satt321, Satt267,
Satt383, Satt295, Satt203, Satt507, SAT.sub.--110, P10620A-1, Satt129,
Satt147, Satt216, SAT.sub.--351, P10621B-2, Satt701, Satt634, Satt558,
Satt266, Satt282, Satt537, Satt506, Satt546, P13072A-1, Satt582, Satt389,
Satt461, Satt311, Satt514, Satt464, Satt662, Satt543, Satt186, Satt413,
Satt672, P13074A-1, P10624A-1, Satt573, Satt598, Satt204, Satt263,
Satt491, Satt602, Satt151, Satt355, Satt452, SAT.sub.--273-DB, Satt146,
Satt193, Satt569, Satt343, Satt586, Satt040, Satt423, Satt348, Satt595,
P10782A-1, P3436A-1, P10598A-1, Satt334, Satt510, Satt510, Satt144,
Satt522, Satt522, P9026A-1, P10646A-1, P5219A-1, P7659A-2, Satt570,
Satt356, Satt130, Satt115, Satt594, Satt533, Satt303, Satt352, Satt566,
Satt199, Satt503, Satt517, Satt191, SAT.sub.--117, Satt353, Satt442,
Satt279, Satt314, Satt142, Satt181, Satt367, Satt127, SCTT012, Satt270,
Satt292, Satt440, P10640A-1, Satt249, SAG1223, SAC1699, SCT.sub.--065,
Satt596, Satt280, Satt406, Satt380, Satt183, Satt529, Satt431, Satt242,
Satt102, Satt441, Satt544, Satt617, Satt240, P10618A-1, Satt475, Satt196,
SAT.sub.--301, Satt523, Satt418, Satt418, Satt398, Satt497, Satt284,
Satt166, Satt448, Satt373, Satt513, P12394A-1, Satt590, Satt567, Satt220,
SAG1048, Satt536, Satt175, Satt677, Satt680, P10615A-1, Satt551, Satt250,
Satt346, Satt336, SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2,
Satt584, SAT.sub.--084, P3050A-2, SAT.sub.--275-DB, Satt387, Satt549,
Satt660, Satt339, Satt255, Satt257, Satt358, P12396A-1, Satt487, Satt259,
Satt259, Satt347, Satt420, Satt576, Satt550, Satt633, Satt262, Satt473,
Satt477, Satt581, P11070A-1, Satt153, Satt243, P8230A-1, P10623A-1,
P10632A-1, P10793A-1, P12391A-1, P13560A-1, P13561A-1, P13561A-1,
P2481A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB, S60149-TB,
S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB, S60361-TB,
S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB, S60519-TB,
S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB, S60812-TB,
SAC1677, SAC1724, SAG1055, Satt040, Satt108, Satt109, Satt111, Satt176,
Satt176, Satt219, Satt299, and Satt512 as enumerated Tables 3 through 12
are detected in a genomic sample of a soybean plant.
[0108] For example, in a breeding program designed to produce soybeans
with increased yield in the Central United States growing region (e.g.,
exemplified by the growing conditions in Iowa), markers can be selected
from the following set of markers: Satt642, Satt042, Satt364, Satt454,
Satt526, Satt300, Satt591, Satt155, Satt385, Satt511, P12390B-1,
Satt632-TB, Satt429, SAT.sub.--261, Satt197, P10641A-1. Satt556, Satt534,
P10638B-2, Satt399. Satt361, P10639A-1, Satt661-TB, Satt190,
SAT.sub.--311-DB, Satt338, Satt640-TB, Satt557, Satt319,
SAT.sub.--142-DB, Satt460, Satt433, Satt357, Satt321, Satt295, Satt203,
Satt507, Satt129, Satt147, SAT.sub.--351, P10621B-2, Satt558, Satt701,
Satt634, Satt582, Satt389, Satt464, Satt662, Satt672, Satt573, Satt598,
Satt263, Satt602, Satt151, SAT.sub.--273-DB, Satt146, Satt193, Satt569,
Satt176, Satt343, Satt586, Satt040, Satt595, P10782A-1, Satt334, Satt144,
Satt522, Satt570, Satt356, Satt533, Satt199, Satt517, Satt191,
SAT.sub.--117, Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223,
SAC1699, SAT.sub.--065, Satt596, Satt406, Satt380, Satt183, Satt529,
Satt242, Satt617, Satt240, SAT.sub.--301, Satt418, Satt398, Satt497,
Satt166, Satt448, Satt373, Satt513, P12394A-1, SAG1048, Satt536, Satt175,
Satt677, Satt680, P10615A-1, Satt551, Satt346, Satt336, SAT.sub.--330-DB,
P13069A-1, P5467A-1, P5467A-2, SAT.sub.--084, SAT.sub.--275-DB, Satt660,
Satt339, P12396A-1, Satt358, Satt487, Satt259, Satt420, Satt576, Satt633,
Satt477, Satt581, Satt153, Satt243, P10793A-1, P12391A-1, P12392A-1,
P13560A-1, P13561A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB,
S60149-TB, S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB,
S60361-TB, S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB,
S60519-TB, S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB,
S60812-TB, SAC1677, SAC1724, SAG1055, Satt040, Satt111, Satt176, Satt219
and Satt299, such as: Satt684, Satt042, Satt364, Satt454, Satt526,
Satt300, Satt591, Satt155, Satt385, Satt632-TB, Satt429, SAT-.sub.--261,
P10641A-1, Satt556, P10638B-2, Satt399, Satt361, Satt661-TB, Satt190,
SAT.sub.--311-DB, Satt338, Satt640-TB, Satt557, Satt319,
SAT.sub.--142-DB, Satt321, Satt203, Satt129, Satt147, SAT.sub.--351,
P10621B-2, Satt701, Satt634, Satt582, Satt389, Satt464, Satt662, Satt672,
Satt573, Satt598, Satt263, Satt151, SAT.sub.--273-DB, Satt146, Satt193,
Satt569, Satt343, Satt586, Satt040, Satt595, Satt334, Satt144, Satt522,
Satt570, Satt356, Satt199, Satt517, Satt191, Sat.sub.--117, Satt279,
Satt181, Satt127, Satt270, Satt292, SAG1223, SAC1699, Sat.sub.--065,
Satt596, Satt406, Satt380, Satt183, Satt529, Satt242, Satt617, Satt240,
SAT.sub.--301, Satt418, Satt398, Satt497, Satt166, Satt448, Satt373,
Satt513, P12394A-1, SAG1048, Satt536, Satt175, Satt677, Satt680,
P10615A-1, Satt551, SAT.sub.--330-DB, P13069A-1, P5467A-1, P5467A-2,
SAT.sub.--084, SAT.sub.--275-DB, Satt660, Satt339, Satt358, Satt487,
Satt487, Satt420, Satt576, Satt633, Satt581, Satt153, Satt243, P10793A-1,
P13560A-1, P13561A-1, S60021-TB, S60048-TB, S60076-TB, S60148-TB,
S60149-TB, S60201-TB, S60243-TB, S60326-TB, S60338-TB, S60350-TB,
S60361-TB, S60422-TB, S60440-TB, S60446-TB, S60505-TB, S60513-TB,
S60519-TB, S60536-TB, S60552-TB, S60585-TB, S60630-TB, S60728-TB,
S60812-TB, SAC1677, SAC1724, SAG1055, Satt111, Satt219 and Satt299. The
following exemplary markers represent the best markers for selection for
yield in the Central Region in each chromosome region: Satt684, Satt526,
Satt591, Satt385, Satt632-TB, Satt429, SAT-.sub.--261, P10641A-1,
Satt556, P10638B-2, Satt190, SAT.sub.--311-DB, Satt338, Satt640-TB,
Satt557, SAT.sub.--142-DB, Satt321, Satt203, Satt129, SAT.sub.--351,
Satt701, Satt582, Satt389, Satt464, Satt672, Satt598, Satt343, Satt595,
Satt334, Satt144, Satt522, Satt570, Satt356, Satt199, Sat.sub.--117,
Satt279, Satt181, Satt127, Satt270, Satt292, SAG1223, Sat.sub.--065,
Satt529, Satt242, Satt617, SAT.sub.--301, Satt398, Satt497, Satt166,
Satt373, SAG1048, Satt680, P10615A-1, SAT.sub.--330-DB, P13069A-1,
SAT.sub.--275-DB, Satt339, Satt487, Satt420, Satt581 and Satt153.
Comparable lists of markers can be compiled from Tables 3 through 12 at
the discretion of the practitioner based on the desired growing region.
[0109] A soybean plant so identified can be utilized in a plant breeding
program to develop lines with improved yield. Similarly, the detection of
favorable (or conversely, non-favorable) allelic forms of the chromosome
segments can be used to trace the flow of alleles in a soybean plant
pedigree to ensure that the desired complement of alleles are included or
excluded in the resulting soybean plant(s).
[0110] After a desired phenotype and a polymorphic chromosomal locus,
e.g., a marker locus or QTL, are determined to segregate together (i.e.,
are determined to be in linkage disequilibrium), alleles corresponding to
the desired phenotype are selected. In brief, a nucleic acid
corresponding to the marker nucleic acid is detected in a biological
sample from a plant to be selected. This detection can take the from of
hybridization of a probe nucleic acid to a marker, e.g., using
allele-specific hybridization, Southern analysis, northern analysis, in
situ hybridization, hybridization of primers followed by PCR
amplification of a product including the marker, or the like. A variety
of procedures for detecting markers are described herein, e.g., in the
section entitled "DETECTION OF MARKER LOCI." After the presence (or
absence) of a particular marker in the biological sample is verified, the
plant is selected and, optionally, crossed to produce progeny plants.
[0111] When a population is segregating for multiple loci affecting one or
multiple traits, e.g., multiple loci involved in resistance to single
disease, or multiple loci each involved in resistance to different
diseases, the efficiency of MAS compared to phenotypic screening becomes
even greater because all the loci can be processed in the lab together
from a single sample of DNA. Thus, use of marker information for each of
the traits in the breeding process is facilitated.
[0112] It will be appreciated that plants positive for a marker of the
invention can be selected and crossed according to any breeding protocol
relevant to the particular breeding program. Accordingly, progeny can be
generated from a selected plant by crossing the selected plant to one or
more additional plants selected on the basis of the same marker or a
different marker, e.g., a different marker correlating with superior
agronomic performance, or a different phentoype of interest, e.g.,
resistance to a particular disease. Alternatively, a selected plant can
be back crossed to one or both parents. Backcrossing is usually done for
the purpose of introgressing one or a few loci from a donor parent, e.g.,
a donor parent comprising exotic germplasm, into an otherwise desirable
genetic background from the recurrent (typically, an elite) parent. The
more cycles of backcrossing that are performed, the greater the genetic
contribution of the recurrent parent to the resulting variety. A selected
plant can also be outcrossed, e.g., to a plant or line not present in its
genealogy. Such a plant can be selected from among a population subject
to a prior round of analysis, or may be introduced into the breeding
program de novo. A plant positive for a desired marker can also be
self-crossed ("selfed") to create a true breeding line with the same
genotype.
[0113] In some instances, even if a marker is close enough to a QTL to
detect breeding bias, the marker may not be close enough for reliable
MAS. If such a marker is far enough away from the QTL of interest, there
may be crossing over between the marker and the QTL leading to repulsion
phase linkages (in the elite population) between the marker allele that
was originally linked in coupling to the favorable QTL allele. With time,
the marker locus and the linked QTL locus could reach "linkage
equilibrium" and this will prevent the use of that marker for reliable
selection of the favorable QTL allele. For highly heritable traits,
linkage phase between marker and QTL in any given parent can be easily
determined through MAS followed by phenotypic characterization. However,
linkage phase determination for traits of low heritability (e.g., yield)
is much more difficult. In fact, the effects of single loci on yield may
be extremely difficult to measure even with highly replicated field
tests. In addition, if yield genes were highly heritable, the continuous
selection for this trait would have "fixed" all of the favorable alleles
quickly and yield progress would have previously reached a plateau. Since
steady yield progress continues in soybean, it does not appear that a
"yield plateau" has been reached yet (Specht et al., 1999, Crop Science
39:1560-1570). Therefore, many of the favorable alleles at the major
yield loci in soybean are not yet fixed within the elite population. The
question remains whether the existing marker loci are still in original
linkage phase with the QTL loci. Fortunately, the results of breeding
bias disclosed here can be used to solve the problem of "imperfect" yield
gene markers several ways: a) linkage phase and QTL effect determination
within specific crosses prior to MAS, b) use of flanking markers to
predict which markers are still linked in coupling, and c) alternating
cycles of MAS and phenotypic selection.
[0114] Linkage Phase and OTL Effect Determination within Specific Crosses:
[0115] Because elite soybean lines are highly related, a relatively small
set of elite lines contains most of the favorable alleles that exist
within the entire elite population. Therefore, determining the linkage
phase between marker and QTL alleles at the loci identified as important
for yield can be accomplished to further increase efficiency of MAS in
the context of a soybean breeding program. Progeny from parents that are
both high yielding and polymorphic for many of the target marker loci are
assessed for yield in a small number of field locations. Using orthoganol
comparisons, one locus at a time, predictions concerning correlations
between marker alleles and phenotype can be made. For example if 40
random homozygous progeny from a cross that is segregating at 10 of the
target loci are field tested, on average, 20 of the progeny will be
homozygous for one of the parental alleles and 20 will be homozygous for
the other parental allele. One can then pool the replicated yield data
for lines containing the same marker allele and determine if the yield of
said group is statistically different from the yield of lines containing
the alternate marker allele. If so, then this marker should be effective
for selection within that cross. This comparison is then done separately
for each of the 10 segregating marker loci to determine which set of
those 10 markers should be effective for selection within that cross.
[0116] Optionally, flanking markers can be used to predict which markers
are linked in coupling. By comparing the genotype of flanking markers
that are linked to the target marker in both elite lines and ancestors,
one can predict which haplotypes have been most conserved during
selection over many cycles. In the event that recombination has occurred
between the target marker and the genetic element contributing to yield,
such that the desired genetic element and the linked target marker locus
are no longer in coupling linkage phase, flanking markers can be utilized
to identify progeny with superior agronomic performance.
[0117] In addition, MAS and phenotypic selection can be alternated to
insure that the allele in coupling phase is detected. Markers identified
by breeding bias are employed for MAS as described above (with or without
the advantages of linkage phase determination within specific crosses)
and replicated yield testing is performed on selected progeny. If enough
replicates and environments are sampled, a reasonable measure of yield
phenotype can be obtained. Progeny that are confirmed as high yielding
can be used as parents in the next cycle of MAS selection. By screening
related populations according to this method, the population will move
toward fixation of the favorable QTL alleles even if the favorable marker
allele is not always in coupling with the QTL allele. Alternatively,
residual polymorphisms at the marker loci described herein can be
detected in near iso-genic lines, and the marker allele in corresponding
to increased yield can be validated.
[0118] Introgression of Flavorable Alleles
[0119] More Efficient Backcrossing of Specific Genes into Elite Lines
[0120] One application of MAS, in the context of the present invention is
to use the "yield gene" markers to increase the efficiency of a
introgression or backcrossing effort. In typical marker assisted
backcrossing of a specific gene(s) from a donor source to an elite
genetic background, one selects among backcross progeny for the donor
trait and then uses markers to reconstitute as much of the elite
background's genome as possible. Prior to the present invention, the
markers used to identify the elite background were of unknown function,
and many of the markers commonly used may be selecting for parts of the
elite genome that do not actually contribute to high yield. Similarly,
prior to the present invention, the major loci that contribute to yield
were largely unknown, so the entire elite genome of the recurrent parent
was selected for with the hopes of including all of the favorable alleles
that it contained. However, the markers identified by breeding bias can
be used to identify only those parts of the elite genome that are most
significant with respect to yield. These markers can be used to
concentrate backcrossing efforts on the most important parts of the elite
genome. The fewer markers needed, the higher the probability of
recapturing the elite phenotype quickly.
[0121] Thus, the markers and methods of the present invention can be
utilized to guide marker assisted selection or breeding of soybean
varieties with the desired complement (i.e., set) of allelic forms of
chromosome segments associated with superior agronomic performance. Each
of the disclosed alleles can be introduced into a soybean line via
introgression, i.e., by means of traditional breeding (or introduced via
transformation, or both) to yield a soybean plant with superior agronomic
performance. The number of alleles associated with superior agronomic
performance that can be introduced or be present in a soybean plant of
the present invention ranges from 1 to the number of alleles disclosed
herein, each integer of which is incorporated herein as if explicitly
recited.
[0122] Exemplary soybean lines including at least one (and typically
several or many) of the favorable allelic forms of the relevant
chromosome segments are provided in Table 1. Without intent to limit the
invention, these include the elite soybean lines: 90A07, 90B11, 90B31,
90B43, 90B72, 90B73, 91B01, 91B12, 91B33, 91B52, 91B53, 91B64, 91B91,
91B92, 92B05, 92B12, 92B23, 92B38, 92B63, 92B74, 92B75, 92B84, 92B95,
93B01, 93B11, 93B15, 93B25, 93B26, 93B41, 93B45, 93B46, 93B66, 93B67,
93B72, 93B82, 93B84, 93B85, 93B86, 93B87, 94B01, 94B23, 94B24, 94B53,
94B54, 94B73, 95B32, 95B33, 95B34, 95B53, 95B95, 95B96, 95B97, 96B21,
96B51, 97B52, 97B61, A1395, A2835, A2943, A3127, A3242, A3431, A4009,
A4138, A4415, A4595, A4715, A5403, A5560, A5843, A5885, A5979, A5980,
A6297, BEDFORD, CM428, CX105, CX232, CX253, CX289, CX394C, CX469C,
D00566D362, ESSEX, EX04C00, EX06A00, EX10F01, EX13P01, EX13Q01, EX15N01,
EX16N00, EX16P01, EX22Y01, EX22Z01, EX39E00, FORREST, G3362, HS93-4118,
HUTCHESON, JIM, KORADA, M015733, MO400644-02, MO413735-11-52,
MO501577-27-23, MO505469-61-89, MP39009, P1677, P9007, P9008, P9041,
P9042, P9061, P9062, P9063, P9071, P9092, P9132, P9151, P9163, P9182,
P9203, P9233, P9244, P9273, P9281, P9305, P9306, P9321, P9392, P9395,
P9481, P9482, P9492, P9521, P9552, P9561, P9584, P9591, P9592, P9631,
P9641, PHARAOH, RA451, R01154R002, S0066, S03W4, S0880, S1550, S1990,
S19T9, S20F8, S22C3, S24L2, S25J5, S32Z3, S33N1, S38T2488, S43B5, S5960,
S6189, S6262, ST0653, ST1073, ST1090, ST1970, ST2250, ST2488, ST2660,
ST2688, ST2870, ST3171, ST3380, ST3630, ST3870, ST3883, TRACY, TRAILL,
X9916, YB03E00, XB03F01, XB07E01, XB10D01, XB15M01, XB20M01, XB22R01,
XB25W01, XB31C01, XB33B, XB34F01, XB35D, XB35W00, XB38A01, XB41M01,
XB42J00, XB42M01, XB48H01, XB54K01, XB55J01, XB58P99, XB63D00, XB67A00,
YB03G01, YB08D01, YB09F01, YB09G01, YB10E01, YB11D0, YB14H01, YB15K99,
YB21F01, YB21G01, YB22S00, YB22V01, YB22W01, YB22X01, YB24Z01, YB25R99,
YB25X00, YB25Y01, YB25Z01, YB27X01, YB27Y01, YB28N01, YB29H01, YB29J01,
YB30J01, YB30N01, YB30P01, YB31E01, YB32K01, YB33K01, YB43H01, YB35C01,
YB36V00, YB39M01, YB40M01, YB40N01, YB41Q01, YB48L01, YB52J00, YB53E00,
YB54H00, YB54J00, YB54L00, YB55H00, YB56E00, YB60N01, and YOUNG. These
lines and progeny derived therefrom, as well as numerous additional elite
lines, are conveniently utilized as breeding material to develop novel
lines with increased numbers of favorable allelic forms of chromosome
segments involved in yield.
[0123] The present invention also extends to a method of making a progeny
soybean plant and these progeny soybean plants, per se. The method
comprises crossing a first parent soybean plant with a second soybean
plant and growing the female soybean plant under plant growth conditions
to yield soybean plant progeny. Methods of crossing and growing soybean
plants are well within the ability of those of ordinary skill in the art.
Such soybean plant progeny can be assayed for the alleles associated with
superior agronomic performance and, thereby, the desired progeny
selected. Such progeny plants or seed can be sold commercially for
soybean production, used for food, processed to obtain a desired
constituent of the soybean, or further utilized in subsequent rounds of
breeding. At least one of the first or second soybean plants is a soybean
plant of the present invention in that it comprises at least one of the
allelic forms of the present invention such that the progeny are capable
of inheriting the allele. Conveniently, the first or second soybean plant
line can be one of the elite lines of Table 1, or a derivative of such a
line (i.e., a descendant or progenitor in that line's pedigree), or any
relative of these elite lines (such as any elite line that was derived
from the ancestors of these elite lines) that retains the same allelic
form as that associated with superior agronomic performance. However, it
will readily be recognized by one of skill in the art that following
characterization essentially any elite line of soybean can be utilized.
[0124] Often, a method of the present invention is applied to at least one
related soybean plant such as from progenitor or descendant lines in the
subject soybean plants pedigree such that inheritance of the desired
allele can be traced. The number of generations separating the soybean
plants being subject to the method of the present invention will
generally be from 1 to 20, commonly 1 to 5, and typically 1, 2, or 3
generations of separation, and quite often a direct descendant or parent
of the soybean plant will be subject to the method (i.e., 1 generation of
separation).
[0125] Incorporation of "Exotic" Germplasm while Maintaining Historical
Progress
[0126] Genetic diversity is important for long term genetic gain in any
breeding program. With limited diversity, genetic gain will eventually
plateau when all of the favorable alleles have been fixed within the
elite population. The challenge is to incorporate diversity into the
elite pool without losing the genetic gain that has already been made and
with the minimum possible investment. Breeding bias results provide an
indication of which genomic regions and which favorable alleles from the
original ancestors have been selected for and conserved over time,
facilitating efforts to incorporate favorable variation from exotic
germplasm sources (parents that are unrelated to the elite gene pool) in
the hopes of finding favorable alleles that do not currently exist in the
elite gene pool.
[0127] For example, the markers of the present invention can be used for
MAS in crosses involving elite x exotic soybean lines by subjecting the
segregating progeny to MAS to maintain the major yield alleles that have
already been "fixed" by decades of selection and leave the rest of the
genome open for contribution from the exotic sources. This would be a
much more efficient system than conventional selection or MAS selection
of elite alleles for which we have no prior information.
[0128] If the donor parent has polymorphic alleles at the elite target
loci as well, the breeder can also relax the backcrossing selection
intensity to allow variation at these loci to slip through. This provides
the opportunity to see if an exotic line has something even more
favorable than what was in the elite gene pool at the elite target loci.
[0129] The methods of the present invention also address another
limitation of conventional backcrossing: that is, as the recurrent parent
is reconstituted with increased cycles of backcrossing, potentially
favorable alleles from the donor parent are excluded along with the
unfavorable alleles from the donor parent. By selectively reconstituting
the recurrent parent's genotype at the loci that have been shown by
breeding bias to be important, one allows favorable alleles to be
introduced from the donor parent at other loci. This allows for the donor
parent to contribute exotic favorable alleles at loci that are not part
of the elite "target genotype." This increases the chances of
transgressive segregation while still retaining the most critical parts
of the recurrent parent's genome. If the donor parent has polymorphic
alleles at the elite target loci as well, the breeder can also relax the
backcrossing selection intensity to allow variation at these loci to slip
through and be tested in the context of a genome that is representative
of the elite gene pool.
[0130] Detection of Marker Loci
[0131] Tables 3 through 12 provide a set of markers and favorable alleles
associated with superior agronomic performance in a variety of geographic
regions with a range of different growing environments. Each of the
markers identifies a chromosome segment that includes one or more genetic
elements, i.e., genes, that influences yield in soybeans. One of skill in
the art will appreciate that the markers provided and discussed herein
are merely exemplary and that numerous other linked markers can be
identified based on genetic linkage and/or physical proximity on a
chromosome to the markers provided herein. Thus, the compositions and
methods of the present invention described herein are not intended to be
limited only to the markers provided in Tables 3 through 12, but also
include additional markers linked thereto. Additionally, while favorable
alleles of the exemplary marker loci are disclosed herein, it will be
readily appreciated by those of skill in the art, that favorable alleles
of additional loci linked to the marker loci described herein can be
determined without undue experimentation and employed in the compositions
and methods of the present invention. Accordingly, any marker locus
linked to the markers described herein, and localized to a chromosome
segment identified by the markers of the invention, can also be used to
identify that chromosome segment, and to define the genotype of a soybean
plant, or to select for favorable allelic forms of a chromosome segment
correlated with superior agronomic performance.
[0132] Although the specific DNA sequences which encode proteins are
generally well-conserved across a species, regions of DNA which are
non-coding, or which encode proteins or portions of proteins which lack
critical function, tend to accumulate mutations, and therefore, are
variable between members of the same species. Such regions provide the
basis for numerous molecular genetic markers. Markers identify
alterations in the genome, which can be insertions, deletions, point
mutations, recombination events, or the presence and sequence of
transposable elements. Many molecular or genetic markers have been
characterized in plant species of interest, including soybean, and are
known to those of skill in the art. For example, a collection of genetic
markers for soybean is publicly available from Linkage Genetics (151 West
2200 South, Suite C, Salt Lake City, Utah 84119, 801-975-1188).
[0133] Molecular markers can be detected by numerous methods,
well-established in the art (e.g., allele specific hybridization (ASH) or
other methods for detecting single nucleotide polymorphisms (SNP),
amplified fragment length polymorphisms (AFLP), amplified variable
sequences, randomly amplified polymorphic DNA (RAPD), restriction
fragment length polymorphisms (RFLP), self-sustained sequence
replication, simple sequence repeat (SSR), single-strand conformation
polymorphisms (SSCP), and isozyme markers). While the exemplary markers
provided in Tables 3 through 12 are either SSR or SNP (ASH) markers, any
of the aforementioned marker types can be employed in the context of the
invention to identify chromosome segments encompassing genetic element
that contribute to superior agronomic performance.
[0134] The majority of genetic markers rely on one or more property of
nucleic acids for their detection. For example, some techniques for
detecting genetic markers utilize hybridization of a probe nucleic acid
to nucleic acids corresponding to the genetic marker. Hybridization
formats including but not limited to, solution phase, solid phase, mixed
phase, or in situ hybridization assays. Among the earliest markers
detected, restriction fragment length polymorphisms (RFLP), are detected
by hybridizing a probe which is typically a sub-fragment (or a synthetic
oligonucleotide corresponding to a sub-fragment) of the nucleic acid to
be detected to restriction digested genomic DNA. The restriction enzyme
is selected to provide restriction fragments of at least two alternative
(or polymorphic) lengths in different individuals, and will often vary
from line to line. Determining a (one or more) restriction enzyme that
produces informative fragments for each cross is a simple procedure, well
known in the art. After separation by length in an appropriate matrix
(e.g., agarose) and transfer to a membrane (e.g., nitrocellulose, nylon),
the labeled probe is hybridized under conditions which result in
equilibrium binding of the probe to the target followed by removal of
excess probe by washing.
[0135] Nucleic acid probes to the marker loci can be cloned and/or
synthesized. Detectable labels suitable for use with nucleic acid probes
include any composition detectable by spectroscopic, radioisotopic,
photochemical, biochemical, immunochemical, electrical, optical or
chemical means. Useful labels include biotin for staining with labeled
streptavidin conjugate, magnetic beads, fluorescent dyes, radiolabels,
enzymes, and colorimetric labels. Other labels include ligands that bind
to antibodies labeled with fluorophores, chemiluminescent agents, and
enzymes. Labeling markers is readily achieved such as by the use of
labeled PCR primers to marker loci.
[0136] The hybridized probe is then detected using, most typically by
autoradiography or other similar detection technique (e.g., fluorography,
liquid scintillation counter, etc.). Examples of specific hybridization
protocols are widely available in the art, see, e.g., Berger, Sambrook,
Ausubel, cited in the section entitled "GENERAL MOLECULAR BIOLOGY
REFERENCES."
[0137] More specifically with respect to certain of the exemplary markers
of the present invention, Allele-specific hybridization (ASH) technology
is based on the stable annealing of a short, single-stranded,
oligonucleotide probe to a completely complementary single-strand target
nucleic acid. Detection is via an isotopic or non-isotopic label attached
to the probe.
[0138] For each polymorphism, two or more different ASH probes are
designed to have identical DNA sequences except at the polymorphic
nucleotides of markers comprising a single nucleotide polymorphism (SNP).
Each probe will have exact homology with one allele sequence so that the
range of probes can distinguish all the known alternative allele
sequences. Each probe is hybridized to the target DNA. With appropriate
probe design and hybridization conditions, a single-base mismatch between
the probe and target DNA will prevent hybridization. In this manner, only
one of the alternative probes will hybridize to a target sample that is
homozygous or homogenous for an allele. Samples that are heterozygous or
heterogeneous for two alleles will hybridize to both of two alternative
probes.
[0139] ASH markers are used as dominant markers where the presence or
absence of only one allele is determined from hybridization or lack of
hybridization by only one probe. The alternative allele may be inferred
from the lack of hybridization. ASH probe and target molecules are
optionally RNA or DNA; the target molecules are any length of nucleotides
beyond the sequence that is complementary to the probe; the probe is
designed to hybridize with either strand of a DNA target; the probe
ranges in size to conform to variously stringent hybridization
conditions, etc.
[0140] PCR allows the target sequence for ASH to be amplified from low
concentrations of nucleic acid in relatively small volumes. Otherwise,
the target sequence from genomic DNA is digested with a restriction
endonuclease and size separated by gel electrophoresis. Hybridizations
typically occur with the target sequence bound to the surface of a
membrane or, as described in U.S. Pat. No. 5,468,613, the ASH probe
sequence may be bound to a membrane.
[0141] In one embodiment, ASH data are obtained by amplifying nucleic acid
fragments (amplicons) from genomic DNA using PCR, transferring the
amplicon target DNA to a membrane in a dot-blot format, hybridizing a
labeled oligonucleotide probe to the amplicon target, and observing the
hybridization dots by autoradiography.
[0142] Other of the exemplary molecular markers provided herein are Simple
sequence repeats (SSR). SSR markers take advantage of high levels of di-,
tri-, or tetra-nucleotide tandem repeats within a genome. Dinucleotide
repeats have been reported to occur in the human genome as many as 50,000
times with n varying from 10 to 60 or more (Jacob et al. (1991) Cell
67:213. Dinucleotide repeats have also been found in higher plants
(Condit and Hubbell (1991) Genome 34:66).
[0143] Briefly, SSR data is generated by hybridizing primers to conserved
regions of the plant genome which flank the SSR sequence. PCR is then
used to amplify the dinucleotide repeats between the primers. The
amplified sequences are then electorphoresed to determine the size and
therefore the number of di-, tri-, and tetra-nucleotide repeats.
[0144] Amplified variable sequences refer to amplified sequences of the
plant genome which exhibit high nucleic acid residue variability between
members of the same species, e.g., microsatellite sequences. All
organisms have variable genomic sequences and each organism (with the
exception of a clone) has a different set of variable sequences. Once
identified, the presence of specific variable sequences can be used to
predict phenotypic traits. Preferably, DNA from the plant serves as a
template for amplification with primers that flank a variable sequence of
DNA. The variable sequence is amplified and then sequenced.
[0145] Randomly amplified polymorphic DNA (RAPD) markers are genomic
sequences amplified by PCR using a single short primer of arbitrary
sequence at low stringency. During amplification at low stringency a
number of PCR products, some of which differ in length (and sequence)
between individuals, are generated from random locations throughout the
genome. Unlike amplified variable sequences, no prior sequence
information is required to identify RAPD markers.
[0146] In vitro amplification techniques are well known in the art.
Examples of techniques sufficient to direct persons of skill through such
in vitro methods, including the polymerase chain reaction (PCR), the
ligase chain reaction (LCR), Q.beta.-replicase amplification and other
RNA polymerase mediated techniques (e.g., NASBA), are found in Berger,
Sambrook and Ausubel as well as Mullis et al. (1987) U.S. Pat. No.
4,683,202; PCR Protocols, A Guide to Methods and Applications (Innis et
al., eds.) Academic Press Inc., San Diego Academic Press Inc. San Diego,
Calif. (1990) (Innis); Arnheim & Levinson (Oct. 1, 1990) C&EN 36-47; The
Journal Of NIH Research (1991) 3, 81-94; (Kwoh et al. (1989) Proc. Natl.
Acad. Sci. USA 86, 1173; Guatelli et al. (1990) Proc. Natl. Acad. Sci.
USA 87, 1874; Lomell et al. (1989) J. Clin. Chem 35, 1826; Landegren et
al., (1988) Science 241, 1077-1080; Van Brunt (1990) Biotechnology 8,
291-294; Wu and Wallace, (1989) Gene 4, 560; Barringer et al. (1990) Gene
89, 117, and Sooknanan and Malek (1995) Biotechnology 13: 563-564.
Improved methods of cloning in vitro amplified nucleic acids are
described in Wallace et al., U.S. Pat. No. 5,426,039. Improved methods of
amplifying large nucleic acids by PCR are summarized in Cheng et al.
(1994) Nature 369: 684, and the references therein, in which PCR
amplicons of up to 40 kb are generated. One of skill will appreciate that
essentially any RNA can be converted into a double stranded DNA suitable
for restriction digestion, PCR expansion and sequencing using reverse
transcriptase and a polymerase. See, Ausubel, Sambrook and Berger.
[0147] Oligonucleotides for use as primers, e.g., in amplification
reactions and for use as nucleic acid sequence probes are typically
synthesized chemically according to the solid phase phosphoramidite
triester method described by Beaucage and Caruthers (1981) Tetrahedron
Lett. 22:1859, or can simply be ordered commercially.
[0148] Alternatively, self-sustained sequence replication can be used to
identify genetic markers. Self-sustained sequence replication refers to a
method of nucleic acid amplification using target nucleic acid sequences
which are replicated exponentially in vitro under substantially
isothermal conditions by using three enzymatic activities involved in
retroviral replication: (1) reverse transcriptase, (2) Rnase H, and (3) a
DNA-dependent RNA polymerase (Guatelli et al. (1990) Proc Natl Acad Sci
USA 87:1874). By mimicking the retroviral strategy of RNA replication by
means of cDNA intermediates, this reaction accumulates cDNA and RNA
copies of the original target.
[0149] Amplified restriction fragment polymorphisms or amplified fragment
length polymorphisms (AFLP) can also be used as genetic markers (Vos et
al. (1995) Nucl Acids Res 23:4407. The phrase "amplified restriction
fragment polymorphism" refers to selected restriction fragments, which
are amplified before or after cleavage by a restriction endonuclease. The
amplification step allows easier detection of specific restriction
fragments. AFLP allows the detection large numbers of polymorphic markers
and has been used for genetic mapping of plants (Becker et al. (1995) Mol
Gen Genet 249:65; and Meksem et al. (1995) Mol Gen Genet 249:74.
[0150] Single nucleotide polymorphisms (SNP) are markers that consist of a
shared sequence differentiated on the basis of a single nucleotide.
Typically, this distinction is detected by differential migration
patterns of an amplicon comprising the SNP on e.g., an acrylamide gel. In
such cases the marker may also be referred to as a single-strand
conformation polymorphism or SSCP. However, alternative modes of
detection, such as hybridization, e.g., ASH, or RFLP analysis are not
excluded.
[0151] Alternatively, isozyme markers are employed as genetic markers.
Isozymes are multiple forms of enzymes that differ from one another in
their amino acid, and therefore their nucleic acid sequences. Some
isozymes are multimeric enzymes containing slightly different subunits.
Other isozymes are either multimeric or monomeric but have been cleaved
from the proenzyme at different sites in the amino acid sequence.
Isozymes can be characterized and analyzed at the protein level, or
alternatively, isozymes that differ at the nucleic acid level can be
determined. In such cases any of the nucleic acid based methods described
herein can be used to analyze isozyme markers.
[0152] In alternative embodiments, in silico methods can be used to detect
the marker loci. For example, the sequence of a nucleic acid comprising
the marker can be stored in a computer. The desired marker locus sequence
or its homolog can be identified using an appropriate nucleic acid search
algorithm as provided by, for example, in such readily available programs
as BLAST.
[0153] Integrated Systems/Computer Assisted Methods
[0154] In some embodiments, the present invention includes an "integrated
system" including an electronic means of storing or transmitting computer
readable data representing or designating the allelic forms determined by
the method of the present invention. The computer readable media includes
cache, main, and storage memory and other electronic data storage means
for storage of computer code. Data representing the allelic forms
determined by the method of the present invention can also be
electronically transmitted in a computer data signal embodied in a
transmission medium over a network such as an intranet or internet or
combinations thereof.
[0155] The phrase "integrated system" in the context of this invention
refers to a system in which data entering a computer corresponds to
physical objects or processes external to the computer, e.g., a marker
allele, and a process that, within a computer, causes a physical
transformation of the input signals to different output signals. In other
words, the input data, e.g., amplification of a particular marker allele
is transformed to output data, e.g., the identification of the allelic
form of a chromosome segment. The process within the computer is a set of
instructions, or "program," by which positive amplification or
hybridization signals are recognized by the integrated system and
attributed to individual samples as a genotype. Additional programs
correlate the identity of individual samples with phenotypic values or
marker alleles, e.g., statistical methods. In addition there are numerous
e.g., C/C++ programs for computing, Delphi and/or Java programs for GUI
interfaces, and productivity tools (e.g., Microsoft Excel and/or
SigmaPlot) for charting. Other useful software tools in the context of
the integrated systems of the invention include statistical packages such
as SAS, Genstat, Matlab, Mathematica, and S-Plus and genetic modeling
packages such as QU-GENE. Furthermore additional programming languages
such as Fortran and the like are also suitably employed in the integrated
systems of the invention.
[0156] For example, marker allele values assigned to a population of
progeny descending from crosses between elite lines are recorded in a
computer readable medium, thereby establishing a database corresponding
allelic forms with unique identifiers for each member of the population
of progeny. Any file or folder, whether custom-made or commercially
available (e.g., from Oracle or Sybase) suitable for recording data in a
computer readable medium is acceptable as a database in the context of
the present invention. Data regarding genotype for one or more molecular
markers, e.g., ASH, SSR, RFLP, RAPD, AFLP, SNP, isozyme markers or other
markers as described herein, are similarly recorded in a computer
accessible database. Optionally, marker data is obtained using an
integrated system that automates one or more aspects of the assay (or
assays) used to determine marker(s) genotype. In such a system, input
data corresponding to genotypes for molecular markers are relayed from a
device, e.g., an array, a scanner, a CCD, or other detection device
directly to files in a computer readable medium accessible to the central
processing unit. A set of instructions (embodied in one or more programs)
encoding the statistical models of the invention is then executed by the
computational device to identify correlations between yield data and
marker genotypes. Typically, the integrated system also includes a user
input device, such as a keyboard, a mouse, a touchscreen, or the like,
for, e.g., selecting files, retrieving data, etc., and an output device
(e.g., a monitor, a printer, etc.) for viewing or recovering the product
of the statistical analysis.
[0157] Thus, in one aspect, the invention provides an integrated system
comprising a computer or computer readable medium comprising set of files
and/or a database with at least one data set that corresponds to
genotypes for genetic markers. The system also includes a user interface
allowing a user to selectively view one or more databases. In addition,
standard text manipulation software such as word processing software
(e.g., Microsoft Word.TM. or Corel Wordperfect.TM.) and database or
spreadsheet software (e.g., spreadsheet software such as Microsoft
Excel.TM., Corel Quattro Pro.TM., or database programs such as Microsoft
Access.TM. or Paradox.TM.) can be used in conjunction with a user
interface (e.g., a GUI in a standard operating system such as a Windows,
Macintosh, Unix or Linux system) to manipulate strings of characters.
[0158] The invention also provides integrated systems for sample
manipulation incorporating robotic devices as previously described. A
robotic liquid control armature for transferring solutions (e.g., plant
cell extracts) from a source to a destination, e.g., from a microtiter
plate to an array substrate, is optionally operably linked to the digital
computer (or to an additional computer in the integrated system). An
input device for entering data to the digital computer to control high
throughput liquid transfer by the robotic liquid control armature and,
optionally, to control transfer by the armature to the solid support is
commonly a feature of the integrated system.
[0159] Integrated systems for molecular marker analysis of the present
invention typically include a digital computer with one or more of
high-throughput liquid control software, image analysis software, data
interpretation software, a robotic liquid control armature for
transferring solutions from a source to a destination operably linked to
the digital computer, an input device (e.g., a computer keyboard) for
entering data to the digital computer to control high throughput liquid
transfer by the robotic liquid control armature and, optionally, an image
scanner for digitizing label signals from labeled probes hybridized,
e.g., to expression products on a solid support operably linked to the
digital computer. The image scanner interfaces with the image analysis
software to provide a measurement of, e.g., differentiating nucleic acid
probe label intensity upon hybridization to an arrayed sample nucleic
acid population, where the probe label intensity measurement is
interpreted by the data interpretation software to show whether, and to
what degree, the labeled probe hybridizes to a label. The data so derived
is then correlated with sample identity, to determine the identity of a
plant with a particular genotype(s) for genetic markers, e.g., to
facilitate marker assisted selection of soybean plants with favorable
allelic forms of chromosome segments involved in agronomic performance.
[0160] Optical images, e.g., hybridization patterns viewed (and,
optionally, recorded) by a camera or other recording device (e.g., a
photodiode and data storage device) are optionally further processed in
any of the embodiments herein, e.g., by digitizing the image and/or
storing and analyzing the image on a computer. A variety of commercially
available peripheral equipment and software is available for digitizing,
storing and analyzing a digitized video or digitized optical image, e.g.,
using PC (Intel .times.86 or pentium chip-compatible DOS.TM., OS2.TM.
WINDOWS.TM., WINDOWS NT.TM. or WINDOWS95.TM. based machines),
MACINTOSH.TM., LINUX, or UNIX based (e.g., SUN.TM. work station)
computers.
[0161] Identification of Additional Markers and OTL Associated with Yield
[0162] Nucleic acids isolated from the chromosome segments of the present
invention, e.g., nucleic acids corresponding to additional marker loci,
nucleic acids corresponding to genetic elements contributing to superior
agronomic performance, are within the scope of the present invention. For
example, in the rare instance in which the markers and alleles of the
present invention are not well suited for MAS, the markers can,
nonetheless, be used to identify additional linked markers that are,
e.g., closer to, or within, the QTL of interest. Based on the markers
disclosed herein, improvement in the efficiency of MAS can be obtained by
saturating the relevant chromosome segments with numerous linked markers.
The breeding bias analysis can be repeated on all markers within the
region to identify those having the highest statistical significance. MAS
can be practiced with all markers in the region followed by phenotypic
characterization (i.e., of yield) to determine which markers are most
efficient.
[0163] Additional markers can be identified in a variety of ways. For
example, additional markers can be identified by evaluating publicly or
privately available markers that have been mapped to the genomic
region(s) of interest, including candidate markers known to encode
proteins that are, at least theoretically, likely to be related to yield.
Alternatively, unmapped markers can be evaluated to determine which
markers map to the same genomic region via independent mapping studies.
[0164] A theoretically optimal marker can be obtained by isolating genetic
elements contributing to superior agronomic performance, such as coding
regions giving rise to expression products that influence yield.
Identification of a QTL underlying superior agronomic performance can be
accomplished by anchoring the marker to a physical DNA map and then
progressing upstream and downstream to identify coding sequences, i.e.,
by "positional gene cloning."
[0165] Positional gene cloning uses the physical proximity of a genetic
marker (such as a marker provided in Tables 3 through 12) to identify a
cloned chromosomal fragment that includes a nucleic acid of interest,
e.g., a QTL contributing to superior agronomic performance. Clones of
nucleic acids linked to the markers of the invention have a variety of
uses, including as additional genetic markers to define the chromosome
segments of the invention and for use in marker assisted selection (MAS).
Markers which are adjacent to an open reading frame (ORF) can hybridize
to a DNA clone, thereby identifying a clone on which an ORF associated
with a trait contributing to yield is located. If the marker is more
distant, a fragment containing the open reading frame is identified by
successive rounds of screening and isolation of clones which together
comprise a contiguous sequence of DNA, a "contig." Protocols sufficient
to guide one of skill through the isolation of clones associated with
linked markers are found in, e.g., in the references cited in the section
entitled "GENERAL MOLECULAR BIOLOGY REFERENCES" below.
[0166] An isolated chromosome fragment can be produced by such well known
methods as digesting chromosomal DNA with one or more restriction
enzymes, or by amplifying a chromosomal region in a polymerase chain
reaction (PCR), or alternative amplification reaction. The digested or
amplified fragment is typically ligated into a vector suitable for
replication, e.g., a plasmid, a cosmid, a phage, an artificial
chromosome, or the like, and, optionally expression, of the inserted
fragment.
[0167] Such chromosome segments can be utilized to identify homologous
nucleic acids, e.g., in other lines or species, and/or can be used in the
production of transgenic plants with desirable phenotypic attributes
related to agronomic performance. A chromosome segment comprising a
nucleic acid contributing to increased yield is isolated, e.g., cloned
via positional cloning methods outlined above. A chromosome segment can
contain one or more ORFs associated with the desired phenotypic trait,
and can be cloned on one or more individual vectors, e.g., depending on
the size of the chromosome interval.
[0168] It will be appreciated that numerous vectors are available in the
art for the isolation and replication of the nucleic acids of the
invention. For example, plasmids, cosmids and phage vectors are well
known in the art, and are sufficient for many applications (e.g., in
applications involving insertion of nucleic acids ranging from less than
1 to about 20 kilobases (kb). In certain applications, it is advantageous
to make or clone large nucleic acids to identify nucleic acids more
distantly linked to a given marker, or to isolate nucleic acids in excess
of 10-20 kb, e.g., up to several hundred kilobases or more, such as the
entire interval between two linked markers, i.e., up to and including one
or more centimorgans (CM), linked to markers as identified herein. In
such cases, a number of vectors capable of accommodating large nucleic
acids are available in the art, these include, yeast artificial
chromosomes (YACs), bacterial artificial chromosomes (BACs), plant
artificial chromosomes (PACs) and the like. For a general introduction to
YACs, BACs, PACs and MACs as artificial chromosomes, see, e.g., Monaco
and Larin (1994) Trends Biotechnol 12:280. In addition, methods for the
in vitro amplification of large nucleic acids linked to genetic markers
are widely available (e.g., Cheng et al. (1994) Nature 369:684, and
references therein). Cloning systems can be created or obtained from
commercially; see, for example, Stratagene (La Jolla, Calif.).
[0169] Vectors, Promoters and Expression Systems
[0170] The present invention includes recombinant constructs incorporating
one or more of the nucleic acid sequences described above. Such
constructs include a vector, for example, a plasmid, a cosmid, a phage, a
virus, a bacterial artificial chromosome (BAC), a yeast artificial
chromosome (YAC), etc., into which one or more polynucleotide sequences
of interest (e.g., a marker or genetic element contributing to yield) has
been inserted, in a forward or reverse orientation. For example, the
inserted nucleic acid can include a chromosomal sequence or cDNA
including a all or part of at least one genetic element or open reading
frame ("ORF") associated with yield. In a preferred embodiment, the
construct further comprises regulatory sequences, including, for example,
a promoter, operably linked to the sequence. Large numbers of suitable
vectors and promoters are known to those of skill in the art, and are
commercially available.
[0171] As desired, the polynucleotides of the present invention, e.g., a
genetic element contributing to superior agronomic performance identified
according to the methods described herein, can be included in any one of
a variety of vectors suitable for generating sense or antisense RNA, and
optionally, polypeptide expression products. Such vectors include
chromosomal, nonchromosomal and synthetic DNA sequences, e.g.,
derivatives of SV40; bacterial plasmids; phage DNA; baculovirus; yeast
plasmids; vectors derived from combinations of plasmids and phage DNA,
viral DNA such as vaccinia, adenovirus, fowl pox virus, pseudorabies,
adenovirus, adeno-associated virus, retroviruses and many others. Any
vector that is capable of introducing genetic material into a cell, and,
if replication is desired, which is replicable in the relevant host can
be used.
[0172] In an expression vector or expression cassette, the polynucleotide
sequence of interest is physically arranged in proximity and orientation
to an appropriate transcription control sequence (promoter, and
optionally, one or more enhancers) to direct mRNA synthesis. That is, the
polynucleotide sequence of interest is "operably linked" to an
appropriate transcription control sequence. Examples of such promoters
include: LTR or SV40 promoter, E. coli lac or trp promoter, phage lambda
P.sub.L promoter, and other promoters known to control expression of
genes in prokaryotic or eukaryotic cells or their viruses. The expression
vector also contains a ribosome binding site for translation initiation,
and a transcription terminator. The vector optionally includes
appropriate sequences for amplifying expression. In addition, the
expression vectors optionally comprise one or more selectable marker
genes to provide a phenotypic trait for selection of transformed host
cells, such as dihydrofolate reductase or neomycin resistance for
eukaryotic cell culture, or such as tetracycline or ampicillin resistance
in E. coli.
[0173] Additional Expression Elements
[0174] Where translation of polypeptide encoded by a nucleic acid
comprising a polynucleotide sequence of the invention is desired,
additional translation specific initiation signals can improve the
efficiency of translation. These signals can include, e.g., an ATG
initiation codon and adjacent sequences. In some cases, for example,
full-length cDNA molecules or chromosomal segments including a coding
sequence incorporating, e.g., a QTL or an ORF associated with a QTL or
QTL marker, a translation initiation codon and associated sequence
elements are inserted into the appropriate expression vector
simultaneously with the polynucleotide sequence of interest. In such
cases, additional translational control signals frequently are not
required. However, in cases where only a polypeptide coding sequence, or
a portion thereof, is inserted, exogenous translational control signals,
including an ATG initiation codon must be provided. Furthermore, the
initiation codon must be in the correct reading frame to ensure
transcription of the polynucleotide sequence of interest. Exogenous
transcriptional elements and initiation codons can be of various origins,
both natural and synthetic. The efficiency of expression can be enhanced
by the inclusion of enhancers appropriate to the cell system in use
(Scharf D et al. (1994) Results Probl Cell Differ 20:125-62; Bittner et
al. (1987) Methods in Enzymol 153:516-544).
[0175] Generation of Transgenic Plants and Cells
[0176] The present invention also relates to host cells and organisms
which are transformed with nucleic acids corresponding to genetic
elements contributing to superior agronomic performance and other genes
identified according to the methods of the invention. For example, such
nucleic acids include chromosome segments, ORFs, and/or cDNAs or
corresponding to a sequence or subsequence included within the identified
chromosome segment or ORF. Additionally, the invention provides for the
production of polypeptides corresponding to such genetic elements by
recombinant nucleic acid (and expression) techniques. Host cells are
genetically engineered (i.e., transduced, transfected or transformed)
with the vectors of this invention (i.e., vectors incorporating genetic
elements contributing to increased yield, or other nucleic acids
identified according to the methods of the invention and as described
above) which are, for example, a cloning vector or an expression vector.
Such vectors include, in addition to those described above, e.g., an
agrobacterium, a virus (such as a plant virus), a naked polynucleotide,
or a conjugated polynucleotide. The vectors are introduced into plant
tissues, cultured plant cells or plant protoplasts by a variety of
standard methods including electroporation (From et al. (1985) Proc.
Natl. Acad. Sci. USA 82;5824), infection by viral vectors such as
cauliflower mosaic virus (CaMV) (Hohn et al. (1982) Molecular Biology of
Plant Tumors (Academic Press, New York, pp. 549-560; Howell U.S. Pat. No.
4,407,956), high velocity ballistic penetration by small particles with
the nucleic acid either within the matrix of small beads or particles, or
on the surface (Klein et al. (1987) Nature 327;70), use of pollen as
vector (WO 85/01856), or use of Agrobacterium tumefaciens or A.
rhizogenes carrying a T-DNA plasmid in which DNA fragments are cloned.
The T-DNA plasmid is transmitted to plant cells upon infection by
Agrobacterium tumefaciens, and a portion is stably integrated into the
plant genome (Horsch et al. (1984) Science 233;496; Fraley et al. (1983)
Proc. Natl. Acad. Sci. USA 80;4803). The method of introducing a nucleic
acid of the present invention into a host cell is not critical to the
instant invention. Thus, any method, e.g., including but not limited to
the above examples, which provides for effective introduction of a
nucleic acid into a cell or protoplast can be employed.
[0177] The engineered host cells can be cultured in conventional nutrient
media modified as appropriate for such activities as, for example,
activating promoters or selecting transformants. These cells can
optionally be cultured into transgenic plants. Plant regeneration from
cultured protoplasts is described in Evans et al. (1983) "Protoplast
Isolation and Culture," Handbook of Plant Cell Cultures 1, 124-176
(MacMillan Publishing Co., N.Y.; Davey (1983) "Recent Developments in the
Culture and Regeneration of Plant Protoplasts," Protoplasts, pp. 12-29,
(Birkhauser, Basel); Dale (1983) "Protoplast Culture and Plant
Regeneration of Cereals and Other Recalcitrant Crops," Protoplasts pp.
31-41, (Birkhauser, Basel); Binding (1985) "Regeneration of Plants,"
Plant Protoplasts, pp. 21-73, (CRC Press, Boca Raton,).
[0178] The present invention also relates to the production of transgenic
organisms, which may be bacteria, yeast, fungi, or plants, transduced
with the nucleic acids, e.g., cloned QTL of the invention. A thorough
discussion of techniques relevant to bacteria, unicellular eukaryotes and
cell culture may be found in references enumerated above and are briefly
outlined as follows. Several well-known methods of introducing target
nucleic acids into bacterial cells are available, any of which may be
used in the present invention. These include: fusion of the recipient
cells with bacterial protoplasts containing the DNA, treatment of the
cells with liposomes containing the DNA, electroporation, projectile
bombardment (biolistics), carbon fiber delivery, and infection with viral
vectors (discussed further, below), etc. Bacterial cells can be used to
amplify the number of plasmids containing DNA constructs of this
invention. The bacteria are grown to log phase and the plasmids within
the bacteria can be isolated by a variety of methods known in the art
(see, for instance, Sambrook). In addition, numerous kits are
commercially available and can be employed according to the manufacturers
instructions for the purification of plasmids from bacteria (and other
cells). For their proper use, follow the manufacturer's instructions
(see, for example, EasyPrep.TM., FlexiPrep.TM., both from Pharmacia
Biotech; StrataClean.TM., from Stratagene; and, QIAprep.TM. from Qiagen).
The isolated and purified plasmids are then further manipulated to
produce other plasmids, used to transfect plant cells or incorporated
into Agrobacterium tumefaciens related vectors to infect plants. Typical
vectors contain transcription and translation terminators, transcription
and translation initiation sequences, and promoters useful for regulation
of the expression of the particular target nucleic acid. The vectors
optionally comprise generic expression cassettes containing at least one
independent terminator sequence, sequences permitting replication of the
cassette in eukaryotes, or prokaryotes, or both, (e.g., shuttle vectors)
and selection markers for both prokaryotic and eukaryotic systems.
Vectors are suitable for replication and integration in prokaryotes,
eukaryotes, or preferably both. See, Giliman & Smith (1979) Gene 8:81;
Roberts et al. (1987) Nature 328:731; Schneider et al. (1995) Protein
Expr. Purif. 6435:10; Ausubel, Sambrook, Berger (all supra). A catalogue
of Bacteria and Bacteriophages useful for cloning is provided, e.g., by
the ATCC, e.g., The ATCC Catalogue of Bacteria and Bacteriophage (1992)
Gherna et al. (eds) published by the ATCC. Additional basic procedures
for sequencing, cloning and other aspects of molecular biology and
underlying theoretical considerations are also found in Watson et al.
(1992) Recombinant DNA, Second Edition, Scientific American Books, NY.
[0179] Transforming Nucleic Acids into Plants.
[0180] Embodiments of the present invention pertain to the production of
transgenic plants comprising the cloned nucleic acids, e.g., chromosome
segments, isolated ORFs, and cDNAs associated with genetic elements
identified by their proximity to the markers of the invention. Techniques
for transforming plant cells with nucleic acids are generally available
and can be adapted to the invention by the use of nucleic acids encoding
or corresponding to chromosome segments, subsequences, e.g., ORFs, and
the like. In addition to Berger, Ausubel and Sambrook (infra), useful
general references for plant cell cloning, culture and regeneration
include Jones (ed) (1995) Plant Gene Transfer and Expression
Protocols--Methods in Molecular Biology, Volume 49 Humana Press Towata
N.J.; Payne et al. (1992) Plant Cell and Tissue Culture in Liquid Systems
John Wiley & Sons, Inc. New York, N.Y. (Payne); and Gamborg and Phillips
(eds) (1995) Plant Cell, Tissue and Organ Culture; Fundamental Methods
Springer Lab Manual, Springer-Verlag (Berlin Heidelberg New York)
(Gamborg). A variety of cell culture media are described in Atlas and
Parks (eds) The Handbook of Microbiological Media (1993) CRC Press, Boca
Raton, Fla. (Atlas). Additional information for plant cell culture is
found in available commercial literature such as the Life Science
Research Cell Culture Catalogue (1998) from Sigma-Aldrich, Inc (St Louis,
Mo.) (Sigma-LSRCCC) and, e.g., the Plant Culture Catalogue and supplement
(1997) also from Sigma-Aldrich, Inc (St Louis, Mo.) (Sigma-PCCS).
Additional details regarding plant cell culture are found in Croy, (ed.)
(1993) Plant Molecular Biology Bios Scientific Publishers, Oxford, U.K.
[0181] The nucleic acid constructs of the invention, e.g., plasmids,
cosmids, artificial chromosomes, DNA and RNA polynucleotides, are
introduced into plant cells, either in culture or in the organs of a
plant by a variety of conventional techniques. Where the sequence is
expressed, the sequence is optionally combined with transcriptional and
translational initiation regulatory sequences which direct the
transcription or translation of the sequence from the exogenous DNA in
the intended tissues of the transformed plant.
[0182] Isolated nucleic acids can be introduced into plants according to
any of a variety of techniques known in the art. Techniques for
transforming a wide variety of higher plant species are well known and
described in the technical, scientific, and patent literature. See, for
example, Weising et al. (1988) Ann. Rev. Genet. 22:421-477.
[0183] For example plasmids, cosmids, phage, naked or variously
conjugated-DNA polynucleotides, (e.g., polylysine-conjugated DNA,
peptide-conjugated DNA, liposome-conjugated DNA, etc.), or artificial
chromosomes, can be introduced directly into the genomic DNA of the plant
cell using techniques such as electroporation and microinjection of plant
cell protoplasts, or the DNA constructs can be introduced directly to
plant cells using ballistic methods, such as DNA particle bombardment.
[0184] Microinjection techniques for injecting e.g., cells, embryos,
callus and protoplasts, are known in the art and well described in the
scientific and patent literature. For example, a number of methods are
described in Jones (ed) (1995) Plant Gene Transfer and Expression
Protocols--Methods in Molecular Biology, Volume 49 Humana Press Towata
N.J., as well as in the other references noted herein and available in
the literature.
[0185] For example, the introduction of DNA constructs using polyethylene
glycol precipitation is described in Paszkowski, et al., EMBO J. 3:2717
(1984). Electroporation techniques are described in Fromm, et al., Proc.
Nat'l. Acad. Sci. USA 82:5824 (1985). Ballistic transformation techniques
are described in Klein, et al., Nature 327:70-73 (1987). Additional
details are found in Jones (1995) and Gamborg and Phillips (1995), supra,
and in U.S. Pat. No. 5,990,387.
[0186] Alternatively, and in some cases preferably, Agrobacterium mediated
transformation is employed to generate transgenic plants.
Agrobacterium-mediated transformation techniques, including disarming and
use of binary vectors, are also well described in the scientific
literature. See, for example Horsch, et al. (1984) Science 233:496; and
Fraley et al. (1984) Proc. Nat'l. Acad. Sci. USA 80:4803 and recently
reviewed in Hansen and Chilton (1998) Current Topics in Microbiology
240:22 and Das (1998) Subcellular Biochemistry 29: Plant Microbe
Interactions pp343-363.
[0187] The DNA constructs may be combined with suitable T-DNA flanking
regions and introduced into a conventional Agrobacterium tumefaciens host
vector. The virulence functions of the Agrobacterium tumefaciens host
will direct the insertion of the construct and adjacent marker into the
plant cell DNA when the cell is infected by the bacteria. See, U.S. Pat.
No. 5,591,616. Although Agrobacterium is useful primarily in dicots,
certain monocots can be transformed by Agrobacterium. For instance,
Agrobacterium transformation of maize is described in U.S. Pat. No.
5,550,318.
[0188] Other methods of transfection or transformation include (1)
Agrobacterium rhizogenes-mediated transformation (see, e.g., Lichtenstein
and Fuller (1987) In: Genetic Engineering, vol. 6, P W J Rigby, Ed.,
London, Academic Press; and Lichtenstein; C. P., and Draper (1985) In:
DNA Cloning, Vol. II, D. M. Glover, Ed., Oxford, IRI Press; WO 88/02405,
published Apr. 7, 1988, describes the use of A. rhizogenes strain A4 and
its Ri plasmid along with A. tumefaciens vectors pARC8 or pARC 16 (2)
liposome-mediated DNA uptake (see, e.g., Freeman et al. (1984) Plant Cell
Physiol. 25:1353), (3) the vortexing method (see, e.g., Kindle (1990)
Proc. Natl. Acad. Sci., (USA) 87:1228.
[0189] DNA can also be introduced into plants by direct DNA transfer into
pollen as described by Zhou et al. (1983) Methods in Enzymology, 101:433;
D. Hess (1987) Intern Rev. Cytol. 107:367; Luo et al. (1988) Plant Mol.
Biol. Reporter 6:165. Expression of polypeptide coding genes can be
obtained by injection of the DNA into reproductive organs of a plant as
described by Pena et al. (1987) Nature 325:274. DNA can also be injected
directly into the cells of immature embryos and the desiccated embryos
rehydrated as described by Neuhaus et al.(1987) Theor. Appl. Genet.
75:30; and Benbrook et al.(1986) in Proceedings Bio Expo Butterworth,
Stoneham, Mass., pp. 27-54. Additionally, a variety of plant viruses that
can be employed as vectors are known in the art and include cauliflower
mosaic virus (CaMV), geminivirus, brome mosaic virus, and tobacco mosaic
virus.
[0190] Regeneration of Transgenic Plants
[0191] Transformed plant cells which are derived by any of the above
transformation techniques can be cultured to regenerate a whole plant
which possesses the transformed genotype and thus the desired phenotype.
Such regeneration techniques rely on manipulation of certain
phytohormones in a tissue culture growth medium, typically relying on a
biocide and/or herbicide marker which has been introduced together with
the desired nucleotide sequences. Plant regeneration from cultured
protoplasts is described in Evans et al. (1983) Protoplasts Isolation and
Culture, Handbook of Plant Cell Culture pp. 124-176, Macmillian
Publishing Company, New York; and Binding (1985) Regeneration of Plants,
Plant Protoplasts pp. 21-73, CRC Press, Boca Raton. Regeneration can also
be obtained from plant callus, explants, somatic embryos (Dandekar et al.
(1989) J. Tissue Cult. Meth. 12:145; McGranahan, et al. (1990) Plant Cell
Rep. 8:512) organs, or parts thereof. Such regeneration techniques are
described generally in Klee et al. (1987)., Ann. Rev. of Plant Phys.
38:467-486. Additional details are found in Payne (1992) and Jones
(1995), both supra, and Weissbach and Weissbach, eds.(1988) Methods for
Plant Molecular Biology Academic Press, Inc., San Diego, Calif. This
regeneration and growth process includes the steps of selection of
transformant cells and shoots, rooting the transformant shoots and growth
of the plantlets in soil. These methods are adapted to the invention to
produce transgenic plants bearing QTLs and other genes isolated according
to the methods of the invention.
[0192] In addition, the regeneration of plants containing the
polynucleotide of the present invention and introduced by Agrobacterium
into cells of leaf explants can be achieved as described by Horsch et al.
(1985) Science 227:1229-1231. In this procedure, transformants are grown
in the presence of a selection agent and in a medium that induces the
regeneration of shoots in the plant species being transformed as
described by Fraley et al. (1983) Proc. Natl. Acad. Sci. (U.S.A.)
80:4803. This procedure typically produces shoots within two to four
weeks and these transformant shoots are then transferred to an
appropriate root-inducing medium containing the selective agent and an
antibiotic to prevent bacterial growth. Transgenic plants of the present
invention may be fertile or sterile.
[0193] In construction of recombinant expression cassettes of the
invention, which include, for example, an ORF associated with a marker or
genetic element contributing to yield, a plant promoter fragment is
optionally employed which directs expression of a nucleic acid in any or
all tissues of a regenerated plant. Examples of constitutive promoters
include the cauliflower mosaic virus (CaMV) 35S transcription initiation
region, the 1'-or 2'-promoter derived from T-DNA of Agrobacterium
tumefaciens, and other transcription initiation regions from various
plant genes known to those of skill. Alternatively, the plant promoter
may direct expression of the polynucleotide of the invention in a
specific tissue (tissue-specific promoters) or may be otherwise under
more precise environmental control (inducible promoters). Examples of
tissue-specific promoters under developmental control include promoters
that initiate transcription only in certain tissues, such as fruit,
seeds, or flowers.
[0194] Any of a number of promoters which direct transcription in plant
cells can be suitable. The promoter can be either constitutive or
inducible. In addition to the promoters noted above, promoters of
bacterial origin which operate in plants include the octopine synthase
promoter, the nopaline synthase promoter and other promoters derived from
native Ti plasmids. See, Herrara-Estrella et al. (1983) , Nature,
303:209. Viral promoters include the 35S and 19S RNA promoters of
cauliflower mosaic virus. See, Odell et al. (1985) Nature, 313:810. Other
plant promoters include the ribulose-1,3-bisphosphate carboxylase small
subunit promoter and the phaseolin promoter. The promoter sequence from
the E8 gene and other genes may also be used. The isolation and sequence
of the E8 promoter is described in detail in Deikman and Fischer (1988)
EMBO J. 7:3315. Many other promoters are in current use and can be
coupled to an exogenous DNA sequence to direct expression of the nucleic
acid.
[0195] If expression of a polypeptide, including those encoded by QTL or
other nucleic acid, is desired, a polyadenylation region at the 3'-end of
the coding region is typically included. The polyadenylation region can
be derived from the natural gene, from a variety of other plant genes, or
from, e.g., T-DNA.
[0196] The vector comprising the sequences (e.g., promoters or coding
regions) from genes encoding expression products and transgenes of the
invention will typically include a nucleic acid subsequence, a marker
gene which confers a selectable, or alternatively, a screenable,
phenotype on plant cells. For example, the marker may encode biocide
tolerance, particularly antibiotic tolerance, such as tolerance to
kanamycin, G418, bleomycin, hygromycin, or herbicide tolerance, such as
tolerance to chlorosluforon, or phosphinothricin (the active ingredient
in the herbicides bialaphos or Basta). See, e.g., Padgette et al. (1996)
In: Herbicide-Resistant Crops (Duke, ed.), pp 53-84, CRC Lewis
Publishers, Boca Raton ("Padgette, 1996"). For example, crop selectivity
to specific herbicides can be conferred by engineering genes into crops
which encode appropriate herbicide metabolizing enzymes from other
organisms, such as microbes. See, Vasil (1996) In: Herbicide-Resistant
Crops (Duke, ed.), pp 85-91, CRC Lewis Publishers, Boca Raton) ("Vasil",
1996).
[0197] One of skill will recognize that after the recombinant expression
cassette is stably incorporated in transgenic plants and confirmed to be
operable, it can be introduced into other plants by sexual crossing. Any
of a number of standard breeding techniques can be used, depending upon
the species to be crossed. In vegetatively propagated crops, mature
transgenic plants can be propagated by the taking of cuttings or by
tissue culture techniques to produce multiple identical plants. Selection
of desirable transgenics is made and new varieties are obtained and
propagated vegetatively for commercial use. In seed propagated crops,
mature transgenic plants can be self crossed to produce a homozygous
inbred plant. The inbred plant produces seed containing the newly
introduced heterologous nucleic acid. These seeds can be grown to produce
plants that would produce the selected phenotype. Parts obtained from the
regenerated plant, such as flowers, seeds, leaves, branches, fruit, and
the like are included in the invention, provided that these parts
comprise cells comprising the isolated nucleic acid of the present
invention. Progeny and variants, and mutants of the regenerated plants
are also included within the scope of the invention, provided that these
parts comprise the introduced nucleic acid sequences.
[0198] Transgenic plants expressing a polynucleotide of the present
invention can be screened for transmission of the nucleic acid of the
present invention by, for example, standard immunoblot and DNA detection
techniques. Expression at the RNA level can be determined initially to
identify and quantitate expression-positive plants. Standard techniques
for RNA analysis can be employed and include PCR amplification assays
using oligonucleotide primers designed to amplify only the heterologous
RNA templates and solution hybridization assays using heterologous
nucleic acid-specific probes. The RNA-positive plants can then analyzed
for protein expression by Western immunoblot analysis using the
specifically reactive antibodies of the present invention. In addition,
in situ hybridization and immunocytochemistry according to standard
protocols can be done using heterologous nucleic acid specific
polynucleotide probes and antibodies, respectively, to localize sites of
expression within transgenic tissue. Generally, a number of transgenic
lines are usually screened for the incorporated nucleic acid to identify
and select plants with the most appropriate expression profiles.
[0199] A preferred embodiment is a transgenic plant that is homozygous for
the added heterologous nucleic acid; i.e., a transgenic plant that
contains two added nucleic acid sequences, one gene at the same locus on
each chromosome of a chromosome pair. A homozygous transgenic plant can
be obtained by sexually mating (selfing) a heterozygous transgenic plant
that contains a single added heterologous nucleic acid, germinating some
of the seed produced and analyzing the resulting plants produced for
altered expression of a polynucleotide of the present invention relative
to a control plant (i.e., native, non-transgenic). Back-crossing to a
parental plant and out-crossing with a non-transgenic plant are also
contemplated.
[0200] General Molecular Biology References
[0201] In the context of the invention, e.g., identifying, monitoring
and/or cloning molecular markers and/or other loci, nucleic acids and/or
proteins are manipulated according to well known molecular biology
techniques. Detailed protocols for numerous such procedures are described
in, e.g., in Ausubel et al. Current Protocols in Molecular Biology
(supplemented through 2000) John Wiley & Sons, New York ("Ausubel");
Sambrook et al. Molecular Cloning--A Laboratory Manual (2nd Ed.), Vol.
1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., 1989
("Sambrook"), and Berger and Kimmel Guide to Molecular Cloning
Techniques, Methods in Enzymology volume 152 Academic Press, Inc., San
Diego, Calif. ("Berger").
[0202] In addition to the above references, protocols for in vitro
amplification techniques, such as the polymerase chain reaction (PCR),
the ligase chain reaction (LCR), Q.beta.-replicase amplification, and
other RNA polymerase mediated techniques (e.g., NASBA), useful e.g., for
amplifying cDNA probes of the invention, are found in Mullis et al.
(1987) U.S. Pat. No. 4,683,202; PCR Protocols A Guide to Methods and
Applications (Innis et al. eds) Academic Press Inc. San Diego, Calif.
(1990) ("Innis"); Arnheim and Levinson (1990) C&EN 36; The Journal Of NIH
Research (1991) 3:81; Kwoh et al. (1989) Proc Natl Acad Sci USA 86, 1173;
Guatelli et al. (1990) Proc Natl Acad Sci USA 87:1874; Lomell et al.
(1989) J Clin Chem 35:1826; Landegren et al. (1988) Science 241:1077; Van
Brunt (1990) Biotechnology 8:291; Wu and Wallace (1989) Gene 4: 560;
Barringer et al. (1990) Gene 89:117, and Sooknanan and Malek (1995)
Biotechnologv 13:563. Additional methods, useful for cloning nucleic
acids in the context of the present invention, inlcude Wallace et al.
U.S. Pat. No. 5,426,039. Improved methods of amplifying large nucleic
acids by PCR are summarized in Cheng et al. (1994) Nature 369:684 and the
references therein.
[0203] Certain polynucleotides of the invention, e.g., oligonucleotides
can be synthesized utilizing various solid-phase strategies involving
mononucleotide- and/or trinucleotide-based phosphoramidite coupling
chemistry. For example, nucleic acid sequences can be synthesized by the
sequential addition of activated monomers and/or trimers to an elongating
polynucleotide chain. See e.g., Caruthers, M. H. et al. (1992) Meth
Enzymol 211:3.
[0204] In lieu of synthesizing the desired sequences, essentially any
nucleic acid can be custom ordered from any of a variety of commercial
sources, such as The Midland Certified Reagent Company (mcrc@oligos.com),
The Great American Gene Company (www.genco.com), ExpressGen, Inc.
(www.expressgen.com), Operon Technologies, Inc. (www.operon.com), and
many others.
[0205] Similarly, commercial sources for nucleic acid and protein
microarrays are available, and include, e.g., Affymetrix, Santa Clara,
Calif. (http://www.affymetrix.com/); and Incyte, Palo Alto, Calif. (on
the world wide web at incyte.com); and Ciphergen Biosciences, Fremont,
Calif. (at ciphergen.com).
[0206] High Throughput Screening
[0207] In one aspect of the invention, the determination of genetic marker
alleles is performed by high throughput screening. High throughput
screening involves providing a library of genetic markers, e.g., SSR
primers, ASH primers and probes, RFLPs, AFLPs, isozymes, specific alleles
and variable sequences, including SSR, RAPD and the like. Such libraries
are then screened against plant genomes to generate a "fingerprint" for
each plant under consideration. In some cases a partial fingerprint
comprising a sub-portion of the markers is generated in an area of
interest. Once the genetic marker alleles of a plant have been
identified, the correspondence between one or several of the marker
alleles and a desired phenotypic trait is determined through statistical
associations based on the methods of this invention.
[0208] High throughput screening can be performed in many different
formats. Hybridization can take place in a 96-, 324-, or a 1524-well
format or in a matrix on a silicon chip or other format.
[0209] A number of well-known robotic systems have been developed for high
throughput screening, particularly in a 96 well format. These systems
inlcude automated worksations like the automated synthesis apparatus
developed by Takeda Chemical Industries, LTD. (Osaka, Japan) and many
robotic systems utilizing robotic arms (Zymate II, Zymark Corporation,
Hopkinton, Mass.; ORCA.TM., Beckman Coulter, Fullerton Calif.). Any of
the above devices are suitable for use with the present invention. The
nature and implementation of modifications to these devices (if any) so
that they can operate as discussed herein will be apparent to persons
skilled in the relevant art.
[0210] In addition, high throughput screening systems themselves are
commercially available (see, e.g., Zymark Corp., Hopkinton, Mass.; Air
Technical Industries, Mentor, Ohio; Beckman Instruments, Inc. Fullerton,
Calif.; Precision Systems, Inc., Natick, Mass., etc.). These systems
typically automate entire procedures including all sample and reagent
pipetting, liquid dispensing, timed incubations, and final readings of
the microplate or membrane in detector(s) appropriate for the assay.
These configurable systems provide high throughput and rapid start up as
well as a high degree of flexibility and customization. The manufacturers
of such systems provide detailed protocols for the use of their products
in high throughput applications.
[0211] In one variation of the invention, solid phase arrays are adapted
for the rapid and specific detection of multiple polymorphic nucleotides.
Typically, a nucleic acid probe is linked to a solid support and a target
nucleic acid is hybridized to the probe. Either the probe, or the target,
or both, can be labeled, typically with a fluorophore. If the target is
labeled, hybridization is evaluated by detecting bound fluorescence. If
the probe is labeled, hybridization is typically detected by quenching of
the label by the bound nucleic acid. If both the probe and the target are
labeled, detection of hybridizaiton is typically performed by monitoring
a color shift resulting from proximity of the two bound labels.
[0212] In one embodiment, an array of probes are synthesized on a solid
support. Using chip masking technologies and photoprotective chemistry,
it is possible to generate ordered arrays of nucleic acid probes. These
arrays, which are known, e.g., as "DNA chips" or as very large scale
immobilized polymer arrays (VLSIPS.TM. arrays) can include millions of
defined probe regions on a substrate having an area of about 1 cm.sup.2
to several cm.sup.2.
[0213] In another embodiment, capillary electrophoresis is used to analyze
polymorphism. This technique works best when the polymorphism is based on
size, for example, SSR and AFLP. This technique is described in detail in
U.S. Pat. Nos. 5,534,123 and 5,728,282. Briefly, capillary
electrophoresis tubes are filled with the separation matrix. The
separation matrix contains hydroxyethyl cellulose, urea and optionally
formamide. The SSR and AFLP samples are loaded onto the capillary tube
and electrophoresed. Because-of the small amount of sample and separation
matrix required by capillary electrophoresis, the run times are very
short. The molecular sizes and therefore, the number of nucleotides
present in the nucleic acid sample is determined by techniques described
herein. In a high throughput format, many capillary tubes are placed in a
capillary electrophoresis apparatus. The samples are loaded onto the
tubes and electrophoresis of the samples is run simultaneously. See,
Mathies and Huang, (1992) Nature 359:167.
EXAMPLES
[0214] It is understood that the examples and embodiments described herein
are for illustrative purposes only and that various modifications or
changes in light thereof will be suggested to persons skilled in the art
and are to be included within the spirit and purview of this application
and scope of the appended claims.
Example 1
Summary of Breeding Bias Data-Favorable Allelic Forms by Geographic Region
[0215] Tables 3 through 9 enumerate favorable allelic forms of chromosome
segments contributing to superior agronomic performance in different
geographic regions and growing environments. In the table, * indicates
95% significance level, while ** indicates a 99% significance level. UM
indicates an unmapped locus. Details regarding SSR and ASH markers and
alleles are provided in Appendices I through IV. The geographic regions
are defined with respect to the following reference points.
[0216] Central Region: Central Midwest United States centered around Des
Moines, Iowa.
[0217] Canada Region: Eastern Canada centered around Chatham, Ontario.
[0218] North Region: Northern Midwest United States centered around
Redwood Falls, Minn.
[0219] Illinois Region: Northern and central Illinois centered around
Champaign, Ill.
[0220] East Region: Eastern Midwest United States centered around
Napoleon, Ohio.
[0221] Mid South Region: Mid Southern United States centered around Hamil,
Ill.
[0222] South Region: Southern United States centered around Memphis, Tenn.
Example 2
Favorable Soybean Alleles and "Target Genotype(s)" for Adaptation to
Environments silimar to Those Found in Iowa USA (Central Region)
[0223] Three separate breeding bias analyses were conducted to identify
favorable alleles that have provided adaptation to geographic regions
that are typical of Iowa, USA. The first analysis used 41 elite lines as
the "elite population" adapted to Iowa, the second analysis included a
larger sample of 71 elite lines (Table 1, Central Region) and the third
analysis, performed following the 2003 growing season, used 86 elite
lines (Table 1, Central Region 2003). In all cases, the elite population
was chosen as a representative sample of elite lines that yield well in
Iowa and/or are good parents for developing elite lines adapted to Iowa.
Each successive analysis was run after more elite line marker data was
available and was therefore considered more rigorous. Despite this fact,
the results of the first analysis were very similar and demonstrated that
the breeding bias method is useful even with smaller datasets. Herein,
results of the second (A) and third (B) analyses are reported due to the
larger sample size of the elite population.
[0224] A computer program described in U.S. Pat. No. 5,437,697 that
simulates the flow of alleles from ancestors to elite lines at each
marker locus was used to determine what percent of the elite lines would
be expected to inherit each marker allele by chance alone. By doing
multiple iterations of the simulation process (10,000 iterations in this
case), a statistical measure of random result variation was obtained. The
average frequency of each allele in the simulated elite population (from
the 10,000 iterations) is herein referred to as the "expected frequency"
(EXP) of each marker allele. The expected frequency was then compared to
the "observed frequency" (OBS) which is simply a count of how many lines
within the elite population actually contain a given allele divided by
the total number of elite lines examined. By comparing the observed
results to the results of the simulation process, one can determine how
often the observed results would be expected to occur by chance alone. If
the observed results would only be expected 5% of the time or less due to
chance (i.e. a LOD score of 1.3 or greater), it is safe to assume that
the observed results did not occur by chance alone. In this case,
selection for grain yield must have biased selection towards one or more
alleles at the locus in question. The allele(s) that occurred at higher
frequency than expected were therefore labeled as "favorable alleles" and
those occurring at a lower frequency than expected were labeled as
"unfavorable alleles." Favorable alleles are those that must have
contributed either directly or indirectly to higher grain yield.
[0225] The statistically significant results of the Iowa (A) analysis are
shown in Table 10. Since only the major ancestors were genotyped in the
analysis, occasionally an allele that was detected in the elite
population was not always found in the ancestral population. This
resulted in an unusually high LOD score since the frequency of the allele
in the ancestral population was assumed to be zero. It is also reasonable
to assume that a favorable allele should have increased in frequency by
some threshold percentage before that allele should be considered
generally "favorable" over the wide range of environments encountered by
soybean breeders over the past century. For these reasons, an allele with
a LOD score of greater than or equal to 1.3 (95% confidence) AND an
observed frequency of at least 25% higher than expected was considered to
be "favorable" and "significant" from both a practical and statistical
view. Alleles with these criteria plus a LOD score of 2.0 or greater (99%
confidence), were considered to be "favorable" and "highly significant."
Alleles with a LOD scores of greater than 1.3 but that did not increase
in frequency by at least 25% over the past century were considered
statistically significant but not enough to be considered generally
favorable over most environments.
[0226] The term "LOD" score refers to the negative inverse log (base 10)
probability that the observed frequency of an allele in the elite
population could be attributed to chance alone. More specifically, the
LOD score is a measure of the number of rounds that the breeding bias
simulation generated an allele frequency at least as extreme as that
observed in the actual elite population of soybean lines. The formula for
a LOD score of a "favorable" allele is: -1.0.times.log10 (f) where:
f=(the number of rounds of simulation where the observed allele frequency
in the elite population was greater than that generated by the simulated
allele frequency) divided by (the total number of rounds of simulation).
[0227] For example, a LOD score of >1.30 is analogous to a probability
of <0.05 that the observed allele frequency in the elite population
was due to chance alone; a LOD score of >2.00 is analogous to a
probability of <0.01 that the observed allele frequency in the elite
population was due to chance alone; a LOD score of >3.00 is analogous
to a probability of <0.001 that the observed allele frequency in the
elite population was due to chance alone; a LOD score of 4.00 is
analogous to a probability of 0.0001 that the observed allele frequency
in the elite population was due to chance alone. Since only 10,000 rounds
of simulation were conducted, the maximum LOD score observed did not go
any higher than 4.00.
[0228] Based on the Iowa analysis, out of 1540 alleles over 309 genetic
marker loci, a total of 94 alleles showed evidence of being favorable by
the aforementioned definition: a LOD score of at least 1.30 (95%
confidence level) and an increase in allele frequency of at least 25%
more than expected by random inheritance. Since some of these favorable
alleles are closely linked (i.e., less than 10 CM apart according to
independent mapping studies), not all 94 alleles are diagnostic of unique
QTL loci. Therefore, the markers were divided into genomic regions with
the assumption that markers on the same chromosome that are approximately
10 CM or greater apart are probably diagnostic of different QTL loci.
[0229] In addition to listing the favorable alleles that span the genome,
Table 10 indicates the favorable allele with the best statistical score
and/or the most marker data within each predefined genomic region. The
"best" marker allele to use for each region can also be chosen based on
which marker works best in the laboratory. In most cases, the marker with
the highest LOD score and highest % difference of expected and observed
allele frequency was considered to "best" marker in its genomic region.
For example, on chromosome C1, there are 4 markers that map to positions
95.8 through 99.0-Satt399, Satt361, P10639A-1, and Satt190. These are all
within 10 CM of each other and therefore were assigned to the same
genomic region #29. Among these 4 markers, markers Satt399 and Satt190
had the highest possible LOD score of 4.0 when 10,000 iterations of a
simulation were done. This means that even after 10,000 iterations were
done, there was not even 1 iteration where the simulation produced
results more extreme than that observed in the actual elite population. A
LOD score of 4.0 is merely the inverse log of the probability (1 in
10,000) that the results obtained at these loci was due to chance alone.
From there, one can decide which of these 2 loci would be the best marker
to identify the favorable allele in that region. In this case, Satt190
was chosen as the best marker in that region because the results were
based on data from 65 as opposed to 61 elite lines. Although a total of
71 elite lines were genotyped for this marker, 6 elite lines had missing
data for this locus.
[0230] Once a desirable marker is identified and the favorable allele of
that marker is determined, selection for that favorable allele in a lab
assay can then be used for MAS to identify plants that have the favorable
allele. The MAS process can be done at multiple loci simultaneously to
select for plants that contain the maximum number of favorable alleles
that span the genome. This genome-wide group of favorable alleles upon
which selection is based is herein referred to as the "Target Genotype."
Table 11 shows the genome-wide Target Genotype for the best locus from
each genomic region that meets the criteria of "significant"--i.e. LOD
score of at least 1.3 and an increase in allele frequency of at least 25%
greater than expected by chance alone. A total of 57 favorable alleles,
each representing what was favored by selection in a different genomic
region, fit these criteria to make the Target Genotype at LOD1.3 (Table
11, *). If one raises the statistical cutoff to a LOD of 2.0, the Target
Genotype focuses in on 30 favorable alleles (Table 11, **). For purposes
of MAS, one would want to select for the favorable alleles with the
highest statistical significance first. Hence, the logical path for
breeding purposes would be to select among segregating progeny for the 30
locus Target Genotype first. Once these loci are fixed for the favorable
alleles, the other 27 loci in the 57-locus Target Genotype could be the
focus of selection. If resources are unlimited, one could conceivably
work with all 57 loci. Loci can also be weighted based on their
statistical significance for selection purposes.
[0231] Following the 2003 growing season, an additional analysis (B) was
performed increasing the number of elite parental lines from 71 to 86,
and evaluating an additional 265 SSR markers. The elite lines used in
this analysis are given in Table 1.
[0232] Significant favorable alleles are given in Table 12. Because more
elite lines were used to define the population, and because some
previously absent data was obtained for the previously employed markers,
some differences in the statistical LOD scores were observed for the
markers in Analysis B as compared to prior Analysis A. Almost all of the
previously identified markers were confirmed in the expanded analysis,
and several new markers in the same genomic regions were found to have
higher LOD scores. Accordingly, these new markers are also deemed to be
useful in defining the target genotype and identifying and tracking
favorable alleles in soybean germplasm.
[0233] The Target Genotype is actually a consensus marker genotype that
the elite population has been moving towards as the result of selection.
Since this genotype is now defined by specific markers and specific
favorable alleles, it is possible to practice selection by genotype
instead of the inefficient and slow process of selection based on
phenotype. The resolution of the consensus genotype is limited only by
the genomic coverage provided by the genetic markers that are available
for MAS.
Example 3
Status of Favorable Alleles in Soybean Variety A3127
[0234] The following example is given to illustrate how the favorable
alleles identified in the previous example came together in the most
famous transgressive segregant in soybean breeding history. Most new
soybean varieties are a small improvement over either of their parents in
terms of yield. Yield progress per cycle (5 to 6 years) of breeding is
commonly a few percent better than either parent. However, in the early
1980's a variety called A3127 was developed that was much better than
either parent (.about.10% better than either parent). In fact, A3127 is
probably one of the few lines that all soybean breeders are familiar with
because it was famous for being the highest yielding variety of it's time
(early 1980's). Prior to commercialization A3127 proved to be much higher
yielding than either of its two parents Williams and Essex. A3127 was so
popular, that it became the most frequently used parent in soybean
breeding history. Since A3127 is adapted to Iowa, we studied the marker
profiles of Williams, Essex, and A3127 at the 30 preferred "yield gene"
loci identified for the Iowa geographic zone. We found that Williams and
Essex differ at 23 out of 30 of these loci (Table 13). Out of the 23
segregating loci, Williams supplied 13 of the favorable alleles and Essex
supplied the other 10 favorable alleles. If these really are the major
yield loci, one would expect that A3127 would have significantly more
favorable alleles than either parent. Amazingly, A3127 had all 23
favorable alleles. Such a segregant would only be expected to happen by
chance in 1 out of >8 million progeny (0.5.sup.23) unless these loci
really are diagnostic of yield and A3127 is truly a unique segregant. The
marker genotype of A3127 is therefore consistent with the hypothesis that
these 30 marker loci are diagnostic of yield.
Example 4
Segregation for Yield in Near-Isogenic Sublines
[0235] Typically, modern soybean varieties originate from a single plant
selected from a partially inbred (commonly F3) population that was
generated from a controlled mating between genetically different parents.
Seed of the new variety is then multiplied by subsequent pooling (or
"bulking") of seed from the self-pollinating progeny of the selected F3
plant. For any locus in the original F3 plant that was heterozygous, the
resulting inbred variety will eventually become a mixture of the two
homozygotes at said locus. For commercial purposes, soybean varieties are
purified for obvious visual traits (e.g., flower color, hilum color,
maturity, and other visual traits that are highly heritable and
controlled by one or a few genes) but often harbor residual genetic
variation that is not obvious to the naked eye. Genetic markers can be
used to detect loci contributing to that variation that are still
segregating within a so-called "pure line." Since the breeding bias
analysis identifies which marker loci have been affected by selection for
seed yield, these markers are ideal tools to identify genetic differences
among plants within the original heterogeneous variety that may translate
into seed yield improvement. These markers can be used to separate the
original heterogeneous variety into "near-isogenic" sublines that differ
at specific genetic loci.
[0236] For example, samples of seed from individual self-pollinated
progeny selected from the variety are genotyped, and seed sharing a
common allele at one or more identified marker loci is pooled to produce
a subline. Such sublines are genetically distinct from one another. In
"blind" sublining (i.e., sublining unassisted by marker data) there is no
guarantee that the generated sublines are genetically distinct. By
pooling seed of many plants with a homogenous marker genotype, enough
seed for controlled replicate trials can be obtained in one generation.
This is preferable to blind sublining in several ways. First, the only
way to attempt genetic homogeneity without markers is to pool the seed of
a single plant that may or may not be genetically distinct from the seed
of other single plants. Second, a single plant can only supply enough
seed for a short row yield test in one environment. Therefore, blind
sublining requires subsequent generations of seed increase to obtain
enough seed for highly replicated yield trials that are necessary for
reliable yield comparisons. Third, even if phenotypic differences are
observed with blind sublining, no genomic information is gained for
future use. By comparing the field performance of such marker-based
sublines in controlled experiments, one can determine the phenotypic
effect (e.g., with respect to yield) of each allele (and the
corresponding genomic region, if the marker is mapped) in a given genetic
background. This is particularly useful for traits, such as yield, in
which gene-by-gene and gene-by-environment interactions play a
substantial role in phenotype. If one subline performs significantly
better than the other, the better subline can be multiplied and released
as an improved version of the original variety.
[0237] Because selection that is based on genotype (e.g., qualitative DNA
polymorphism) and then confirmed with a phenotypic difference is more
reliable and heritable than selection based on phenotype alone (i.e.,
blind sublining), the improvement in phenotype in a blind subline is less
likely to be heritable, and unlikely to be repeated in subsequent
generations. In contrast, marker-based sublining not only provides useful
genomic information, but it also improves the heritability and
reproducibility of selected traits. Thus, marker-based "sublining" can be
a powerful tool for both product development and to determine the
phenotypic effect of individual loci in a given genetic background.
[0238] In accordance with this method, the genetic markers identified
through breeding bias were shown to be effective tools to select within
elite lines for residual yield gain. When genotyping elite lines with
genetic markers, 8 random plants from each elite line are routinely
sampled and bulked. If the elite line is a 50:50 mixture of two
homozygotes at a given locus, a random 8-plant sample will detect both
alleles >99% of the time. Using this sampling procedure, segregation
within commercial soybean lines is detected at an average of about 4% of
the marker loci assayed (e.g., when assaying the "best" marker loci for
each chromosomal region).
[0239] In the following exemplary trial, six elite lines were examined.
Two of the elite lines (91B91 and 92M70) were shown to be segregating at
two marker loci each while the other 4 elite lines (92B05, 93B01, 93M80,
and 93M90) were segregating at one marker locus as indicated in Table 14.
[0240] To develop the sublines, leaf tissue from individual plants of each
of the above elite lines was genotyped with the marker(s) segregating in
the originating line. Progeny seed from individual homozygous plants of
the same marker genotype were then pooled to obtain enough seed of each
subline to conduct replicated yield trials. The number of plant used to
create each subline is shown in Table 15.
[0241] To determine the relative seed yield, sublines derived from the
same elite line were planted in a split plot field design at between 5
and 14 locations, treating each location as an individual replication.
Locations were chosen to span the soybean growing region of appropriate
maturity zone for the lines being tested in the Midwestern United States.
Sublines derived from a given elite line were randomly assigned to split
plots within each main plot. Each split plot consisted of two 12-foot
long rows of a given subline that were spaced 30 inches apart. Seed yield
was measured at maturity and converted to bushels per acre (1 bushel-60
pounds).
[0242] Significant yield differences between isolines were detected in 3
of the 6 isoline tests. Since the isolines tested above were derived from
pooling many plants of similar genotype, one can reasonably assume that
the possibility of residual segregation at other independent loci was
randomized and not the source of the yield difference between isolines.
The magnitude of the significant yield differences (5.4 to 6.2% between
sublines or 2.7 to 3.1% better than the original mixed variety) is of
similar magnitude as yield improvements that can typically be obtained
using much more exhaustive breeding efforts. New soybean varieties
developed without the aid of yield gene markers can easily require
hundreds to thousands of yield plots to identify a new variety that is 2
or 3% better than it's best parent. This method can be used to identify
yield gains of similar magnitude with very limited resources (2
isolines.times.14 replications=28 plots per test). In addition, by basing
selection on a real genetic difference at a locus showing historical
breeding bias, the confidence that the yield differences detected are
genetically based (as opposed to environmental or experimental error) is
substantially increased.
[0243] Additionally, these results confirm that the effects of epistasis,
gene-by-environment interactions and/or recombination between the marker
allele identified by breeding bias and the genetic element underlying
yield improvements, while prevalent, do not impair selection of improved
soybean varieties, especially if care is taken to identify residual
variation and select appropriate sublines. For example, Satt591 was used
to select sublines from two different elite lines (93B01 and 93M90).
Breeding Bias analysis alone indicated that Satt591 allele 3 was the one
favored by breeders over time. In the case of 93B01, allele 3 was the
favorable allele since it was the genotype of the better-yielding
subline. In contrast, in the case of 93M90, marker allele 1 was the
favorable allele.
[0244] For example, while the Breeding Bias analysis identifies marker
loci linked to genetic elements which have been favorable on most genetic
backgrounds in a variety of growing environments, epistatis and other
non-additive interactions influence which allele is "favorable" within
specific populations, or for particular environments. In addition,
disease resistance genes, which contribute to higher relative yield when
the disease is prevalent, have been documented to result in lower yield
in the absence of disease pressure.
[0245] Recombination between a marker locus and the linked genetic element
contributing to improved yield can also reduce efficiency of marker
assisted. An accepted and proven genetic principle is that the frequency
of crossing over between two genetic loci, e.g., a marker locus and a
quantitative trait locus, is a function of genetic distance between the
two loci. The only way to avoid such phase reversals is to develop
"perfect" markers that are diagnostic of the DNA polymorphism that is
responsible for the phenotypic difference controlled by the QTL. That is,
recombination can only be eliminated by cloning the QTL, and identifying
the mutation causally determining the difference in phenotype.
Development of perfect markers is possible but is not a trivial exercise.
It requires DNA sequencing of the surrounding genomic region and
exhaustive sequence-phenotype association to determine conclusively which
DNA polymorphism is always associated with the desired phenotype. This is
an expensive and time-consuming endeavor, the benefits of which can, in
large part, be achieved using the methods and marker loci of the present
invention, without the expense and delay of cloning each significant
yield QTL associated with a marker locus identified using breeding bias.
By periodically confirming marker and phenotype association, using the
methods of the present invention, breeders can still reap the benefits of
linked (but non-perfect) markers.
[0246] Breeding Bias is an effective method for identifying genomic
regions that have undergone directional selection. If a marker is close
enough (typically within about 10 CM) to an important QTL in a given
ancestor, the marker allele originally linked in coupling to the
favorable QTL allele will remain in coupling phase for a sufficient
period of selection under standard breeding procedures to detect that
selection is occurring in the genomic region including the marker. Thus,
the marker loci enumerated herein are associated with, and stand as
proxies for, QTL contributing to increased yield. Although, with repeated
cycles of recombination among members of a given gene pool, genetic
crossovers between the marker locus and the QTL will tend to accumulate
and eventually result in a state of "linkage equilibrium" between the
marker alleles and the QTL, periodic reassessment using the near isogenic
subline procedures described herein can insure that selection proceeds
for the allele in linkage phase with the desired QTL allele, despite the
potential for recombination.
[0247] The above subline experiments indicate that marker-based sublining
is an effective method for purification and improvement of elite soybean
lines. If the number of markers segregating within a given line or
population is small, non-additive effects and linkage phase (coupling or
repulsion) do not pose a problem, as it is fairly inexpensive to field
test all possible recombinants and identify those with the optimal
phenotype.
Example 5
Allele Confirmation using Random Crosses
[0248] To increase efficiency of marker assisted selection for improved
yield using, e.g., the marker loci enumerated herein, the allele of the
marker locus segregating with yield can be confirmed. Following
identification of marker loci by Breeding Bias, a limited number of
crosses is performed between the highest yielding elite parents for a
particular geographic zone or growing environment. Preferably, the
parents should be as polymorphic as possible at the identified marker
loci. Progeny (F1) from these elite by elite crosses is inbred to
generate a large population (e.g., between about 200-5000, typically at
least about 1000) F3-derived lines. If desired, inbreeding to later
generations can also be done to increase genetic variation among lines.
[0249] A subset of "tester lines" is randomly selected from among the
inbred lines derived from each cross. For example, between about 10 and
500 lines can be selected. Typically, between about 50 and 100 inbred
lines are randomly selected, and enough seed to conduct a reliable yield
trial is produced. Several (i.e., between at least 5 and 12, e.g., 8)
plants from each inbred line are genotyped at marker loci segregating in
the elite parents from which the line was founded, to determine whether
the line is segregating or fixed (homozygous) with respect to the
relevant marker. The remaining lines ("remnant population") can be stored
under conditions that preserve seed viability. If desired, additional
lines can be selected for testing, or genotyped for presence of the
alleles confirmed to be in coupling linkage phase.
[0250] For each cross, a replicated yield trial of each tester line is
performed. The test is replicated in enough environments to adequately
sample the geographic region of interest and to gain a reliable measure
of phenotype. The effect on yield for each marker locus within each cross
is determined by comparing the mean yield of lines with a first allele to
the mean yield of lines with the alternate allele. If the difference in
yield is not significant, the marker can be eliminated in that cross. In
contrast, if the difference in yield is significant, the "favorable"
allele is confirmed and the locus is used for subsequent marker assisted
selection for yield.
[0251] Confirmation of the favorable alleles also permits identification
of a "target genotype" including all of the favorable alleles across all
segregating loci in a particular elite by elite cross. As indicated
above, the entire remnant population can be screened with the subset of
confirmed markers to identify those segregants that have the highest
number of favorable alleles. Typically, at least 5% of the remnant lines
that most closely approach the target genotype will be selected, although
additional lines can be included at the breeder's discretion. The
selected lines can then be evaluated in highly replicated yield tests to
identify which crosses perform better than either elite parent under a
variety of environments and growing conditions.
[0252] While the foregoing invention has been described in some detail for
purposes of clarity and understanding, it will be clear to one skilled in
the art from a reading of this disclosure that various changes in form
and detail can be made without departing from the true scope of the
invention. For example, all the techniques and apparatus described above
can be used in various combinations. All publications, patents, patent
applications, and/or other documents cited in this application are
incorporated by reference in their entirety for all purposes to the same
extent as if each individual publication, patent, patent application,
and/or other document were individually indicated to be incorporated by
reference for all purposes.
1TABLE 1
Elite Soybean Lines by Geographic Region
Central Iowa RF SJ NP HL ME
(Iowa) (2003) Canada (North)
(Illinois) (East) (MidSouth) (South)
91B91 92B05 90A07
90B73 92B38 92B52 93B66 94B73
92B05 92B12 90B11 91B12 92B63 91B91
93B67 94B74
92B38 92B38 90B31 91B33 92B74 92B05 93B82 94M70
92B52 92B52 90B43 91B53 93B01 92B38 93B84 94M90
92B63 92B63 90B72
91B91 93B11 92B63 93B85 95B32
92B74 92B74 90B73 91B92 93B15 92B74
93B86 95B33
92B75 92M30 91B01 92B05 93B41 93B01 93B87 95B34
92B84 92M31 91B12 92B12 93B66 93B46 94B01 95B42
93B01 92M70 91B33
92B23 93B67 93B67 94B23 95B43
93B41 92M71 91B52 92B38 93B82 93B82
94B24 95B53
93B45 92M72 91B53 92B63 93B85 93B86 94B53 95B95
93B47 92M80 91B64 92B74 93B86 93B87 94B54 95B96
93B66 92M91 92B95
A1395 93B87 A3431 94B73 95B97
93B67 93B09 93B25 CX105 94B53 CX289
A4009 95M80
93B72 93B36 93B26 D00566D362 94B54 CX394C A4138 96B21
93B82 93B41 A1395 EX04C00 94B73 EX39E00 A4415 96B32
93B85
93B45 CX105 EX06A00 CM428 G3362 A4595 96B51
93B86 93B66 EX10F01
EX10F01 MO15733 MO15733 A4715 96M20
93B87 93B67 P1677 EX13P01
MP39009 P9273 CM428 97B52
94B23 93B68 P9007 EX13Q01 P9306 P9281
CX469C 97B61
94B54 93B82 P9008 EX15N01 P9395 P9306 P9395 A5403
A2835 93B85 P9041 EX16N00 S32Z3 P9392 P9481 A5560
A2943 93B86
P9042 EX16P01 S33N1 P9395 P9482 A5843
A3127 93B87 P9061 EX22Y01
S38T8 S3911 P9492 A5885
A3242 93M10 P9062 EX22Z01 S42H1 ST2660
S3911 A5979
CX232 93M30 P9063 JIM ST2660 ST3380 S4260 A5980
CX253 93M40 P9071 KORADA ST2870 ST3883 S42H1 A6297
HS93-4118 93M50
P9092 MO400644-02 ST3171 XB31C01 S43B5 BEDFORD
P9273 93M60 P9132
MO413735- ST3630 XB33B ST3870 CLIFFORD
11-52
P9281 93M80
P9141 MO501577- ST3883 XB34F01 ST3883 CM428
27-23
P9305
93M90 P9163 MO505469- XB31C01 XB35D XB41M01 DAVIS
61-89
P9306 93M92 P9182 P9007 XB34F01 XB35W00 XB42J00 ESSEX
P9321 93M93
P9203 P9151 XB38A01 XB38A01 XB42M01 EX53F03
P9341 A2722 P9244
P9233 XB42M01 YB25Y01 XB48H01 EX56H03
P9395 A3237 R01154 S03W4
XB48H01 YB25Z01 YB40M01 EX61E03
R002
S22C3 A3322 S0066
S0880 YB28N01 YB27X01 YB40N01 FORREST
ST1970 A3431 S0880 S19T9
YB29H01 YB27Y01 YB41Q01 FOWLER
ST2250 A4138 S1550 S20F8 YB29J01
YB28N01 YB48L01 HARTWIG
ST2488 EX23B03 S1990 S24L2 YB30J01 YB29H01
HOLLADAY
ST2688 EX34T03 S19T9 S25J5 YB30N01 YB29J01 HOOD
ST2870 EX35F03 ST1073 ST0653 YB35C01 YB30J01 HUTCHESON
ST3171
EX36Y01 XB03F01 ST1090 YB36V00 YB30P01 LEE
ST3380 EX40T03 XB07E01
ST1970 YB39M01 YB33K01 MANOKIN
ST3630 EX44V03 TRAILL YB40M01
YB35C01 P9481
ST3870 S25J5 X9916 YB40N01 YB36V00 P9482
ST3883 S32Z3 XB03F01 YB48L01 YB40N01 P9492
XB22R01 ST1570
XB10D01 P9521
XB25W01 ST1690 XB15M01 P9552
XB31C01
ST1970 XB20M01 P9561
XB34F01 ST2488 XB22R01 P9584
XB35W00 ST2686 XB25W01 P9591
XB38A01 ST2788 YB03E00 P9592
XB42M01 ST2870 YB03G01 P9593
YB22W01 ST3171 YB08D01
P9594
YB25R99 ST3630 YB09F01 P9611
YB25Y01 ST3660
YB09G01 P9631
YB27X01 ST3870 YB10E01 P9641
YB28N01
ST3883 YB11D01 P5960
YB29H01 XB19U04 YB14H01 PHARAOH
YB29J01 XB22C04 YB15K99 RA451
YB30J01 XB23W03 YB21F01
S5960
YB30P01 XB23Y02 YB21G01 S6189
YB31E01 XB25E02
YB22S00 S6262
YB32K01 XB25L04 YB22V01 TRACY
YB33K01
XB25X04 YB22W01 XB48H01
YB34H01 XB26L04 YB22X01 XB48T04
YB37A01 XB27P04 YB24Z01 XB53J04
YB39M01 XB29A04 YB25R99
XB54K01
YB40M01 XB29D01 YB25X00 XB55J01
YB40N01
XB29K04 YB25Y01 XB55K04
YB41Q01 XB29L04 YB25Z01 XB57M04
XB30E04 YB27Y01 XB58P99
XB31R04 XB58Y02
XB34D04 XB58Z04
XB35L04 XB59U04
YB25R03
XB63D00
YB27L03 XB64C04
YB27S00 XB67A00
YB28A03 YB48L01
YB29T04 YB51D00
YB34R03
YB52J00
YB34S03 YB53C00
YB36E03 YB53D00
YB38E03 YB53E00
YB38G03 YB53G03
YB39V03
YB53H03
YB53K03
YB54H00
YB54J00
YB54L00
YB55G00
YB55H00
YB56E00
YB56G03
YB57L03
YB58B04
YB59T03
YB59V03
YB60N01
YB60P03
YB63E00
YB65C00
YOUNG
[0253]
2TABLE 2
Exemplary Target Progeny from Segregating
Cross.
Target
Marker Parent 1 Parent 2 Progeny
1 + - +
2 - + +
3 + - +
4 - + +
5 + - +
6 - + +
7 + - +
8 - + +
9 + - +
10 - + +
where
+ = contains the most
favorable allele
- = contains some other allele
[0254]
3TABLE 3
Favorable Allelic Forms: Central Region
Region Chromosome Position Locus Allele Sig.
2 A1 19.1
SATT042 3 **
2 A1 19.1 SATT364 3 **
2 A1 19.1 SATT454 1 **
2 A1 19.1 SATT526 1 **
3 A1 27.1 SATT300 3 **
3 A1
27.1 SATT591 3 **
3 A1 28.5 SATT155 2 **
4 A1 69.9 SATT385
1 *
5 A1 87.3 SATT236 1 *
5 A1 87.3 SATT511 1 *
6 A2
0.0 P12390B-1 1 **
13 A2 184.0 SATT429 4 **
15 B1 39.0
SATT197 5 *
17 B1 74.1 SATT583 1 *
20 B2 20.0 P12105A-1 1 *
21 B2 45.0 P10641A-1 2 **
23 B2 86.1 SATT556 2 *
23
B2 88.6 SATT020 2 *
25 B2 110.9 SATT534 7 *
26 B2 120.0
P10638B-2 1 *
29 C1 95.8 SATT399 3 **
29 C1 96.5 SATT361 2
**
29 C1 98.0 P10639A-1 1 **
29 C1 99.0 SATT190 4 **
31 C1 173.0 SATT338 3 *
36 C2 140.9 SATT557 5 **
36 C2
145.8 SATT319 1 *
37 C2 165.8 SATT460 5 *
37 C2 170.5
SATT433 3 *
38 C2 193.5 SATT357 1 *
40 D1a 54.7 SATT321 2 *
41 D1a 68.6 SATT383 3 *
41 D1a 69.8 SATT295 2 *
42
D1a 80.3 SATT507 3 *
43 D1a 129.2 SATT129 5 **
43 D1a 129.2
SATT147 1 *
44 D1b 0.0 SATT216 4 *
45 D1b 20.0 P10621B-2 2
*
46 D1b 39.8 SATT558 3 *
49 D1b 76.1 SATT546 4 *
51
D1b 120.0 P13072A-1 2 *
53 D2 66.9 SATT582 2 **
54 D2 93.8
SATT389 6 **
55 D2 110.4 SATT464 1 **
57 D2 148.3 SATT413 3
*
60 E 40.0 P13074A-1 1 *
62 E 98.0 SATT573 2 *
62 E
98.0 SATT598 1 **
62 E 100.6 SATT263 2 *
62 E 101.0 SATT602
4 *
62 E 102.2 SATT151 4 *
62 E 102.2 SATT355 3 *
62
E 102.2 SATT452 2 *
64 F 0.0 SATT146 1 **
64 F 0.0 SATT193
3 **
64 F 0.0 SATT569 3 **
64 F 1.7 SATT343 3 **
64
F 1.9 SATT586 5 **
66 F 57.5 SATT595 1 *
67 F 85.0
P10782A-1 1 **
67 F 95.0 SATT334 3 *
70 F 157.8 SATT144 1 *
71 F 165.8 SATT522 5 *
73 G 8.5 SATT356 2 *
75 G
66.1 SATT533 1 *
77 G 100.8 SATT199 1 **
77 G 103.2 SATT503
3 *
77 G 103.2 SATT517 4 *
79 G 137.0 SATT191 2 *
79
G 138.4 SAT_117 1 *
81 H 0.0 SATT353 2 *
85 H 125.3 SATT181
5 **
86 I 15.5 SATT127 2 *
87 I 57.9 SATT270 5 *
92
J 61.3 SCT_065 2 **
92 J 63.6 SATT596 4 **
92 J 68.0
SATT406 2 **
92 J 70.8 SATT380 3 **
92 J 72.6 SATT183 1 **
92 J 74.4 SATT529 1 **
94 K 17.9 SATT242 4 **
96 K
70.9 SATT441 2 *
96 K 72.8 SATT544 2 *
97 K 85.8 SATT240 2
*
101 L 34.6 SATT398 4 **
102 L 42.3 SATT497 3 *
102
L 47.7 SATT284 2 *
103 L 77.1 SATT166 3 **
103 L 78.0
SATT448 4 *
104 L 118.0 SATT373 3 **
104 L 118.6 SATT513 11
*
104 L 124.5 P12394A-1 2 **
108 M 87.6 SATT536 4 **
108 M 91.1 SATT175 5 *
109 M 115.0 P10615A-1 1 *
110 M
143.5 SATT346 3 *
111 M 173.5 SATT336 4 *
112 N 20.0
P13069A-1 3 **
112 N 25.0 P5467A-1 2 **
115 N 88.6 SATT339
5 *
118 O 2.4 P12396A-1 1 *
118 O 6.3 SATT487 4 **
119 O 37.7 SATT259 4 **
120 O 60.5 SATT420 2 **
120 O 65.1
SATT576 4 *
122 O 103.8 SATT477 3 *
123 O 125.0 SATT581 2
**
124 O 153.3 SATT153 3 **
124 O 155.1 SATT243 1 **
128 UM # P10793A-1 2 *
129 UM # P12391A-1 1 *
129 UM #
P12392A-1 1 *
131 UM # P13560A-1 1 *
131 UM # P13561A-1 2
**
133 UM # SAC1677 3 **
134 UM # SATT040 4 **
139
UM # SATT111 3 **
140 UM # SATT176 3 **
143 UM # SATT299 5
*
[0255]
4TABLE 4
Favorable Allelic Forms: Canada
Region Chromosome Position Locus Allele Sig.
4 A1 69.9
SATT385 6 *
5 A1 87.3 SATT225 1 *
9 A2 108.7 SATT508 2 **
11 A2 136.0 P10635A-1 1 *
15 B1 39.0 SATT197 5 *
17 B1
68.1 SATT597 3 *
17 B1 71.6 SCT_026 1 *
17 B1 73.3 SATT415
3 *
17 B1 74.1 SATT583 1 *
18 B1 80.0 P12198A-1 1 *
18 B1 92.1 SATT359 1 *
23 B2 86.1 SATT556 2 *
29 C1 95.8
SATT399 3 **
29 C1 96.5 SATT361 2 *
29 C1 98.0 P10639A-1 1
**
29 C1 99.0 SATT190 4 **
31 C1 173.0 SATT338 3 **
32 C2 25.4 SATT227 1 *
33 C2 48.1 SATT422 4 *
36 C2 145.8
SATT319 3 *
41 D1a 68.6 SATT267 1 **
42 D1a 80.3 SATT507 3
**
42 D1a 85.0 P10620A-1 1 *
43 D1a 129.2 SATT129 2 *
46 D1b 39.8 SATT558 3 *
48 D1b 65.9 SATT506 3 *
53 D2 66.9
SATT582 2 **
55 D2 110.4 SATT464 1 **
62 E 100.6 SATT204 4
*
62 E 100.6 SATT263 2 *
62 E 101.0 SATT491 3 **
62
E 102.2 SATT151 4 *
62 E 102.2 SATT355 3 *
62 E 102.2
SATT452 2 *
64 F 0.0 SATT146 1 **
64 F 0.0 SATT193 3 *
64 F 0.0 SATT569 3 *
64 F 1.7 SATT343 3 *
64 F 1.7
SATT343 5 *
64 F 1.9 SATT586 5 **
67 F 85.0 P10782A-1 1 **
67 F 90.0 P10598A-1 1 *
67 F 95.0 SATT334 3 **
68 F
114.8 SATT510 3 *
72 F 175.0 P9026A-1 2 *
75 G 53.0 SATT115
1 *
85 H 125.3 SATT181 5 **
86 I 9.8 SATT367 2 *
86
I 15.5 SATT127 2 **
86 I 16.6 SCTT012 2 *
87 I 57.9 SATT270
5 *
89 I 114.2 SATT440 2 *
92 J 68.0 SATT406 2 *
92
J 70.8 SATT380 3 *
92 J 72.6 SATT183 1 **
92 J 74.4 SATT529
1 *
96 K 70.9 SATT441 3 *
97 K 85.8 SATT240 2 *
101
L 32.4 SATT523 1 *
103 L 77.1 SATT166 3 *
103 L 78.0
SATT448 1 *
104 L 124.5 P12394A-1 2 **
108 M 87.6 SATT536 4
*
110 M 143.5 SATT346 3 *
112 N 20.0 P13069A-1 2 **
113 N 35.4 SATT584 3 *
113 N 40.0 P3050A-2 2 *
114 N 61.2
SATT387 4 *
118 O 2.4 P12396A-1 1 *
118 O 6.3 SATT487 4 **
119 O 37.7 SATT259 4 *
119 O 39.0 SATT347 2 *
120 O
69.2 SATT262 5 *
123 O 130.0 P11070A-1 1 **
131 UM #
P13560A-1 1 **
131 UM # P13561A-1 2 **
133 UM # SAC1677 3 *
134 UM # SATT040 4 *
138 UM # SATT109 3 **
139 UM #
SATT111 3 **
140 UM # SATT176 3 *
140 UM # SATT176 5 *
145 UM # SATT512 5 **
[0256]
5TABLE 5
Favorable Allelic Forms: North Region
Region Chromosome Position Locus Allele Sig.
2 A1 19.1
SATT454 1 *
2 A1 19.1 SATT526 1 *
3 A1 27.1 SATT300 3 *
3 A1 27.1 SATT591 3 *
6 A2 0.0 P12390B-1 1 *
9 A2 108.7
SATT327 2 *
9 A2 108.7 SATT508 2 *
12 A2 154.7 SATT409 5 **
13 A2 184.0 SATT429 4 **
14 B1 22.5 SATT426 5 *
14
B1 26.7 SATT509 1 **
15 B1 39.0 SATT197 5 **
17 B1 73.3
SATT415 3 *
17 B1 74.1 SATT583 1 *
18 B1 80.0 P12198A-1 1
**
18 B1 85.0 P8584A-1 1 *
18 B1 92.1 SATT359 1 *
21
B2 45.0 P10641A-1 2 *
23 B2 86.1 SATT556 2 *
23 B2 86.1
SATT556 4 *
23 B2 88.6 SATT020 2 *
25 B2 110.9 SATT534 7 *
29 C1 95.8 SATT399 3 **
29 C1 96.5 SATT361 2 **
29
C1 98.0 P10639A-1 1 **
29 C1 99.0 SATT190 4 **
31 C1 173.0
SATT338 3 **
32 C2 25.4 SATT227 1 **
33 C2 53.4 SATT457 1 *
36 C2 140.9 SATT557 5 *
37 C2 168.3 P13073A-1 2 *
37
C2 170.5 SATT433 3 *
41 D1a 68.6 SATT267 1 *
42 D1a 80.3
SATT507 3 **
42 D1a 82.0 SAT_110 3 *
43 D1a 129.2 SATT129 5
*
46 D1b 39.8 SATT558 3 *
48 D1b 65.9 SATT506 3 **
51 D1b 120.0 P13072A-1 2 *
53 D2 66.9 SATT582 2 **
54 D2
93.8 SATT389 6 **
54 D2 99.6 SATT461 1 *
55 D2 110.4
SATT464 1 **
62 E 98.0 SATT573 2 *
62 E 98.0 SATT598 1 *
62 E 100.6 SATT204 3 *
64 F 0.0 SATT146 1 **
64 F 0.0
SATT193 3 **
64 F 0.0 SATT569 3 **
64 F 1.7 SATT343 3 **
64 F 1.9 SATT586 5 **
65 F 18.0 SATT348 2 *
67 F 85.0
P10782A-1 1 **
67 F 90.0 P10598A-1 1 *
67 F 95.0 SATT334 3
*
70 F 157.8 SATT144 1 *
71 F 165.8 SATT522 4 *
75 G
53.0 SATT115 3 *
75 G 61.3 SATT594 5 *
76 G 71.6 SATT303 6
*
76 G 72.4 SATT566 1 *
77 G 100.8 SATT199 1 *
77 G
103.2 SATT517 2 *
81 H 0.0 SATT353 2 *
85 H 125.3 SATT181 5
**
86 I 15.5 SATT127 2 **
87 I 57.9 SATT270 5 **
89
I 114.2 SATT440 2 **
90 J 10.5 SATT249 4 *
92 J 61.3
SCT_065 2 *
92 J 63.6 SATT596 4 *
92 J 68.0 SATT406 2 **
92 J 70.8 SATT380 3 **
92 J 72.6 SATT183 1 **
92 J 74.4
SATT529 1 **
94 K 17.9 SATT242 4 *
101 L 32.4 SATT523 1 *
101 L 34.6 SATT398 4 **
102 L 42.3 SATT497 3 *
103 L
77.1 SATT166 3 **
103 L 78.0 SATT448 4 *
104 L 118.6
SATT513 11 *
104 L 124.5 P12394A-1 2 **
108 M 87.6 SATT536
4 **
108 M 91.1 SATT175 5 *
110 M 143.5 SATT346 3 *
111 M 173.5 SATT336 4 *
112 N 20.0 P13069A-1 3 **
113 N
38.3 SAT_084 2 *
115 N 88.6 SATT339 5 *
116 N 92.2 SATT255
3 *
118 O 2.4 P12396A-1 1 **
118 O 6.3 SATT487 4 **
119 O 37.7 SATT259 4 **
120 O 60.5 SATT420 2 *
120 O 65.1
SATT576 4 *
123 O 130.0 P11070A-1 1 *
129 UM # P12391A-1 1
*
131 UM # P13560A-1 1 **
131 UM # P13561A-1 2 **
133 UM # SAC1677 3 **
134 UM # SATT040 4 **
138 UM #
SATT109 3 *
139 UM # SATT111 3 **
140 UM # SATT176 3 **
[0257]
6TABLE 6
Favorable Allelic Forms: Illinois
Region Chromosome Position Locus Allele Sig.
2 A1 19.1
SATT042 3 **
2 A1 19.1 SATT364 3 **
2 A1 19.1 SATT454 1 **
2 A1 19.1 SATT526 1 **
3 A1 27.1 SATT300 3 **
3 A1
27.1 SATT591 3 **
3 A1 28.5 SATT155 2 **
4 A1 69.9 SATT385
1 *
5 A1 87.3 SATT511 1 *
6 A2 0.0 P12390B-1 1 **
13
A2 184.0 SATT429 4 **
15 B1 39.0 SATT197 5 *
16 B1 56.6
SATT519 1 *
21 B2 45.0 P10641A-1 2 **
23 B2 86.1 SATT556 2
*
23 B2 87.0 SATT272 1 *
23 B2 88.6 SATT020 2 *
24
B2 97.3 SATT066 3 *
26 B2 120.0 P10638B-2 1 *
29 C1 95.8
SATT399 3 **
29 C1 96.5 SATT361 2 **
29 C1 98.0 P10639A-1 1
**
29 C1 99.0 SATT190 4 **
31 C1 173.0 SATT338 3 *
36 C2 140.9 SATT557 5 **
36 C2 145.8 SATT319 1 *
37 C2
165.8 SATT460 5 *
37 C2 170.5 SATT433 3 *
38 C2 193.5
SATT357 1 *
40 D1a 54.7 SATT321 2 *
42 D1a 73.1 SATT203 4 *
43 D1a 129.2 SATT129 5 **
43 D1a 129.2 SATT147 1 *
44 D1b 0.0 SATT216 4 *
45 D1b 20.0 P10621B-2 2 *
46 D1b
39.8 SATT558 3 *
54 D2 93.8 SATT389 6 *
55 D2 110.4 SATT464
1 **
60 E 40.0 P13074A-1 1 *
62 E 98.0 SATT573 2 *
62 E 98.0 SATT598 1 *
62 E 100.6 SATT263 2 *
62 E 101.0
SATT602 4 *
62 E 102.2 SATT151 4 *
62 E 102.2 SATT355 3 *
62 E 102.2 SATT452 2 *
64 F 0.0 SATT146 1 *
64 F 0.0
SATT193 3 **
64 F 0.0 SATT569 3 **
64 F 1.7 SATT343 3 **
64 F 1.9 SATT586 5 **
66 F 57.5 SATT595 1 **
67 F 85.0
P10782A-1 1 *
67 F 95.0 SATT334 3 *
70 F 157.8 SATT144 1 *
71 F 165.8 SATT522 5 *
73 G 4.0 P7659A-2 2 *
73 G
7.7 SATT570 3 **
73 G 8.5 SATT356 2 *
75 G 53.0 SATT115 1
**
75 G 66.1 SATT533 1 *
77 G 100.8 SATT199 1 **
77
G 103.2 SATT503 3 *
77 G 103.2 SATT517 4 *
79 G 137.0
SATT191 2 **
79 G 138.4 SAT_117 1 **
83 H 77.3 SATT279 6 *
85 H 125.3 SATT181 5 **
86 I 15.5 SATT127 2 *
92 J
61.3 SCT_065 2 **
92 J 63.6 SATT596 4 *
92 J 68.0 SATT406 2
**
92 J 70.8 SATT380 3 **
92 J 72.6 SATT183 1 **
92
J 74.4 SATT529 1 **
94 K 17.9 SATT242 4 **
97 K 85.8
SATT240 2 *
101 L 34.6 SATT398 4 **
102 L 42.3 SATT497 3 *
103 L 77.1 SATT166 3 **
103 L 78.0 SATT448 4 *
104 L
118.0 SATT373 3 **
104 L 124.5 P12394A-1 2 **
105 M 12.4
SATT590 17 *
106 M 41.0 SATT567 3 *
107 M 78.9 SATT220 3 *
108 M 87.6 SATT536 4 *
108 M 91.1 SATT175 5 *
109 M
115.0 P10615A-1 1 *
112 N 20.0 P13069A-1 3 **
112 N 25.0
P5467A-1 2 **
112 N 25.0 P5467A-2 1 *
113 N 38.3 SAT_084 2
*
115 N 88.6 SATT339 5 *
118 O 2.4 P12396A-1 1 *
118
O 6.3 SATT487 4 **
119 O 37.7 SATT259 4 *
120 O 60.5
SATT420 2 **
120 O 65.1 SATT576 4 *
122 O 103.8 SATT477 3 *
123 O 125.0 SATT581 2 **
123 O 130.0 P11070A-1 1 *
124 O 153.3 SATT153 3 **
124 O 155.1 SATT243 1 *
127 UM #
P10632A-1 2 *
128 UM # P10793A-1 2 *
129 UM # P12391A-1 1 *
129 UM # P12392A-1 1 *
131 UM # P13560A-1 1 *
131 UM
# P13561A-1 2 **
133 UM # SAC1677 3 **
134 UM # SATT040 4
**
139 UM # SATT111 3 **
140 UM # SATT176 3 **
141
UM # SATT219 1 *
143 UM # SATT299 5 *
[0258]
7TABLE 7
Favorable Allelic Forms: East Region
Region Chromosome Position Locus Allele Sig.
2 A1 19.1
SATT042 3 **
2 A1 19.1 SATT364 3 **
2 A1 19.1 SATT454 1 **
2 A1 19.1 SATT526 1 **
3 A1 27.1 SATT300 3 **
3 A1
27.1 SATT591 3 **
3 A1 28.5 SATT155 2 **
4 A1 69.9 SATT385
1 *
5 A1 87.3 SATT236 1 *
5 A1 87.3 SATT511 1 *
6 A2
0.0 P12390B-1 1 **
12 A2 154.7 SATT409 5 *
13 A2 184.0
SATT429 4 **
15 B1 39.0 SATT197 5 *
17 B1 74.1 SATT583 1 *
21 B2 45.0 P10641A-1 2 **
23 B2 86.1 SATT556 2 *
23
B2 88.6 SATT020 2 *
26 B2 120.0 P10638B-2 1 *
29 C1 95.8
SATT399 3 **
29 C1 96.5 SATT361 2 **
29 C1 98.0 P10639A-1 1
**
29 C1 99.0 SATT190 4 **
31 C1 173.0 SATT338 3 *
36 C2 140.9 SATT557 5 **
36 C2 145.8 SATT319 1 *
37 C2
170.5 SATT433 3 *
38 C2 193.5 SATT357 1 *
40 D1a 54.7
SATT321 2 *
41 D1a 68.6 SATT383 3 *
43 D1a 129.2 SATT129 5
**
43 D1a 129.2 SATT147 1 *
45 D1b 20.0 P10621B-2 2 *
46 D1b 39.8 SATT558 3 *
53 D2 66.9 SATT582 2 *
54 D2 93.8
SATT389 6 *
55 D2 110.4 SATT464 1 **
60 E 40.0 P13074A-1 1
*
62 E 98.0 SATT573 2 *
62 E 98.0 SATT598 1 *
62 E
100.6 SATT263 2 *
62 E 101.0 SATT602 4 *
62 E 102.2 SATT151
4 *
62 E 102.2 SATT355 1 *
62 E 102.2 SATT452 2 *
64
F 0.0 SATT146 1 **
64 F 0.0 SATT193 3 **
64 F 0.0 SATT569 3
**
64 F 1.7 SATT343 3 **
64 F 1.9 SATT586 5 **
66 F
57.5 SATT595 1 *
67 F 85.0 P10782A-1 1 **
67 F 95.0 SATT334
3 *
70 F 157.8 SATT144 1 *
73 G 8.5 SATT356 2 *
75 G
53.0 SATT115 1 **
75 G 66.1 SATT533 1 *
77 G 100.8 SATT199
1 **
77 G 103.2 SATT503 3 *
77 G 103.2 SATT517 4 **
79 G 137.0 SATT191 2 *
79 G 138.4 SAT_117 1 *
83 H 77.3
SATT279 6 *
85 H 125.3 SATT181 5 **
86 I 15.5 SATT127 2 *
92 J 61.3 SCT_065 2 **
92 J 63.6 SATT596 4 **
92 J
68.0 SATT406 2 **
92 J 70.8 SATT380 3 **
92 J 72.6 SATT183
1 **
92 J 74.4 SATT529 1 **
94 K 17.9 SATT242 4 **
97 K 85.8 SATT240 2 *
103 L 77.1 SATT166 3 **
104 L 118.0
SATT373 3 **
104 L 124.5 P12394A-1 2 **
108 M 87.6 SATT536
4 **
108 M 91.1 SATT175 5 *
109 M 115.0 P10615A-1 1 *
110 M 143.5 SATT346 3 *
111 M 173.5 SATT336 4 *
112 N 20.0
P13069A-1 3 **
112 N 25.0 P5467A-1 2 **
112 N 25.0 P5467A-2
1 *
113 N 38.3 SAT_084 2 *
115 N 88.6 SATT339 5 *
118 O 2.4 P12396A-1 1 *
118 O 6.3 SATT487 4 **
119 O 37.7
SATT259 4 **
120 O 60.5 SATT420 2 **
120 O 65.1 SATT576 4 *
120 O 69.2 SATT473 2 *
122 O 103.8 SATT477 3 *
123 O
125.0 SATT581 2 **
123 O 130.0 P11070A-1 1 *
124 O 153.3
SATT153 3 **
124 O 155.1 SATT243 1 **
128 UM # P10793A-1 2
*
129 UM # P12391A-1 1 *
129 UM # P12392A-1 1 *
131
UM # P13560A-1 1 *
131 UM # P13561A-1 2 **
133 UM # SAC1677
3 **
134 UM # SATT040 4 **
139 UM # SATT111 3 **
140
UM # SATT176 3 **
141 UM # SATT219 9 **
143 UM # SATT299 5
*
[0259]
8TABLE 8
Favorable Allelic Forms: Mid South Region
Region Chromosome Position Locus Allele Sig.
2 A1
19.1 SATT042 3 *
2 A1 19.1 SATT364 3 **
2 A1 19.1 SATT454 1
**
2 A1 19.1 SATT526 1 **
3 A1 27.1 SATT300 3 **
3
A1 27.1 SATT591 3 **
3 A1 28.5 SATT155 2 **
5 A1 87.3
SATT511 1 *
6 A2 0.0 P12390B-1 1 *
8 A2 96.2 SATT233 6 *
11 A2 136.0 P10635A-1 1 *
12 A2 154.7 SATT409 2 *
12 A2
161.8 SATT228 4 **
13 A2 184.0 SATT429 4 **
15 B1 39.0
SATT197 5 *
18 B1 92.1 SATT359 3 **
20 B2 20.0 P12105A-1 1
*
21 B2 45.0 P10641A-1 2 **
23 B2 86.1 SATT556 2 *
23 B2 87.0 SATT272 1 *
23 B2 88.6 SATT020 2 *
29 C1 95.8
SATT399 3 **
29 C1 96.5 SATT361 2 *
29 C1 98.0 P10639A-1 1
*
29 C1 99.0 SATT190 4 **
31 C1 173.0 SATT338 3 *
36
C2 140.9 SATT557 5 **
36 C2 145.8 SATT319 1 *
37 C2 165.8
SATT460 5 *
40 D1a 54.7 SATT321 2 **
42 D1a 73.1 SATT203 4
*
43 D1a 129.2 SATT129 5 **
44 D1b 0.0 SATT216 4 *
45 D1b 20.0 P10621B-2 2 **
46 D1b 39.8 SATT558 3 *
54 D2
93.8 SATT389 6 *
55 D2 110.4 SATT464 1 **
62 E 98.0 SATT598
1 *
62 E 100.6 SATT263 2 *
62 E 102.2 SATT151 4 *
62
E 102.2 SATT355 3 *
62 E 102.2 SATT452 2 *
64 F 0.0 SATT146
1 **
64 F 0.0 SATT193 3 **
64 F 0.0 SATT569 3 **
64
F 1.7 SATT343 3 **
64 F 1.9 SATT586 5 **
65 F 16.4 SATT423
3 *
65 F 18.0 SATT348 4 *
66 F 57.5 SATT595 1 *
67 F
85.0 P10782A-1 1 **
67 F 95.0 SATT334 3 *
73 G 2.0
P10646A-1 2 **
73 G 3.0 P5219A-1 2 *
73 G 4.0 P7659A-2 2 *
73 G 7.7 SATT570 3 **
73 G 8.5 SATT356 2 *
75 G 53.0
SATT115 1 *
75 G 66.1 SATT533 1 *
77 G 100.8 SATT199 1 *
77 G 103.2 SATT503 3 *
77 G 103.2 SATT517 4 *
79 G
137.0 SATT191 2 *
79 G 138.4 SAT_117 1 **
84 H 116.9
SATT142 3 *
85 H 125.3 SATT181 5 **
86 I 9.8 SATT367 6 *
86 I 15.5 SATT127 2 *
89 I 114.2 SATT440 3 *
92 J 61.3
SCT_065 2 **
92 J 63.6 SATT596 4 *
92 J 68.0 SATT406 2 **
92 J 70.8 SATT380 3 **
92 J 72.6 SATT183 1 **
92 J
74.4 SATT529 1 **
94 K 17.9 SATT242 4 *
96 K 72.8 SATT544 2
*
97 K 85.8 SATT240 2 *
101 L 34.6 SATT398 4 **
102
L 42.3 SATT497 3 *
103 L 77.1 SATT166 3 **
103 L 78.0
SATT448 4 *
104 L 124.5 P12394A-1 2 *
107 M 78.9 SATT220 3
*
108 M 87.6 SATT536 4 *
109 M 115.0 P10615A-1 1 *
110 M 143.5 SATT346 3 *
115 N 88.6 SATT339 5 *
118 O 2.4
P12396A-1 1 *
118 O 6.3 SATT487 4 **
119 O 37.7 SATT259 4 *
120 O 60.5 SATT420 2 *
120 O 65.1 SATT576 4 *
122 O
103.8 SATT477 3 *
123 O 125.0 SATT581 2 **
124 O 153.3
SATT153 3 **
124 O 155.1 SATT243 1 *
129 UM # P12392A-1 1 *
131 UM # P13560A-1 1 *
131 UM # P13561A-1 2 **
133
UM # SAC1677 3 *
134 UM # SATT040 4 **
139 UM # SATT111 3
**
140 UM # SATT176 3 **
141 UM # SATT219 9 **
143
UM # SATT299 5 *
[0260]
9TABLE 9
Favorable Allelic Forms: South Region
Region Chromosome Position Locus Allele Sig.
2 A1 14.6
SATT165 2 *
2 A1 19.1 SATT042 3 *
2 A1 19.1 SATT042 4 *
2 A1 19.1 SATT364 4 *
2 A1 19.1 SATT526 1 **
3 A1 27.1
SATT300 3 **
3 A1 27.1 SATT591 3 *
3 A1 28.5 SATT155 2 **
4 A1 69.9 SATT385 1 *
5 A1 87.3 SATT236 1 *
5 A1 87.3
SATT511 1 *
6 A2 0.0 P12390B-1 1 *
7 A2 20.0 SATT480 1 *
9 A2 108.7 SATT329 2 *
14 B1 26.7 SATT509 6 *
17 B1
71.6 SCT_026 2 **
17 B1 74.1 SATT583 3 *
17 B1 74.8 SATT430
1 *
18 B1 92.1 SATT359 3 **
19 B1 120.0 P10648A-1 2 *
21 B2 55.8 SATT168 2 *
23 B2 86.1 SATT556 2 *
23 B2 87.0
SATT272 2 *
29 C1 95.8 SATT399 3 *
29 C1 99.0 SATT190 4 *
33 C2 53.4 SATT457 2 *
37 C2 168.3 SATT307 2 *
37 C2
168.3 SCT_028 2 *
37 C2 170.5 SATT433 4 *
38 C2 193.5
SATT357 1 *
40 D1a 54.7 SATT321 2 *
46 D1b 39.8 SATT558 3 *
47 D1b 51.6 SATT266 1 *
48 D1b 64.3 SATT282 4 *
48
D1b 65.1 SATT537 8 *
54 D2 93.8 SATT389 6 *
54 D2 99.6
SATT461 1 *
55 D2 106.6 SATT311 2 *
55 D2 107.8 SATT514 1 *
55 D2 110.4 SATT464 1 **
55 D2 112.1 SATT543 4 **
56
D2 139.1 SATT186 1 *
60 E 40.0 P13074A-1 1 *
61 E 80.0
P10624A-1 2 *
62 E 100.6 SATT204 1 *
62 E 101.0 SATT491 3 *
62 E 102.2 SATT151 3 *
62 E 102.2 SATT355 3 *
62 E
102.2 SATT452 2 **
64 F 0.0 SATT146 4 **
64 F 0.0 SATT569 3
*
64 F 1.7 SATT343 5 *
64 F 1.9 SATT586 2 *
67 F
87.5 P3436A-1 2 **
67 F 95.0 SATT334 3 *
68 F 114.8 SATT510
5 **
70 F 157.8 SATT144 1 *
73 G 2.0 P10646A-1 2 **
73 G 3.0 P5219A-1 2 *
73 G 4.0 P7659A-2 2 *
73 G 7.7
SATT570 3 **
73 G 8.5 SATT356 2 *
74 G 13.5 SATT130 2 *
75 G 61.3 SATT594 6 **
75 G 66.1 SATT533 1 *
76 G 71.6
SATT303 7 *
76 G 71.6 SATT303 9 **
76 G 72.4 SATT352 10 **
76 G 72.4 SATT566 3 *
77 G 100.8 SATT199 1 **
77 G
103.2 SATT503 3 **
77 G 103.2 SATT517 4 *
79 G 137.0
SATT191 2 *
79 G 138.4 SAT_117 1 **
81 H 0.0 SATT353 2 *
82 H 42.3 SATT442 4 **
83 H 77.3 SATT279 5 **
83 H 77.3
SATT314 1 **
86 I 9.8 SATT367 6 **
86 I 15.5 SATT127 2 *
87 I 57.9 SATT270 5 *
89 I 115.0 P10640A-1 1 *
92 J
67.2 SATT280 5 **
92 J 70.8 SATT380 3 **
92 J 72.6 SATT183
2 **
93 J 118.0 SATT431 3 **
94 K 17.9 SATT242 1 **
95 K 44.0 SATT102 2 *
98 K 144.0 P10618A-1 1 *
98 K 144.3
SATT475 1 *
99 K 164.8 SATT196 4 *
101 L 32.4 SATT523 3 *
101 L 33.9 SATT418 4 *
103 L 77.1 SATT166 1 **
103 L
78.0 SATT448 4 *
104 L 118.0 SATT373 17 *
104 L 118.6
SATT513 1 **
104 L 124.5 P12394A-1 2 **
105 M 12.4 SATT590
2 *
108 M 87.6 SATT536 4 *
110 M 139.4 SATT250 1 *
113 N 35.4 SATT584 2 *
114 N 61.2 SATT387 5 **
115 N 80.2
SATT549 5 *
117 N 113.0 SATT257 2 *
118 O 2.4 P12396A-1 1 *
118 O 6.3 SATT487 1 **
119 O 37.7 SATT259 3 **
119 O
39.0 SATT347 2 *
120 O 65.1 SATT576 4 *
120 O 67.5 SATT550
1 *
124 O 155.1 SATT243 2 *
125 O 175.7 P8230A-1 2 **
126 UM # P10623A-1 2 *
129 UM # P12391A-1 2 *
131 UM #
P13561A-1 1 *
132 UM # p2481A-1 1 *
133 UM # SAC1677 3 **
136 UM # SATT108 1 *
139 UM # SATT111 3 *
140 UM #
SATT176 8 **
141 UM # SATT219 4 **
143 UM # SATT299 5 *
145 UM # SATT512 3 *
[0261]
10TABLE 10
Favorable Alleles: Iowa (A)
Best
Marker in
Genomic Genomic LOD CHG
Region Chromosome Position Locus Allele Region Sig. Iowa EXP Iowa OBS
Iowa Iowa #ELI Iowa
2 A1 19.1 Satt042 3 ** 2.68 0.13 0.75
0.62 67
2 A1 19.1 Satt364 3 ** 3.22 0.05 0.72 0.67 68
2 A1
19.1 Satt454 1 ** 3.70 0.05 0.74 0.69 69
2 A1 19.1 Satt526 1 Best
** 4.00 0.06 0.79 0.72 68
3 A1 27.1 Satt300 3 ** 3.22 0.08 0.77
0.69 64
3 A1 27.1 Satt591 3 Best ** 3.22 0.13 0.82 0.70 67
3 A1 28.5 Satt155 2 ** 2.96 0.18 0.82 0.64 66
5 A1 87.3 Satt511 1
Best * 1.40 0.14 0.57 0.44 61
6 A2 0.0 P12390B-1 1 Best ** 2.59
0.32 0.89 0.57 66
13 A2 184.0 Satt429 4 Best ** 4.00 0.02 0.63
0.61 53
15 B1 39.0 Satt197 5 Best * 1.66 0.22 0.64 0.41 48
21 B2 45.0 P10641A-1 2 Best ** 3.22 0.34 0.94 0.60 70
25 B2 110.9
Satt534 7 Best * 1.47 0.49 0.89 0.40 61
26 B2 120.0 P10638B-2 1
Best * 1.91 0.62 0.94 0.33 70
29 C1 95.8 Satt399 3 ** 4.00 0.08
0.89 0.81 61
29 C1 96.5 Satt361 2 ** 3.05 0.15 0.80 0.65 68
29 C1 98.0 P10639A-1 1 ** 3.22 0.19 0.86 0.66 63
29 C1 99.0
Satt190 4 Best ** 4.00 0.05 0.79 0.73 65
31 C1 173.0 Satt338 3
Best * 1.55 0.25 0.71 0.46 63
36 C2 140.9 Satt557 5 Best ** 3.52
0.58 1.00 0.42 70
36 C2 145.8 Satt319 1 * 1.90 0.74 1.00 0.26 67
37 C2 165.8 Satt460 5 * 1.43 0.63 0.97 0.34 44
37 C2 170.5
Satt433 3 Best * 1.59 0.15 0.48 0.34 61
38 C2 193.5 Satt357 1 Best
* 1.52 0.68 0.97 0.29 66
40 D1a 54.7 Satt321 2 Best * 1.82 0.19
0.69 0.50 66
41 D1a 69.8 Satt295 2 Best * 1.67 0.70 0.98 0.28 50
42 D1a 80.3 Satt507 3 Best * 1.37 0.20 0.63 0.43 66
43 D1a
129.2 Satt129 5 Best ** 2.89 0.31 0.93 0.61 61
43 D1a 129.2
Satt147 1 * 1.46 0.69 0.97 0.28 62
46 D1b 39.8 Satt558 3 Best *
1.76 0.29 0.81 052 56
53 D2 66.9 Satt582 2 ** 2.89 0.01 0.28 0.27
50
54 D2 93.8 Satt389 6 Best ** 2.46 0.05 0.53 0.49 63
55
D2 110.4 Satt464 1 Best ** 3.40 0.05 0.67 0.62 66
62 E 98.0
Satt573 2 * 1.72 0.46 0.91 0.45 62
62 E 98.0 Satt598 1 Best **
2.28 0.38 0.92 0.54 66
62 E 100.6 Satt263 2 * 1.88 0.07 0.53 0.46
59
62 E 101.0 Satt602 4 * 1.36 0.12 0.43 0.31 61
62 E
102.2 Satt151 4 * 1.78 0.05 0.44 0.40 60
64 F 0.0 Satt146 1 **
2.89 0.20 0.86 0.66 50
64 F 0.0 Satt193 3 ** 3.70 0.10 0.84 0.74
62
64 F 0.0 Sat569 3 ** 3.16 0.15 0.86 0.71 69
64 F 1.7
Satt176 3 ** 4.00 0.09 0.85 0.76 71
64 F 1.7 Satt343 3 Best **
4.00 0.09 0.89 0.80 66
64 F 1.9 Satt586 5 ** 4.00 0.09 0.84 0.75
55
64 F 2.5 Satt040 4 ** 3.52 0.09 0.81 0.73 67
67 F 85.0
P10782A-1 1 Best ** 3.16 0.27 0.90 0.64 70
70 F 157.8 Satt144 1
Best * 1.79 0.45 0.90 0.45 67
71 F 165.8 Satt522 5 Best * 1.48
0.04 0.32 0.27 63
73 G 8.5 Satt356 2 Best * 1.42 0.69 0.97 0.28 68
75 G 66.1 Satt533 1 Best * 1.94 0.43 0.90 0.46 68
77 G
100.8 Satt199 1 Best ** 3.22 0.44 0.98 0.54 65
77 G 103.2 Satt517
4 * 1.54 0.10 0.54 0.44 64
79 G 137.0 Satt191 2 * 1.65 0.15 0.62
0.47 65
79 G 138.4 Sat_117 1 Best * 1.70 0.20 0.68 0.48 66
85 H 125.3 Satt181 5 Best ** 2.14 0.05 0.61 0.56 61
86 I 15.5
Satt127 2 Best * 1.99 0.62 0.99 0.37 58
92 J 61.3 Sct_065 2 **
3.70 0.26 0.91 0.65 58
92 J 63.6 Satt596 4 ** 2.47 0.14 0.79 0.65
65
92 J 68.0 Satt406 2 Best ** 3.52 0.18 0.87 0.69 66
92 J
70.8 Satt380 3 ** 3.70 0.28 0.94 0.66 63
92 J 72.6 Satt183 1 **
3.30 0.45 0.97 0.53 69
92 J 74.4 Satt529 1 ** 3.52 0.38 0.97 0.59
61
94 K 17.9 Satt242 4 Best ** 3.05 0.07 0.71 0.64 65
97 K
85.8 Satt240 2 Best * 1.49 0.11 0.52 0.41 54
101 L 34.6 Satt398 4
Best ** 2.00 0.33 0.83 0.50 51
103 L 77.1 Satt166 3 Best ** 4.00
0.10 0.90 0.80 61
103 L 78.0 Satt448 4 * 1.46 0.22 0.68 0.46 67
104 L 118.0 Satt373 3 ** 3.70 0.00 0.37 0.36 52
104 L 118.6
Satt513 11 * 1.47 0.07 0.33 0.26 40
104 L 124.5 P12394A-1 2 Best
** 3.05 0.66 1.00 0.35 71
108 M 87.6 Satt536 4 Best ** 3.70 0.14
0.83 0.70 66
108 M 91.1 Satt175 5 * 1.99 0.19 0.77 0.58 68
109 M 115.0 P10615A-1 1 Best * 1.75 0.10 0.54 0.44 70
110 M 143.5
Satt346 3 Best * 1.30 0.16 0.61 0.45 61
111 M 173.5 Satt336 4 Best
* 1.63 0.25 0.71 0.46 44
112 N 20.0 P13069A-1 3 Best ** 4.00 0.00
0.31 0.31 61
112 N 25.0 P5467A-1 2 ** 4.00 0.00 0.26 0.26 62
115 N 88.6 Satt339 5 Best * 1.96 0.19 0.73 0.54 60
118 O 2.4
P12396A-1 1 * 1.89 0.64 0.97 0.33 66
118 O 6.3 Satt487 4 Best **
4.00 0.37 0.97 0.60 64
119 O 37.7 Satt259 4 Best ** 3.30 0.37 0.92
0.55 63
120 O 60.5 Satt420 2 Best ** 2.28 0.30 0.85 0.54 68
120 O 65.1 Satt576 4 * 1.80 0.22 0.77 0.55 44
122 O 103.8 Satt477
3 Best * 1.32 0.43 0.83 0.40 53
123 O 125.0 Satt581 2 Best ** 2.70
0.35 0.88 0.53 69
124 O 153.3 Satt153 3 Best ** 2.96 0.49 0.94
0.46 68
124 O 155.1 Satt243 1 ** 2.17 0.60 0.96 0.36 56
129 UM P12391A-1 1 Best * 1.84 0.39 0.84 0.45 69
129 UM
P12392A-1 1 Best * 1.30 0.55 0.88 0.33 69
131 UM P13560A-1 1 Best
* 1.71 0.23 0.64 0.41 70
131 UM P13561A-1 2 Best ** 3.10 0.63
1.00 0.37 67
133 UM Sac1677 3 Best ** 4.00 0.15 0.90 0.75 70
139 UM Satt111 3 Best ** 4.00 0.08 0.81 0.74 67
143 UM Satt299
5 Best * 1.59 0.17 0.65 0.48 61
* 95% significance level
** 99% significance level
[0262]
11TABLE 11
Significant Alleles at the "Best" marker
locus in each genomic region.
SIGNIF LOD EXP OBS CHG #ELI
Region Chromosome Position Locus Allele Iowa Iowa Iowa Iowa Iowa Iowa
2 A1 19.1 Satt526 1 ** 4.00 0.06 0.79 0.72 68
3 A1
27.1 Satt591 3 ** 3.22 0.13 0.82 0.70 67
5 A1 87.3 Satt511 1 *
1.40 0.14 0.57 0.44 61
6 A2 0.0 P12390B-1 1 ** 2.59 0.32 0.89 0.57
66
13 A2 184.0 Satt429 4 ** 4.00 0.02 0.63 0.61 53
15 B1
39.0 Satt197 5 * 1.66 0.22 0.64 0.41 48
21 B2 45.0 P10641A-1 2 **
3.22 0.34 0.94 0.60 70
25 B2 110.9 Satt534 7 * 1.47 0.49 0.89 0.40
61
26 B2 120.0 P10638B-2 1 * 1.91 0.62 0.94 0.33 70
29 C1
99.0 Satt190 4 ** 4.00 0.05 0.79 0.73 65
31 C1 173.0 Satt338 3 *
1.55 0.25 0.71 0.46 63
36 C2 140.9 Satt557 5 ** 3.52 0.58 1.00
0.42 70
37 C2 170.5 Satt433 3 * 1.59 0.15 0.48 0.34 61
38
C2 193.5 Satt357 1 * 1.52 0.68 0.97 0.29 66
40 D1a 54.7 Satt321 2
* 1.82 0.19 0.69 0.50 66
41 D1a 69.8 Satt295 2 * 1.67 0.70 0.98
0.28 50
42 D1a 80.3 Satt507 3 * 1.37 0.20 0.63 0.43 66
43
D1a 129.2 Satt129 5 ** 2.89 0.31 0.93 0.61 61
46 D1b 39.8 Satt558
3 * 1.76 0.29 0.81 0.52 56
54 D2 93.8 Satt389 6 ** 2.46 0.05 0.53
0.49 63
55 D2 110.4 Satt464 1 ** 3.40 0.05 0.67 0.62 66
62
E 98.0 Satt598 1 ** 2.28 0.38 0.92 0.54 66
64 F 1.7 Satt343 3 **
4.00 0.09 0.89 0.80 66
67 F 85.0 P10782A-1 1 ** 3.16 0.27 0.90
0.64 70
70 F 157.8 Satt144 1 * 1.79 0.45 0.90 0.45 67
71 F
165.8 Satt522 5 * 1.48 0.04 0.32 0.27 63
73 G 8.5 Satt356 2 * 1.42
0.69 0.97 0.28 68
75 G 66.1 Satt533 1 * 1.94 0.43 0.90 0.46 68
77 G 100.8 Satt199 1 ** 3.22 0.44 0.98 0.54 65
79 G 138.4
Sat_117 1 * 1.70 0.20 0.68 0.48 66
85 H 125.3 Satt181 5 ** 2.14
0.05 0.61 0.56 61
86 I 15.5 Satt127 2 * 1.99 0.62 0.99 0.37 58
92 J 68.0 Satt406 2 ** 3.52 0.18 0.87 0.69 66
94 K 17.9 Satt242
4 ** 3.05 0.07 0.71 0.64 65
97 K 85.8 Satt240 2 * 1.49 0.11 0.52
0.41 54
101 L 34.6 Satt398 4 ** 2.00 0.33 0.83 0.50 51
103
L 77.1 Satt166 3 ** 4.00 0.10 0.90 0.80 61
104 L 124.5 P12394A-1 2
** 3.05 0.66 1.00 0.35 71
108 M 87.6 Satt536 4 ** 3.70 0.14 0.83
0.70 66
109 M 115.0 P10615A-1 1 * 1.75 0.10 0.54 0.44 70
110 M 143.5 Satt346 3 * 1.30 0.16 0.61 0.45 61
111 M 173.5 Satt336
4 * 1.63 0.25 0.71 0.46 44
112 N 20.0 P13069A-1 3 ** 4.00 0.00
0.31 0.31 61
115 N 88.6 Satt339 5 * 1.96 0.19 0.73 0.54 60
118 O 6.3 Satt487 4 ** 4.00 0.37 0.97 0.60 64
119 O 37.7 Satt259 4
** 3.30 0.37 0.92 0.55 63
120 O 60.5 Satt420 2 ** 2.28 0.30 0.85
0.54 68
122 O 103.8 5att477 3 * 1.32 0.43 0.83 0.40 53
123
O 125.0 Satt581 2 ** 2.70 0.35 0.88 0.53 69
124 O 153.3 Satt153 3
** 2.96 0.49 0.94 0.46 68
129 UM P12391A-1 1 * 1.84 0.39 0.84
0.45 69
129 UM P12392A-1 1 * 1.30 0.55 0.88 0.33 69
131 UM
P13560A-1 1 * 1.71 0.23 0.64 0.41 70
131 UM P13561A-1 2 ** 3.10
0.63 1.00 0.37 67
133 UM Sac1677 3 ** 4.00 0.15 0.90 0.75 70
139 UM Satt111 3 ** 4.00 0.08 0.81 0.74 67
143 UM Satt299 5 *
1.59 0.17 0.65 0.48 61
* 95% significance level
**
99% significance level
[0263]
12TABLE 12
Favorable Alleles: Iowa (B)
New Best
Marker Market
in in
Genomic
Genomic Genomic LOD EXP OBS CHG #ELI
Region Chromosome
Position Locus Allele Region Region Sig. Iowa Iowa Iowa Iowa Iowa
1 A1 1.4 Satt684 3 NEW Best * 1.84 0.03 0.36 0.36 79
2 A1
19.1 Satt042 3 ** 2.66 0.14 0.77 0.64 82
2 A1 19.1 Satt364 3
** 3.16 0.05 0.70 0.65 65
2 A1 19.1 Satt454 1 ** 3.40 0.06 0.71
0.64 66
2 A1 19.1 Satt526 1 Best ** 4.00 0.07 0.77 0.70 86
3 A1 27.1 Satt300 3 ** 3.30 0.11 0.79 0.68 82
3 A1 27.1 Satt591
3 Best ** 3.40 0.15 0.83 0.69 81
3 A1 28.5 Satt155 2 ** 2.89
0.18 0.83 0.65 81
4 A1 69.9 Satt385 1 Best * 1.99 0.44 0.91 0.48
80
7.5 A2 41.8 Satt632-TB 4 NEW Best * 1.88 0.66 1.00 0.34 41
13 A2 184.0 Satt429 4 Best ** 4.00 0.04 0.80 0.76 76
14 B1 27.6
SAT_261 1 NEW Best ** 2.42 0.49 1.00 0.51 28
21 B2 45.0 P10641A-1
2 Best ** 3.70 0.34 0.99 0.65 86
23 B2 86.1 Satt556 2 Best *
1.73 0.16 0.75 0.59 76
26 B2 120.0 P10638B-2 1 Best * 1.30 0.63
0.93 0.30 85
29 C1 95.8 Satt399 3 ** 4.00 0.07 0.89 0.82 77
29 C1 96.5 Satt361 2 ** 3.30 0.16 0.83 0.67 64
29 C1 99.0
SATT661-TB 2 NEW * 1.68 0.39 0.94 0.55 59
29 C1 99.0 Satt190 4
Best ** 4.00 0.05 0.83 0.78 79
30 C1 117.6 SAT_311-DB 3 NEW Best *
1.72 0.45 0.92 0.47 56
31 C1 173.0 Satt338 3 Best * 1.49 0.24
0.67 0.43 47
32 C2 28.6 SATT640-TB 3 NEW Best * 1.50 0.57 0.98
0.41 45
36 C2 140.9 Satt557 5 Best ** 2.68 0.61 1.00 0.28 84
36 C2 145.8 Satt319 1 * 1.66 0.72 1.00 0.28 84
36.5 C2 153.5
SAT_142-DB 3 NEW Best ** 2.44 0.58 1.00 0.42 62
40 D1a 54.7
Satt321 2 Best * 1.52 0.17 0.64 0.47 82
42 D1a 73.1 Satt203 4
Best * 1.54 0.07 0.42 0.35 76
43 D1a 129.2 Satt129 5 Best ** 3.52
0.36 0.99 0.63 75
43 D1a 129.2 Satt147 1 * 1.85 0.70 1.00 0.30
63
45 D1b 19.1 SAT_351 3 NEW Best ** 2.08 0.14 0.73 0.59 66
45 D1b 20.0 P10621B-2 2 NEW * 1.51 0.60 0.96 0.36 86
46 D1b 36.7
Satt701 2 NEW Best * 1.97 0.15 0.59 0.44 27
46 D1b 39.8 Satt634 3
NEW * 1.40 0.44 0.89 0.45 27
53 D2 66.9 Satt582 2 Best * 1.33
0.06 0.37 0.31 78
54 D2 93.8 Satt389 6 Best * 1.35 0.11 0.48 0.37
81
55 D2 110.4 Satt464 1 Best ** 4.00 0.05 0.75 0.70 83
55
D2 111.8 Satt662 1 NEW ** 2.08 0.06 0.65 0.59 27
57 D2 149.1
Satt672 1 NEW Best * 1.64 0.15 0.57 0.42 27
62 E 98.0 Satt573 2
* 1.83 0.45 0.92 0.47 61
62 E 98.0 Satt598 1 Best * 1.92 0.49
0.95 0.46 73
62 E 100.6 Satt263 2 * 1.42 0.11 0.49 0.38 58
62 E 102.2 Satt151 4 * 1.39 0.08 0.44 0.36 79
62 E 103.7
SAT_273-DB 1 NEW * 1.59 0.11 0.59 0.48 62
64 F 0.0 Satt146 1 **
3.10 0.20 0.89 0.70 75
64 F 0.0 Satt193 3 ** 3.70 0.09 0.86 0.77
62
64 F 0.0 Satt569 3 ** 3.30 0.18 0.91 0.73 64
64 F 1.7
Satt343 3 Best ** 3.70 0.08 0.89 0.81 74
64 F 1.9 Satt586 5 **
3.10 0.08 0.82 0.74 70
64 F 2.5 Satt040 4 ** 3.16 0.08 0.81 0.73
62
66 F 57.5 Satt595 1 NEW Best ** 2.35 0.69 1.00 0.31 75
67 F 95.0 Satt334 3 NEW Best * 1.55 0.15 0.63 0.48 73
70 F 157.8
Satt144 1 Best * 1.77 0.46 0.93 0.47 84
71 F 165.8 Satt522 5
Best * 1.32 0.06 0.37 0.31 77
73 G 7.7 Satt570 3 NEW Best ** 3.70
0.00 0.27 0.27 82
73 G 8.5 Satt356 2 Best * 1.30 0.68 0.98 0.30
62
77 G 100.8 Satt199 1 Best * 1.61 0.45 0.89 0.44 81
77 G
103.2 Satt517 4 * 1.59 0.07 0.51 0.44 58
79 G 137.0 Satt191 2
* 1.39 0.18 0.63 0.44 80
79 G 138.4 Sat_117 1 Best * 1.89 0.23
0.77 0.54 77
83 H 77.3 Satt279 6 NEW Best * 2.02 0.40 0.90 0.50 63
85 H 125.3 Satt181 5 Best ** 3.06 0.05 0.72 0.66 76
86 I
15.5 Satt127 2 Best ** 2.36 0.64 1.00 0.36 77
87 I 57.9 Satt270
12 NEW Best ** 2.28 0.00 0.04 0.04.dagger. 82
88 I 77.4 Satt292 3
NEW Best * 1.84 0.06 0.56 0.50 64
90 J 19.6 SAG1223 4 NEW Best **
2.59 0.22 0.89 0.67 27
90 J 26.4 SAC1699 3 NEW ** 1.96 0.04 0.62
0.58 74
91.5 J 61.3 Sat_065 2 Best ** 4.00 0.23 0.96 0.73 65
91.5 J 63.6 Satt596 4 * 1.77 0.24 0.80 0.56 64
92 J 68.0
Satt406 2 ** 3.05 0.19 0.89 0.70 77
92 J 70.8 Satt380 3 **
2.80 0.29 0.91 0.63 69
92 J 72.6 Satt183 1 ** 2.52 0.43 0.96
0.53 84
92 J 74.4 Satt529 1 Best ** 3.22 0.43 0.99 0.56 73
94 K 17.9 Satt242 4 Best ** 3.05 0.07 0.71 0.64 65
97 K 83.7
Satt617 6 NEW Best * 1.68 0.08 0.60 0.53 24
97 K 85.8 Satt240 2
* 1.57 0.05 0.60 0.45 74
100 L 15.6 SAT_301 5 NEW Best * 1.97 0.22
0.83 0.61 78
101 L 33.9 Satt418 2 NEW * 1.44 0.56 0.94 0.37 63
101 L 34.6 Satt398 4 Best ** 3.22 0.34 0.94 0.61 71
102 L
42.3 Satt497 3 NEW Best * 1.66 0.60 0.96 0.36 78
103 L 77.1
Satt166 3 Best ** 3.22 0.11 0.86 0.75 80
103 L 78.0 Satt448 4 *
1.80 0.22 0.74 0.52 65
104 L 118.0 Satt373 3 Best ** 4.00 0.02
0.44 0.42 79
104 L 118.6 Satt513 5 ** 2.51 0.02 0.24
0.22.dagger. 64
104 L 124.5 P12394A-1 2 ** 2.64 0.68 1.00 0.32
85
107 M 84.2 SAG1048 2 NEW Best ** 4.00 0.01 0.69 0.68 76
108 M 87.6 Satt536 4 ** 2.12 0.12 0.69 0.58 83
108 M 91.1
Satt175 5 * 1.76 0.18 0.75 0.57 78
108 M 98.1 Satt677 2 NEW *
1.34 0.56 0.96 0.40 28
108 M 100.5 Satt680 2 NEW Best ** 3.22 0.13
0.86 0.73 71
109 M 115.0 P10615A-1 1 Best * 1.96 0.09 0.58 0.50
85
109 M 127.3 Satt551 3 NEW * 1.62 0.04 0.49 0.45 59
111
M 180.5 SAT_330-DB 1 NEW Best ** 3.00 0.16 0.85 0.69 61
112 N 20.0
P13069A-1 3 Best ** 4.00 0.00 0.44 0.44 84
112 N 25.0 P5467A-1 2
NEW ** 4.00 0.00 0.45 0.45 83
112 N 25.0 P5467A-2 1 * 1.72 0.21
0.63 0.42 86
113 N 38.3 SAT_084 2 NEW * 1.37 0.13 0.55 0.42 68
113 N 40.2 SAT_275-DB 2 NEW Best * 1.89 0.17 0.73 0.55 62
115
N 83.4 Satt660 2 NEW ** 2.34 0.10 0.77 0.67 28
115 N 88.6 Satt339
5 Best ** 2.44 0.17 0.77 0.60 80
118 O 2.4 Satt358 2 ** 2.35
0.03 0.44 0.41 75
118 O 6.3 Satt487 1 ** 2.33 0.00 0.02
0.02.dagger. 84
118 O 6.3 Satt487 4 Best ** 2.30 0.37 0.89 0.52
84
120 O 60.5 Satt420 2 Best * 1.95 0.33 0.85 0.52 84
120
O 65.1 Satt576 4 * 1.49 0.20 0.74 0.54 54
120 O 68.9 Satt633 1
NEW * 1.40 0.25 0.72 0.47 27
123 O 125.0 Satt581 2 Best ** 2.31
0.35 0.89 0.54 85
124 O 153.3 Satt153 3 Best ** 2.75 0.49 0.96
0.47 84
124 O 155.1 Satt243 1 * 1.82 0.61 0.97 0.36 68
UM
UM P10793A-1 2 * 1.40 0.10 0.52 0.43 86
UM UM P13560A-1 1 *
1.42 0.22 0.60 0.38 86
UM UM P13561A-1 2 ** 2.51 0.64 1.00 0.36
82
UM UM S60021-TB 1 NEW * 1.32 0.53 0.95 0.41 28
UM UM
S60048-TB 2 NEW ** 2.00 0.60 1.00 0.40 28
UM UM S60076-TB 1 NEW
* 1.35 0.59 0.93 0.34 28
UM UM S60148-TB 2 NEW ** 2.22 0.52 0.96
0.45 28
UM UM S60149-TB 1 NEW * 1.46 0.36 0.86 0.50 28
UM
UM S60201-TB 1 NEW ** 2.35 0.24 0.89 0.65 28
UM UM S60243-TB 3
NEW ** 2.92 0.27 0.93 0.66 28
UM UM S60326-TB 2 NEW ** 2.11
0.43 0.96 0.54 27
UM UM S60338-TB 1 NEW ** 2.60 0.51 1.00 0.49
25
UM UM S60350-TB 3 NEW ** 2.64 0.08 0.75 0.67 26
UM UM
S60361-TB 2 NEW ** 3.22 0.14 0.88 0.74 25
UM UM S60422-TB 1 NEW
** 2.17 0.52 1.00 0.48 27
UM UM S60440-TB 2 NEW * 1.63 0.06 0.59
0.53 28
UM UM S60446-TB 2 NEW ** 2.80 0.51 1.00 0.49 28
UM UM S60505-TB 1 NEW ** 2.03 0.51 1.00 0.49 26
UM UM S60513-TB
2 NEW * 1.70 0.52 0.96 0.45 28
UM UM S60519-TB 2 NEW ** 3022
0.44 1.00 0.56 28
UM UM 560536-TB 1 NEW ** 2.64 0.43 1.00 0.57
28
UM UM S60552-TB 2 NEW * 1.69 0.31 0.88 0.56 28
UM UM
S60585-TB 4 NEW * 1.45 0.56 0.96 0.40 27
UM UM S60630-TB 3 NEW
* 1.48 0.32 0.89 0.57 26
UM UM S60728-TB 3 NEW * 1.46 0.45 0.91
0.46 28
UM UM S60812-TB 1 NEW * 1.51 0.40 0.87 0.47 27
UM
UM SAC1677 3 ** 2.85 0.16 0.82 0.66 84
UM UM SAC1724 3 NEW *
1.49 .64 1.00 0.36 27
UM UM SAG1055 5 NEW * 1.62 0.65 1.00 0.35
28
UM UM Satt111 3 ** 4.00 .40 0.90 .80 83
UM UM
Satt219 4 NEW * 1.42 0.05 0.42 0.37 80
UM UM Satt299 10 NEW *
1.38 0.65 0.35 0.30 79
* 95% significance level
**
99% significance level
.dagger.LOD greater than 2.0, increase in
frequency less than 25%.
[0264]
13TABLE 13
Favorable Alleles in A3127
Probability of
Parent 1 Parent 2 inheriting "+"
Chromosome Position Allele WILLIAMS ESSEX Progeny A3127 allele
A1 19.1 1 - + + 0.5
A1 27.1 3 + - + 0.5
A2 0.0 1 - + +
0.5
A2 184.0 4 + - + 0.5
B2 45.0 2 + - + 0.5
C1 99.0
4 - + + 0.5
C2 140.9 5 + - + 0.5
D1a 129.2 5 + - + 0.5
D2 93.8 6 - + 0.5
D2 110.4 1 - + + 0.5
E 98.0 1 - + +
0.5
F 1.7 3 + - + 0.5
F 85.0 1 + - + 0.5
G 100.8 1 -
+ + 0.5
H 125.3 5 + - + 0.5
J 68.0 2 not segregating
K 17.9 4 + - + 0.5
L 34.6 4 not segregating
L 77.1 3 + -
+ 0.5
L 124.5 2 not segregating
M 87.6 4 - + + 0.5
N 20.0 3 not segregating
O 6.3 4 not segregating
O 37.7 4
not segregating
O 60.5 2 not segregating
O 125.0 2 + - +
0.5
O 153.3 3 + - + 0.5
UM # 2 + - + 0.5
UM # 3 - +
+ 0.5
UM # 3 - + + 0.5
# of "+" 13 10 all 23 0.000000119
alleles .fwdarw.
chance of this happening by chance = 1
in 8,388,608
[0265]
14TABLE 14
# of
SATT591 SATT429
SATT144 SATT270 SATT529 SATT166 sublines
Elite line segregating
alleles at the above marker loci
91B91 1 and 2 3 and 5
4
92M70 1 and 2 2 and 5 4
92B05 3 and 4 2
93B01 1 and 3 2
93M80 5 and 12 2
93M90 1 and 3
2
[0266]
15TABLE 15
Isoline seed yield means from field
tests
Statistical
grouping
# of plants Mean
Yield
Genotype (allele) bulked to Seed advantage of
Original Subline at indicated marker form # of reps Yield better subline
Elite Line Name locus Isoline (locations) (bu/ac) (bu/ac and %)
SATT529 SATT166 LSD
(0.05) = 2.30
91B91 91B91-13 1 3 42 7 37.5 A
91B91 91B91-15 1 5 24 7 38.0 A
91B91 91B91-23 2 3 14 7 38.8 A
91B91 91B91-25 2 5 7 7 37.2 A
SATT144 SATT270 LSD
(0.05) = 3.48
92M70
92M70-12 1 2 14 10 43.4 A
92M70 92M70-15 1 5 46 10 44.4 A
92M70 92M70-22 2 2 13 10 43.0 A
92M70 92M70-25 2 5 15 10 43.8 A
SATT429 LSD
(0.05) = 2.01
92B05 92B05-3 3
175 5 37.0 A
92B05 92B05-4 4 116 5 37.0 A
SATT591
LSD
(0.05) = 1.71
93B01 93B01-1 1 51 13 38.0 B
93B01 93B01-3 3 28 13 40.1 A 2.1 bu = 5.4%
SATT270 LSD
(0.05) = 2.54
93M80 93M80-12 12 5 14 48.5 B
93M80 93M80-5 5 4 14 51.3 A 2.8 bu = 5.6%
SATT591 LSD
(0.05) = 2.16
93M90 93M90-1 1 38 14 50.2 A 3.0 bu = 6.2%
93M90 93M90-3 3 47 14 47.2 B
[0267]
16APPENDIX I
Allele Definition Table
Marker
Allele Size Range bp
Sac1006 1 91.54 92.48
Sac1006 2 93.67 94.41
Sac1006 3 105.49 106.22
Sac1677 1
191.96 192.46
Sac1677 2 193.94 194.93
Sac1677 3 205.90
206.91
Sac1677 4 332.86 333.35
Sac1677 5 190.20 190.40
Sac1677 6 204.40 204.60
Sat_084 1 151.69 152.30
Sat_084 2 153.28 154.30
Sat_084 3 157.85 158.85
Sat_084 4
163.84 164.82
Sat_084 5 172.09 172.56
Sat_084 6 160.26
160.46
Sat_090 1 336.52 337.36
Sat_090 2 349.84 350.92
Sat_090 3 355.55 356.29
Sat_090 4 338.64 339.23
Sat_090 5 340.79 340.09
Sat_090 6 348.04 348.57
Sat_090 7
351.54 352.37
Sat_090 8 353.79 354.12
Sat_090 9 357.22
357.86
Sat_104 1 266.36 267.50
Sat_104 2 284.72 285.73
Sat_110 1 165.79 166.96
Sat_110 2 179.85 180.32
Sat_110 3 181.88 183.02
Sat_110 4 183.72 184.58
Sat_110 5
185.65 186.71
Sat_110 6 187.88 188.05
Sat_117 1 210.34
211.45
Sat_117 2 218.15 219.41
Sat_117 3 220.13 221.05
Sat_117 4 226.19 226.96
Sat_117 5 232.64 232.99
Sat_117 6 212.76 213.12
Sat_117 7 216.86 217.25
Sat_117 8
222.62 222.99
Sat_117 9 224.83 224.93
Sat_117 10 242.75
242.85
Sat_117 11 252.02 252.23
Sat_117 12 255.97 256.17
Sat_117 13 203.20 203.35
Sat_117 14 208.93 209.13
Sat_117 15 234.70 234.90
Satt020 1 147.56 148.62
Satt020
2 160.10 160.87
Satt040 1 317.25 318.26
Satt040 2 323.30
324.31
Satt040 3 326.83 327.81
Satt040 4 329.75 330.66
Satt040 5 332.71 333.35
Satt040 6 320.28 321.05
Satt040 7 335.83 336.33
Satt040 8 308.03 308.53
Satt040 9
338.89 339.09
Satt040 10 314.39 314.69
Satt042 1 148.26
149.12
Satt042 2 157.59 158.45
Satt042 3 160.69 161.67
Satt042 4 163.71 164.64
Satt042 5 154.82 155.22
Satt042 6 160.26 160.46
Satt042 7 166.96 167.56
Satt042 8
135.78 135.98
Satt042 9 145.41 145.61
Satt050 1 221.60
222.79
Satt050 2 224.66 225.89
Satt050 3 239.80 240.68
Satt050 4 252.17 253.84
Satt050 5 255.27 255.57
Satt066 1 160.87 161.87
Satt066 2 105.74 106.69
Satt066 3
166.76 167.07
Satt066 4 169.95 171.12
Satt066 5 110.96
112.11
Satt066 6 151.75 152.78
Satt066 7 155.32 155.84
Satt066 8 158.20 158.86
Satt066 9 164.34 164.82
Satt066 10 168.42 169.19
Satt066 11 173.11 174.03
Satt066
12 176.16 176.93
Satt066 13 148.64 149.67
Satt066 14
117.63 118.13
Satt092 1 340.69 341.13
Satt092 2 349.07
350.92
Satt092 3 332.36 333.35
Satt092 4 338.79 339.25
Satt092 5 341.43 342.56
Satt092 6 353.17 353.29
Satt102 1 160.36 161.37
Satt102 2 175.43 176.43
Satt102 3
172.56 173.53
Satt102 4 178.87 179.35
Satt102 5 181.79
182.25
Satt102 6 184.58 185.65
Satt108 1 169.96 171.12
Satt108 2 173.06 174.00
Satt108 3 175.96 177.00
Satt108 4 167.15 167.92
Satt108 5 161.77 161.97
Satt108 6
149.12 149.22
Satt109 1 146.99 148.06
Satt109 2 164.62
165.54
Satt109 3 167.65 168.42
Satt109 4 162.27 162.47
Satt109 5 144.05 144.33
Satt111 1 254.76 255.57
Satt111 2 257.79 258.83
Satt111 3 260.76 261.78
Satt111 4
263.82 264.54
Satt111 5 278.98 280.09
Satt111 6 285.22
285.73
Satt111 7 288.24 288.75
Satt111 8 266.88 267.90
Satt111 9 270.13 270.33
Satt111 10 276.28 276.85
Satt111 11 248.80 249.00
Satt115 1 218.67 219.88
Satt115
2 236.35 237.83
Satt115 3 239.29 240.78
Satt115 4 242.65
243.20
Satt115 5 221.82 222.22
Satt115 6 238.31 238.51
Satt122 1 234.40 235.37
Satt122 2 282.65 283.26
Satt122 3 291.75 292.40
Satt127 1 241.61 242.25
Satt127 2
244.22 245.31
Satt127 3 232.44 233.10
Satt127 4 238.81
239.29
Satt127 5 247.60 248.20
Satt127 6 250.71 251.22
Satt129 1 120.97 122.06
Satt129 2 133.16 134.38
Satt129 3 136.28 137.33
Satt129 4 149.22 150.17
Satt129 5
152.23 153.28
Satt129 6 155.84 156.34
Satt129 8 127.42
128.08
Satt129 9 140.02 140.52
Satt129 10 143.25 143.75
Satt129 11 132.05 132.25
Satt130 1 142.17 143.25
Satt130 2 147.44 148.62
Satt130 3 150.62 151.75
Satt130 4
154.30 154.82
Satt131 1 157.85 159.25
Satt131 2 173.06
174.03
Satt131 3 175.96 176.93
Satt131 4 179.35 179.85
Satt131 5 182.16 182.75
Satt131 6 148.62 149.67
Satt133 1 163.80 164.82
Satt133 2 181.99 182.85
Satt133 3
167.36 167.56
Satt133 4 176.06 176.26
Satt133 5 172.96
173.41
Satt138 1 226.66 227.82
Satt138 2 229.84 230.75
Satt138 3 232.85 233.82
Satt138 4 235.30 236.56
Satt138 5 241.66 242.45
Satt138 6 231.47 231.87
Satt138 7
238.91 239.11
Satt138 8 214.97 215.17
Satt142 1 122.55
123.25
Satt142 2 137.89 138.69
Satt142 3 141.10 142.17
Satt142 4 144.33 145.41
Satt142 5 156.34 156.94
Satt142 6 148.06 148.16
Satt142 7 108.04 108.20
Satt142 8
129.00 129.20
Satt144 1 210.72 212.56
Satt144 2 227.62
228.58
Satt144 3 231.25 231.40
Satt146 1 291.11 292.36
Satt146 2 294.26 295.32
Satt146 3 309.55 310.57
Satt146 4 313.02 314.06
Satt146 5 316.11 317.50
Satt146 7
306.71 307.71
Satt146 8 298.10 298.30
Satt146 9 319.97
320.38
Satt146 10 322.80 323.30
Satt146 11 329.37 329.87
Satt146 12 331.75 332.66
Satt146 13 334.84 335.83
Satt147 1 176.73 177.70
Satt147 2 182.75 183.52
Satt147 3
193.45 195.03
Satt147 4 179.95 180.52
Satt147 5 207.01
208.72
Satt147 6 211.35 211.55
Satt147 7 213.90 214.77
Satt147 8 216.95 217.55
Satt147 9 199.29 199.49
Satt150 1 246.11 248.00
Satt150 2 249.20 249.70
Satt150 3
267.70 267.90
Satt150 4 270.95 271.05
Satt150 5 282.55
283.21
Satt150 6 234.18 234.60
Satt150 7 237.33 237.63
Satt151 1 169.04 170.15
Satt151 2 178.06 179.35
Satt151 3 184.04 185.15
Satt151 4 187.08 188.05
Satt151 5
205.40 206.40
Satt151 6 166.29 166.76
Satt151 7 175.46
175.96
Satt151 8 198.89 199.39
Satt153 1 198.39 199.49
Satt153 2 201.39 202.09
Satt153 3 213.47 214.57
Satt153 4 231.30 232.17
Satt153 5 220.13 220.53
Satt155 1
151.75 152.78
Satt155 2 160.87 162.37
Satt155 3 163.84
165.02
Satt155 4 167.11 167.72
Satt155 5 158.35 158.45
Satt155 6 148.82 149.12
Satt155 7 197.50 197.90
Satt156 1 326.12 326.84
Satt156 2 329.20 330.02
Satt156 3
343.89 344.85
Satt156 4 349.65 350.00
Satt156 5 346.68
347.11
Satt159 1 268.90 269.53
Satt159 2 269.73 270.33
Satt159 3 271.94 272.78
Satt159 4 287.23 287.74
Satt159 5 290.25 290.95
Satt165 1 210.44 211.45
Satt165 2
213.37 214.63
Satt165 3 219.41 220.48
Satt165 4 216.55
216.95
Satt165 5 204.70 204.90
Satt165 6 225.69 225.99
Satt166 1 187.08 188.10
Satt166 2 220.48 221.35
Satt166 3 226.44 227.62
Satt166 4 229.48 230.24
Satt166 5
232.37 233.67
Satt166 6 223.39 223.76
Satt166 7 217.35
217.75
Satt168 1 345.94 346.71
Satt168 2 359.98 360.77
Satt168 3 362.95 363.65
Satt168 4 365.87 367.16
Satt168 5 368.83 369.34
Satt168 6 335.26 336.00
Satt168 7
311.10 311.30
Satt168 8 329.57 329.77
Satt172 1 155.84
156.89
Satt172 2 167.72 168.52
Satt172 3 170.62 171.52
Satt172 4 179.35 180.32
Satt172 5 182.25 183.22
Satt175 1 160.21 161.37
Satt175 2 163.31 164.54
Satt175 3
166.29 166.96
Satt175 4 169.19 170.50
Satt175 5 178.30
179.55
Satt175 6 193.45 194.74
Satt175 7 172.76 173.26
Satt175 8 175.61 176.26
Satt175 9 188.05 188.60
Satt175 10 190.98 191.96
Satt175 11 181.69 181.89
Satt176
1 138.39 139.15
Satt176 2 141.60 142.77
Satt176 3 160.36
162.37
Satt176 4 169.68 170.77
Satt176 5 172.49 173.68
Satt176 6 187.65 188.85
Satt176 7 148.16 149.02
Satt176 8 163.55 164.67
Satt176 9 175.56 176.63
Satt176
10 181.62 182.75
Satt181 1 324.31 325.37
Satt181 2 346.28
347.16
Satt181 3 354.62 355.60
Satt181 4 357.67 358.46
Satt181 5 363.01 364.30
Satt181 6 351.94 352.47
Satt181 7 360.53 361.17
Satt181 8 366.01 367.16
Satt181 9
349.07 349.55
Satt183 1 122.04 123.05
Satt183 2 128.08
129.10
Satt183 3 144.83 145.91
Satt183 4 148.16 148.62
Satt183 5 131.43 131.85
Satt183 6 135.26 135.78
Satt183 7 138.49 138.95
Satt183 8 141.70 141.90
Satt186 1
361.81 363.07
Satt186 2 364.72 365.94
Satt186 3 367.87
368.83
Satt186 4 339.25 341.09
Satt186 5 359.30 359.73
Satt186 6 371.22 371.72
Satt186 7 373.74 374.61
Satt186 8 376.96 377.46
Satt186 9 353.39 353.59
Satt190 1
187.01 188.06
Satt190 2 193.35 194.10
Satt190 3 196.42
197.40
Satt190 4 225.99 227.06
Satt190 5 181.17 182.03
Satt190 6 184.18 184.68
Satt190 7 190.50 190.98
Satt190 8 220.38 220.85
Satt190 9 214.07 214.57
Satt191 1
319.67 320.28
Satt191 2 334.84 336.02
Satt191 3 337.81
339.02
Satt191 4 349.07 350.00
Satt191 5 352.14 352.77
Satt191 6 354.62 355.65
Satt191 7 360.69 361.17
Satt191 8 346.68 346.71
Satt193 1 165.79 166.76
Satt193 2
168.94 169.65
Satt193 3 174.59 175.96
Satt193 4 180.77
181.79
Satt193 5 186.61 187.75
Satt193 6 189.97 190.60
Satt193 7 192.95 193.75
Satt193 8 198.89 199.89
Satt193 9 202.19 203.40
Satt193 10 183.72 185.15
Satt193
11 171.79 173.06
Satt193 12 160.11 161.37
Satt193 13
177.85 179.08
Satt193 14 163.36 163.84
Satt195 1 214.77
216.11
Satt195 2 217.93 218.91
Satt195 3 221.65 222.22
Satt196 1 230.04 230.50
Satt196 2 232.94 233.52
Satt196 3 239.29 239.79
Satt196 4 241.76 243.00
Satt196 5
253.84 255.27
Satt196 6 236.17 236.45
Satt196 7 257.51
257.81
Satt196 8 263.52 263.92
Satt197 1 177.50 179.35
Satt197 2 184.18 185.15
Satt197 3 187.08 188.55
Satt197 4 190.00 191.48
Satt197 5 192.95 194.44
Satt197 6
205.65 206.00
Satt197 7 170.35 170.72
Satt197 8 175.44
175.96
Satt197 9 166.96 167.71
Satt197 10 181.59 181.79
Satt199 1 161.84 162.86
Satt199 2 202.22 204.40
Satt199 3 166.29 166.76
Satt199 4 168.22 168.69
Satt199 5
174.03 174.99
Satt199 6 178.37 178.87
Satt199 7 220.85
221.35
Satt199 8 176.01 176.16
Satt199 9 170.20 170.60
Satt202 1 306.48 307.51
Satt202 2 309.55 310.72
Satt202 3 287.33 287.79
Satt202 4 313.03 313.66
Satt202 5
285.22 285.93
Satt203 1 223.29 224.06
Satt203 2 229.53
230.04
Satt203 3 249.20 250.00
Satt203 4 258.33 259.34
Satt203 5 205.40 205.90
Satt203 6 212.56 212.76
Satt203 7 219.61 219.81
Satt203 8 226.51 227.82
Satt203 9
231.30 231.67
Satt204 1 234.88 235.87
Satt204 2 240.78
241.76
Satt204 3 247.00 248.20
Satt204 4 250.00 250.76
Satt204 5 253.03 253.79
Satt204 6 238.13 238.31
Satt204 7 256.59 256.79
Satt209 1 311.80 312.63
Satt209 2
329.37 330.48
Satt209 3 310.77 311.10
Satt212 1 347.13
348.73
Satt212 2 355.42 356.49
Satt212 3 358.66 358.80
Satt213 1 326.34 327.35
Satt213 2 318.46 318.66
Satt216 1 195.43 196.42
Satt216 2 198.79 199.39
Satt216 3
222.79 223.29
Satt216 4 225.69 226.66
Satt216 5 210.95
211.45
Satt216 6 216.95 217.45
Satt216 7 220.18 220.38
Satt218 1 341.13 342.16
Satt218 2 344.12 344.85
Satt219 1 126.38 127.07
Satt219 2 141.95 142.67
Satt219 3
157.69 158.55
Satt219 4 175.46 176.58
Satt219 5 120.38
120.77
Satt219 6 154.82 155.12
Satt219 7 160.97 161.37
Satt219 8 172.80 173.46
Satt219 9 178.78 179.55
Satt219 10 182.09 182.45
Satt220 1 337.71 338.29
Satt220
2 340.39 341.23
Satt220 3 343.24 344.42
Satt220 4 346.28
346.71
Satt220 5 334.54 334.94
Satt220 6 354.62 355.02
Satt220 7 360.23 360.63
Satt220 8 352.52 352.57
Satt221 1 228.12 229.08
Satt221 2 234.12 234.90
Satt221 3
237.33 238.31
Satt221 4 246.50 247.50
Satt221 5 231.20
231.47
Satt225 1 99.95 101.41
Satt225 2 102.87 103.93
Satt225 3 109.05 110.01
Satt227 1 327.28 328.73
Satt227 2 333.35 334.55
Satt228 1 293.26 294.26
Satt228 2
298.30 299.06
Satt228 3 310.00 310.57
Satt228 4 323.40
324.31
Satt228 5 326.44 327.35
Satt228 6 329.61 330.88
Satt228 7 317.45 317.65
Satt228 8 341.53 341.73
Satt230 1 226.14 227.26
Satt230 2 229.33 230.50
Satt230 3
231.97 232.99
Satt230 4 214.97 215.47
Satt230 5 223.39
224.26
Satt233 1 90.83 91.34
Satt233 4 91.64 92.40
Satt233 5 97.75 98.46
Satt233 6 103.73 105.25
Satt233 7
112.77 113.66
Satt234 1 175.32 176.21
Satt234 2 178.35
179.35
Satt234 3 172.96 173.06
Satt236 1 250.00 250.81
Satt236 2 255.77 256.79
Satt236 3 259.08 259.84
Satt236 4 262.16 262.95
Satt236 5 265.46 265.66
Satt236 6
271.35 271.56
Satt236 7 253.39 253.84
Satt240 1 310.07
311.30
Satt240 2 313.16 314.63
Satt240 3 316.44 317.86
Satt240 4 319.60 320.48
Satt240 5 326.34 326.54
Satt242 1 182.25 183.22
Satt242 2 191.48 192.46
Satt242 3
197.40 198.39
Satt242 4 200.39 201.52
Satt242 5 203.25
204.40
Satt242 6 209.43 210.44
Satt242 7 212.91 213.47
Satt242 8 167.72 168.22
Satt242 9 179.85 180.32
Satt242 10 188.55 189.02
Satt242 11 194.93 195.43
Satt242
12 206.91 207.41
Satt242 13 185.65 186.11
Satt242 14
171.94 172.29
Satt242 15 175.09 175.34
Satt242 16 181.17
181.22
Satt242 17 196.12 196.27
Satt243 1 334.69 335.83
Satt243 2 343.48 344.47
Satt243 3 363.37 364.48
Satt243 4 366.24 366.91
Satt243 5 357.96 358.16
Satt243 6
337.76 338.69
Satt243 7 346.58 347.06
Satt243 8 349.47
349.60
Satt247 1 363.62 365.08
Satt247 2 369.29 370.70
Satt247 3 375.25 376.51
Satt247 4 366.63 367.67
Satt247 5 373.15 373.35
Satt247 6 381.11 382.25
Satt247 7
378.30 378.50
Satt247 8 355.25 355.45
Satt249 1 217.90
218.73
Satt249 2 220.85 221.65
Satt249 3 251.19 252.23
Satt249 4 257.31 258.33
Satt249 5 254.61 255.06
Satt249 6 260.96 261.38
Satt249 7 236.75 236.95
Satt250 1
189.02 190.00
Satt250 2 207.16 208.02
Satt250 3 228.68
228.98
Satt250 4 213.87 213.97
Satt250 5 219.41 219.61
Satt251 1 215.92 216.95
Satt251 2 218.91 219.41
Satt251 3 254.26 255.27
Satt251 4 257.81 259.08
Satt251 5
266.48 267.50
Satt251 6 209.77 210.95
Satt251 7 260.86
261.38
Satt251 8 212.97 213.97
Satt251 9 272.98 273.18
Satt255 1 236.85 237.43
Satt255 2 242.75 243.33
Satt255 3 245.71 246.70
Satt255 4 248.70 249.30
Satt256 1
338.99 340.19
Satt256 2 348.27 349.50
Satt256 3 351.28
352.27
Satt257 1 255.20 256.29
Satt257 2 267.33 268.52
Satt257 3 242.25 243.15
Satt257 4 249.20 250.41
Satt257 5 258.48 259.74
Satt257 6 261.78 262.18
Satt258 1
156.62 157.69
Satt258 2 159.71 160.71
Satt258 3 161.37
161.87
Satt258 4 162.82 163.84
Satt258 5 165.79 166.76
Satt258 6 168.22 168.42
Satt259 1 187.08 188.15
Satt259 2 193.45 194.44
Satt259 3 205.40 206.50
Satt259 4
207.65 209.58
Satt259 5 211.82 212.64
Satt262 1 231.97
232.94
Satt262 2 244.38 245.21
Satt262 3 247.20 248.20
Satt262 4 250.21 251.22
Satt262 5 256.29 257.31
Satt262 6 241.41 242.06
Satt262 7 253.42 254.04
Satt262 8
235.67 235.87
Satt263 1 249.53 250.21
Satt263 2 252.43
253.44
Satt263 3 267.70 268.72
Satt263 4 276.75 278.02
Satt263 5 289.18 290.10
Satt263 6 255.72 256.29
Satt263 7 265.04 265.46
Satt263 8 271.56 271.76
Satt263 9
246.44 247.20
Satt264 1 216.38 217.45
Satt264 2 219.35
220.38
Satt264 3 228.41 229.54
Satt264 4 237.33 238.46
Satt264 5 246.65 247.70
Satt264 6 243.63 244.72
Satt265 1 264.64 265.96
Satt265 2 267.88 268.52
Satt265 3
285.98 286.93
Satt265 4 289.35 289.55
Satt265 5 258.93
259.54
Satt265 6 262.18 262.40
Satt265 7 283.11 283.41
Satt266 1 321.79 323.05
Satt266 2 330.98 332.32
Satt266 3 303.20 303.93
Satt267 1 232.44 233.42
Satt267 2
241.46 242.25
Satt267 3 250.61 251.32
Satt267 4 244.82
245.21
Satt267 5 247.80 248.00
Satt270 1 110.46 110.96
Satt270 2 119.10 119.98
Satt270 3 140.97 141.60
Satt270 4 141.86 142.67
Satt270 5 144.05 144.98
Satt270 6
150.65 151.28
Satt270 7 116.20 116.71
Satt270 8 125.06
125.56
Satt270 9 128.60 129.10
Satt270 10 147.56 148.06
Satt270 11 154.31 155.32
Satt270 12 156.61 157.34
Satt272 1 223.76 225.23
Satt272 2 229.94 231.00
Satt272 3
226.94 228.12
Satt272 4 233.42 233.92
Satt272 5 242.25
242.75
Satt272 6 247.20 247.70
Satt274 1 195.33 196.32
Satt274 2 198.38 199.44
Satt274 3 201.34 202.44
Satt274 4 204.40 205.40
Satt274 5 180.25 180.52
Satt274 6
189.42 189.62
Satt274 7 207.45 208.72
Satt279 1 174.83
175.76
Satt280 1 230.04 231.05
Satt280 2 180.92 181.79
Satt280 2 233.19 233.92
Satt280 3 236.17 236.90
Satt280 4 202.69 203.00
Satt280 5 224.29 224.98
Satt282 1
314.04 314.79
Satt282 2 317.12 318.31
Satt282 3 326.34
327.85
Satt282 4 192.95 193.65
Satt282 4 332.51 334.05
Satt282 5 335.68 336.92
Satt282 6 347.84 348.04
Satt282 7 323.45 324.31
Satt282 8 329.67 330.58
Satt282 9
338.88 339.75
Satt282 10 310.67 311.70
Satt284 1 115.70
116.66
Satt284 2 124.56 125.61
Satt284 3 127.95 128.70
Satt284 4 119.20 119.30
Satt284 5 112.86 113.06
Satt284 6 198.89 199.89
Satt284 6 129.40 129.61
Satt285 1
208.93 210.19
Satt285 2 231.97 232.95
Satt285 3 235.51
236.36
Satt285 4 241.26 242.50
Satt285 5 243.73 245.41
Satt285 6 206.40 206.80
Satt285 7 187.28 187.48
Satt285 7 229.32 230.34
Satt287 1 203.38 204.40
Satt287 2
233.55 234.40
Satt287 3 197.70 198.09
Satt287 4 243.15
243.53
Satt287 9 205.60 205.70
Satt292 1 221.35 221.82
Satt292 2 224.26 225.23
Satt292 3 239.29 240.28
Satt292 4 242.66 243.23
Satt292 5 251.70 252.23
Satt292 6
229.18 229.48
Satt292 7 232.24 232.44
Satt292 8 211.25
211.55
Satt292 9 254.76 255.06
Satt292 10 257.91 258.21
Satt295 1 220.38 221.25
Satt295 2 223.29 224.26
Satt295 3 226.57 227.16
Satt295 4 232.44 233.14
Satt295 5
244.62 245.21
Satt295 6 256.73 257.41
Satt295 7 264.94
265.96
Satt295 8 208.82 209.03
Satt295 9 217.15 217.75
Satt295 11 235.62 236.02
Satt295 12 238.81 239.01
Satt295 13 241.66 241.91
Satt295 14 253.84 254.04
Satt299
1 258.01 258.21
Satt299 2 283.51 283.61
Satt299 3 284.01
284.97
Satt299 4 285.12 286.27
Satt299 5 291.25 292.30
Satt299 6 303.70 304.30
Satt299 7 306.88 307.51
Satt299 8 313.66 314.69
Satt299 9 318.76 319.47
Satt299
10 328.65 329.87
Satt299 11 331.87 332.81
Satt299 12
342.56 343.49
Satt299 13 293.26 293.36
Satt300 1 238.81
239.79
Satt300 2 245.20 245.81
Satt300 3 254.26 255.27
Satt300 4 266.48 267.23
Satt300 5 269.53 270.55
Satt300 6 242.25 242.85
Satt300 7 257.81 258.11
Satt300 8
263.57 263.82
Satt303 1 128.60 129.10
Satt303 2 131.59
132.15
Satt303 3 134.68 135.34
Satt303 4 137.82 138.39
Satt303 5 140.96 141.70
Satt303 6 144.13 144.88
Satt303 7 153.65 154.41
Satt303 8 156.67 157.49
Satt303 9
119.59 120.08
Satt303 10 122.75 123.05
Satt303 11 147.66
147.86
Satt303 12 159.96 160.16
Satt304 1 98.35 99.16
Satt304 2 162.75 163.84
Satt304 3 165.99 166.59
Satt304 4 168.81 169.65
Satt307 1 336.32 337.51
Satt307 2
350.42 351.44
Satt307 3 355.99 356.92
Satt307 4 358.93
360.82
Satt307 5 362.00 362.67
Satt307 6 365.33 365.84
Satt307 7 367.97 368.48
Satt311 1 152.78 153.81
Satt311 2 164.82 165.79
Satt311 3 175.96 176.43
Satt311 4
156.24 156.34
Satt311 5 192.37 192.47
Satt311 6 207.41
207.51
Satt311 7 201.44 201.89
Satt311 8 210.14 210.85
Satt311 9 181.29 181.69
Satt314 1 241.26 242.25
Satt314 2 244.22 245.21
Satt314 3 247.50 247.80
Satt319 1
173.31 174.59
Satt319 2 176.48 177.50
Satt319 3 179.47
180.52
Satt319 4 218.91 219.88
Satt319 5 237.16 238.13
Satt319 6 240.78 241.26
Satt319 7 183.22 183.32
Satt319 8 192.27 192.47
Satt319 9 210.44 210.95
Satt319
10 234.90 235.00
Satt321 1 231.97 233.25
Satt321 2 235.25
236.23
Satt321 3 238.31 239.26
Satt321 4 244.22 244.72
Satt322 1 313.03 313.81
Satt322 2 312.13 312.78
Satt326 1 323.68 324.81
Satt326 2 326.69 327.85
Satt326 3
329.87 330.88
Satt326 4 342.06 342.76
Satt326 5 320.83
321.39
Satt327 1 251.22 252.23
Satt327 2 254.26 255.28
Satt327 3 245.41 245.81
Satt327 4 248.55 248.80
Satt327 5 233.62 234.02
Satt327 6 263.72 263.92
Satt327 7
242.42 242.80
Satt327 8 257.51 257.66
Satt328 1 248.20
249.20
Satt328 2 251.22 252.23
Satt329 1 238.31 238.81
Satt329 2 268.52 269.53
Satt329 3 272.58 273.38
Satt329 4 275.63 276.65
Satt329 5 279.69 280.70
Satt329 6
247.20 247.70
Satt329 7 254.26 254.76
Satt329 8 257.31
258.33
Satt329 9 267.50 268.00
Satt329 10 277.25 277.45
Satt329 11 279.28 279.48
Satt329 12 274.00 274.10
Satt329 13 270.13 270.23
Satt330 1 308.03 308.78
Satt330
2 342.41 343.49
Satt330 3 348.04 348.72
Satt330 4 355.55
355.99
Satt330 5 364.02 364.78
Satt330 6 344.72 344.85
Satt330 7 345.50 345.98
Satt330 8 346.71 346.91
Satt330 9 316.53 316.93
Satt331 1 233.42 234.40
Satt331 2
239.67 240.28
Satt331 3 251.56 252.43
Satt331 4 236.95
237.15
Satt332 1 161.69 162.37
Satt332 2 176.57 177.90
Satt332 3 179.65 180.32
Satt332 4 182.85 183.20
Satt332 5 173.78 174.13
Satt333 1 180.32 180.82
Satt333 2
195.43 196.47
Satt333 3 198.89 199.39
Satt333 4 201.89
202.39
Satt333 5 204.74 205.40
Satt333 6 171.79 171.99
Satt333 7 189.82 190.00
Satt333 8 177.75 178.20
Satt333 9 192.76 193.10
Satt334 1 202.39 202.90
Satt334 2
207.92 208.93
Satt334 3 210.95 211.96
Satt334 4 214.33
215.77
Satt334 5 215.96 216.76
Satt334 6 217.93 218.70
Satt334 7 220.38 221.35
Satt334 8 196.12 196.32
Satt334 9 230.34 230.50
Satt335 1 145.31 146.49
Satt335 2
154.82 156.09
Satt335 3 164.34 165.27
Satt335 4 167.51
168.22
Satt335 5 170.55 171.27
Satt336 1 176.43 177.40
Satt336 2 182.53 183.22
Satt336 3 185.50 186.61
Satt336 4 191.48 192.46
Satt338 1 229.08 230.04
Satt338 2
232.54 233.92
Satt338 3 253.24 254.37
Satt338 4 265.46
266.68
Satt338 5 274.62 275.57
Satt338 6 277.67 278.68
Satt338 7 238.31 238.71
Satt338 8 280.90 281.90
Satt339 1 234.90 235.38
Satt339 2 240.47 241.97
Satt339 3
243.73 244.72
Satt339 4 255.91 257.31
Satt339 5 264.88
266.26
Satt339 6 268.14 269.07
Satt339 7 206.70 206.91
Satt339 8 252.83 253.92
Satt339 9 271.45 271.56
Satt339 10 250.21 250.61
Satt339 11 262.28 262.40
Satt339
12 259.13 259.35
Satt343 1 138.39 138.95
Satt343 2 141.50
142.37
Satt343 3 160.14 161.87
Satt343 4 169.19 169.87
Satt343 5 172.09 173.06
Satt343 6 187.08 188.05
Satt343 7 148.16 148.62
Satt343 8 181.12 181.99
Satt343 9
163.26 164.00
Satt343 10 175.46 175.86
Satt343 11 154.51
154.71
Satt346 1 321.29 322.30
Satt346 2 327.35 328.36
Satt346 3 336.32 337.41
Satt346 4 339.25 340.29
Satt346 5 318.20 319.27
Satt346 6 324.40 325.32
Satt347 1
345.25 346.28
Satt347 2 347.94 349.07
Satt347 3 351.02
351.84
Satt348 1 308.03 308.72
Satt348 2 333.01 333.85
Satt348 3 341.83 342.66
Satt348 4 344.67 345.35
Satt348 5 330.27 330.58
Satt348 6 326.84 327.35
Satt348 7
339.55 339.85
Satt352 1 334.54 335.83
Satt352 2 341.13
341.63
Satt352 3 346.15 347.21
Satt352 4 351.72 352.87
Satt352 5 354.62 355.55
Satt352 6 340.59 340.89
Satt352 7 343.79 343.92
Satt352 8 369.49 369.69
Satt352 9
349.17 349.80
Satt352 10 337.41 338.54
Satt352 11 360.53
360.63
Satt353 1 144.20 145.03
Satt353 2 159.86 160.87
Satt353 3 169.19 169.75
Satt353 4 174.94 175.96
Satt353 5 184.18 185.08
Satt353 6 178.37 178.97
Satt353 7
165.84 166.96
Satt353 8 154.30 154.50
Satt355 1 248.20
249.20
Satt355 2 257.22 258.33
Satt355 3 272.58 273.43
Satt355 4 236.35 237.33
Satt355 5 254.26 254.86
Satt355 6 269.53 270.03
Satt355 7 275.88 276.33
Satt355 8
278.88 279.28
Satt355 9 260.76 260.96
Satt356 1 322.24
323.30
Satt356 2 325.15 326.72
Satt357 1 171.69 173.06
Satt357 2 275.63 277.15
Satt357 3 278.60 280.19
Satt357 4 183.22 184.18
Satt357 5 198.70 199.39
Satt357 6
262.40 262.90
Satt357 7 269.53 270.03
Satt357 8 286.73
287.23
Satt358 1 194.93 196.42
Satt358 2 206.91 208.17
Satt358 3 209.94 210.95
Satt358 4 203.90 205.10
Satt358 5 164.82 166.29
Satt358 6 198.39 199.39
Satt358 7
162.85 163.84
Satt359 1 182.58 183.65
Satt359 2 184.68
186.00
Satt359 3 197.75 198.64
Satt359 4 200.89 201.39
Satt359 5 189.12 189.42
Satt359 6 203.80 204.50
Satt359 7 206.75 207.36
Satt359 8 164.82 165.27
Satt361 1
271.81 273.08
Satt361 2 274.95 275.93
Satt361 3 269.02
269.93
Satt361 4 278.07 278.78
Satt364 1 232.45 233.40
Satt364 2 235.38 236.18
Satt364 3 241.26 242.55
Satt364 4 244.22 245.41
Satt364 5 238.61 238.91
Satt364 6
229.54 229.74
Satt367 1 191.96 193.00
Satt367 2 212.97
214.07
Satt367 3 215.96 217.15
Satt367 4 221.82 223.09
Satt367 5 225.13 226.19
Satt367 6 228.12 229.18
Satt367 7 231.25 232.07
Satt367 8 195.58 195.83
Satt367 9
189.07 189.47
Satt367 10 234.70 234.80
Satt367 11 196.72
197.10
Satt367 12 210.34 210.64
Satt369 1 234.60 236.18
Satt369 2 236.65 237.33
Satt369 3 239.55 240.78
Satt369 4 242.51 243.73
Satt369 5 218.43 219.06
Satt369 6
227.53 228.02
Satt369 7 231.00 231.20
Satt369 8 245.63
246.11
Satt372 1 336.98 338.29
Satt372 2 340.05 341.13
Satt372 3 345.78 346.28
Satt372 4 348.57 349.17
Satt372 5 351.34 352.27
Satt372 6 334.15 334.54
Satt372 7
350.52 350.62
Satt372 8 306.08 306.58
Satt373 1 143.25
143.75
Satt373 2 146.49 147.09
Satt373 3 149.52 150.27
Satt373 4 158.75 159.51
Satt373 5 179.85 180.32
Satt373 6 182.70 183.72
Satt373 7 212.97 213.47
Satt373 8
152.78 153.28
Satt373 9 155.72 156.54
Satt373 10 173.73
174.43
Satt373 11 178.20 178.58
Satt373 12 185.85 186.11
Satt373 13 191.97 192.37
Satt373 14 194.93 195.33
Satt373 15 198.10 198.40
Satt373 16 215.96 216.71
Satt373
17 218.80 219.66
Satt373 18 207.11 207.31
Satt378 1
138.70 139.45
Satt378 2 151.75 152.78
Satt378 3 145.61
145.81
Satt378 4 148.82 149.22
Satt378 5 159.86 160.26
Satt380 1 147.94 148.82
Satt380 2 149.22 149.82
Satt380 3 151.31 152.15
Satt380 4 152.35 152.78
Satt383 1
341.04 341.89
Satt383 2 345.72 346.86
Satt383 3 348.45
349.60
Satt383 4 351.32 352.42
Satt383 5 340.29 340.49
Satt383 6 344.22 344.42
Satt384 1 125.06 125.76
Satt384 2 153.68 154.45
Satt384 3 156.80 157.34
Satt385 1
100.37 101.41
Satt385 2 115.70 116.30
Satt385 3 128.08
128.60
Satt385 4 140.32 141.60
Satt385 5 143.45 144.53
Satt385 6 149.91 151.11
Satt385 7 146.97 148.06
Satt385 8 156.29 157.34
Satt385 9 160.87 161.37
Satt385
10 163.84 164.34
Satt385 11 173.06 174.29
Satt385 12
153.18 153.38
Satt387 1 171.12 171.59
Satt387 2 177.90
178.37
Satt387 3 200.35 201.14
Satt387 4 204.90 206.40
Satt387 5 212.97 213.97
Satt387 6 216.16 216.36
Satt389 1 145.91 146.49
Satt389 2 148.87 149.67
Satt389 3
153.28 154.21
Satt389 4 173.49 174.62
Satt389 5 176.43
177.45
Satt389 6 179.45 180.94
Satt389 7 147.96 148.26
Satt389 8 156.44 156.84
Satt389 9 182.45 183.15
Satt389 10 140.52 140.82
Satt389 11 168.22 168.42
Satt390
1 358.94 359.73
Satt390 2 361.62 362.77
Satt390 3 356.49
356.92
Satt390 4 364.73 365.44
Satt390 5 350.67 350.87
Satt391 1 168.69 169.65
Satt391 2 183.67 184.68
Satt391 3 181.12 181.29
Satt391 4 187.28 187.48
Satt393 1
325.13 326.20
Satt393 2 328.15 329.16
Satt393 3 322.09
322.50
Satt393 4 331.47 331.67
Satt398 1 151.61 152.78
Satt398 2 154.70 155.84
Satt398 3 157.71 158.86
Satt398 4 166.76 167.92
Satt398 5 169.85 171.12
Satt398 6
160.79 161.87
Satt398 7 163.89 164.82
Satt398 8 176.43
176.93
Satt398 9 179.85 180.32
Satt398 10 182.25 183.72
Satt398 11 148.62 148.82
Satt399 1 312.51 313.76
Satt399 2 325.00 326.12
Satt399 3 328.05 329.37
Satt399 4
331.31 332.12
Satt406 1 161.57 162.57
Satt406 2 239.21
240.38
Satt406 3 242.25 243.33
Satt406 4 245.41 246.20
Satt406 5 158.73 159.46
Satt406 6 233.42 233.82
Satt406 7 230.34 230.44
Satt406 8 215.47 215.57
Satt406 9
236.28 237.26
Satt406 10 227.26 227.46
Satt409 1 166.29
166.76
Satt409 2 169.19 170.15
Satt409 3 174.99 176.16
Satt409 4 184.18 185.20
Satt409 5 187.08 188.35
Satt409 6 193.68 194.44
Satt409 7 199.69 200.39
Satt409 8
196.72 196.90
Satt409 9 157.24 157.54
Satt409 10 163.56
163.66
Satt409 11 202.79 202.90
Satt409 12 190.51 191.28
Satt409 13 172.29 172.39
Satt411 1 97.95 99.06
Satt411 2 100.93 102.21
Satt411 3 93.77 94.17
Satt412 1
248.70 249.57
Satt412 2 254.76 255.78
Satt412 3 257.79
258.83
Satt412 4 260.81 261.88
Satt412 5 263.87 264.94
Satt412 6 251.91 252.73
Satt412 7 267.23 268.00
Satt413 1 333.87 334.84
Satt413 2 356.86 357.96
Satt413 3
359.66 360.72
Satt413 4 369.64 370.75
Satt413 5 342.16
342.36
Satt413 6 342.96 342.99
Satt413 7 366.61 366.81
Satt413 8 368.63 369.49
Satt414 1 277.87 279.18
Satt414 2 317.75 319.16
Satt414 3 321.22 322.30
Satt414 4
324.15 325.32
Satt414 5 327.35 328.36
Satt414 7 336.62
337.31
Satt414 8 302.69 303.40
Satt414 9 315.21 316.23
Satt414 10 339.70 340.19
Satt414 11 342.66 342.86
Satt414 12 330.78 331.28
Satt415 1 290.25 291.40
Satt415
2 297.19 298.60
Satt415 3 300.33 301.87
Satt415 4 303.75
304.96
Satt415 5 310.07 310.97
Satt415 6 323.10 323.50
Satt415 7 292.05 292.35
Satt415 8 307.61 307.96
Satt415 9 288.66 289.45
Satt416 1 241.96 242.16
Satt416 2
253.32 254.26
Satt416 3 256.56 257.91
Satt416 4 262.64
263.42
Satt416 5 265.86 266.48
Satt416 6 280.82 282.30
Satt416 7 244.34 244.77
Satt416 8 259.66 260.05
Satt417 1 299.06 300.33
Satt417 2 318.26 319.27
Satt417 3
321.44 322.30
Satt417 4 339.40 341.13
Satt417 5 342.44
343.92
Satt417 6 336.96 337.51
Satt418 1 207.40 208.42
Satt418 2 210.36 211.45
Satt418 3 213.40 214.47
Satt418 4 216.35 217.45
Satt418 5 219.41 219.88
Satt418 6
189.53 190.00
Satt418 7 192.47 193.45
Satt418 8 198.60
199.39
Satt418 9 204.50 204.85
Satt420 1 249.67 250.71
Satt420 2 260.72 261.97
Satt420 3 266.98 268.00
Satt420 4 274.62 275.12
Satt420 5 252.88 253.74
Satt420 6
256.08 256.29
Satt420 7 264.22 264.44
Satt421 1 135.14
136.43
Satt421 2 144.83 146.49
Satt421 3 148.55 149.22
Satt422 1 311.60 312.77
Satt422 2 321.29 321.79
Satt422 3 330.38 331.74
Satt422 4 333.35 334.34
Satt422 5
315.31 315.61
Satt422 6 318.36 319.30
Satt422 7 336.92
337.12
Satt422 8 327.85 328.05
Satt423 1 241.76 242.75
Satt423 2 262.90 263.92
Satt423 3 266.06 266.98
Satt423 4 340.05 340.69
Satt423 6 321.79 321.89
Satt423 7
248.60 248.80
Satt426 1 198.39 199.59
Satt426 2 200.39
200.89
Satt426 3 202.39 203.40
Satt426 4 203.90 204.40
Satt426 5 205.34 206.40
Satt426 6 217.55 218.03
Satt426 7 192.46 192.66
Satt426 8 223.59 223.76
Satt426 9
196.90 197.00
Satt429 1 263.42 264.22
Satt429 2 266.48
267.50
Satt429 3 269.53 270.65
Satt429 4 272.54 273.65
Satt429 5 245.21 246.11
Satt429 6 248.20 248.85
Satt429 7 254.46 254.76
Satt429 8 275.83 276.43
Satt429 9
257.31 257.71
Satt430 1 241.76 242.75
Satt430 2 244.72
245.41
Satt430 3 248.10 248.70
Satt430 4 254.26 254.46
Satt430 5 257.31 257.51
Satt430 6 239.21 239.30
Satt431 1 305.46 306.38
Satt431 2 318.20 318.96
Satt431 3
321.29 322.30
Satt431 4 324.31 324.81
Satt431 5 342.51
343.29
Satt431 6 348.11 349.07
Satt431 7 315.41 315.51
Satt431 8 331.37 331.57
Satt431 9 328.05 328.25
Satt431 10 340.15 340.29
Satt431 11 345.50 345.88
Satt431
12 351.44 351.54
Satt432 1 255.78 256.29
Satt432 2 258.53
259.50
Satt432 3 261.72 262.40
Satt432 4 264.89 265.46
Satt433 1 125.56 126.50
Satt433 2 134.74 135.78
Satt433 3 144.33 145.41
Satt433 4 153.80 154.98
Satt433 5
166.14 167.20
Satt433 6 156.64 156.94
Satt433 7 138.19
138.49
Satt433 8 159.55 159.86
Satt436 1 187.58 188.75
Satt436 2 208.93 209.94
Satt436 3 244.72 245.76
Satt436 4 226.96 227.26
Satt436 5 247.70 248.85
Satt436 6
250.71 251.92
Satt436 7 203.30 203.60
Satt436 8 254.11
254.76
Satt440 1 286.23 287.23
Satt440 2 307.74 308.88
Satt440 3 320.28 320.98
Satt440 4 338.74 339.58
Satt440 5 287.33 288.04
Satt440 6 323.70 324.36
Satt440 7
335.80 336.32
Satt440 8 317.45 317.65
Satt441 1 161.80
162.77
Satt441 2 164.82 165.79
Satt441 3 173.68 174.69
Satt441 4 176.69 177.70
Satt441 5 179.78 180.62
Satt441 6 191.90 192.86
Satt441 7 207.02 207.92
Satt441 8
171.07 171.59
Satt441 10 168.42 168.52
Satt441 11 136.72
137.43
Satt441 12 152.88 153.48
Satt441 13 158.95 159.75
Satt441 14 155.84 156.44
Satt441 15 195.03 195.70
Satt441 16 185.85 186.39
Satt441 17 189.52 189.72
Satt442
1 240.28 240.78
Satt442 2 246.20 247.30
Satt442 3 249.20
250.21
Satt442 4 252.23 253.24
Satt442 5 261.68 262.40
Satt442 6 264.44 265.46
Satt442 7 258.63 259.03
Satt442 8 243.66 244.13
Satt442 9 267.80 268.20
Satt442
10 237.63 238.03
Satt442 11 255.72 256.29
Satt442 12
283.01 283.31
Satt442 13 270.75 271.00
Satt444 1 134.84
136.28
Satt444 2 137.94 138.95
Satt445 1 166.18 166.96
Satt445 2 169.02 169.85
Satt445 3 184.28 185.15
Satt445 4 193.35 194.54
Satt445 5 196.52 197.40
Satt445 6
215.17 215.47
Satt445 7 163.11 163.76
Satt445 8 190.20
190.80
Satt445 9 178.78 179.08
Satt445 10 187.38 187.83
Satt445 11 217.45 217.90
Satt448 1 340.69 341.63
Satt448 2 347.47 348.19
Satt448 3 350.42 350.92
Satt448 4
355.86 356.92
Satt448 5 361.62 362.12
Satt448 6 358.66
359.34
Satt448 7 364.68 365.08
Satt448 8 343.59 343.92
Satt448 9 332.07 332.27
Satt448 10 324.81 325.01
Satt451 1 325.32 326.49
Satt451 2 351.84 352.37
Satt451 3
354.42 355.25
Satt451 4 357.52 357.72
Satt451 5 319.67
319.92
Satt452 1 256.74 257.81
Satt452 2 259.68 260.86
Satt452 3 263.42 263.92
Satt452 4 265.81 266.98
Satt452 5 274.98 275.83
Satt452 6 296.79 296.99
Satt452 7
254.14 254.36
Satt452 8 302.57 303.40
Satt452 9 272.46
272.78
Satt452 10 309.45 309.55
Satt454 1 158.70 159.60
Satt454 2 170.40 171.59
Satt454 3 173.53 174.49
Satt454 4 176.63 177.50
Satt454 5 161.87 162.37
Satt454 6
182.95 183.15
Satt455 1 273.08 274.10
Satt455 2 276.03
277.15
Satt455 3 279.31 280.19
Satt457 1 356.79 357.42
Satt457 2 368.33 369.29
Satt457 3 324.81 325.11
Satt457 4 336.67 337.12
Satt457 5 354.12 354.32
Satt457 6
365.44 366.09
Satt457 7 371.42 371.82
Satt460 1 113.80
114.50
Satt460 2 116.66 117.63
Satt460 3 135.24 136.08
Satt460 4 144.83 145.61
Satt460 5 160.36 161.17
Satt460 6 163.46 164.24
Satt460 7 129.40 129.61
Satt461 1
128.60 129.61
Satt461 2 131.65 132.67
Satt461 3 138.39
138.95
Satt461 4 144.83 145.41
Satt461 5 154.30 154.82
Satt461 6 122.95 124.05
Satt461 7 153.28 153.80
Satt461 8 134.94 135.24
Satt464 1 144.77 146.31
Satt464 2
164.65 165.79
Satt464 3 174.13 175.55
Satt464 4 168.02
168.42
Satt464 5 177.90 178.10
Satt466 1 165.27 166.29
Satt466 2 168.32 169.19
Satt466 3 177.80 178.77
Satt467 1 274.79 275.52
Satt467 2 294.84 295.98
Satt469 1
245.81 246.90
Satt469 2 276.65 277.20
Satt469 3 279.69
280.24
Satt469 4 225.63 225.89
Satt469 5 249.50 249.80
Satt470 1 263.62 264.94
Satt470 2 266.48 268.00
Satt471 1 233.92 234.60
Satt471 2 245.71 246.70
Satt471 3
248.70 249.70
Satt473 1 307.51 308.53
Satt473 2 310.57
311.60
Satt473 3 313.86 314.69
Satt473 4 317.25 317.75
Satt473 5 295.27 295.78
Satt473 6 298.30 298.81
Satt473 7 301.35 302.37
Satt473 8 304.43 305.46
Satt473 9
292.20 292.35
Satt475 1 150.47 151.46
Satt475 2 159.86
160.87
Satt475 3 171.92 173.06
Satt475 4 175.46 175.66
Satt475 5 178.10 178.30
Satt476 1 343.49 344.02
Satt476 2 346.43 347.21
Satt476 3 354.62 355.55
Satt476 4
357.52 358.36
Satt476 5 365.94 366.91
Satt476 6 349.17
349.37
Satt476 7 351.94 352.27
Satt476 8 360.53 360.67
Satt476 9 363.37 363.72
Satt476 10 325.62 325.92
Satt477 1 141.60 142.67
Satt477 2 157.34 158.35
Satt477 3
163.62 164.44
Satt477 4 154.71 155.42
Satt477 5 161.07
161.27
Satt478 1 194.36 195.23
Satt478 2 227.16 227.82
Satt478 3 232.95 234.12
Satt478 4 236.28 236.86
Satt478 5 239.30 240.15
Satt478 6 242.25 243.03
Satt478 7
254.26 255.11
Satt478 8 257.31 258.06
Satt478 9 224.41
224.73
Satt478 10 230.40 230.70
Satt478 11 251.42 251.82
Satt478 12 248.40 248.75
Satt478 13 221.56 221.66
Satt478 14 221.26 221.46
Satt479 1 114.30 115.40
Satt479
2 117.63 118.50
Satt479 3 142.52 143.25
Satt479 4 145.41
146.49
Satt479 5 151.75 152.83
Satt479 6 111.76 112.01
Satt480 1 137.81 138.95
Satt480 2 140.99 142.17
Satt480 3 144.33 145.41
Satt487 1 114.30 114.90
Satt487 2
117.16 118.13
Satt487 3 120.28 121.07
Satt487 4 123.05
124.20
Satt487 5 126.31 127.07
Satt487 6 111.36 111.91
Satt487 7 132.67 132.87
Satt487 8 135.78 135.98
Satt487 9 129.61 129.91
Satt488 1 295.57 295.78
Satt488 2
311.35 312.33
Satt488 3 314.61 315.41
Satt488 4 323.85
324.71
Satt488 5 327.29 327.75
Satt491 1 195.86 196.52
Satt491 2 201.78 202.49
Satt491 3 207.41 208.42
Satt491 4 216.95 217.35
Satt491 5 219.98 220.28
Satt492 1
235.72 237.13
Satt492 2 238.81 239.97
Satt492 3 227.46
227.62
Satt493 1 266.41 267.50
Satt493 2 269.83 270.55
Satt493 3 281.93 282.71
Satt493 4 279.23 279.38
Satt495 1 231.00 231.97
Satt495 2 243.11 244.22
Satt495 3
237.33 237.83
Satt497 1 268.82 269.53
Satt497 2 271.96
272.58
Satt497 3 274.62 276.23
Satt497 4 277.67 279.03
Satt497 5 296.18 297.49
Satt497 6 299.31 300.43
Satt497 7 281.10 281.90
Satt497 8 265.86 266.06
Satt497 9
262.80 263.20
Satt503 1 226.19 227.00
Satt503 2 228.90
230.09
Satt503 3 231.92 233.15
Satt503 4 221.83 222.55
Satt503 5 211.15 211.65
Satt503 6 224.93 225.23
Satt503 7 234.95 235.59
Satt503 8 223.00 223.10
Satt506 1
280.19 281.35
Satt506 2 283.21 284.01
Satt506 3 292.26
293.61
Satt506 4 289.75 290.15
Satt507 1 230.60 231.47
Satt507 2 233.42 234.40
Satt507 3 236.36 237.43
Satt507 4 227.62 228.02
Satt507 5 239.70 239.90
Satt508 1
333.06 334.15
Satt508 2 336.15 337.31
Satt508 3 339.25
339.65
Satt509 1 192.47 193.55
Satt509 2 195.43 196.52
Satt509 3 198.40 199.39
Satt509 4 201.39 202.59
Satt509 5 204.78 205.40
Satt509 6 236.86 238.41
Satt509 7
249.20 250.41
Satt509 8 240.68 241.18
Satt509 9 243.79
244.22
Satt509 10 184.18 184.48
Satt509 11 252.83 253.13
Satt510 1 109.05 109.55
Satt510 2 127.07 127.58
Satt510 3 136.08 137.33
Satt510 4 139.35 140.52
Satt510 5
96.44 97.45
Satt510 6 121.07 122.06
Satt510 7 111.91
112.86
Satt510 8 130.11 130.31
Satt511 1 255.83 256.79
Satt511 2 262.02 262.90
Satt511 3 265.03 266.01
Satt511 4 268.06 269.07
Satt511 5 277.87 278.07
Satt511 6
259.23 259.55
Satt512 1 316.16 317.25
Satt512 2 319.37
320.28
Satt512 3 325.53 326.44
Satt512 4 328.71 329.57
Satt512 5 331.77 332.86
Satt512 6 310.07 310.62
Satt512 7 313.03 313.68
Satt513 1 117.63 118.13
Satt513 2
130.11 130.63
Satt513 3 142.17 143.25
Satt513 4 173.06
173.83
Satt513 5 181.79 182.75
Satt513 6 185.05 185.65
Satt513 7 170.15 170.62
Satt513 8 164.34 164.44
Satt513 9 167.26 167.63
Satt513 10 133.07 133.37
Satt513
11 139.14 139.65
Satt513 12 148.77 148.92
Satt513 13
153.38 153.58
Satt513 14 145.81 146.11
Satt514 1 314.34
315.21
Satt514 2 316.63 317.25
Satt514 3 325.76 326.34
Satt514 4 351.18 351.94
Satt514 5 328.96 329.26
Satt514 6 329.77 330.38
Satt514 7 331.97 332.36
Satt514 8
337.12 337.71
Satt514 9 340.15 340.39
Satt514 10 328.25
328.56
Satt514 11 345.78 346.18
Satt514 12 359.93 360.13
Satt514 13 362.87 363.17
Satt514 14 354.22 354.62
Satt514 15 334.25 334.34
Satt515 1 341.13 342.06
Satt515
2 352.77 353.69
Satt515 3 355.05 356.44
Satt517 1 159.30
160.36
Satt517 2 162.12 163.56
Satt517 3 165.13 166.49
Satt517 4 173.15 174.50
Satt517 5 176.21 177.23
Satt517 6 179.45 180.15
Satt517 7 149.89 150.91
Satt517 8
170.05 171.02
Satt519 1 354.60 355.55
Satt519 2 357.42
358.36
Satt519 3 360.33 361.17
Satt519 4 363.27 363.47
Satt519 5 372.02 372.18
Satt522 1 231.47 232.34
Satt522 2 240.78 241.26
Satt522 3 255.78 256.89
Satt522 4
258.83 259.90
Satt522 5 283.31 284.36
Satt522 6 237.53
238.13
Satt522 7 252.83 253.39
Satt522 8 280.29 280.70
Satt523 1 171.58 172.39
Satt523 2 174.43 175.34
Satt523 3 189.52 190.40
Satt523 4 192.76 193.45
Satt523 5
184.02 184.18
Satt523 6 199.04 199.39
Satt524 1 168.22
168.94
Satt524 2 171.12 172.29
Satt524 3 177.40 177.60
Satt524 4 174.59 174.89
Satt526 1 322.80 324.22
Satt526 2 332.02 333.44
Satt529 1 214.47 215.47
Satt529 2
220.38 221.35
Satt529 3 217.85 218.08
Satt529 4 229.67
230.24
Satt529 5 223.96 224.06
Satt532 1 170.15 171.37
Satt532 2 172.86 174.48
Satt532 3 179.35 180.37
Satt532 4 182.50 183.32
Satt532 5 154.40 154.82
Satt532 6
175.31 176.55
Satt532 7 176.58 177.40
Satt532 8 167.48
168.17
Satt533 1 169.19 170.15
Satt533 2 174.99 175.96
Satt533 3 193.94 194.44
Satt533 4 196.72 196.90
Satt533 5 172.23 172.96
Satt533 6 178.37 178.57
Satt533 7
193.15 193.55
Satt534 1 165.79 166.90
Satt534 2 172.01
173.06
Satt534 3 177.90 178.87
Satt534 4 180.82 181.84
Satt534 5 183.72 185.15
Satt534 6 187.08 188.55
Satt534 7 190.00 191.28
Satt534 8 192.95 194.44
Satt534 9
195.92 197.05
Satt534 10 163.36 163.84
Satt534 11 160.36
160.76
Satt534 12 169.18 169.85
Satt534 13 157.34 157.74
Satt534 14 175.04 175.96
Satt534 15 199.89 200.19
Satt534 16 150.71 151.11
Satt536 1 220.76 221.83
Satt536
2 223.76 224.26
Satt536 3 226.66 227.72
Satt536 4 229.38
230.70
Satt536 5 197.20 197.50
Satt536 6 232.95 234.22
Satt536 7 236.18 236.61
Satt537 1 292.58 293.76
Satt537 2 295.78 296.79
Satt537 3 311.28 312.63
Satt537 4
317.75 318.36
Satt537 5 320.67 321.79
Satt537 6 323.80
324.81
Satt537 7 301.35 302.62
Satt537 8 308.03 309.10
Satt537 9 280.43 281.20
Satt537 10 289.75 290.25
Satt537 11 299.31 299.83
Satt537 12 305.46 305.98
Satt537
13 314.69 315.21
Satt537 14 333.35 333.85
Satt537 15
283.61 283.91
Satt540 1 175.14 176.06
Satt540 2 181.17
182.25
Satt540 3 190.28 191.48
Satt540 4 192.71 194.19
Satt540 5 199.39 200.39
Satt540 6 210.59 211.96
Satt540 7 213.37 213.85
Satt540 8 169.65 170.15
Satt540 9
184.68 185.15
Satt540 10 187.58 188.05
Satt540 12 195.73
196.12
Satt540 13 196.56 197.20
Satt540 14 204.70 204.90
Satt543 1 171.12 172.39
Satt543 2 174.39 175.09
Satt543 3 175.29 176.26
Satt543 4 195.43 196.52
Satt543 5
198.83 199.49
Satt543 6 207.41 208.42
Satt543 7 210.95
211.45
Satt543 8 213.65 214.47
Satt543 9 210.34 210.54
Satt543 10 216.56 217.05
Satt543 11 178.15 178.78
Satt544 1 116.66 117.63
Satt544 2 147.56 149.12
Satt544 3
154.30 155.84
Satt544 4 160.56 161.37
Satt544 5 163.56
164.34
Satt544 6 166.54 167.72
Satt544 7 169.59 170.62
Satt544 8 132.15 132.45
Satt544 9 151.41 151.61
Satt544 10 157.54 157.79
Satt544 11 172.44 173.53
Satt545
1 242.25 242.85
Satt545 2 254.11 254.66
Satt545 3 260.29
260.86
Satt545 4 275.32 276.23
Satt545 5 278.35 279.48
Satt545 6 281.59 282.30
Satt545 7 284.46 285.32
Satt545 8 290.55 292.10
Satt545 9 293.64 294.56
Satt545
10 266.36 266.78
Satt545 11 272.58 272.98
Satt545 12
287.84 288.04
Satt545 13 296.79 297.29
Satt545 14 269.53
269.98
Satt546 1 332.22 333.26
Satt546 2 341.33 342.06
Satt546 3 344.12 344.85
Satt546 4 355.29 356.49
Satt546 5 358.46 358.66
Satt548 1 352.27 352.77
Satt548 2
359.73 361.17
Satt548 3 362.57 364.12
Satt548 4 365.63
366.41
Satt548 5 348.57 348.97
Satt548 6 351.34 351.54
Satt548 7 368.83 368.93
Satt549 1 225.08 225.33
Satt549 2 233.92 234.80
Satt549 3 240.10 240.38
Satt549 4
243.05 243.23
Satt549 5 245.94 246.80
Satt549 6 249.00
250.00
Satt549 7 255.38 255.88
Satt550 1 223.78 224.73
Satt550 2 226.66 227.46
Satt550 3 241.76 242.85
Satt550 4 244.72 245.71
Satt550 5 229.84 230.24
Satt550 6
220.91 221.36
Satt550 7 232.80 233.35
Satt550 8 238.96
239.16
Satt551 1 230.50 231.57
Satt551 2 236.85 237.63
Satt551 3 242.75 243.73
Satt552 1 160.66 161.67
Satt552 2 175.66 176.68
Satt552 3 181.73 182.80
Satt552 4
184.98 185.75
Satt552 5 169.80 170.62
Satt555 1 249.45
250.30
Satt555 2 271.56 272.75
Satt555 3 274.87 275.78
Satt555 4 277.96 278.68
Satt556 1 135.95 136.83
Satt556 2 148.62 149.94
Satt556 3 151.96 152.78
Satt556 4
184.68 186.12
Satt556 5 197.30 197.99
Satt556 6 155.32
155.84
Satt556 7 164.34 164.82
Satt556 8 168.22 168.69
Satt556 9 176.93 177.90
Satt556 10 188.45 188.65
Satt557 1 201.89 202.90
Satt557 2 204.90 205.90
Satt557 3
211.20 212.01
Satt557 4 214.17 214.97
Satt557 5 220.29
221.36
Satt557 6 208.42 208.93
Satt557 7 193.45 193.75
Satt558 1 220.38 221.46
Satt558 2 229.54 230.55
Satt558 3 235.38 236.66
Satt558 4 238.68 239.70
Satt558 6
233.05 233.42
Satt558 7 241.86 242.45
Satt558 8 245.21
245.41
Satt560 1 231.06 231.97
Satt560 2 237.11 238.91
Satt560 3 243.15 244.52
Satt560 4 246.20 247.40
Satt560 5 276.80 277.67
Satt560 6 286.02 286.83
Satt560 7
273.70 274.30
Satt560 8 271.25 271.56
Satt560 9 283.06
283.41
Satt560 10 250.10 250.21
Satt560 11 253.13 253.24
Satt563 1 166.45 167.82
Satt563 2 169.65 170.82
Satt563 3 236.85 237.83
Satt563 4 173.53 174.49
Satt563 5
181.49 182.95
Satt563 6 212.36 213.47
Satt563 8 160.42
160.99
Satt563 9 164.34 164.54
Satt563 10 206.30 207.41
Satt563 11 185.15 185.55
Satt563 12 209.63 209.83
Satt565 1 299.47 300.33
Satt565 2 302.62 304.63
Satt565 3
327.31 328.46
Satt565 4 351.34 351.84
Satt565 5 330.48
331.06
Satt566 1 356.49 357.42
Satt566 2 359.44 360.23
Satt566 3 362.12 363.07
Satt566 4 365.18 365.94
Satt566 5 374.11 375.06
Satt566 6 376.96 377.90
Satt566 7
367.97 368.33
Satt566 8 379.97 380.17
Satt566 9 385.90
386.30
Satt567 1 97.45 97.95
Satt567 2 112.41 113.36
Satt567 3 115.25 116.20
Satt567 4 118.56 119.10
Satt567 5 110.01 110.51
Satt567 6 98.05 98.15
Satt568 1
345.38 346.08
Satt568 2 350.62 351.84
Satt568 3 357.02
357.22
Satt568 4 354.12 354.42
Satt569 1 103.33 104.23
Satt569 2 115.60 116.30
Satt569 3 121.47 122.55
Satt569 4 124.56 125.31
Satt569 5 127.58 128.38
Satt569 6
130.63 131.38
Satt569 7 133.70 134.10
Satt569 8 106.59
106.99
Satt570 1 108.50 109.05
Satt570 2 111.41 112.41
Satt570 3 114.48 115.25
Satt570 4 126.57 127.58
Satt570 5 120.72 120.97
Satt570 6 130.01 130.21
Satt570 7
132.94 133.17
Satt572 1 172.56 174.08
Satt572 2 175.86
176.53
Satt572 3 178.87 179.50
Satt572 4 169.89 170.45
Satt573 1 167.36 167.56
Satt573 2 163.84 164.82
Satt573 3 175.96 176.53
Satt576 1 261.38 261.88
Satt576 2
288.75 290.25
Satt576 3 342.10 342.99
Satt576 4 350.48
351.84
Satt576 5 361.83 363.07
Satt576 6 365.11 365.94
Satt576 7 353.64 354.12
Satt576 8 356.69 356.89
Satt576 9 359.09 359.59
Satt577 1 312.82 313.96
Satt577 2
316.01 317.25
Satt578 1 178.37 179.35
Satt578 2 181.29
182.25
Satt578 3 199.89 200.39
Satt578 4 202.39 203.00
Satt578 5 199.49 199.79
Satt578 6 184.78 184.98
Satt581 1 138.39 139.45
Satt581 2 148.06 149.22
Satt581 3
151.21 152.43
Satt581 4 161.07 161.27
Satt582 1 333.83
334.34
Satt582 2 336.72 337.31
Satt582 3 339.75 340.19
Satt582 4 318.76 319.16
Satt583 1 122.06 123.05
Satt583 2 146.99 147.86
Satt583 3 150.17 151.21
Satt583 4
180.82 181.64
Satt583 5 125.56 125.76
Satt583 6 131.58
131.95
Satt583 7 137.89 138.09
Satt583 8 141.10 141.30
Satt583 9 144.15 144.60
Satt583 10 113.46 113.66
Satt583 11 116.50 116.66
Satt583 12 153.80 154.00
Satt583
13 156.84 157.04
Satt583 14 159.96 160.16
Satt583 15
168.99 169.19
Satt583 16 198.89 199.19
Satt583 17 202.09
202.39
Satt583 18 222.89 223.39
Satt584 1 171.59 172.56
Satt584 2 174.49 175.46
Satt584 3 180.77 181.32
Satt584 4 195.43 196.42
Satt584 5 183.72 184.68
Satt584 6
186.91 188.85
Satt584 7 198.89 199.89
Satt586 1 318.76
319.77
Satt586 2 337.12 338.01
Satt586 3 342.99 343.79
Satt586 4 348.57 349.50
Satt586 5 354.14 355.05
Satt586 6 331.18 331.77
Satt586 7 334.15 334.54
Satt586 8
340.05 340.84
Satt586 9 351.34 351.94
Satt587 1 168.02
169.19
Satt587 2 170.97 172.20
Satt587 3 173.93 175.05
Satt587 4 176.93 177.90
Satt587 5 129.10 129.61
Satt587 6 155.84 156.34
Satt590 1 317.18 318.26
Satt590 2
323.30 324.81
Satt590 3 332.86 333.55
Satt590 4 335.78
336.87
Satt590 6 339.25 339.75
Satt590 7 266.88 268.52
Satt590 8 272.88 274.62
Satt590 9 264.22 264.44
Satt590 12 314.28 315.21
Satt590 13 304.83 305.16
Satt590
14 320.48 321.06
Satt590 15 300.73 301.25
Satt590 16
311.10 312.00
Satt590 17 326.64 327.60
Satt590 18 329.87
330.88
Satt591 1 137.43 138.44
Satt591 2 140.52 141.60
Satt591 3 150.27 151.59
Satt591 4 144.63 144.83
Satt591 5 119.20 119.50
Satt592 1 236.85 237.43
Satt592 2
242.75 243.53
Satt592 3 245.71 246.85
Satt592 4 254.76
255.87
Satt594 1 122.50 123.25
Satt594 2 134.74 135.44
Satt594 3 141.10 141.60
Satt594 4 144.33 145.61
Satt594 5 147.56 148.82
Satt594 6 166.22 167.36
Satt594 7
172.56 173.63
Satt594 8 157.04 157.64
Satt594 9 160.06
160.46
Satt594 10 178.20 179.13
Satt594 11 138.09 138.29
Satt594 12 169.19 169.59
Satt595 1 340.05 341.13
Satt595 2 342.99 344.85
Satt595 4 345.98 346.38
Satt596 1
257.73 258.83
Satt596 2 260.72 261.88
Satt596 3 266.68
267.70
Satt596 4 272.86 273.90
Satt596 5 275.88 277.15
Satt596 6 281.80 282.91
Satt596 7 264.17 264.64
Satt596 8 270.10 270.65
Satt596 9 279.43 279.58
Satt596
10 251.92 252.12
Satt597 1 144.68 145.51
Satt597 2 147.86
148.82
Satt597 3 160.26 161.37
Satt597 4 157.34 157.90
Satt598 1 163.36 164.37
Satt598 2 175.46 176.43
Satt601 1 344.86 345.93
Satt601 2 347.84 348.57
Satt601 3
342.36 342.66
Satt601 4 351.02 351.22
Satt601 5 356.39
356.59
Satt601 6 336.72 336.92
Satt602 1 336.21 336.82
Satt602 2 339.14 341.14
Satt602 4 342.86 343.92
Sct_010 1 107.29 108.10
Sct_010 2 99.45 99.95
Sct_010 3
105.44 105.64
Sct_026 1 328.36 329.37
Sct_026 2 330.38
331.37
Sct_026 3 326.64 326.84
Sct_026 4 333.80 334.25
Sct_026 5 317.50 317.65
Sct_028 1 236.07 237.33
Sct_028 2 244.12 245.21
Sct_028 3 248.10 249.20
Sct_028 4
242.10 243.23
Sct_034 1 126.67 127.58
Sct_034 2 128.83
129.61
Sct_034 3 130.63 131.65
Sct_046 1 260.66 262.28
Sct_046 2 267.69 268.52
Sct_046 3 275.88 276.65
Sct_046 4 277.89 278.68
Sct_046 5 284.15 285.73
Sct_046 6
286.23 287.83
Sct_046 7 288.24 288.75
Sct_046 8 292.89
293.76
Sct_046 9 298.60 299.83
Sct_046 10 290.05 290.75
Sct_046 11 270.03 270.55
Sct_046 12 274.10 274.62
Sct_046 13 282.71 283.71
Sct_065 1 163.26 164.40
Sct_065
2 165.27 166.40
Sct_065 3 171.12 172.18
Sct_137 1 269.32
270.03
Sct_137 2 273.08 274.10
Sct_147 1 130.11 131.03
Sct_147 2 136.28 136.93
Sct_186 1 97.45 97.95
Sct_186 2 105.24 105.74
Sct_186 3 111.36 111.91
Sct_186 4
113.36 113.80
Sct_186 5 115.35 115.70
Sct_186 6 117.36
117.63
Sct_187 1 313.34 314.19
Sct_187 2 311.40 311.90
Sct_187 3 315.21 317.35
Sct_188 1 242.25 243.23
Sct_188 2 252.23 253.24
Sct_188 3 254.76 254.96
Sctt008 1
159.36 159.86
Sctt008 2 162.36 163.36
Sctt008 3 165.42
165.62
Sctt008 4 147.56 147.76
Sctt009 1 213.93 214.97
Sctt009 2 219.71 220.86
Sctt012 1 343.92 344.72
Sctt012 2 351.84 353.02
Sctt012 3 355.05 355.55
[0268]
17APPENDIX II
SSR Markers: Table of Primers
Marker Re-
Name Left Primer Sequence Right Primer Sequence
Pigtail peat
Sac1006 CAATCAGGTTAGTGGTCCTACC SEQ ID NO: 1
CAAAAGGTTTTCAGTGGTGG SEQ ID NO: 285 GTTTCTT 2
Sac1677
AAGTTCACAGCACCTGGACCAT SEQ ID NO: 2 CCAATCCCTACCCTGATTGCAC SEQ ID NO: 286
GTTTCTT 2
Sat_084 GAAACAAACACCCCCAAGGTGA SEQ ID NO: 3
GGAAGTGGATTTGGAGTTGGGA SEQ ID NO: 287 GTTTCTT 2
Sat_090
CCAATCGTGTCTTATCCTCGGG SEQ ID NO: 4 AACCCATAACCTGCTCGTCTCA SEQ ID NO: 288
GTTTCTT 2
Sat_104 TTCCAGACAGAACCCAAGTAGCC SEQ ID NO: 5
AGGCGGGTTTGGAGCTGTTTAC SEQ ID NO: 289 GTTTCTT 2
Sat_110
AACATTTTTCATCGCTTTTCTTAG SEQ ID NO: 6 TCTTCTCAGGAACTTGAATTACTCA SEQ ID
NO: 290 GTTTCTT 2
Sat_117 AAAGCATTTTTGGCAGTTTCTTGT SEQ ID
NO: 7 GGAATGTCCCAAGTGTCAGCAA SEQ ID NO: 291 GTTTCTT 2
Satt020 TGGACAGAATGGAGAAAGAAATGTG SEQ ID NO: 8 TGAAAATAAGTGGAAAGAAAAAGGAA
SEQ ID NO: 292 GTTTCTT 3
AA
Satt040
CCAAGCCAAACAAAGAATCACA SEQ ID NO: 9 GTCCCCGTTCCTCAAACACCTT SEQ ID NO: 293
GTTTCTT 3
Satt042 GCTTGCTATGATTGATTGATTGATTG SEQ ID NO:
10 TGCACACTCACTTGGTCTTACACA SEQ ID NO: 294 GTTTCTT 3
A
Satt050 AGAACGGTGTTAAGATAGAATAGTT SEQ ID NO: 11
CTGTTGGAAAATGATGCGTGGC SEQ ID NO: 295 GTTTCTT 3
Satt066
GGGAAGCTTAATAATGAAAATGACAC SEQ ID NO: 12 TTGATCACTTCTGTAACATTC SEQ ID NO:
296 GTTTCTT 3
Satt092 AACCGATGCTTTTTTCGCCTTT SEQ ID NO:
13 CTTTGTTGTTTGGTCAAGGCCC SEQ ID NO: 297 GTTTCTT 3
Satt102 GAAGAAGGCTCAGCAACACCTTG SEQ ID NO: 14 TGCCGAAATAAGTGAGAGCATAGAA
SEQ ID NO: 298 GTTTCTT 3
Satt108 CACCCAATCTTGCCTTTGAAACA
SEQ ID NO: 15 TGTTAGGTATGGGATTTAGGGTTTTG SEQ ID NO: 299 GTTTCTT 3
A
Satt109 TAGACACTTTCAGGTTAAAATATAAC SEQ ID NO: 16
TCCACTCACTGTATGTCTTCCCTTG SEQ ID NO: 300 GTTTCTT 3
Satt111 CAACTTGTCACTTACACATAGTTTAG SEQ ID NO: 17 TTGAGTGCTCTTGGTGTTTTCCT
SEQ ID NO: 301 GTTTCTT 3
Satt115 ATTTCGCTCGCAAACACAAGGT
SEQ ID NO: 18 CCAAACACCCCAGTATAAAAAATGGA SEQ ID NO: 302 GTTTCTT 3
Satt122 TCAAGAAATACAAGTGCAAGAAAGAC SEQ ID NO: 19
GGATTTTGAGTTGCTCCAAGGC SEQ ID NO: 303 GTTTCTT 3
C
Satt127 AATTTTCGCTTGTGAACCCTGC SEQ ID NO: 20 TGTATGATCCATCCTCTGAAACCG SEQ
ID NO: 304 GTTTCTT 3
Satt129 TTCAGTACAAGTCGGGTGAATAATAA
SEQ ID NO: 21 TCACATGTTCGGGACTTAAGGTAT SEQ ID NO: 305 GTTTCTT 3
TAA
Satt130 TGGGACTCTCACACACGGAAAA SEQ ID NO: 22
TGAATGGCTAAAAACGTGATTTGGA SEQ ID NO: 306 GTTTCTT 3
Satt131 TGGGAGTCAATTTCCCATTATCA SEQ ID NO: 23 GGACCTTCCCTTTCCCATGACT SEQ
ID NO: 307 GTTTCTT 3
Satt133 TGGATTTGAAACCACAAATAACAACA
SEQ ID NO: 24 AGCGATGGTTGAAGAAAGGGTC SEQ ID NO: 308 GTTTCTT 3
AC
Satt138 AAAAGGGGGACATTTTTCCACG SEQ ID NO: 25
AGAGAACGGGCGATTTATGGCT SEQ ID NO: 309 GTTTCTT 3
Satt142
CATTAGGGACAACAACAGCGTTT SEQ ID NO: 26 ATGTCGCCACTAGGCCAATCAG SEQ ID NO:
310 GTTTCTT 3
Satt144 CGTCGCCATCACTATGAGAA SEQ ID NO: 27
CCATCTTGAGCAGAGTTTGAAGTT SEQ ID NO: 311 GTTTCTT 3
Satt146
AAGGGATCCCTCAACTGACTG SEQ ID NO: 28 GTGGTGGTGGTGAAAACTATTAGAA SEQ ID NO:
312 GTTTCTT 3
Satt147 CCATCCCTTCCTCCAAATAGAT SEQ ID NO:
29 CTTCCACACCCTAGTTTAGTGACAA SEQ ID NO: 313 GTTTCTT 3
Satt150 TGACGAAGCTTGAGGTTATTCG SEQ ID NO: 30 TCAAGCTTATGCTCTATAGGCT SEQ
ID NO: 314 GTTTCTT 3
Satt151 GCCAAGAAGATAACAAGCTCGGC SEQ
ID NO: 31 GACCAAAATTCAAGGCAGTGACAA SEQ ID NO: 315 GTTTCTT 3
Satt153 GGGTTATATCAGTTTTTCTTTTTGTT SEQ ID NO: 32 CCATCCTCGTTAGCATCTAT
SEQ ID NO: 316 GTTTCTT 3
Satt155 AGATCCAACACCTGGCCTAAT
SEQ ID NO: 33 GCTGCACAATTCATTCCATTT SEQ ID NO: 317 GTTTCTT 3
Satt156 AACAAAACTAGCCCATAGAAACATTG SEQ ID NO: 34
CCCAGGGACTTACTTTTTTCAGTTT SEQ ID NO: 318 GTTTCTT 3
A
Satt159 GAAATGCCCAGAAAAACCTAATAAC SEQ ID NO: 35
TGAAGCAACAAAATAGAGGAATAGA SEQ ID NO: 319 GTTTCTT 3
Satt165 GCCGTGAAGACAGTTGATCGTT SEQ ID NO: 36 CATTACTAGGCGTGTGTTGTTTCAA
SEQ ID NO: 320 GTTTCTT 3
Satt166 CAGTTGATTTTTGTTTTTCGGCA
SEQ ID NO: 37 CACGCGCATCAGCTTTGTAGAG SEQ ID NO: 321 GTTTCTT 3
Satt168 TTGTCCAAACTTGCAGGGAACA SEQ ID NO: 38 TCTTTTCACCATTCTCCAACCTCA
SEQ ID NO: 322 GTTTCTT 3
Satt172 TGGATGAGTTCTATTGGGGTAGTC-
G SEQ ID NO: 39 TCCTTTCTCCCATTTTTTTTGGG SEQ ID NO: 323 GTTTCTT 3
Satt175 GACCTCGCTCTCTGTTTCTCAT SEQ ID NO: 40
GGTGACCACCCCTATTCCTTAT SEQ ID NO: 324 GTTTCTT 3
Satt176
CGCAATCCCAATTCACCTCTTC SEQ ID NO: 41 AATTGTTAGGGCGCGAGAAACA SEQ ID NO:
325 GTTTCTT 3
Satt181 GAACCCCGTTTCAACATTTTATGA SEQ ID NO:
42 CTAGCCAAGGGAGACAGGAGCA SEQ ID NO: 326 GTTTCTT 3
Satt183 GGTCGTTAAGCCCACTTTGAGA SEQ ID NO: 43 CCTCACACCAACCAGCACAAAA SEQ
ID NO: 327 GTTTCTT 3
Satt186 TGCAGCTTTCACTAATCGTCAGAA SEQ
ID NO: 44 CAATTTTATGATTTGCTTTTGAAGGG SEQ ID NO: 328 GTTTCTT 3
A
Satt190 GGGAGTGTGAACTTACATTGTCT SEQ ID NO: 45
GGGCCTTGAATTTTGTGCTAT SEQ ID NO: 329 GTTTCTT 3
Satt191
GCGATCATGTCTCTGCCATCAG SEQ ID NO: 46 CCTCTTGAAACCGTGAAACCGT SEQ ID NO:
330 GTTTCTT 3
Satt193 GAAGGGATATGGAGAGTGAGAAATTA SEQ ID
NO: 47 TCTTTTTCTTCATTTTTGTTCGCA SEQ ID NO: 331 GTTTCTT 3
GA
Satt195 AAAAATTGTGTAAACGAAAGATGGGA SEQ ID NO: 48
AAACTGCTCTGAAGTGCGACACA SEQ ID NO: 332 GTTTCTT 3
Satt196
TGAGCCCAACCTCCACATCTTT SEQ ID NO: 49 TGTGAATAAAAGAAATCCCCATTGA SEQ ID NO:
333 GTTTCTT 3
Satt197 CACTGCTTTTTCCCCTCTCT SEQ ID NO: 50
AAGATACCCCCAACATTATTTGTAA SEQ ID NO: 334 GTTTCTT 3
Satt199 GCGGTAAATGGTGAAAATCATTTATG SEQ ID NO: 51 GCGTTTTCATACGGTGTTTTGCCT-
AT SEQ ID NO: 335 GTTTCTT 3
GTT
Satt202
GGAATGCATGAGTATTAACCTCTTAT SEQ ID NO: 52 GGGCTAACGAACATGTAACTTATCAA SEQ
ID NO: 336 GTTTCTT 3
C
Satt203
TGTCTTCCCAATCCATCTAATCTAAT SEQ ID NO: 53 GCACTACAATATGTGCATGAATTTTT SEQ
ID NO: 337 GTTTCTT 3
C CT
Satt204
CCAATTATGTTTCTATGCCATCTTGT SEQ ID NO: 54 TCTCCTTACTCACTCCATTGGCA SEQ ID
NO: 338 GTTTCTT 3
T
Satt209
TGTGGATAAAAGCCATCTCTAACAAA SEQ ID NO: 55 TCCATGTCACCAACCACAAAAA SEQ ID
NO: 339 GTTTCTT 3
Satt212 TCGATACCAATCATCCAATCCAAA SEQ ID
NO: 56 CGTATCCCTTTCCCATACGTGG SEQ ID NO: 340 GTTTCTT 3
Satt213 TCCTTAGACCTCTCGCGCAAAC SEQ ID NO: 57 ATGACGGGATGAAAGAGCCAAA SEQ
ID NO: 341 GTTTCTT 3
Satt216 TACCCTTAATCACCGGACAA SEQ ID
NO: 58 AGGGAACTAACACATTTAATCATCA SEQ ID NO: 342 GTTTCTT 3
Satt218 ACGTATGCGAAGGAGGGAGATG SEQ ID NO: 59 CGGAAAAGATATACCGAGTGAGAAAA
SEQ ID NO: 343 GTTTCTT 3
Satt219 TTGGCTCTTGTGTAAGTGGCCC
SEQ ID NO: 60 TTCTAAGGATTTGTGGTAATCGGC SEQ ID NO: 344 GTTTCTT 3
Satt220 TTCAGTCCCTTGGTGGTTCCAA SEQ ID NO: 61
CATGGAGAAAAGAAGAGGAGGGA SEQ ID NO: 345 GTTTCTT 3
Satt221
CATGGTCCTTGGGTGCAATTTA SEQ ID NO: 62 AAGCAAACCATTATCTTCATTGGTG SEQ ID NO:
346 GTTTCTT 3
Satt225 AAAAATGTGTTAGAGCTTGTGTTGTT SEQ ID
NO: 63 GCCACACTATTCCAGCCACTAC SEQ ID NO: 347 GTTTCTT 3
A
Satt227 ATGGTGCAGTGTTGCAGGTTGT SEQ ID NO: 64
AGTAGTCCAAACCGAAACGCCA SEQ ID NO: 348 GTTTCTT 3
Satt228
GACCACAACTTCTTTTTTGTGAATGG SEQ ID NO: 65 TCAATTCGGTCAAAAGGCTTGA SEQ ID
NO: 349 GTTTCTT 3
Satt230 CCGTCACCGTTAATAAAATAGCAT SEQ ID
NO: 66 CTCCCCCAAATTTAACCTTAAAGA SEQ ID NO: 350 GTTTCTT 3
Satt233 AGAAGCATACTCGTCGTAACACTATC SEQ ID NO: 67 AATGGATGGACCTGTCAGTCTGC
SEQ ID NO: 351 GTTTCTT 3
C
Satt234
GAGCAGGACATTTTTTTTATCCTTGA SEQ ID NO: 68 TGCTTCCATTAGTCTCTCATCCTCC SEQ ID
NO: 352 GTTTCTT 3
Satt236 TGCTTGAGGTTGAAGGAAATGC SEQ ID
NO: 69 TGGAAAAAGATAACGTGTGTTTGCAG SEQ ID NO: 353 GTTTCTT 3
Satt240 TCCTTGGGGTGAATTGTTTTTCA SEQ ID NO: 70 CCTTCTTTTGACATGGGGCCTA SEQ
ID NO: 354 GTTTCTT 3
Satt242 GCGTTGATCAGGTCGATTTTTATTTG
SEQ ID NO: 71 GCGAGTGCCAACTAACTACTTTTATG SEQ ID NO: 355 GTTTCTT 3
T A
Satt243 AATGCTTTGGTCGTTGCATTG SEQ ID NO: 72
TGGTATCGGGAGATTTTTTCAGC SEQ ID NO: 356 GTTTCTT 3
Satt247
CAGCGTCTGCATGATAGCGTTT SEQ ID NO: 73 TTTTTCGTCAGCACATTATTACACAT SEQ ID
NO: 357 GTTTCTT 3
TT
Satt249
TGCAAATTGTTATTGTGAGACTGAAT SEQ ID NO: 74 GGCCAGTGTTGAGGGATTTAGAA SEQ ID
NO: 358 GTTTCTT 3
GA
Satt250 CGCCAGCTAGCTAGTCTCAT
SEQ ID NO: 75 AATTTGCTCCAGTGTTTTAAGTTT SEQ ID NO: 359 GTTTCTT 3
Satt251 CCTCCACCCCCTTCCCACCCAAAA SEQ ID NO: 76
GGTGATATCGCGCTAAAATTA SEQ ID NO: 360 GTTTCTT 3
Satt255
AGCGTCGTCTGGCTAGGTCTGT SEQ ID NO: 77 GGAAACCCTGTCATTTTCGTGC SEQ ID NO:
361 GTTTCTT 3
Satt256 GTCATTGGTCTCAAACAATCTTCAT SEQ ID
NO: 78 AGAGTTCTCACTCCGCCAGCTC SEQ ID NO: 362 GTTTCTT 3
Satt257 GCGACTTTCTTTTCAATTTCACTCC SEQ ID NO: 79 GCGCAATTGTCACCAACACAT SEQ
ID NO: 363 GTTTCTT 3
Satt258 CACTTTTTCACTGTCTCCCCCC SEQ
ID NO: 80 CCCAAACCAACAAGCAACAACA SEQ ID NO: 364 GTTTCTT 3
Satt259 TGGGCCATTTGGGCAGCTCGACT SEQ ID NO: 81 ATTCACACGCATCTGGAATAATA SEQ
ID NO: 365 GTTTCTT 3
Satt262 GCGCCCCATTAATGTTAACACA SEQ
ID NO: 82 GCGGAGTTCAACGCATTCACCTT SEQ ID NO: 366 GTTTCTT 3
Satt263 GGAGAGAATCCATATATATTGAATTG SEQ ID NO: 83 TGAAAGCAAACGAGACTCATGGA
SEQ ID NO: 367 GTTTCTT 3
C
Satt264
CCTTTTGACAATTATGGCATATA SEQ ID NO: 84 GCATAGAAGGGCATCATTCAGAT SEQ ID NO:
368 GTTTGTT 3
Satt265 CCTGTCAAATTGGCTGATGCAA SEQ ID NO:
85 GCTAGATGAGCAGACCATTGCACTT SEQ ID NO: 369 GTTTCTT 3
Satt266 TTTTACCAAACAAATTAAACTGCGTC SEQ ID NO: 86 CAAGAGGTTGTTGTAAGAGTGATC-
TC SEQ ID NO: 370 GTTTCTT 3
T G
Satt267
CCGGTCTGACCTATTCTCAT SEQ ID NO: 87 CACGGCGTATTTTTATTTTG SEQ ID NO: 371
GTTTCTT 3
Satt270 GGTGTTTCAAGTTTCAACACCAT SEQ ID NO: 88
CAGTGCATGGTTTTCTCATGTACC SEQ ID NO: 372 GTTTCTT 3
Satt272
ATGACAAGGAAAAATCAATCAAC SEQ ID NO: 89 GCTGCTGTTAAGACTGTTTG SEQ ID NO: 373
GTTTCTT 3
Satt274 GCGGGGTCAATTAGTTTTCGTCAGTT SEQ ID NO:
90 GCGCACGGTATATAATCGAACCTAT SEQ ID NO: 374 GTTTCTT 3
Satt279 GCGCAAAAGGACGCCCACCAATAG SEQ ID NO: 91 GCGGTGATCGGATGTTATAGTTTCAG
SEQ ID NO: 375 GTTTCTT 3
Satt280 TCTGCTTATTCATTGTGTGCGTG
SEQ ID NO: 92 TGTCTCCATGCTGTAACACGTCAA SEQ ID NO: 376 GTTTCTT 3
Satt282 TGGTATATGTTTTTGCGGGACAA SEQ ID NO: 93
CGCCAAAGATGCAACACACTTG SEQ ID NO: 377 GTTTCTT 3
Satt284
AGGTGGGCTAGGAGTGACCACA SEQ ID NO: 94 TGTAATTGCTGTTTTGGTTTCATTTC SEQ ID
NO: 378 GTTTCTT 3
Satt285 GCGACATATTGCATTAAAAACATACT SEQ
ID NO: 95 GCGGACTAATTCTATTTTACACCAAC SEQ ID NO: 379 GTTTCTT 3
T
AAC
Satt287 GGGGTGAATGAATGTCAAGATGA SEQ ID NO: 96
GCGCGAGGTATCAACACAATTACT SEQ ID NO: 380 GTTTCTT 3
Satt292
GCGGAATTAGAACTCCAGTAAAGA SEQ ID NO: 97 GCGAGGCCAACATTGAAAAGT SEQ ID NO:
381 GTTTCTT 3
Satt295 TTAGTGGATCTACACTAAACTAATCC SEQ ID
NO: 98 CGTACCATTGGACAACTCTGTAATTC SEQ ID NO: 382 GTTTCTT 3
G AA
Satt299 AGGCATTTCTGGCAAGGGTATG SEQ ID NO: 99
TGGTGAATCAATCTCCTCTAAGTGC SEQ ID NO: 383 GTTTCTT 3
Satt300 GCGCCCACACAACCTTTAATCTT SEQ ID NO: 100 GCGGCGACTGTTAACGTGTC SEQ
ID NO: 384 GTTTCTT 3
Satt303 ATGGGCTATGGGAGGAGGTGTT SEQ
ID NO: 101 CCACACGGGACTTTCCATTTTC SEQ ID NO: 385 GTTTCTT 3
Satt304 CCAGTGCAGTTTTACATGAACTT SEQ ID NO: 102 TATATGTAATGACCCCCATCATG
SEQ ID NO: 386 GTTTCTT 3
Satt307 GCTGGCCTTTAGAACGTCTGACT
SEQ ID NO: 103 CGTTGGATTCGACTTTTTGGGA SEQ ID NO: 387 GTTTCTT 3
Satt311 CCACAAAAAGATGAAACAAAATAG SEQ ID NO: 104
TTGAAGCTCAGGCTGTGATGAAT SEQ ID NO: 388 GTTTCTT 3
Satt314
GCGGAGATTGGAACCTACTCATTC SEQ ID NO: 105 GCGGGGACCAAAAATTCAAAA SEQ ID NO:
389 GTTTCTT 3
Satt319 AACAATTTGATGGTCGGGGTTG SEQ ID NO:
106 TAGGGGAACCGATTTGGTGAAA SEQ ID NO: 390 GTTTCTT 3
Satt321 CACCGTCGTAAAAACTGTGTCGT SEQ ID NO: 107 GCGTGTCAAAGAGTTTTAGACATC
SEQ ID NO: 391 GTTTCTT 3
Satt322 GAAGTGCTAGTTCTCACGGTGTGG
SEQ ID NO: 108 TTGGAGGGAGGTTTTGGAAGGT SEQ ID NO: 392 GTTTCTT 3
Satt326 TTCGCTGCATAATTTTTAGCATCA SEQ ID NO: 109
CCAATCTTTTTGTTAGTTCACCTTCC SEQ ID NO: 393 GTTTCTT 3
A
Satt327 AAAGAGACACCCAAAAGATAACAAAC SEQ ID NO: 110
TTCGTAGCAATGTCACCACCTTGT SEQ ID NO: 394 GTTTCTT 3
A
Satt328 TGACCACCATGAGTTCATT SEQ ID NO: 111 GGGGGTGGCTTTTAGATTC SEQ ID
NO: 395 GTTTCTT 3
Satt329 GCGGGACGCAAAATTGGATTTAGT SEQ ID
NO: 112 GCGCCGAATAAAACGTGAGAACTG SEQ ID NO: 396 GTTTCTT 3
Satt330 GTGACCCTCCATTCCACAACAA SEQ ID NO: 113 TCCTTGCCTTTTAGTTGTTCGGT SEQ
ID NO: 397 GTTTCTT 3
Satt331 GCAGAGTCCCCCCTAAATATAG SEQ
ID NO: 114 CGGGAACAACCACACTCTCCATT SEQ ID NO: 398 GTTTCTT 3
Satt332 GTGAATCATCCAGGGCTTGC SEQ ID NO: 115 TCCTTCACTTTCAAAAACAAAAACAA
SEQ ID NO: 399 GTTTCTT 3
Satt333 GCGAATGGTTTTTGCTGGAAAGTA
SEQ ID NO: 116 GCGCAACGACATTTTCACGAAGTT SEQ ID NO: 400 GTTTCTT 3
Satt334 GCGTTAAGAATGCATTTATGTTTAGT SEQ ID NO: 117
GCGAGTTTTTGGTTGGATTGAGTTG SEQ ID NO: 401 GTTTCTT 3
C
Satt335 CAAGCTCAAGCCTCACACAT SEQ ID NO: 118 TGACCAGAGTCCAAAGTTCATC SEQ
ID NO: 402 GTTTCTT 3
Satt336 AATTGGAGTGGGTCACAC SEQ ID
NO: 119 TTCCCGGAAAGAAAGAAA SEQ ID NO: 403 GTTTCTT 3
Satt338 GCGCCCAAGTATTATGAGATATTTGA SEQ ID NO: 120 GCGATAATTTTAAAACTGGACCA
SEQ ID NO: 404 GTTTCTT 3
T
Satt339
CTTTGTTTGGTTGGTGATAAGTTTCT SEQ ID NO: 121 AAGCAGTTCCTCTCATCACGTAACA SEQ
ID NO: 405 GTTTCTT 3
A
Satt343
AATTGTTAGGGCGCGAGAAACA SEQ ID NO: 122 CGCAATCCCAATTCACCTCTTC SEQ ID NO:
406 GTTTCTT 3
Satt346 GTTCGGAGGGAGGAAAGTGTTG SEQ ID NO:
123 CCATAAAACATAGCAACTGTCGTCTC SEQ ID NO: 407 GTTTCTT 3
Satt347 TGTTCAATCGACAAATAAGGGTGC SEQ ID NO: 124 TCCCAGGGTAAAAGTTCAAGTTCA
SEQ ID NO: 408 GTTTCTT 3
Satt348 CTTTACTTAGTAATGGTTCCCACA-
G SEQ ID NO: 125 TCGATGTTTCTCCCTCCCTTAGA SEQ ID NO: 409 GTTTCTT 3
Satt352 CCAGATTTTACGTTAATGTTTTAGTT SEQ ID NO: 126
TTGCACATGGTCCTTGGTTGAT SEQ ID NO: 410 GTTTCTT 3
TTA
Satt353 CATACACGCATTGCCTTTCCTGAA SEQ ID NO: 127 GCGAATGGGAATGCCTTCTTATT-
CTA SEQ ID NO: 411 GTTTCTT 3
Satt355
GCGTCCCAGGACATCATCATCATC SEQ ID NO: 128 GCGTAGCGTGTTATTTTGTGTTTG SEQ ID
NO: 412 GTTTCTT 3
Satt356 TGCTGCTTGTGTTTGGTTGATCT SEQ ID
NO: 129 CATATCCTGCCCCCCCAATTAT SEQ ID NO: 413 GTTTCTT 3
Satt357 AATCCCCAAACAAACGCACAGT SEQ ID NO: 130 TGACATCTAAGTCCAGAAATCAAAGC
SEQ ID NO: 414 GTTTCTT 3
A
Satt358
GCGGCGCTTTATGTAACAATACGATT SEQ ID NO: 131 GCGAGTAAAAGCAGAGTGCGGAGTA SEQ
ID NO: 415 GTTTCTT 3
T
Satt359
GCGAGAAAATAATCCTGCTCAAG SEQ ID NO: 132 GCGTTTAAGTCCAATAACAAAGATAA SEQ ID
NO: 416 GTTTCTT 3
C
Satt361
TCGGGAGACACTAAAGGCACTG SEQ ID NO: 133 TCGTTGACACACAAAAAAAGCGA SEQ ID NO:
417 GTTTCTT 3
Satt364 GCGGCATAAGTTTTCATCCCATC SEQ ID NO:
134 ATCGGGTCATGACTTTTGAAGA SEQ ID NO: 418 GTTTCTT 3
Satt367 GCGGATATGCCACTTCTCTCGTGAC SEQ ID NO: 135 GCGGAATAGTTGCCAAACAATAAT-
C SEQ ID NO: 419 GTTTCTT 3
Satt369 GGAGAAAAACATCCAAAGAAATG-
TG SEQ ID NO: 136 CAAGTGGATTGACACACTAAGGTTTG SEQ ID NO: 420 GTTTCTT 3
A
Satt372 GGATAATTTTTTCATCATCACAATTT SEQ ID NO: 137
CACAAAAGACAGGAGATGTGAGCAA SEQ ID NO: 421 GTTTCTT 3
ATC
Satt373 TGGAAATACTGAATGAAAGCATATTG SEQ ID NO: 138
TTGTAGACGACTTGTGGTTCGATTC SEQ ID NO: 422 GTTTCTT 3
G
Satt378 CCATTGGGATCGAGAATAATTGATG SEQ ID NO: 139
AATGAAGAAAATGTGAATTTGAAACC SEQ ID NO: 423 GTTTCTT 3
A
Satt380 AGAGTAATGGCTCCTGCTCCGA SEQ ID NO: 140
TTCCCTTTTTCTAACTCTCCTTTTTC SEQ ID NO: 424 GTTTCTT 3
A
Satt383 CGATCTAACACGCATATTTCCTCTGA SEQ ID NO: 141
TGTCTTGGTGCAATACCTGACATTT SEQ ID NO: 425 GTTTCTT 3
Satt384 TGGGGGTCAATTTTAATTTGTGC SEQ ID NO: 142 ATTTCCCTTTCACCCACCTCTGTTT
SEQ ID NO: 426 GTTTCTT 3
Satt385 GATTCTCTTCATTCTAATACTCGT-
TT SEQ ID NO: 143 CAACGATCCCAGCTCACAGTTT SEQ ID NO: 427 GTTTCTT 3
Satt387 GCGTTACGTTTCACTATTTATTTAAC SEQ ID NO: 144
GCGGCAGGCTAGCTACATCAAGAG SEQ ID NO: 428 GTTTCTT 3
AT
Satt389 GCTGGTGTATGGTGAAATCAAATTAC SEQ ID NO: 145
TTTCAAACAAGGAAGAAACCTCTTTT SEQ ID NO: 429 GTTTCTT 3
T T
Satt390 TGATATTGTTTTGTGTGAAAGATGCA SEQ ID NO: 146
AAGTAACACTGTGGCGGCATCC SEQ ID NO: 430 GTTTCTT 3
C
Satt391 TGCTCAAAGGGTCAATTTCTTTCC SEQ ID NO: 147 TGTGTAATTTCTATCACCTTATTGT-
G SEQ ID NO: 431 GTTTCTT 3
CC
Satt393
CCAAGCCCATAAACGAAATAAAACA SEQ ID NO: 148 TCCTTTGGCTCGGCCTATGTAA SEQ ID
NO: 432 GTTTCTT 3
Satt398 CAGTGCTCATATCAAATTAAAGTGG SEQ
ID NO: 149 CGCGGACTCAGTTAAACCGTAT SEQ ID NO: 433 GTTTCTT 3
Satt399 TTTCAACCACCAAGCCAACCTT SEQ ID NO: 150 CGTTCAATAGTTCCTATGATGGACGA
SEQ ID NO: 434 GTTTCTT 3
Satt406 TGCAGCATGTGTTTTAGGCTTTC
SEQ ID NO: 151 GCATTGCACGTCGATTTTAGGG SEQ ID NO: 435 GTTTCTT 3
Satt409 CCTTAGACCATGAATGTCTCGAAGAT SEQ ID NO: 152
CTTAAGGACACGTGGAAGATGACTAC SEQ ID NO: 436 GTTTCTT 3
A
Satt411 TGGCCATGTCAAACCATAACAACA SEQ ID NO: 153
GCGTTGAAGCCGCCTACAAATATAAT SEQ ID NO: 437 GTTTCTT 3
Satt412 ACTGGCGCTGACCTTAAATTGC SEQ ID NO: 154 TCCTTTTAATTCTAACATTGAGACAG
SEQ ID NO: 438 GTTTCTT 3
CA
Satt413
TGTTTTTAAGTAATCCGGTGAAATAG SEQ ID NO: 155 TCTGTCCCAAAAAAGAAAGAAGATAT SEQ
ID NO: 439 GTTTCTT 3
CA G
Satt414
TCACATCACAAGTTTCATAAATGCTG SEQ ID NO: 156 CAATCTTTAATGCTCTGGAGTTTGAG SEQ
ID NO: 440 GTTTCTT 3
A
Satt415
GCGTCTCCCTTAATCTTCAAGC SEQ ID NO: 157 GCGTGTGACGGTTCAAAATGATAGTT SEQ ID
NO: 441 GTTTCTT 3
Satt416 AGATTAAGAGAAGACGAGAGTTTTA SEQ
ID NO: 158 GGGTAATTTATCTGTGTGATTGTTC SEQ ID NO: 442 GTTTCTT 3
Satt417 GCCAGGTGCTCACTTCTTGCTA SEQ ID NO: 159 TTGCTTGGGATTTTCATTTTATT-
ATA SEQ ID NO: 443 GTTTCTT 3
GG
Satt418
CAGGAGAAAAAAGGAAAAGAAAAGCA SEQ ID NO: 160 GGTCAAAGAATAAGGGATTTGCCTC SEQ
ID NO: 444 GTTTCTT 3
Satt420 AGGTTTTGCTTCTTGAAAGTGAAGAG
SEQ ID NO: 161 TGGGACTTTGATTTTTGGAATACCC SEQ ID NO: 445 GTTTCTT 3
Satt421 GTCTCCGTTCAAAGCTTCTTCTTC SEQ ID NO: 162
CCAGAGAAATTATTGGAGTGGCAA SEQ ID NO: 446 GTTTCTT 3
Satt422
GAGGGGAGGTAAAAAGTCGGGA SEQ ID NO: 163 GCGCAAAGAAATTCCCATCCTA SEQ ID NO:
447 GTTTCTT 3
Satt423 CGCTTGGGTTCAGTTACTTGGTG SEQ ID NO:
164 GGGAGAGGTTCAGTTGGGGAAT SEQ ID NO: 448 GTTTCTT 3
Satt426 GCGCCCATTATTATTTTCCTTCAATT SEQ ID NO: 165 GCGAATGCCTAATGTGATGATAA-
AAA SEQ ID NO: 449 GTTTCTT 3
GT AATA
Satt429
GCGACCATCATCTAATCACAATCTAC SEQ ID NO: 166 TCCCCATCATTTATCGAAAATAATAA SEQ
ID NO: 450 GTTTCTT 3
TA TT
Satt430
AATGCAAGAAGCPATCAGCAAGA SEQ ID NO: 167 AATAATGGAGTGACCCGCTGCT SEQ ID NO:
451 GTTTCTT 3
Satt431 TGGCACCCTTGATAAATAAGAGAAGG SEQ ID
NO: 168 TGGTACGAGTGGCACGAAATGT SEQ ID NO: 452 GTTTCTT 3
Satt432 AATTGAACCACTCAGCCAAGCC SEQ ID NO: 169 GCCATCCTTTCCTTTCAACCAA SEQ
ID NO: 453 GTTTCTT 3
Satt433 GTCTAAGATTTTAATTAGGGTGATTC
SEQ ID NO: 170 TGTAGGCTATTGGAAGGGTGCG SEQ ID NO: 454 GTTTCTT 3
Satt436 GCGTATAAAGAAAAACGAGCATATCA SEQ ID NO: 171
GCGCTTATAAAGGCTTGTGAAAGACA SEQ ID NO: 455 GTTTCTT 3
T CT
Satt440 CAAATTGAAAAACACAAGAAAACACA SEQ ID NO: 172
TGGTTAGTGCAATCTTGGCGGT SEQ ID NO: 456 GTTTCTT 3
A
Satt441 AGCAACTAACCTTGGGCTTCAGTAA SEQ ID NO: 173 TGCACCCATCAATCACATTTTTG
SEQ ID NO: 457 GTTTCTT 3
Satt442 CCTGGACTTGTTTGCTCATCAA
SEQ ID NO: 174 GCGGTTCAAGGCTTCAAGTAGTCAC SEQ ID NO: 458 GTTTCTT 3
Satt444 AATTCCTTTGGCATTTTTGTTGC SEQ ID NO: 175
TTTTTTCTCACACCCATGCCGT SEQ ID NO: 459 GTTTCTT 3
Satt445
TTTCACTTTTGAATCATTGCATCG SEQ ID NO: 176 CCGGTTGGCGTAATGTGTCTTT SEQ ID NO:
460 GTTTCTT 3
Satt448 CACCACTCGTATCCTTCACAAGAGC SEQ ID
NO: 177 GCCAGCAGCCTGTTCAGTTTTT SEQ ID NO: 461 GTTTCTT 3
Satt451 GCGCAATTAAAAGGATAACTTATATC SEQ ID NO: 178 CCCCTCTTTGGCCCTCACACCTT-
CTC SEQ ID NO: 462 GTTTCTT 3
Satt452
AAAATTCATGTCGCTGCGTTCA SEQ ID NO: 179 ATTTGAAGCTCTTGGTATCTTGGC SEQ ID NO:
463 GTTTCTT 3
Satt454 TGAAAACCATGTCAAAGTAATGGCA SEQ ID
NO: 180 CAACCATGATAAATGTGAGTGAGCTT SEQ ID NO: 464 GTTTCTT 3
G
Satt455 GGAAAGTTTTGTTACATGCCGGA SEQ ID NO: 181
GTCACAATTTCACGATCCCAAA SEQ ID NO: 465 GTTTCTT 3
Satt457
TTTTAACTGGAGAAACCTGAGGGA SEQ ID NO: 182 TCCATTTTCCCTTTAGTCCAACG SEQ ID
NO: 466 GTTTCTT 3
Satt460 GCGCGATGGGCTGTTGGTTTTTAT SEQ ID
NO: 183 GCGCATACGATTTGGCATTTTTCTAT SEQ ID NO: 467 GTTTCTT 3
TG
Satt461 TTGCTTGCTGCACATGACTGAG SEQ ID NO: 184
AATTTCTTACGTTTCCATAGATTTCT SEQ ID NO: 468 GTTTCTT 3
CG
Satt464 TTGAAATAGTGGTGGGGTTGGG SEQ ID NO: 185
TGCCCTCTTCCCAAACTAGGGT SEQ ID NO: 469 GTTTCTT 3
Satt466
TGTGTTGTGTGGTGTGGTGGTC SEQ ID NO: 186 CAACCACTGATTCAAGCCAACAA SEQ ID NO:
470 GTTTCTT 3
Satt467 CAAAGTCCCCTTTCACACCTTTT SEQ ID NO:
187 CAATTTAAGCACGGTCATATTTTCTC SEQ ID NO: 471 GTTTCTT 3
A
Satt469 AAAGGGAAAGGAAGAATAAACCGA SEQ ID NO: 188
CGTGATGCAGTGAATTTTTTTCG SEQ ID NO: 472 GTTTCTT 3
Satt470
TGCTTTTTCTCTTTGGCAACCC SEQ ID NO: 189 GGGTTACTTTTCTTTATCCTCCTCCA SEQ ID
NO: 473 GTTTCTT 3
Satt471 GCGCCCAAAACTATCTAGTAATTCTT SEQ
ID NO: 190 GGGCTATCAAATTGACTAAAGCCAAA SEQ ID NO: 474 GTTTCTT 3
Satt473 CCAACAACCAAATCAATCACTGC SEQ ID NO: 191
ACACTTGAATCATCGAGAGTTGCTAA SEQ ID NO: 475 GTTTCTT 3
Satt475 GCTCCGGTCCTTCAACTGACCT SEQ ID NO: 192 TGTGCTGCTTGCTTCAATTTGG SEQ
ID NO: 476 GTTTCTT 3
Satt476 ATGTGGGTATGTTGCAGGCAGA SEQ
ID NO: 193 TGGCCTGTCTTATATTACCGAACCAA SEQ ID NO: 477 GTTTCTT 3
Satt477 GTTGGGAAAAGGTTACTACCATATC SEQ ID NO: 194
GGTCCGTATGCAATTCTTGACTAATA SEQ ID NO: 478 GTTTCTT 3
Satt478 CAGCCAAGCAAAAGATAAATAATA SEQ ID NO: 195 TCCCCCACAAGAGAACAAGAAGGT
SEQ ID NO: 479 GTTTCTT 3
Satt479 GCGCTTTCAAAAAGTAACAATTAA-
TG SEQ ID NO: 196 GCGGGAATTGGTTAATCTCATCGTGA SEQ ID NO: 480 GTTTCTT 3
AAA C
Satt480 GACCTTTCATCATCGTCCCCAC SEQ ID NO: 197
TGCGAAAAAGCAGAGTGACCAA SEQ ID NO: 481 GTTTCTT 3
Satt487
TCAATTCATCACGGACCAGTTCA SEQ ID NO: 198 TGAGCATATTTTGATCCGATGCC SEQ ID NO:
482 GTTTCTT 3
Satt488 GTGAGTTTCGGTGCTGTATTCC SEQ ID NO:
199 GTTGCTTTGTTATGTAATGGAAGTC SEQ ID NO: 483 GTTTCTT 3
Satt491 GCTAAATTGATGAAAGGATACAAG SEQ ID NO: 200 GCCCCACAAATATTCAGAAGGTAA
SEQ ID NO: 484 GTTTCTT 3
Satt492 GTATCGTTCGCGTCTTGAGTC
SEQ ID NO: 201 GCAGCGGTGTAGTTCGTTCTTTCT SEQ ID NO: 485 GTTTCTT 3
Satt493 TTAACGGGAAAAAATTAAACCTACGA SEQ ID NO: 202
AAATCGTCTTTTGTGGCTGCCT SEQ ID NO: 486 GTTTCTT 3
Satt495
TGGAGATTTAATATAGATGCCGCGA SEQ ID NO: 203 GCACCATGTTCTTTTTCCATCAAA SEQ ID
NO: 487 GTTTCTT 3
Satt497 GCGGTTTTGGATTGACTTTGTTGA SEQ ID
NO: 204 GGCTCAATTAGAGCATGCAACATC SEQ ID NO: 488 GTTTCTT 3
Satt503 CCGTGACTTTTGTTATCCTGAGTTCC SEQ ID NO: 205 CATGTTAAACGTCCACCCACCA
SEQ ID NO: 489 GTTTCTT 3
Satt506 GCGAATTGGCATACATAGTACC
SEQ ID NO: 206 GCGTGAATTCGCCTAAGTTTAT SEQ ID NO: 490 GTTTCTT 3
Satt507 TGCACCACTAATGTCCTCAGCC SEQ ID NO: 207
TCCCTACTCTCGTGTCGTTAGTTATT SEQ ID NO: 491 GTTTCTT 3
TT
Satt508 GCAATGGGTATTGATCGTGTCA SEQ ID NO: 208
AGTTACATTATTTTTGTCTTTCTGCC SEQ ID NO: 492 GTTTCTT 3
GT
Satt509 GCGCTACCGTGTGGTGGTGTGCTACC SEQ ID NO: 209
GCGCAAGTGGCCAGCTCATCTATT SEQ ID NO: 493 GTTTCTT 3
T
Satt510 GCGAGTTTCGCCGTTACCACCTCAGC SEQ ID NO: 210
CCCTCTTATTTCACCCTAAGACCTAC SEQ ID NO: 494 GTTTCTT 3
TT AA
Satt511 TGCGACTTTACTGAAAACCTGGAA SEQ ID NO: 211
GGAAATGCTTCAAACCAACAAACA SEQ ID NO: 495 GTTTCTT 3
Satt512
AACGTCTTCAAGTCAAGTGCCTACA SEQ ID NO: 212 GCCCACATAGTTTTCATTTTTCTCCA SEQ
ID NO: 496 GTTTCTT 3
Satt513 GCGCATCACAAGTTTTATAGATGCTG
SEQ ID NO: 213 GAGGTCTAGTGCTTTGGTAAGGTT SEQ ID NO: 497 GTTTCTT 3
A
Satt514 GGGTACATTTTATTAAAAGTAAACAC SEQ ID NO: 214
TGTCACACAACCAGTGTCTCAAAATC SEQ ID NO: 498 GTTTCTT 3
ACC
Satt515 TGAACCTTGTCTGTTGATTTTTTTAT SEQ ID NO: 215
CACACCCCAGGACCCATTAAGA SEQ ID NO: 499 GTTTCTT 3
GT
Satt517 TCTCCTACTTCTCTTTCTCCCGTTCA SEQ ID NO: 216
AAAGCGCACACAATGCAAATACA SEQ ID NO: 500 GTTTCTT 3
Satt519
CCTGATTATATGTCTAGACAAACAT SEQ ID NO: 217 CAAGGTTACGAACTGCTCGAATAAG SEQ ID
NO: 501 GTTTCTT 3
Satt522 GAGATCACATCAAAGTCAAAACTGCC SEQ
ID NO: 218 TGAGGAGGCAAGATGATCCAAA SEQ ID NO: 502 GTTTCTT 3
Satt523 GCGATTTCTTCCTTGAAGAATTTTCT SEQ ID NO: 219
GCGCTTTTTCGGCTGTTATTTTTAAC SEQ ID NO: 503 GTTTCTT 3
G T
Satt524 GCGAATTATCCAAAGATACACTTAGT SEQ ID NO: 220
GCGGGTCTTACGAACGTGTCACATTA SEQ ID NO: 504 GTTTCTT 3
C T
Satt526 ATATCGAAAATCGCGCATCTGG SEQ ID NO: 221
CGAACCCAAACCACAAAGCATA SEQ ID NO: 505 GTTTCTT 3
Satt529
GCGCATTAAGGCATAAAAAAGGATA SEQ ID NO: 222 GCACAATGACAATCACATACA SEQ ID NO:
506 GTTTCTT 3
Satt532 GCGCCAATATTATCATGCTTTATGT SEQ ID
NO: 223 GCGTGTAAAAATCTTTGAATCTTGA SEQ ID NO: 507 GTTTCTT 3
Satt533 AGTGGTCGTCACAACACTATCATAT SEQ ID NO: 224 CACCCATTATTGAAAATACAAGG-
ACC SEQ ID NO: 508 GTTTCTT 3
A
Satt534
CTCCTCCTGCGCAACAACAATA SEQ ID NO: 225 GGGGGATCTAGGCCATGAC SEQ ID NO: 509
GTTTCTT 3
Satt536 GCGTGGAATGAGACGTAACCAA SEQ ID NO: 226
GCATAATGGTCTAATAAAAGTGGAGA SEQ ID NO: 510 GTTTCTT 3
CC
Satt537 CAAAACTTATGTGCAACACGACTTCA SEQ ID NO: 227
TGCTTTTGGAGGAACTTTGTCTCA SEQ ID NO: 511 GTTTCTT 3
Satt540
AATGTAGCAATTTGACTGGCGAA SEQ ID NO: 228 CATTCAACCGTGATTGCGAAGA SEQ ID NO:
512 GTTTCTT 3
Satt543 GCGGATCTAAGGATAATTCATTAA SEQ ID NO:
229 GGGAGCGGATCATTCGGTGAAA SEQ ID NO: 513 GTTTCTT 3
Satt544 GCTATGGGAAAAGGATGTGTG SEQ ID NO: 230 GAGCTACCCGAGATGATACTC SEQ ID
NO: 514 GTTTCTT 3
Satt545 ATGTGATGGCATGTGAAATGGT SEQ ID
NO: 231 GGATCAAATTGGGAAACACAAAGG SEQ ID NO: 515 GTTTCTT 3
Satt546 CAGGGTATAGTTCAATTCAGTGAGCG SEQ ID NO: 232 CTCACATACATGGCAGCCGTAA
SEQ ID NO: 516 GTTTCTT 3
Satt548 CCTCTTTGTTGGTGGTTAAGTCTC-
C SEQ ID NO: 233 TGCCTTTAGCTGGTGGGAAAAA SEQ ID NO: 517 GTTTCTT 3
Satt549 GCGGCAAAACTTTGGAGTATTGCAA SEQ ID NO: 234
GCGCGCAACAATCACTAGTACG SEQ ID NO: 518 GTTTCTT 3
Satt550
TCGTCAATTAAGCAAAAATGTGAGA SEQ ID NO: 235 TTAGAGGTTTTCGGATGAGCGTG SEQ ID
NO: 519 GTTTCTT 3
Satt551 GAATATCACGCGAGAATTTTAC SEQ ID
NO: 236 TATATGCGAACCCTCTTACAAT SEQ ID NO: 520 GTTTCTT 3
Satt552 ACAAAAAGAAATCGAACCGGCA SEQ ID NO: 237 GTTTTGGTTGATCCGCATTGGT SEQ
ID NO: 521 GTTTCTT 3
Satt555 TGGCTTTGATGATGTTTGAGACAA SEQ
ID NO: 238 TTTCATTACCGCATGTTCTTGGA SEQ ID NO: 522 GTTTCTT 3
Satt556 CCCAGATACAGACAATAAAACCCGA SEQ ID NO: 239
TTATGTTCGTTCATCTCTGAAGCCT SEQ ID NO: 523 GTTTCTT 3
Satt557 TCCACCATGTAATATGTGAAGTGGAT SEQ ID NO: 240 TTCTGTCCATTCTAGCTCACTAA-
CCC SEQ ID NO: 524 GTTTCTT 3
Satt558 CTCACACCCTTTCATTATCTA
SEQ ID NO: 241 AAATCGCGCATCTAAATTTAC SEQ ID NO: 525 GTTTCTT 3
Satt560 ACTCTATTTTATTATCGTGCAAGAA SEQ ID NO: 242
ACAATAACTTGTTTTGCACACTATT SEQ ID NO: 526 GTTTCTT 3
Satt563 GATGACAACGTAGGCTAAAAA SEQ ID NO: 243 GCGCCCACATGATTTTGTACTGAT SEQ
ID NO: 527 GTTTCTT 3
Satt565 GATTCTATATCCATCGTGTTGCT SEQ
ID NO: 244 TATGGTAAATATTAACCATTGTCCT SEQ ID NO: 528 GTTTCTT 3
Satt566 CCCACTGTATCCTTAGTGTGCCA SEQ ID NO: 245 CCTCGTTTTATTCCGAAAGCCG
SEQ ID NO: 529 GTTTCTT 3
Satt567 GGCTAACCCGCTCTATGT SEQ
ID NO: 246 GGGCCATGCACCTGCTACT SEQ ID NO: 530 GTTTCTT 3
Satt568 CATTAACTAATAAGTTGTTGGTAGC SEQ ID NO: 247 TTAGATTCGGACACCGGTCTACT
SEQ ID NO: 531 GTTTCTT 3
Satt569 ACCAAATTGCTTCACGCATCC
SEQ ID NO: 248 TCTTAATTTTTTTAGGAATGGCATCA SEQ ID NO: 532 GTTTCTT 3
A
Satt570 TGCTCATGTGGTCCTACCCAGA SEQ ID NO: 249
CGCTATCCCTTTGTATTTTCTTTTGC SEQ ID NO: 533 GTTTCTT 3
Satt572 GCGGAGCATGTAAATCCAGCCTATTG SEQ ID NO: 250 GCGGGCTAACTTATGTTACTAAA-
CAA SEQ ID NO: 534 GTTTCTT 3
A T
Satt573
GCGGATTTCGATTTGAATATACTTAC SEQ ID NO: 251 CCTGTGGCTGTTATACTATGCATATA SEQ
ID NO: 535 GTTTCTT 3
Satt576 TGGACACACACAAACACCTACAGAA
SEQ ID NO: 252 GGGTGGCGTTGACAATGTTTTA SEQ ID NO: 536 GTTTCTT 3
Satt577 AGCAAGTCTTGAGTCTTTTGTCT SEQ ID NO: 253
TTATTATCTAAACTTATATGTGCAT SEQ ID NO: 537 GTTTCTT 3
Satt578 TCCCACGTCATATCCACTGCTC SEQ ID NO: 254 GCCTCCTAAGTCCGTACACAGCAT
SEQ ID NO: 538 GTTTCTT 3
Satt581 CCAAAGCTGAGCAGCTGATAACT
SEQ ID NO: 255 CCCTCACTCCTAGATTATTTGTTGT SEQ ID NO: 539 GTTTCTT 3
Satt582 CCGGTGATACTCCATACCAATAACA SEQ ID NO: 256
GGATTTGGTTTCTGTGTGCTGTG SEQ ID NO: 540 GTTTCTT 3
Satt583
CCGAGCTAACAAAGGCGACCAAAT SEQ ID NO: 257 GGGGCACAAGCCACACTT SEQ ID NO: 541
GTTTCTT 3
Satt584 GCGCCCAAACCTATTAAGGTATGAAC SEQ ID NO:
258 GCGGGTCAGAAGATGCTACCAAACTC SEQ ID NO: 542 GTTTCTT 3
A T
Satt586 ATGGCCGTCTCAAAAGAACTGG SEQ ID NO: 259
TGGGCACTTGCAGTCCAAATAG SEQ ID NO: 543 GTTTCTT 3
Satt587
GCGAATGGTTGCTCAAATAATC SEQ ID NO: 260 GCGCAAACCGCACAAGTTTATGT SEQ ID NO:
544 GTTTCTT 3
Satt590 GCGCGCATTTTTTAAGTTAATGTTCT SEQ ID
NO: 261 GCGCGAGTTAGCGAATTATTTGTC SEQ ID NO: 545 GTTTCTT 3
Satt591 GGCAGACTCGTAGAGCAATTTA SEQ ID NO: 262 TGTTGAAATTGACCAAAATTCCCA
SEQ ID NO: 546 GTTTCTT 3
Satt592 GCGAAGATTGGTCTTTTATGTCAA-
AT SEQ ID NO: 263 GCGGAGGAATACAAGTCTCTATTCAA SEQ ID NO: 547 GTTTCTT 3
G
Satt594 GCGGTAACTCCTCGAGTCCCTCTCAA SEQ ID NO: 264
GCGCCGCTAACAGACATCCAATA SEQ ID NO: 548 GTTTCTT 3
T
Satt595 TGGTGATGGGAAGCAAACAAGA SEQ ID NO: 265 TGGATTTCACCCAAGAAAAAAGC
SEQ ID NO: 549 GTTTCTT 3
Satt596 CCATCCCTTCGTCCACCAAATA
SEQ ID NO: 266 TCGACTACCCGTCGATTCCGTA SEQ ID NO: 550 GTTTCTT 3
Satt597 TGCTGCAGCGTGTCTGTAGTATAATT SEQ ID NO: 267
GGCACAACCATCACCACCTTATT SEQ ID NO: 551 GTTTCTT 3
T
Satt598 CGATTTGAATATACTTACCGTCTATA SEQ ID NO: 268
CACAATACCTGTGGCTGTTATACTAT SEQ ID NO: 552 GTTTCTT 3
Satt601 GTAACATTGGTTGTCATCTTTGTCTA SEQ ID NO: 269 ATCGAACTGTGACCGTCCCTTC
SEQ ID NO: 553 GTTTCTT 3
Satt602 GGAGGTTATCTAGTGGTATAGATG-
GT SEQ ID NO: 270 AAGGGAAGAGAGTCGTGCTTCTTT SEQ ID NO: 554 GTTTCTT 3
Sct_010 CCAAAAGCATTGAGAGTGGGGA SEQ ID NO: 271
CCAGCAAACCCCCAGGTAAAG SEQ ID NO: 555 GTTTCTT 2
Sct_026
GAAACCCGAAACGCAAAATCTC SEQ ID NO: 272 GAAGAAAACOCGAATAACCCCA SEQ ID NO:
556 GTTTCTT 2
Sct_028 CTCTCGCCGGTACAAAACACCT SEQ ID NO:
273 GCACGCAGACTCAAGTTCATTCA SEQ ID NO: 557 GTTTCTT 2
Sct_034 TCACTCTCACAACTTCAATCTCTTTC SEQ ID NO: 274 AATAGTTGGGTCGTCGAAGGGG
SEQ ID NO: 558 GTTTCTT 2
TC
Sct_046
CACGACTCTTCCTCTTCCTCCG SEQ ID NO: 275 TCCAACTTAACACAAGATCAGCGAA SEQ ID
NO: 559 GTTTCTT 2
Sct_065 CCCTGTGTTTCCCTCT SEQ ID NO: 276
GAAAAGTTTTATGTTCTGAGTG SEQ ID NO: 560 GTTTCTT 2
Sct_137
GTGTTGCTCTTGGGAATCTGGC SEQ ID NO: 277 CCACACACACTGACACAGTAAAGCA SEQ ID
NO: 561 GTTTCTT 2
Sct_147 TCTCGACTCACGACTCA SEQ ID NO:
278 CCAAGGTCTCTCAGAGG SEQ ID NO: 562 GTTTCTT 2
Sct_186
AAAATGAAAACACACAGAGAGAGAGA SEQ ID NO: 279 ACGGAGGCACTTCCCATTGTTA SEQ ID
NO: 563 GTTTCTT 2
GA
Sct_187
AGCGAAATGTGTGGTCCAAGGT SEQ ID NO: 280 CAAAACGACATGACAAGGAAACTTCA SEQ ID
NO: 564 GTTTCTT 2
Sct_188 CGTCGAGAAAGGAAGAGAGGCA SEQ ID
NO: 281 TCCTCCATAAAAATTAAAAACATTGG SEQ ID NO: 565 GTTTCTT 2
AA
Sctt008 GCGGAAACCATTCTGACGGATA SEQ ID NO: 282
GCCCCCAGACACAACATAATCA SEQ ID NO: 566 GTTTCTT 3
Sctt009
GGAGGAACTTGGAAGGCATATCA SEQ ID NO: 283 CGAGAGAAGCAGAAGCAGAGGC SEQ ID NO:
567 GTTTCTT 3
Sctt012 ACGGACAACGCTGGCACTAAG SEQ ID NO:
284 GAGAAAGGTGACGATGGACGCT SEQ ID NO: 568 GTTTCTT 3
[0269]
18APPENDIX III
ASH marker primer sequences
Primer 1 Primer 1 sequence
Primer 2 Primer 2 sequence
Marker Name Primer 3 Primer 3 sequence
P10355B-1 19392
CATGGTTTCTCTTATCTTAT(AG)ACATTGTTGCCAAG SEQ ID NO: 569
19393 CAATTCATGGTTTCTCTTAT(AG)ACATTGTTGCCAAG SEQ ID NO: 570
25400 CACTGTCCCTGCTCCTGTTTCAAGTATC SEQ ID NO: 571
P10598A-1 14507 CAATTCTTGTGGGTTGAAGCCTTGTTCTGAC SEQ ID NO: 572
14505 GGAATCAACTTCTTCGTGAGTGGGTTGTTC SEQ ID NO: 573
P10615A-1 1125 TGGTGGCTATGGAAATCTCATGTGTGGA SEQ ID NO: 574
4186 CTCTCATTTACCAAACTCCAACATTTGATCACC SEQ ID NO: 575
P10618A-1 14492 CACACTGGTAGATGGGAAGCAAGAATAGG SEQ ID NO: 576
14490 GAAGAAGATTCCACCCAGATCATCATCAGTAG SEQ ID NO: 577
P10620A-1 12099 GCTTGTGCAGCTCCAATCGGTGTAAC SEQ ID NO: 578
12101 GTGCAATCCAAGACATCTGGTTCGGAC SEQ ID NO: 579
P10621B-2 12158 CAGCTAAACCTTACAAGGATGATTGGTCAAG SEQ ID NO: 580
12159 CCCTGGACTGAAGTTGCCATAATGTATC SEQ ID NO: 581
P10623A-1 12153 GCTGGTTGGGAGAAAGCACTTCC SEQ ID NO: 582
12154 GAATCTAACATTACGCTCTGCTGGAGTATC SEQ ID NO: 583
P10624A-1 14328 GAGGGACTATGTGAAATGGAGAGGAGTG SEQ ID NO: 584
14330 GTATGCTAAAAGAGGAGACTTGACTGGTGAG SEQ ID NO: 585
P10632A-1 12105 GATGAAGGAACCAACACTTGCATAACAATTTG SEQ ID NO: 586
12106 TGTCAGCACTCCTCACTCATTTGCCGA SEQ ID NO: 587
P10633A-1 4187 CACTTAACAGGAGTGCTCCTGATCACCAG SEQ ID NO: 588
1183 GAGAACAAGGACAAATCAATAGGTGAGACGAAGAAA SEQ ID NO: 589
P10634A-1 12600 CTCATCTGCTCAGAACCTTCAGTCAGTC SEQ ID NO: 590
12601 CGGATCATGTCTAGTACATTAGAGATGCTTGTG SEQ ID NO: 591
P10635A-1 12779 CTACACTTCTAATGCCTATTTAGGTGTGCTTG SEQ ID NO: 592
12780 GTCATATCTAGGGAGATTTCTAACCAGTTGTC SEQ ID NO: 593
P10636A-1 15252 TAGGCAGCGTGACAAACTGAGCATAGG SEQ ID NO: 594
15258 GGGTTGATGTCCGATGGGTAAATGAAGTTG SEQ ID NO: 595
P10637A-1 14136 GCACTAATACTTTAGTTGACTTTTGAGGTGGTTGAG SEQ ID NO: 596
14138 GCTATGTGGTAGAAGTATATGAAAAGGTAGATGACAG SEQ ID NO: 597
P10638B-2 15290 GAATATACTAGCTTGATGCCTATTTGTTTCTAAACCC SEQ
ID NO: 598
15040 AGCAGTCATACAATGCTCTTTATTGTGGTGAAG SEQ
ID NO: 599
P10639A-1 15345 GCTCATAGCCTGCTTCTTAATCTTGTTATT-
CTG SEQ ID NO: 600
15347 CTATTTGTTTCAGGAGTTTCACAACCATCTCA-
AGT SEQ ID NO: 601
P10640A-1 15280 CATCTTGACCAGTCACCAATCTG-
AGTACAG SEQ ID NO: 602
15281 GGCTTCAGTGAGAAAGGTGGATCAAATG-
GA SEQ ID NO: 603
P10641A-1 15349 CAACGTGTAAAATCAAGAGATTGA-
GCTTCTGG SEQ ID NO: 604
15350 TCGGTGTGTTCAACTATGACTTTGGTT-
CTG SEQ ID NO: 605
P10646A-1 5642 CCCCCAACAAAACTAAAAATAGA-
ACCCTCAACAACC SEQ ID NO: 606
6803
GGTCCAAGAACATTATGATCTTGAACAATCTTCAC SEQ ID NO: 607
P10648A-1 14436 TGGCTAGAGCATCAACACATCTATACCTTC SEQ ID NO: 608
14333 CATTGGGCATGATTCTTGAATAGCCTTTTTACC SEQ ID NO: 609
P10649C-3 12171 GAGGGCTATGTTTTCTTCTCCAGATGTGAG SEQ ID NO: 610
12173 AAGGTCGGCTTGGTGGTTAAAGGCAG SEQ ID NO: 611
P10651A-1 10414 GCCTTTTATGCACATTTTTCCTGGGGATCTAAC SEQ ID NO: 612
10415 CTCAATGTCATGGGATCAATTTGGAAATTCAATGACC SEQ ID NO: 613
P10782A-1 15294 GATTACATTAATTACCGCTATGACTATATCTTGGGAC SEQ ID NO:
614
15296 GCTACCTTCTCCATTGCTTCTATGTATTGGTC SEQ ID NO:
615
P10783A-1 15771 GGGCATCACATACATGAAAACAACTACACTTG SEQ
ID NO: 616
15772 CAACAGCTCTCTTCCACCACAATCCTG SEQ ID NO:
617
P10792A-1 15805 GAGATTGGAAATTGTAGCTCTCTTTACTTGCTG SEQ
ID NO: 618
15807 CTTTGAGGACTTATTTGGTTGTTATAGGCATTTGG SEQ
ID NO: 619
P10793A-1 15621 GTGTTTCCCTCCATTTTTGCCAAAAGACAG
SEQ ID NO: 620
15814 TATACACACTAAGAATTCGCTCGCTGTACAA SEQ
ID NO: 621
P11070A-1 16188 GGTCTAGACTTTCACTCAGACAAGGAAC
SEQ ID NO: 622
17490 CAAAATACTACAGACCTAATTTGTAACTAATTGCT-
CCC SEQ ID NO: 623
P11347A-1 9642 GGGAAGAAGAAGAACACTCGGTA-
CAGTAG SEQ ID NO: 624
7693 AAGCTAGGAAATCCACACTCAAATTATCG-
ACTTGTGT SEQ ID NO: 625
P12105A-1 23538
GACCTGAAGCAAAACGCCACCATTTCC SEQ ID NO: 626
29311
CGTTGGAGACGACGCCGTTTGATTAC SEQ ID NO: 627
P12198A-1 39113
CAGTCGACACGTCTTCTACTCC SEQ ID NO: 628
39114
TGGAAGGCATGTCGGAACTTG SEQ ID NO: 629
P12390B-1 21397
CCTTTTTGCCCTCACTTCATGCCTTCTATG SEQ ID NO: 630
23525
GCATAACCCAAGAGCTGGACTGACAAG SEQ ID NO: 631
P12391A-1
21215 CAAGCTCTGCTGCCAGGTTAAGTGTTTC SEQ ID NO: 631
21216
GACTAGAACAAATTGGGGCTAGTGTGTTTGAG SEQ ID NO: 633
P12392A-1
21494 TGGACTTGCGGGACTATGCCTTAGAG SEQ ID NO: 634
21495
GCAGGAACACGTTCGTAACCATCAACTG SEQ ID NO: 635
P12394A-1
22161 GTCCCTTTCTGAACCACTTAAAGAGTCAACAG SEQ ID NO: 636
22163 GGCATAGTGAGTTGAATACCAGGAGGAATC SEQ ID NO: 637
P12396A-1 23679 ATTGAAGGGTGGGCGTTACCAGGTTAC SEQ ID NO: 638
23680 GAGTTAATGGGGCTATGCTATTGGCTATTCAC SEQ ID NO: 639
P13069A-1 24588 GTCACTATATGGAGTCAAGGTAATTATTGTGTTCAC SEQ ID NO: 640
24590 GGTCTAGAGTTTGAATATTAGTAATGACTTGTATTGAC SEQ ID NO: 641
24591 GCATATTGTGCCATAGAGAGAGAAAATGTAGTAAG SEQ ID NO: 642
P13070A-1 24508 CGCTACTGCAAGTTATCAGTCAAGAGATTATTC SEQ ID NO:
643
24510 GCCGTGTAAGCGTGTTTACCAATCTAGTTG SEQ ID NO: 644
P13071A-1 24795 CAAAACTGAGCGAAACTTGTGTTGGGAGAAAG SEQ ID
NO: 645
25395 GTGGGGACTCTTTATTCGAAGTTTGCTGAAC SEQ ID NO:
646
P13072A-1 24499 CTCATGTAACCAACTCTCTATGAAGTTTGAGATCCA
SEQ ID NO: 647
24500 CTCTAATCGGATTTGGTGTTTCACTTCGGTAAG
SEQ ID NO: 648
P13073A-1 24503 GATGGCTGTCATTGCTACAGAGGAGT-
ATC SEQ ID NO: 649
24505 GTGACTCCAAAGGAAAGAGAAATGTTTCTTAA-
ATCATC SEQ ID NO: 650
P13074A-1 25287
GGATAGCAAGTCAATTTCATGCCTTGTGATAG SEQ ID NO: 651
25288
GCAGGACATGAAGATGTACTTAGTGAATGTGAAG SEQ ID NO: 652
P13158A-1 24713 ACTGGAAGAGGGTGCTTAGGGAATCTG SEQ ID NO: 653
24715 GAGAATCTAGTCTACCACCATACCACGAAC SEQ ID NO: 654
P13560A-1 25671 GGGACTCTCTTTATATTGGAAGGTATAACTCAGTG SEQ ID NO: 655
25672 GTTGGACCCTTGTAATTAGACCCGAAACAAATG SEQ ID NO: 656
P13561A-1 26537 ATCTGTCCAACGATCTCTCCATGTTCATTC SEQ ID NO: 657
27121 CGAATAAGAAGTTGGGTATCACTTACACGTTGG SEQ ID NO: 658
P2447B-2 8815 GATGGGGTTCTAGACTGGGATCTGGAT SEQ ID NO: 659
8816 CTTTTCCTACAGGATTGTCAGGCTTATCGTCA SEQ ID NO: 660
P2481A-1 10468 GAAGAGTAACAGAGTCTACGCACCGAC SEQ ID NO: 661
10470 GTCAACGAACATACTATGCATGATGATTTCTGATTAG SEQ ID NO: 662
P2636C-2 21190 CATATGCAGACAGCAGGCTAAGGAACTC SEQ ID NO: 663
15824 GCAGCAATATAACCAGGATTCAGAATTAATCTAGTTAG SEQ ID NO: 664
P3050A-2 16940 CGATGGGGTTGACTTAGAAATGGCATATAC SEQ ID NO: 665
16860 GCACCAATTCCTCAAGCATACTCCAAAC SEQ ID NO: 666
P3436A-1 10660 CTGACAAGGTGTTTTGGTAGGGAGAGATTC SEQ ID NO: 667
10663 GCATCCTCCGTTACTCCAATCAGAGTTTCCAT SEQ ID NO: 668
P3436A-7 10660 CTGACAAGGTGTTTTGGTAGGGAGAGATTC SEQ ID NO: 669
10663 GCATCCTCCGTTACTCCAATCAGAGTTTCCAT SEQ ID NO: 670
P5219A-1 11913 CACACTATCAACACCTATTGGTGACCATTGTA SEQ ID NO: 671
8395 GGAGGGTGCTTATGTAAATGATGTAAAGACCAT SEQ ID NO: 672
P5219A-2 11917 GATGACCATTTTGATTCCCTCATGCTATTAGTACC SEQ ID NO:
673
8396 CCAATAAGTTAGCAGCATGTGGATCACAGTGTA SEQ ID NO:
674
P5467A-1 19156 GTCTCTCGGAGTTGCTTCAATTGCTCATAC SEQ ID
NO: 675
19864 GAGAGTGTGGAATTGTA(AG)TCATTGATTGAAAACTC SEQ
ID NO: 676
P5467A-2 19156 GTCTCTCGGAGTTGCTTCAATTGCTCATAC
SEQ ID NO: 677
19864 GAGAGTGTGGAATTGTA(AG)TCATTGATTGAAAA-
CTC SEQ ID NO: 678
P6181A-2 17279 GCAGAAGGAGCATTGAGGCTTTCC-
AG SEQ ID NO: 679
9372 GAAAAGGTTTGTTATGCTTCGTACTCTGTCTC
SEQ ID NO: 680
P7659A-1 7847 CATGAAGCTCCACCATTTGCTAGTACA-
TGAAAC SEQ ID NO: 681
10878 CCAGAGTTACCAAACCATCTGTGAGAAAT-
ATCC SEQ ID NO: 682
P7659A-2 7847 CATGAAGCTCCACCATTTGCTAG-
TACATGAAAC SEQ ID NO: 683
10878 CCAGAGTTACCAAACCATCTGTGAG-
AAATATCC SEQ ID NO: 684
P8230A-1 13958
CAAACGCTCCCAACAGCTTCAGAATCTC SEQ ID NO: 685
13959
TTGAAGGTTGTAAGAGTCTCGGTCGTCG SEQ ID NO: 686
P8584A-1
15081 CCCATTCTTCATGTACTCATACACCAAGAG SEQ ID NO: 687
15086 GCAGCCACCAATAATTTCTCATTTGACAACAAG SEQ ID NO: 688
P8584A-2 19660 CTCTGTATATGAYATATGTGCTCAGTGCCTC SEQ ID NO: 689
19389 GACTTACCAAATGAGTTTGACCAGGTTTTACC SEQ ID NO: 690
P9026A-1 14494 AGGATTCAACCTCTAGCCATGATGATGTTG SEQ ID NO: 691
14493 CCAAGCTCTTTCCGTGTGTATCAATCTG SEQ ID NO: 692
[0270]
19APPENDIX IV
ASH marker: probes
Marker
Probe Probe Probe
Name Allele1 1 sequence Allele2 2 sequence
Allele3 3 sequence
P10355B-1 18119 GTTTCAGATAAC SEQ ID
18118 GTTTCTGATAAC SEQ ID
NO: 693 NO: 754
P10598A-1 13358 GAATGACTTTGA SEQ ID 13360 GAATGATTTTGAC SEQ ID
NO: 694 NO: 755
P10615A-1 1630 TCATTCTTTCATG SEQ ID
1629 TCATTCATTCATG SEQ ID
NO: 695 NO: 756
P10618A-1 13392 TCTCCAGAAACA SEQ ID 13394 GTTTCGGGAG SEQ ID
NO:
696 NO: 757
P10620A-1 13919 GTGATCCGTG SEQ ID 11141
ATCACGAATCAC SEQ ID
NO: 697 NO: 758
P10621B-2
11150 ATCTTTTCAGGTT SEQ ID 12304 TATGGAGTAATTG SEQ ID
NO: 698
NO: 759
P10623A-1 11360 GGATTACATACTA SEQ ID 11361
TGGATTTTATACTA SEQ ID
NO: 699 NO: 760
P10624A-1 13361 TTTGAAGCTTTAT SEQ ID 13362 TTTGAATCTTTATC SEQ ID
NO: 700 NO: 761
P10632A-1 10697 AAGAATCTTCCTA SEQ ID
10698 AAAGAATATTCCTA SEQ ID
NO: 701 NO: 762
P10633A-1 15132 AATCTAAAATTTAG SEQ ID 15131 ATCTAACATTTAGT SEQ ID
T NO: 702 NO: 763
P10634A-1 11987 ACTAAATTTATACC SEQ
ID 11988 GGTATACATTTAG SEQ ID
NO: 703 NO: 764
P10635A-1 12562 ATTAGGGGCAG SEQ ID 12563 ATTAGGGGGCA SEQ ID
NO:
704 NO: 765
P10636A-1 13678 ACTATAGTTCGC SEQ ID 13679
ACTATACTTCGC SEQ ID
NO: 705 NO: 766
P10637A-1
13428 AACCTTTCTGTC SEQ ID 13430 ACCTTGCTGTC SEQ ID
NO: 706
NO: 767
P10638B-2 12041 TTGAGGATTTAG SEQ ID 12042
TTGAGCATTTAG SEQ ID
NO: 707 NO: 768
P10639A-1
15144 TCTCAACTTGGA SEQ ID 15145 TCTCAATTTGGAA SEQ ID
NO: 708
NO: 769
P10640A-1 15137 TTCAGTCAAACC SEQ ID 20442
TTCAGTTAAACC SEQ ID
NO: 709 NO: 770
P10641A-1
13673 TTCTTTTGTGACA SEQ ID 13675 TTCTTTGGTGAC SEQ ID
NO: 710
NO: 771
P10646A-1 6223 TGTGACAACCGA SEQ ID 6224
GTGACACAACC SEQ ID
NO: 711 NO: 772
P10648A-1
14935 AATCTTTTTTAAAG SEQ ID 13666 AATCTTCTTTAAAG SEQ ID
NO: 712
NO: 773
P10649C-3 11356 TCATCTGTGATAA SEQ ID 11358
TCATGTGTGATAA SEQ ID 11357 TCATCTCTGATAA SEQ ID
NO: 713 NO:
774 NO: 815
P10651A-1 8002 AAGAGAAGGCTA SEQ ID 8003
AAGAGATGGCTA SEQ ID
NO: 714 NO: 775
P10782A-1
15302 ATATAAGTAAGGG SEQ ID 15301 AATATAAATAAGGG SEQ ID
NO: 715
NO: 776
P10783A-1 15599 GCATGTCGAC SEQ ID 15600
GCATCTCGAC SEQ ID
NO: 716 NO: 777
P10792A-1
16759 TTGGAAGTTATAC SEQ ID 15707 TTGGAAGATATAC SEQ ID 15706
TTGGAATATATACT SEQ ID
NO: 717 NO: 778 NO: 816
P10793A-1 15624 GGCATGTGAGT SEQ ID 15625 GGCTTGCGAG SEQ ID
NO: 718 NO: 779
P11070A-1 14956 ATTAACAGTAAAGT SEQ ID
14957 TATTAACATTAAAG SEQ ID
NO: 719 T NO: 780
P11347A-1 15073 TATCTATGTATATT SEQ ID 17277 TATCTATATATATT SEQ ID
A NO: 720 AA NO: 781
P12105A-1 23530 TGGGAATGATG SEQ ID
21919 ATCATCCCCAA SEQ ID
NO: 721 NO: 782
P12198A-1 23164 TAAATAAATAAGAT SEQ ID 23165 AAATAAGTAAGATG SEQ ID
G NO: 722 NO: 783
P12390B-1 21475 AAAAAAAATGAGG SEQ ID
23524 AAAAAATATGAGG SEQ ID
NO: 723 NO: 784
P12391A-1 23527 TCAATGTTGGAT SEQ ID 21219 TCAATGATGGATA SEQ ID
NO: 724 NO: 785
P12392A-1 21493 TTGTGACCAATAT SEQ ID
23437 ATATTGATCACAA SEQ ID
NO: 725 NO: 786
P12394A-1 23401 ATCAAGCCCAA SEQ ID 23528 TGGGTTTGATC SEQ ID
NO:
726 NO: 787
P12396A-1 23681 AGAAGCTCGTG SEQ ID 24579
GAAGGTCGTG SEQ ID
NO: 727 NO: 788
P13069A-1
24584 GAAAAAGAAAGG SEQ ID 24585 GAAAAAAAAAGGA SEQ ID 29603 AGATGTTAGAGTTA
SEQ ID
NO: 723 NO: 789 NO: 817
P13070A-1
24509 ACATATAATAGTAG SEQ ID 25336 ACATATAGTAGTA SEQ ID
NO: 729
NO: 790
P13071A-1 24796 TTTTGTATCTGTAT SEQ ID 24798
TTTGTGGCTGTA SEQ ID
NO: 730 NO: 791
P13072A-1
25337 TTAACTTGCCAG SEQ ID 25338 TTAACTAGCCAG SEQ ID
NO: 731
NO: 792
P13073A-1 24504 AATGATAATTTAGT SEQ ID 24506
AATGATCATTTAG SEQ ID
NO: 732 NO: 793
P13074A-1 25306 GAATGAATTTTTC SEQ ID 25307 AATGAACTTTTTC SEQ ID
NO: 733 NO: 794
P13158A-1 25339 ACACTGCTTAC SEQ ID
25700 TTTTTGCTAGAG SEQ ID
NO: 734 NO: 795
P13560A-1 25673 ACAACTAATAAGG SEQ ID 25674 TACAACTAAGGTA SEQ ID
NO: 735 NO: 796
P13561A-1 26309 TTCTGATAAAAAAA SEQ ID
26310 TCTGATGAAAAAA SEQ ID
NO: 736 NO: 797
P2447B-2 11878 TGTAATGCGTG SEQ ID 8378 TGTAACGCGTG SEQ ID 12003
TGTAACGCATGT SEQ ID
NO: 737 NO: 798 NO: 818
P2481A-1 5486 GACAATCTAAAAA SEQ ID 7441 GACAATTTAAAAA SEQ ID
NO: 738 NO: 799
P2636C-2 14678 TAATAATAATTGTG SEQ ID
15823 AATAATGATTGTG SEQ ID
T NO: 739 NO: 800
P3050A-2 16760 TTCCTTCTTTTTTT SEQ ID 16761 TCCTTCCTTTTTT SEQ ID
NO: 740 NO: 801
P3436A-1 10239 GGAACGTTACC SEQ ID 10240
TGGAACATTACC SEQ ID
NO: 741 NO: 802
P3436A-7
18078 AATTTTTTAGTATG SEQ ID 14278 AATTTTTGAGTATG SEQ ID 14617
ATTTTTTGGTATG SEQ ID
NO: 742 NO: 803 NO: 819
P5219A-1 7700 TTATAGACACTTG SEQ ID 7699 TATAGGCACTTG SEQ ID
NO: 743 NO: 804
P5219A-2 7944 ATAAACCATATATG SEQ ID
7943 ATAAACCTTATATG SEQ ID
NO: 744 NO: 805
P5467A-1 18980 TTAATTACCTTAAG SEQ ID 18981 TTAATTATCTTAAG SEQ ID
NO: 745 NO: 806
P5467A-2 20239 ATGCCATTTTG SEQ ID
19197 ATGCCCTTTTGT SEQ ID
NO: 746 NO: 807
P6181A-2 9448 TAATATCTTATGCA SEQ ID 9292 AATATCGTATGCA SEQ ID
NO: 747 NO: 808
P7659A-1 7858 AGTGCGATGAAA SEQ ID
7859 AGTGCACTGAAA SEQ ID
NO: 748 NO: 809
P7659A-2 10390 GAGGAGATGTAG SEQ ID 7845 AGAGGAGATGTA SEQ ID
NO: 749 NO: 810
P8230A-1 10586 TAAAATTGTTGGTT SEQ ID
14933 TAAAATTATTGGTT SEQ ID
NO: 750 NO: 811
P8584A-1 14786 TGAAGAAAAATATG SEQ ID 14787 GAACAAAAAGATG SEQ ID
NO: 751 NO: 812
P8584A-2 20517 CTGTCCACTAA SEQ ID 20358
ACTGTCAACTAA SEQ ID
NO: 752 NO: 813
P9026A-1
14340 AGAGGCAGTGA SEQ ID 14341 AGAGCCAGTGA SEQ ID
NO: 753 NO:
814
[0271]
Sequence CWU
1
1
819 1 22 DNA Artificial oligonucleotide primer 1 caatcaggtt agtggtccta cc
22 2 22 DNA Artificial
oligonucleotide primer 2 aagttcacag cacctggacc at
22 3 22 DNA Artificial oligonucleotide primer 3
gaaacaaaca cccccaaggt ga 22
4 22 DNA Artificial oligonucleotide primer 4 ccaatcgtgt cttatcctcg gg
22 5 23 DNA Artificial
oligonucleotide primer 5 ttccagacag aacccaagta gcc
23 6 24 DNA Artificial oligonucleotide primer 6
aacatttttc atcgcttttc ttag 24
7 24 DNA Artificial oligonucleotide primer 7 aaagcatttt tggcagtttc ttgt
24 8 25 DNA Artificial
oligonucleotide primer 8 tggacagaat ggagaaagaa atgtg
25 9 22 DNA Artificial oligonucleotide primer 9
ccaagccaaa caaagaatca ca 22
10 27 DNA Artificial oligonucleotide primer 10 gcttgctatg attgattgat
tgattga 27 11 25 DNA Artificial
oligonucleotide primer 11 agaacggtgt taagatagaa tagtt
25 12 26 DNA Artificial oligonucleotide primer 12
gggaagctta ataatgaaaa tgacac 26
13 22 DNA Artificial oligonucleotide primer 13 aaccgatgct tttttcgcct tt
22 14 23 DNA Artificial
oligonucleotide primer 14 gaagaaggct cagcaacacc ttg
23 15 23 DNA Artificial oligonucleotide primer 15
cacccaatct tgcctttgaa aca 23
16 26 DNA Artificial oligonucleotide primer 16 tagacacttt caggttaaaa
tataac 26 17 26 DNA Artificial
oligonucleotide primer 17 caacttgtca cttacacata gtttag
26 18 22 DNA Artificial oligonucleotide primer 18
atttcgctcg caaacacaag gt 22
19 27 DNA Artificial oligonucleotide primer 19 tcaagaaata caagtgcaag
aaagacc 27 20 22 DNA Artificial
oligonucleotide primer 20 aattttcgct tgtgaaccct gc
22 21 28 DNA Artificial oligonucleotide primer 21
ttcagtacaa gtcgggtgaa taataata 28
22 22 DNA Artificial oligonucleotide primer 22 tgggactctc acacacggaa aa
22 23 23 DNA Artificial
oligonucleotide primer 23 tgggagtcaa tttcccatta tca
23 24 28 DNA Artificial oligonucleotide primer 24
tggatttgaa accacaaata acaacaac 28
25 22 DNA Artificial oligonucleotide primer 25 aaaaggggga catttttcca cg
22 26 23 DNA Artificial
oligonucleotide primer 26 cattagggac aacaacagcg ttt
23 27 20 DNA Artificial oligonucleotide primer 27
cgtcgccatc actatgagaa 20
28 21 DNA Artificial oligonucleotide primer 28 aagggatccc tcaactgact g
21 29 22 DNA Artificial
oligonucleotide primer 29 ccatcccttc ctccaaatag at
22 30 22 DNA Artificial oligonucleotide primer 30
tgacgaagct tgaggttatt cg 22
31 23 DNA Artificial oligonucleotide primer 31 gccaagaaga taacaagctc ggc
23 32 26 DNA Artificial
oligonucleotide primer 32 gggttatatc agtttttctt tttgtt
26 33 21 DNA Artificial oligonucleotide primer 33
agatccaaca cctggcctaa t 21
34 27 DNA Artificial oligonucleotide primer 34 aacaaaacta gcccatagaa
acattga 27 35 25 DNA Artificial
oligonucleotide primer 35 gaaatgccca gaaaaaccta ataac
25 36 22 DNA Artificial oligonucleotide primer 36
gccgtgaaga cagttgatcg tt 22
37 23 DNA Artificial oligonucleotide primer 37 cagttgattt ttgtttttcg gca
23 38 22 DNA Artificial
oligonucleotide primer 38 ttgtccaaac ttgcagggaa ca
22 39 25 DNA Artificial oligonucleotide primer 39
tggatgagtt ctattggggt agtcg 25
40 22 DNA Artificial oligonucleotide primer 40 gacctcgctc tctgtttctc at
22 41 22 DNA Artificial
oligonucleotide primer 41 cgcaatccca attcacctct tc
22 42 24 DNA Artificial oligonucleotide primer 42
gaaccccgtt tcaacatttt atga 24
43 22 DNA Artificial oligonucleotide primer 43 ggtcgttaag cccactttga ga
22 44 24 DNA Artificial
oligonucleotide primer 44 tgcagctttc actaatcgtc agaa
24 45 23 DNA Artificial oligonucleotide primer 45
gggagtgtga acttacattg tct 23
46 22 DNA Artificial oligonucleotide primer 46 gcgatcatgt ctctgccatc ag
22 47 28 DNA Artificial
oligonucleotide primer 47 gaagggatat ggagagtgag aaattaga
28 48 26 DNA Artificial oligonucleotide primer 48
aaaaattgtg taaacgaaag atggga 26
49 22 DNA Artificial oligonucleotide primer 49 tgagcccaac ctccacatct tt
22 50 20 DNA Artificial
oligonucleotide primer 50 cactgctttt tcccctctct
20 51 29 DNA Artificial oligonucleotide primer 51
gcggtaaatg gtgaaaatca tttatggtt 29
52 26 DNA Artificial oligonucleotide primer 52 ggaatgcatg agtattaacc
tcttat 26 53 28 DNA Artificial
oligonucleotide primer 53 tgtcttccca atccatctaa tctaatca
28 54 27 DNA Artificial oligonucleotide primer 54
ccaattatgt ttctatgcca tcttgtt 27
55 26 DNA Artificial oligonucleotide primer 55 tgtggataaa agccatctct
aacaaa 26 56 24 DNA Artificial
oligonucleotide primer 56 tcgataccaa tcatccaatc caaa
24 57 22 DNA Artificial oligonucleotide primer 57
tccttagacc tctcgcgcaa ac 22
58 20 DNA Artificial oligonucleotide primer 58 tacccttaat caccggacaa
20 59 22 DNA Artificial
oligonucleotide primer 59 acgtatgcga aggagggaga tg
22 60 22 DNA Artificial oligonucleotide primer 60
ttggctcttg tgtaagtggc cc 22
61 22 DNA Artificial oligonucleotide primer 61 ttcagtccct tggtggttcc aa
22 62 22 DNA Artificial
oligonucleotide primer 62 catggtcctt gggtgcaatt ta
22 63 27 DNA Artificial oligonucleotide primer 63
aaaaatgtgt tagagcttgt gttgtta 27
64 22 DNA Artificial oligonucleotide primer 64 atggtgcagt gttgcaggtt gt
22 65 26 DNA Artificial
oligonucleotide primer 65 gaccacaact tcttttttgt gaatgg
26 66 24 DNA Artificial oligonucleotide primer 66
ccgtcaccgt taataaaata gcat 24
67 27 DNA Artificial oligonucleotide primer 67 agaagcatac tcgtcgtaac
actatcc 27 68 26 DNA Artificial
oligonucleotide primer 68 gagcaggaca ttttttttat ccttga
26 69 22 DNA Artificial oligonucleotide primer 69
tgcttgaggt tgaaggaaat gc 22
70 23 DNA Artificial oligonucleotide primer 70 tccttggggt gaattgtttt tca
23 71 27 DNA Artificial
oligonucleotide primer 71 gcgttgatca ggtcgatttt tatttgt
27 72 21 DNA Artificial oligonucleotide primer 72
aatgctttgg tcgttgcatt g 21
73 22 DNA Artificial oligonucleotide primer 73 cagcgtctgc atgatagcgt tt
22 74 28 DNA Artificial
oligonucleotide primer 74 tgcaaattgt tattgtgaga ctgaatga
28 75 20 DNA Artificial oligonucleotide primer 75
cgccagctag ctagtctcat 20
76 24 DNA Artificial oligonucleotide primer 76 cctccacccc cttcccaccc
aaaa 24 77 22 DNA Artificial
oligonucleotide primer 77 agcgtcgtct ggctaggtct gt
22 78 25 DNA Artificial oligonucleotide primer 78
gtcattggtc tcaaacaatc ttcat 25
79 25 DNA Artificial oligonucleotide primer 79 gcgactttct tttcaatttc
actcc 25 80 22 DNA Artificial
oligonucleotide primer 80 cactttttca ctgtctcccc cc
22 81 23 DNA Artificial oligonucleotide primer 81
tgggccattt gggcagctcg act 23
82 22 DNA Artificial oligonucleotide primer 82 gcgccccatt aatgttaaca ca
22 83 27 DNA Artificial
oligonucleotide primer 83 ggagagaatc catatatatt gaattgc
27 84 23 DNA Artificial oligonucleotide primer 84
ccttttgaca attatggcat ata 23
85 22 DNA Artificial oligonucleotide primer 85 cctgtcaaat tggctgatgc aa
22 86 27 DNA Artificial
oligonucleotide primer 86 ttttaccaaa caaattaaac tgcgtct
27 87 20 DNA Artificial oligonucleotide primer 87
ccggtctgac ctattctcat 20
88 23 DNA Artificial oligonucleotide primer 88 ggtgtttcaa gtttcaacac cat
23 89 23 DNA Artificial
oligonucleotide primer 89 atgacaagga aaaatcaatc aac
23 90 26 DNA Artificial oligonucleotide primer 90
gcggggtcaa ttagttttcg tcagtt 26
91 24 DNA Artificial oligonucleotide primer 91 gcgcaaaagg acgcccacca
atag 24 92 23 DNA Artificial
oligonucleotide primer 92 tctgcttatt cattgtgtgc gtg
23 93 23 DNA Artificial oligonucleotide primer 93
tggtatatgt ttttgcggga caa 23
94 22 DNA Artificial oligonucleotide primer 94 aggtgggcta ggagtgacca ca
22 95 27 DNA Artificial
oligonucleotide primer 95 gcgacatatt gcattaaaaa catactt
27 96 23 DNA Artificial oligonucleotide primer 96
ggggtgaatg aatgtcaaga tga 23
97 24 DNA Artificial oligonucleotide primer 97 gcggaattag aactccagta
aaga 24 98 27 DNA Artificial
oligonucleotide primer 98 ttagtggatc tacactaaac taatccg
27 99 22 DNA Artificial oligonucleotide primer 99
aggcatttct ggcaagggta tg 22
100 23 DNA Artificial oligonucleotide primer 100 gcgcccacac aacctttaat
ctt 23 101 22 DNA Artificial
oligonucleotide primer 101 atgggctatg ggaggaggtg tt
22 102 23 DNA Artificial oligonucleotide primer
102 ccagtgcagt tttacatgaa ctt
23 103 23 DNA Artificial oligonucleotide primer 103 gctggccttt
agaacgtctg act 23 104 24 DNA
Artificial oligonucleotide primer 104 ccacaaaaag atgaaacaaa atag
24 105 24 DNA Artificial
oligonucleotide primer 105 gcggagattg gaacctactc attc
24 106 22 DNA Artificial oligonucleotide primer
106 aacaatttga tggtcggggt tg
22 107 23 DNA Artificial oligonucleotide primer 107 caccgtcgta
aaaactgtgt cgt 23 108 24 DNA
Artificial oligonucleotide primer 108 gaagtgctag ttctcacggt gtgg
24 109 24 DNA Artificial
oligonucleotide primer 109 ttcgctgcat aatttttagc atca
24 110 27 DNA Artificial oligonucleotide primer
110 aaagagacac ccaaaagata acaaaca
27 111 19 DNA Artificial oligonucleotide primer 111 tgaccaccat gagttcatt
19 112 24 DNA Artificial
oligonucleotide primer 112 gcgggacgca aaattggatt tagt
24 113 22 DNA Artificial oligonucleotide primer
113 gtgaccctcc attccacaac aa
22 114 22 DNA Artificial oligonucleotide primer 114 gcagagtccc
ccctaaatat ag 22 115 20 DNA
Artificial oligonucleotide primer 115 gtgaatcatc cagggcttgc
20 116 24 DNA Artificial
oligonucleotide primer 116 gcgaatggtt tttgctggaa agta
24 117 27 DNA Artificial oligonucleotide primer
117 gcgttaagaa tgcatttatg tttagtc
27 118 20 DNA Artificial oligonucleotide primer 118 caagctcaag
cctcacacat 20 119 18 DNA
Artificial oligonucleotide primer 119 aattggagtg ggtcacac
18 120 27 DNA Artificial
oligonucleotide primer 120 gcgcccaagt attatgagat atttgat
27 121 27 DNA Artificial oligonucleotide primer
121 ctttgtttgg ttggtgataa gtttcta
27 122 22 DNA Artificial oligonucleotide primer 122 aattgttagg
gcgcgagaaa ca 22 123 22 DNA
Artificial oligonucleotide primer 123 gttcggaggg aggaaagtgt tg
22 124 24 DNA Artificial
oligonucleotide primer 124 tgttcaatcg acaaataagg gtgc
24 125 25 DNA Artificial oligonucleotide primer
125 ctttacttag taatggttcc cacag
25 126 29 DNA Artificial oligonucleotide primer 126 ccagatttta
cgttaatgtt ttagtttta 29 127 24 DNA
Artificial oligonucleotide primer 127 catacacgca ttgcctttcc tgaa
24 128 24 DNA Artificial
oligonucleotide primer 128 gcgtcccagg acatcatcat catc
24 129 23 DNA Artificial oligonucleotide primer
129 tgctgcttgt gtttggttga tct
23 130 22 DNA Artificial oligonucleotide primer 130 aatccccaaa
caaacgcaca gt 22 131 27 DNA
Artificial oligonucleotide primer 131 gcggcgcttt atgtaacaat acgattt
27 132 23 DNA Artificial
oligonucleotide primer 132 gcgagaaaat aatcctgctc aag
23 133 22 DNA Artificial oligonucleotide primer
133 tcgggagaca ctaaaggcac tg
22 134 23 DNA Artificial oligonucleotide primer 134 gcggcataag
ttttcatccc atc 23 135 25 DNA
Artificial oligonucleotide primer 135 gcggatatgc cacttctctc gtgac
25 136 25 DNA Artificial
oligonucleotide primer 136 ggagaaaaac atccaaagaa atgtg
25 137 29 DNA Artificial oligonucleotide primer
137 ggataatttt ttcatcatca caatttatc
29 138 27 DNA Artificial oligonucleotide primer 138 tggaaatact
gaatgaaagc atattgg 27 139 25 DNA
Artificial oligonucleotide primer 139 ccattgggat cgagaataat tgatg
25 140 22 DNA Artificial
oligonucleotide primer 140 agagtaatgg ctcctgctcc ga
22 141 26 DNA Artificial oligonucleotide primer
141 cgatctaaca cgcatatttc ctctga
26 142 23 DNA Artificial oligonucleotide primer 142 tgggggtcaa
ttttaatttg tgc 23 143 26 DNA
Artificial oligonucleotide primer 143 gattctcttc attctaatac tcgttt
26 144 28 DNA Artificial
oligonucleotide primer 144 gcgttacgtt tcactattta tttaacat
28 145 27 DNA Artificial oligonucleotide primer
145 gctggtgtat ggtgaaatca aattact
27 146 27 DNA Artificial oligonucleotide primer 146 tgatattgtt
ttgtgtgaaa gatgcac 27 147 24 DNA
Artificial oligonucleotide primer 147 tgctcaaagg gtcaatttct ttcc
24 148 25 DNA Artificial
oligonucleotide primer 148 ccaagcccat aaacgaaata aaaca
25 149 25 DNA Artificial oligonucleotide primer
149 cagtgctcat atcaaattaa agtgg
25 150 22 DNA Artificial oligonucleotide primer 150 tttcaaccac
caagccaacc tt 22 151 23 DNA
Artificial oligonucleotide primer 151 tgcagcatgt gttttaggct ttc
23 152 27 DNA Artificial
oligonucleotide primer 152 ccttagacca tgaatgtctc gaagata
27 153 24 DNA Artificial oligonucleotide primer
153 tggccatgtc aaaccataac aaca
24 154 22 DNA Artificial oligonucleotide primer 154 actggcgctg
accttaaatt gc 22 155 28 DNA
Artificial oligonucleotide primer 155 tgtttttaag taatccggtg aaatagca
28 156 26 DNA Artificial
oligonucleotide primer 156 tcacatcaca agtttcataa atgctg
26 157 22 DNA Artificial oligonucleotide primer
157 gcgtctccct taatcttcaa gc
22 158 25 DNA Artificial oligonucleotide primer 158 agattaagag
aagacgagag tttta 25 159 22 DNA
Artificial oligonucleotide primer 159 gccaggtgct cacttcttgc ta
22 160 26 DNA Artificial
oligonucleotide primer 160 caggagaaaa aaggaaaaga aaagca
26 161 26 DNA Artificial oligonucleotide primer
161 aggttttgct tcttgaaagt gaagag
26 162 24 DNA Artificial oligonucleotide primer 162 gtctccgttc
aaagcttctt cttc 24 163 22 DNA
Artificial oligonucleotide primer 163 gaggggaggt aaaaagtcgg ga
22 164 23 DNA Artificial
oligonucleotide primer 164 cgcttgggtt cagttacttg gtg
23 165 28 DNA Artificial oligonucleotide primer
165 gcgcccatta ttattttcct tgaattgt
28 166 28 DNA Artificial oligonucleotide primer 166 gcgaccatca
tctaatcaca atctacta 28 167 23 DNA
Artificial oligonucleotide primer 167 aatgcaagaa gcaatcagca aga
23 168 26 DNA Artificial
oligonucleotide primer 168 tggcaccctt gataaataag agaagg
26 169 22 DNA Artificial oligonucleotide primer
169 aattgaacca ctcagccaag cc
22 170 26 DNA Artificial oligonucleotide primer 170 gtctaagatt
ttaattaggg tgattc 26 171 27 DNA
Artificial oligonucleotide primer 171 gcgtataaag aaaaacgagc atatcat
27 172 27 DNA Artificial
oligonucleotide primer 172 caaattgaaa aacacaagaa aacacaa
27 173 25 DNA Artificial oligonucleotide primer
173 agcaactaac cttgggcttc agtaa
25 174 22 DNA Artificial oligonucleotide primer 174 cctggacttg
tttgctcatc aa 22 175 23 DNA
Artificial oligonucleotide primer 175 aattcctttg gcatttttgt tgc
23 176 24 DNA Artificial
oligonucleotide primer 176 tttcactttt gaatcattgc atcg
24 177 25 DNA Artificial oligonucleotide primer
177 caccactcgt atccttcaca agagc
25 178 26 DNA Artificial oligonucleotide primer 178 gcgcaattaa
aaggataact tatatc 26 179 22 DNA
Artificial oligonucleotide primer 179 aaaattcatg tcgctgcgtt ca
22 180 25 DNA Artificial
oligonucleotide primer 180 tgaaaaccat gtcaaagtaa tggca
25 181 23 DNA Artificial oligonucleotide primer
181 ggaaagtttt gttacatgcc gga
23 182 24 DNA Artificial oligonucleotide primer 182 ttttaactgg
agaaacctga ggga 24 183 24 DNA
Artificial oligonucleotide primer 183 gcgcgatggg ctgttggttt ttat
24 184 22 DNA Artificial
oligonucleotide primer 184 ttgcttgctg cacatgactg ag
22 185 22 DNA Artificial oligonucleotide primer
185 ttgaaatagt ggtggggttg gg
22 186 22 DNA Artificial oligonucleotide primer 186 tgtgttgtgt
ggtgtggtgg tc 22 187 23 DNA
Artificial oligonucleotide primer 187 caaagtcccc tttcacacct ttt
23 188 24 DNA Artificial
oligonucleotide primer 188 aaagggaaag gaagaataaa ccga
24 189 22 DNA Artificial oligonucleotide primer
189 tgctttttct ctttggcaac cc
22 190 26 DNA Artificial oligonucleotide primer 190 gcgcccaaaa
ctatctagta attctt 26 191 23 DNA
Artificial oligonucleotide primer 191 ccaacaacca aatcaatcac tgc
23 192 22 DNA Artificial
oligonucleotide primer 192 gctccggtcc ttcaactgac ct
22 193 22 DNA Artificial oligonucleotide primer
193 atgtgggtat gttgcaggca ga
22 194 25 DNA Artificial oligonucleotide primer 194 gttgggaaaa
ggttactacc atatc 25 195 24 DNA
Artificial oligonucleotide primer 195 cagccaagca aaagataaat aata
24 196 29 DNA Artificial
oligonucleotide primer 196 gcgctttcaa aaagtaacaa ttaatgaaa
29 197 22 DNA Artificial oligonucleotide primer
197 gacctttcat catcgtcccc ac
22 198 23 DNA Artificial oligonucleotide primer 198 tcaattcatc
acggaccagt tca 23 199 22 DNA
Artificial oligonucleotide primer 199 gtgagtttcg gtgctgtatt cc
22 200 24 DNA Artificial
oligonucleotide primer 200 cctaaattga tgaaaggata caag
24 201 21 DNA Artificial oligonucleotide primer
201 gtatcgttcg cgtcttgagt c
21 202 26 DNA Artificial oligonucleotide primer 202 ttaacgggaa
aaaattaaac ctacga 26 203 25 DNA
Artificial oligonucleotide primer 203 tggagattta atatagatgc cgcga
25 204 24 DNA Artificial
oligonucleotide primer 204 gcggttttgg attgactttg ttga
24 205 26 DNA Artificial oligonucleotide primer
205 ccgtgacttt tgttatcctg agttcc
26 206 22 DNA Artificial oligonucleotide primer 206 gcgaattggc
atacatagta cc 22 207 22 DNA
Artificial oligonucleotide primer 207 tgcaccacta atgtcctcag cc
22 208 22 DNA Artificial
oligonucleotide primer 208 gcaatgggta ttgatcgtgt ca
22 209 27 DNA Artificial oligonucleotide primer
209 gcgctaccgt gtggtggtgt gctacct
27 210 28 DNA Artificial oligonucleotide primer 210 gcgagtttcg
ccgttaccac ctcagctt 28 211 24 DNA
Artificial oligonucleotide primer 211 tgcgacttta ctgaaaacct ggaa
24 212 25 DNA Artificial
oligonucleotide primer 212 aacgtcttca agtcaagtgc ctaca
25 213 27 DNA Artificial oligonucleotide primer
213 gcgcatcaca agttttatag atgctga
27 214 29 DNA Artificial oligonucleotide primer 214 gggtacattt
tattaaaagt aaacacacc 29 215 28 DNA
Artificial oligonucleotide primer 215 tgaaccttgt ctgttgattt ttttatgt
28 216 26 DNA Artificial
oligonucleotide primer 216 tctcctactt ctctttctcc cgttca
26 217 25 DNA Artificial oligonucleotide primer
217 cctgattata tgtctagaca aacat
25 218 26 DNA Artificial oligonucleotide primer 218 gagatcacat
caaagtcaaa actgcc 26 219 27 DNA
Artificial oligonucleotide primer 219 gcgatttctt ccttgaagaa ttttctg
27 220 27 DNA Artificial
oligonucleotide primer 220 gcgaattatc caaagataca cttagtc
27 221 22 DNA Artificial oligonucleotide primer
221 atatcgaaaa tcgcgcatct gg
22 222 25 DNA Artificial oligonucleotide primer 222 gcgcattaag
gcataaaaaa ggata 25 223 25 DNA
Artificial oligonucleotide primer 223 gcgccaatat tatcatgctt tatgt
25 224 25 DNA Artificial
oligonucleotide primer 224 agtggtcgtc acaacactat catat
25 225 22 DNA Artificial oligonucleotide primer
225 ctcctcctgc gcaacaacaa ta
22 226 22 DNA Artificial oligonucleotide primer 226 gcgtggaatg
agacgtaacc aa 22 227 26 DNA
Artificial oligonucleotide primer 227 caaaacttat gtgcaacacg acttca
26 228 23 DNA Artificial
oligonucleotide primer 228 aatgtagcaa tttgactggc gaa
23 229 24 DNA Artificial oligonucleotide primer
229 gcggatctaa ggataattca ttaa
24 230 21 DNA Artificial oligonucleotide primer 230 gctatgggaa
aaggatgtgt g 21 231 22 DNA
Artificial oligonucleotide primer 231 atgtgatggc atgtgaaatg gt
22 232 26 DNA Artificial
oligonucleotide primer 232 cagggtatag ttcaattcag tgagcg
26 233 25 DNA Artificial oligonucleotide primer
233 cctctttgtt ggtggttaag tctcc
25 234 25 DNA Artificial oligonucleotide primer 234 gcggcaaaac
tttggagtat tgcaa 25 235 25 DNA
Artificial oligonucleotide primer 235 tcgtcaatta agcaaaaatg tgaga
25 236 22 DNA Artificial
oligonucleotide primer 236 gaatatcacg cgagaatttt ac
22 237 22 DNA Artificial oligonucleotide primer
237 acaaaaagaa atcgaaccgg ca
22 238 24 DNA Artificial oligonucleotide primer 238 tggctttgat
gatgtttgag acaa 24 239 25 DNA
Artificial oligonucleotide primer 239 cccagataca gacaataaaa cccga
25 240 26 DNA Artificial
oligonucleotide primer 240 tccaccatgt aatatgtgaa gtggat
26 241 21 DNA Artificial oligonucleotide primer
241 ctcacaccct ttcattatct a
21 242 25 DNA Artificial oligonucleotide primer 242 actctatttt
attatcgtgc aagaa 25 243 21 DNA
Artificial oligonucleotide primer 243 gatgacaacg taggctaaaa a
21 244 23 DNA Artificial
oligonucleotide primer 244 gattctatat ccatcgtgtt gct
23 245 23 DNA Artificial oligonucleotide primer
245 cccactgtat ccttagtgtg cca
23 246 18 DNA Artificial oligonucleotide primer 246 ggctaacccg ctctatgt
18 247 25 DNA Artificial
oligonucleotide primer 247 cattaactaa taagttgttg gtagc
25 248 21 DNA Artificial oligonucleotide primer
248 accaaattgc ttcacgcatc c
21 249 22 DNA Artificial oligonucleotide primer 249 tgctcatgtg
gtcctaccca ga 22 250 27 DNA
Artificial oligonucleotide primer 250 gcggagcatg taaatccagc ctattga
27 251 26 DNA Artificial
oligonucleotide primer 251 gcggatttcg atttgaatat acttac
26 252 25 DNA Artificial oligonucleotide primer
252 tggacacaca caaacaccta cagaa
25 253 23 DNA Artificial oligonucleotide primer 253 agcaagtctt
gagtcttttg tct 23 254 22 DNA
Artificial oligonucleotide primer 254 tcccacgtca tatccactgc tc
22 255 23 DNA Artificial
oligonucleotide primer 255 ccaaagctga gcagctgata act
23 256 25 DNA Artificial oligonucleotide primer
256 ccggtgatac tccataccaa taaca
25 257 24 DNA Artificial oligonucleotide primer 257 ccgagctaac
aaaggcgacc aaat 24 258 27 DNA
Artificial oligonucleotide primer 258 gcgcccaaac ctattaaggt atgaaca
27 259 22 DNA Artificial
oligonucleotide primer 259 atggccgtct caaaagaact gg
22 260 22 DNA Artificial oligonucleotide primer
260 gcgaatggtt gctcaaataa tc
22 261 26 DNA Artificial oligonucleotide primer 261 gcgcgcattt
tttaagttaa tgttct 26 262 22 DNA
Artificial oligonucleotide primer 262 ggcagactcg tagagcaatt ta
22 263 27 DNA Artificial
oligonucleotide primer 263 gcgaagattg gtcttttatg tcaaatg
27 264 27 DNA Artificial oligonucleotide primer
264 gcggtaactc ctcgagtccc tctcaat
27 265 22 DNA Artificial oligonucleotide primer 265 tggtgatggg
aagcaaacaa ga 22 266 22 DNA
Artificial oligonucleotide primer 266 ccatcccttc gtccaccaaa ta
22 267 27 DNA Artificial
oligonucleotide primer 267 tgctgcagcg tgtctgtagt ataattt
27 268 26 DNA Artificial oligonucleotide primer
268 cgatttgaat atacttaccg tctata
26 269 26 DNA Artificial oligonucleotide primer 269 gtaacattgg
ttgtcatctt tgtcta 26 270 26 DNA
Artificial oligonucleotide primer 270 ggaggttatc tagtggtata gatggt
26 271 22 DNA Artificial
oligonucleotide primer 271 ccaaaagcat tgagagtggg ga
22 272 22 DNA Artificial oligonucleotide primer
272 gaaacccgaa acgcaaaatc tc
22 273 22 DNA Artificial oligonucleotide primer 273 ctctcgccgg
tacaaaacac ct 22 274 28 DNA
Artificial oligonucleotide primer 274 tcactctcac aacttcaatc tctttctc
28 275 22 DNA Artificial
oligonucleotide primer 275 cacgactctt cctcttcctc cg
22 276 16 DNA Artificial oligonucleotide primer
276 ccctgtgttt ccctct
16 277 22 DNA Artificial oligonucleotide primer 277 gtgttgctct
tgggaatctg gc 22 278 17 DNA
Artificial oligonucleotide primer 278 tctcgactca cgactca
17 279 28 DNA Artificial
oligonucleotide primer 279 aaaatgaaaa cacacagaga gagagaga
28 280 22 DNA Artificial oligonucleotide primer
280 agcgaaatgt gtggtccaag gt
22 281 22 DNA Artificial oligonucleotide primer 281 cgtcgagaaa
ggaagagagg ca 22 282 22 DNA
Artificial oligonucleotide primer 282 gcggaaacca ttctgacgga ta
22 283 23 DNA Artificial
oligonucleotide primer 283 ggaggaactt ggaaggcata tca
23 284 21 DNA Artificial oligonucleotide primer
284 acggacaacg ctggcactaa g
21 285 20 DNA Artificial oligonucleotide primer 285 caaaaggttt
tcagtggtgg 20 286 22 DNA
Artificial oligonucleotide primer 286 ccaatcccta ccctgattgc ac
22 287 22 DNA Artificial
oligonucleotide primer 287 ggaagtggat ttggagttgg ga
22 288 22 DNA Artificial oligonucleotide primer
288 aacccataac ctgctcgtct ca
22 289 22 DNA Artificial oligonucleotide primer 289 aggcgggttt
ggagctgttt ac 22 290 25 DNA
Artificial oligonucleotide primer 290 tcttctcagg aacttgaatt actca
25 291 22 DNA Artificial
oligonucleotide primer 291 ggaatgtccc aagtgtcagc aa
22 292 28 DNA Artificial oligonucleotide primer
292 tgaaaataag tggaaagaaa aaggaaaa
28 293 22 DNA Artificial oligonucleotide primer 293 gtccccgttc
ctcaaacacc tt 22 294 24 DNA
Artificial oligonucleotide primer 294 tgcacactca cttggtctta caca
24 295 22 DNA Artificial
oligonucleotide primer 295 ctgttggaaa atgatgcgtg gc
22 296 21 DNA Artificial oligonucleotide primer
296 ttgatcactt ctgtaacatt c
21 297 22 DNA Artificial oligonucleotide primer 297 ctttgttgtt
tggtcaaggc cc 22 298 25 DNA
Artificial oligonucleotide primer 298 tgccgaaata agtgagagca tagaa
25 299 27 DNA Artificial
oligonucleotide primer 299 tgttaggtat gggatttagg gttttga
27 300 25 DNA Artificial oligonucleotide primer
300 tccactcact gtatgtcttc ccttg
25 301 23 DNA Artificial oligonucleotide primer 301 ttgagtgctc
ttggtgtttt cct 23 302 26 DNA
Artificial oligonucleotide primer 302 ccaaacaccc cagtataaaa aatgga
26 303 22 DNA Artificial
oligonucleotide primer 303 ggattttgag ttgctccaag gc
22 304 24 DNA Artificial oligonucleotide primer
304 tgtatgatcc atcctctgaa accg
24 305 24 DNA Artificial oligonucleotide primer 305 tcacatgttc
gggacttaag gtat 24 306 25 DNA
Artificial oligonucleotide primer 306 tgaatggcta aaaacgtgat ttgga
25 307 22 DNA Artificial
oligonucleotide primer 307 ggaccttccc tttcccatga ct
22 308 22 DNA Artificial oligonucleotide primer
308 agcgatggtt gaagaaaggg tc
22 309 22 DNA Artificial oligonucleotide primer 309 agagaacggg
cgatttatgg ct 22 310 22 DNA
Artificial oligonucleotide primer 310 atgtcgccac taggccaatc ag
22 311 24 DNA Artificial
oligonucleotide primer 311 ccatcttgag cagagtttga agtt
24 312 25 DNA Artificial oligonucleotide primer
312 gtggtggtgg tgaaaactat tagaa
25 313 25 DNA Artificial oligonucleotide primer 313 cttccacacc
ctagtttagt gacaa 25 314 22 DNA
Artificial oligonucleotide primer 314 tcaagcttat gctctatagg ct
22 315 24 DNA Artificial
oligonucleotide primer 315 gaccaaaatt caaggcagtg acaa
24 316 20 DNA Artificial oligonucleotide primer
316 ccatcctcgt tagcatctat
20 317 21 DNA Artificial oligonucleotide primer 317 gctgcacaat
tcattccatt t 21 318 25 DNA
Artificial oligonucleotide primer 318 cccagggact tacttttttc agttt
25 319 26 DNA Artificial
oligonucleotide primer 319 tgaagcaaca aaatagagga atagag
26 320 25 DNA Artificial oligonucleotide primer
320 cattactagg cgtgtgttgt ttcaa
25 321 22 DNA Artificial oligonucleotide primer 321 cacgcgcatc
agctttgtag ag 22 322 24 DNA
Artificial oligonucleotide primer 322 tcttttcacc attctccaac ctca
24 323 23 DNA Artificial
oligonucleotide primer 323 tcctttctcc catttttttt ggg
23 324 22 DNA Artificial oligonucleotide primer
324 ggtgaccacc cctattcctt at
22 325 22 DNA Artificial oligonucleotide primer 325 aattgttagg
gcgcgagaaa ca 22 326 22 DNA
Artificial oligonucleotide primer 326 ctagccaagg gagagaggag ca
22 327 22 DNA Artificial
oligonucleotide primer 327 cctcacacca accagcacaa aa
22 328 27 DNA Artificial oligonucleotide primer
328 caattttatg atttgctttt gaaggga
27 329 21 DNA Artificial oligonucleotide primer 329 gggccttgaa
ttttgtgcta t 21 330 22 DNA
Artificial oligonucleotide primer 330 cctcttgaaa ccgtgaaacc gt
22 331 24 DNA Artificial
oligonucleotide primer 331 tctttttctt catttttgtt cgca
24 332 23 DNA Artificial oligonucleotide primer
332 aaactgctct gaagtgcgac aca
23 333 25 DNA Artificial oligonucleotide primer 333 tgtgaataaa
agaaatcccc attga 25 334 25 DNA
Artificial oligonucleotide primer 334 aagatacccc caacattatt tgtaa
25 335 26 DNA Artificial
oligonucleotide primer 335 gcgttttcat acggtgtttt gcctat
26 336 27 DNA Artificial oligonucleotide primer
336 gggctaacga acatgtaact tatcaac
27 337 28 DNA Artificial oligonucleotide primer 337 gcactacaat
atgtgcatga atttttct 28 338 23 DNA
Artificial oligonucleotide primer 338 tctccttact cactccattg gca
23 339 22 DNA Artificial
oligonucleotide primer 339 tccatgtcac caaccacaaa aa
22 340 22 DNA Artificial oligonucleotide primer
340 cgtatccctt tcccatacgt gg
22 341 22 DNA Artificial oligonucleotide primer 341 atgacgggat
gaaagagcca aa 22 342 25 DNA
Artificial oligonucleotide primer 342 agggaactaa cacatttaat catca
25 343 26 DNA Artificial
oligonucleotide primer 343 cggaaaagat ataccgagtg agaaaa
26 344 24 DNA Artificial oligonucleotide primer
344 ttctaaggat ttgtggtaat cggc
24 345 23 DNA Artificial oligonucleotide primer 345 catggagaaa
agaagaggag gga 23 346 25 DNA
Artificial oligonucleotide primer 346 aagcaaacca ttatcttcat tggtg
25 347 22 DNA Artificial
oligonucleotide primer 347 gccacactat tccagccact ac
22 348 22 DNA Artificial oligonucleotide primer
348 agtagtccaa accgaaacgc ca
22 349 22 DNA Artificial oligonucleotide primer 349 tcaattcggt
caaaaggctt ga 22 350 24 DNA
Artificial oligonucleotide primer 350 ctcccccaaa tttaacctta aaga
24 351 23 DNA Artificial
oligonucleotide primer 351 aatggatgga cctgtcagtc tgc
23 352 25 DNA Artificial oligonucleotide primer
352 tgcttccatt agtctctcat cctcc
25 353 26 DNA Artificial oligonucleotide primer 353 tggaaaaaga
taacgtgtgt ttgcag 26 354 22 DNA
Artificial oligonucleotide primer 354 ccttcttttg acatggggcc ta
22 355 27 DNA Artificial
oligonucleotide primer 355 gcgagtgcca actaactact tttatga
27 356 23 DNA Artificial oligonucleotide primer
356 tggtatcggg agattttttc agc
23 357 28 DNA Artificial oligonucleotide primer 357 tttttcgtca
gcacattatt acacattt 28 358 23 DNA
Artificial oligonucleotide primer 358 ggccagtgtt gagggattta gaa
23 359 24 DNA Artificial
oligonucleotide primer 359 aatttgctcc agtgttttaa gttt
24 360 21 DNA Artificial oligonucleotide primer
360 ggtgatatcg cgctaaaatt a
21 361 22 DNA Artificial oligonucleotide primer 361 ggaaaccctg
tcattttcgt gc 22 362 22 DNA
Artificial oligonucleotide primer 362 agagttctca gtccgccagc tc
22 363 21 DNA Artificial
oligonucleotide primer 363 gcgcaattgt caccaacaca t
21 364 22 DNA Artificial oligonucleotide primer
364 cccaaaccaa caagcaacaa ca
22 365 23 DNA Artificial oligonucleotide primer 365 attcacacgc
atctggaata ata 23 366 23 DNA
Artificial oligonucleotide primer 366 gcggagttca acgcattcac ctt
23 367 23 DNA Artificial
oligonucleotide primer 367 tgaaagcaaa cgagactcat gga
23 368 23 DNA Artificial oligonucleotide primer
368 gcatagaagg gcatcattca gat
23 369 25 DNA Artificial oligonucleotide primer 369 gctagatgag
cagaccattg cactt 25 370 27 DNA
Artificial oligonucleotide primer 370 caagaggttg ttgtaagagt gatctcg
27 371 20 DNA Artificial
oligonucleotide primer 371 cacggcgtat ttttattttg
20 372 24 DNA Artificial oligonucleotide primer
372 cagtgcatgg ttttctcatg tacc
24 373 20 DNA Artificial oligonucleotide primer 373 gctgctgtta
agagtgtttg 20 374 25 DNA
Artificial oligonucleotide primer 374 gcgcacggta tataatcgaa cctat
25 375 26 DNA Artificial
oligonucleotide primer 375 gcggtgatcg gatgttatag tttcag
26 376 24 DNA Artificial oligonucleotide primer
376 tgtctccatg ctgtaacacg tcaa
24 377 22 DNA Artificial oligonucleotide primer 377 cgccaaagat
gcaacacact tg 22 378 26 DNA
Artificial oligonucleotide primer 378 tgtaattgct gttttggttt catttc
26 379 29 DNA Artificial
oligonucleotide primer 379 gcggactaat tctattttac accaacaac
29 380 24 DNA Artificial oligonucleotide primer
380 gcgcgaggta tcaacacaat tact
24 381 21 DNA Artificial oligonucleotide primer 381 gcgaggccaa
cattgaaaag t 21 382 28 DNA
Artificial oligonucleotide primer 382 cgtaccattg gacaactctg taattcaa
28 383 25 DNA Artificial
oligonucleotide primer 383 tggtgaatca atctcctcta agtgc
25 384 20 DNA Artificial oligonucleotide primer
384 gcggcgactg ttaacgtgtc
20 385 22 DNA Artificial oligonucleotide primer 385 ccacacggga
ctttccattt tc 22 386 23 DNA
Artificial oligonucleotide primer 386 tatatgtaat gacccccatc atg
23 387 22 DNA Artificial
oligonucleotide primer 387 cgttggattc gactttttgg ga
22 388 23 DNA Artificial oligonucleotide primer
388 ttgaagctca ggctgtgatg aat
23 389 21 DNA Artificial oligonucleotide primer 389 gcggggacca
aaaattcaaa a 21 390 22 DNA
Artificial oligonucleotide primer 390 taggggaacc gatttggtga aa
22 391 24 DNA Artificial
oligonucleotide primer 391 gcgtgtcaaa gagttttaga catc
24 392 22 DNA Artificial oligonucleotide primer
392 ttggagggag gttttggaag gt
22 393 27 DNA Artificial oligonucleotide primer 393 ccaatctttt
tgttagttca ccttcca 27 394 24 DNA
Artificial oligonucleotide primer 394 ttcgtagcaa tgtcaccacc ttgt
24 395 19 DNA Artificial
oligonucleotide primer 395 gggggtggct tttagattc
19 396 24 DNA Artificial oligonucleotide primer
396 gcgccgaata aaacgtgaga actg
24 397 23 DNA Artificial oligonucleotide primer 397 tccttgcctt
ttagttgttc ggt 23 398 23 DNA
Artificial oligonucleotide primer 398 cgggaacaac cacactctcc att
23 399 26 DNA Artificial
oligonucleotide primer 399 tccttcactt tcaaaaacaa aaacaa
26 400 24 DNA Artificial oligonucleotide primer
400 gcgcaacgac attttcacga agtt
24 401 25 DNA Artificial oligonucleotide primer 401 gcgagttttt
ggttggattg agttg 25 402 22 DNA
Artificial oligonucleotide primer 402 tgaccagagt ccaaagttca tc
22 403 18 DNA Artificial
oligonucleotide primer 403 ttcccggaaa gaaagaaa
18 404 23 DNA Artificial oligonucleotide primer
404 gcgataattt taaaactgga cca
23 405 25 DNA Artificial oligonucleotide primer 405 aagcagttcc
tctcatcacg taaca 25 406 22 DNA
Artificial oligonucleotide primer 406 cgcaatccca attcacctct tc
22 407 26 DNA Artificial
oligonucleotide primer 407 ccataaaaca tagcaactgt cgtctc
26 408 24 DNA Artificial oligonucleotide primer
408 tcccagggta aaagttcaag ttca
24 409 23 DNA Artificial oligonucleotide primer 409 tcgatgtttc
tccctccctt aga 23 410 22 DNA
Artificial oligonucleotide primer 410 ttgcacatgg tccttggttg at
22 411 26 DNA Artificial
oligonucleotide primer 411 gcgaatggga atgccttctt attcta
26 412 24 DNA Artificial oligonucleotide primer
412 gcgtagcgtg ttattttgtg tttg
24 413 22 DNA Artificial oligonucleotide primer 413 catatcctgc
ccccccaatt at 22 414 27 DNA
Artificial oligonucleotide primer 414 tgacatctaa gtccagaaat caaagca
27 415 25 DNA Artificial
oligonucleotide primer 415 gcgagtaaaa gcagagtgcg gagta
25 416 27 DNA Artificial oligonucleotide primer
416 gcgtttaagt ccaataacaa agataac
27 417 23 DNA Artificial oligonucleotide primer 417 tcgttgacac
acaaaaaaag cga 23 418 22 DNA
Artificial oligonucleotide primer 418 atcgggtcat gacttttgaa ga
22 419 25 DNA Artificial
oligonucleotide primer 419 gcggaatagt tgccaaacaa taatc
25 420 27 DNA Artificial oligonucleotide primer
420 caagtggatt gacacactaa ggtttga
27 421 25 DNA Artificial oligonucleotide primer 421 cacaaaagac
aggagatgtg agcaa 25 422 25 DNA
Artificial oligonucleotide primer 422 ttgtagacga cttgtggttc gattc
25 423 27 DNA Artificial
oligonucleotide primer 423 aatgaagaaa atgtgaattt gaaacca
27 424 27 DNA Artificial oligonucleotide primer
424 ttcccttttt ctaactctcc tttttca
27 425 25 DNA Artificial oligonucleotide primer 425 tgtcttggtg
caatacctga cattt 25 426 25 DNA
Artificial oligonucleotide primer 426 atttcccttt cacccacctc tgttt
25 427 22 DNA Artificial
oligonucleotide primer 427 caacgatccc agctcacagt tt
22 428 24 DNA Artificial oligonucleotide primer
428 gcggcaggct agctacatca agag
24 429 27 DNA Artificial oligonucleotide primer 429 tttcaaacaa
ggaagaaacc tcttttt 27 430 22 DNA
Artificial oligonucleotide primer 430 aagtaacact gtggcggcat cc
22 431 28 DNA Artificial
oligonucleotide primer 431 tgtgtaattt ctatcacctt attgtgcc
28 432 22 DNA Artificial oligonucleotide primer
432 tcctttggct cggcctatgt aa
22 433 22 DNA Artificial oligonucleotide primer 433 cgcggactca
gttaaaccgt at 22 434 26 DNA
Artificial oligonucleotide primer 434 cgttcaatag ttcctatgat ggacga
26 435 22 DNA Artificial
oligonucleotide primer 435 gcattgcacg tcgattttag gg
22 436 26 DNA Artificial oligonucleotide primer
436 cttaaggaca cgtggaagat gactac
26 437 26 DNA Artificial oligonucleotide primer 437 gcgttgaagc
cgcctacaaa tataat 26 438 28 DNA
Artificial oligonucleotide primer 438 tccttttaat tctaacattg agacagca
28 439 27 DNA Artificial
oligonucleotide primer 439 tctgtcccaa aaaagaaaga agatatg
27 440 27 DNA Artificial oligonucleotide primer
440 caatctttaa tgctctggag tttgaga
27 441 26 DNA Artificial oligonucleotide primer 441 gcgtgtgacg
gttcaaaatg atagtt 26 442 25 DNA
Artificial oligonucleotide primer 442 gggtaattta tctgtgtgat tgttc
25 443 28 DNA Artificial
oligonucleotide primer 443 ttgcttggga ttttcatttt attatagg
28 444 25 DNA Artificial oligonucleotide primer
444 ggtcaaagaa taagggattt gcctc
25 445 25 DNA Artificial oligonucleotide primer 445 tgggactttg
atttttggaa taccc 25 446 24 DNA
Artificial oligonucleotide primer 446 ccagagaaat tattggagtg gcaa
24 447 22 DNA Artificial
oligonucleotide primer 447 gcgcaaagaa attcccatcc ta
22 448 22 DNA Artificial oligonucleotide primer
448 gggagaggtt cagttgggga at
22 449 30 DNA Artificial oligonucleotide primer 449 gcgaatgcct
aatgtgatga taaaaaaata 30 450 28 DNA
Artificial oligonucleotide primer 450 tccccatcat ttatcgaaaa taataatt
28 451 22 DNA Artificial
oligonucleotide primer 451 aataatggag tgacccgctg ct
22 452 22 DNA Artificial oligonucleotide primer
452 tggtacgagt ggcacgaaat gt
22 453 22 DNA Artificial oligonucleotide primer 453 gccatccttt
cctttcaacc aa 22 454 22 DNA
Artificial oligonucleotide primer 454 tgtaggctat tggaagggtg cg
22 455 28 DNA Artificial
oligonucleotide primer 455 gcgcttataa aggcttgtga aagacact
28 456 22 DNA Artificial oligonucleotide primer
456 tggttagtgc aatcttggcg gt
22 457 23 DNA Artificial oligonucleotide primer 457 tgcacccatc
aatcacattt ttg 23 458 25 DNA
Artificial oligonucleotide primer 458 gcggttcaag gcttcaagta gtcac
25 459 22 DNA Artificial
oligonucleotide primer 459 ttttttctca cacccatgcc gt
22 460 22 DNA Artificial oligonucleotide primer
460 ccggttggcg taatgtgtct tt
22 461 22 DNA Artificial oligonucleotide primer 461 gccagcagcc
tgttcagttt tt 22 462 26 DNA
Artificial oligonucleotide primer 462 cccctctttg gccctcacac cttctc
26 463 24 DNA Artificial
oligonucleotide primer 463 atttgaagct cttggtatct tggc
24 464 27 DNA Artificial oligonucleotide primer
464 caaccatgat aaatgtgagt gagcttg
27 465 22 DNA Artificial oligonucleotide primer 465 gtcacaattt
cacgatccca aa 22 466 23 DNA
Artificial oligonucleotide primer 466 tccattttcc ctttagtcca acg
23 467 28 DNA Artificial
oligonucleotide primer 467 gcgcatacga tttggcattt ttctattg
28 468 28 DNA Artificial oligonucleotide primer
468 aatttcttac gtttccatag atttctcg
28 469 22 DNA Artificial oligonucleotide primer 469 tgccctcttc
ccaaactagg gt 22 470 23 DNA
Artificial oligonucleotide primer 470 caaccactga ttcaagccaa caa
23 471 27 DNA Artificial
oligonucleotide primer 471 caatttaagc acggtcatat tttctca
27 472 23 DNA Artificial oligonucleotide primer
472 cgtgatgcag tgaatttttt tcg
23 473 26 DNA Artificial oligonucleotide primer 473 gggttacttt
tctttatcct cctcca 26 474 26 DNA
Artificial oligonucleotide primer 474 gggctatcaa attgactaaa gccaaa
26 475 26 DNA Artificial
oligonucleotide primer 475 acacttgaat catcgagagt tgctaa
26 476 22 DNA Artificial oligonucleotide primer
476 tgtgctgctt gcttcaattt gg
22 477 26 DNA Artificial oligonucleotide primer 477 tggcctgtct
tatattaccg aaccaa 26 478 26 DNA
Artificial oligonucleotide primer 478 ggtccgtatg caattcttga ctaata
26 479 24 DNA Artificial
oligonucleotide primer 479 tcccccacaa gagaacaaga aggt
24 480 27 DNA Artificial oligonucleotide primer
480 gcgggaattg gttaatctca tcgtgac
27 481 22 DNA Artificial oligonucleotide primer 481 tgcgaaaaag
cagagtgacc aa 22 482 23 DNA
Artificial oligonucleotide primer 482 tgagcatatt ttgatccgat gcc
23 483 25 DNA Artificial
oligonucleotide primer 483 gttgctttgt tatgtaatgg aagtc
25 484 24 DNA Artificial oligonucleotide primer
484 gccccacaaa tattcagaag gtaa
24 485 24 DNA Artificial oligonucleotide primer 485 gcagcggtgt
agttcgttct ttct 24 486 22 DNA
Artificial oligonucleotide primer 486 aaatcgtctt ttgtggctgc ct
22 487 24 DNA Artificial
oligonucleotide primer 487 gcaccatgtt ctttttccat caaa
24 488 24 DNA Artificial oligonucleotide primer
488 ggctcaatta gagcatgcaa catc
24 489 22 DNA Artificial oligonucleotide primer 489 catgttaaac
gtccacccac ca 22 490 22 DNA
Artificial oligonucleotide primer 490 gcgtgaattc gcctaagttt at
22 491 28 DNA Artificial
oligonucleotide primer 491 tccctactct cgtgtcgtta gttatttt
28 492 28 DNA Artificial oligonucleotide primer
492 agttacatta tttttgtctt tctgccgt
28 493 24 DNA Artificial oligonucleotide primer 493 gcgcaagtgg
ccagctcatc tatt 24 494 28 DNA
Artificial oligonucleotide primer 494 ccctcttatt tcaccctaag acctacaa
28 495 24 DNA Artificial
oligonucleotide primer 495 ggaaatgctt caaaccaaca aaca
24 496 26 DNA Artificial oligonucleotide primer
496 gcccacatag ttttcatttt tctcca
26 497 24 DNA Artificial oligonucleotide primer 497 gaggtctagt
gctttggtaa ggtt 24 498 26 DNA
Artificial oligonucleotide primer 498 tgtcacacaa ccagtgtctc aaaatc
26 499 22 DNA Artificial
oligonucleotide primer 499 cacaccccag gacccattaa ga
22 500 23 DNA Artificial oligonucleotide primer
500 aaagcgcaca caatgcaaat aca
23 501 25 DNA Artificial oligonucleotide primer 501 caaggttacg
aactgctcga ataag 25 502 22 DNA
Artificial oligonucleotide primer 502 tgaggaggca agatgatcca aa
22 503 27 DNA Artificial
oligonucleotide primer 503 gcgctttttc ggctgttatt tttaact
27 504 27 DNA Artificial oligonucleotide primer
504 gcgggtctta cgaacgtgtc acattat
27 505 22 DNA Artificial oligonucleotide primer 505 cgaacccaaa
ccacaaagca ta 22 506 21 DNA
Artificial oligonucleotide primer 506 gcacaatgac aatcacatac a
21 507 25 DNA Artificial
oligonucleotide primer 507 gcgtgtaaaa atctttgaat cttga
25 508 27 DNA Artificial oligonucleotide primer
508 cacccattat tgaaaataca aggacca
27 509 19 DNA Artificial oligonucleotide primer 509 gggggatcta ggccatgac
19 510 28 DNA Artificial
oligonucleotide primer 510 gcataatggt ctaataaaag tggagacc
28 511 24 DNA Artificial oligonucleotide primer
511 tgcttttgga ggaactttgt ctca
24 512 22 DNA Artificial oligonucleotide primer 512 cattcaaccg
tgattgcgaa ga 22 513 22 DNA
Artificial oligonucleotide primer 513 gggagcggat cattcggtga aa
22 514 21 DNA Artificial
oligonucleotide primer 514 gagctacccg agatgatact c
21 515 24 DNA Artificial oligonucleotide primer
515 ggatcaaatt gggaaacaca aagg
24 516 22 DNA Artificial oligonucleotide primer 516 ctcacataca
tggcagccgt aa 22 517 22 DNA
Artificial oligonucleotide primer 517 tgcctttagc tggtgggaaa aa
22 518 22 DNA Artificial
oligonucleotide primer 518 gcgcgcaaca atcactagta cg
22 519 23 DNA Artificial oligonucleotide primer
519 ttagaggttt tcggatgagc gtg
23 520 22 DNA Artificial oligonucleotide primer 520 tatatgcgaa
ccctcttaca at 22 521 22 DNA
Artificial oligonucleotide primer 521 gttttggttg atccgcattg gt
22 522 23 DNA Artificial
oligonucleotide primer 522 tttcattacc gcatgttctt gga
23 523 25 DNA Artificial oligonucleotide primer
523 ttatgttcgt tcatctctga agcct
25 524 26 DNA Artificial oligonucleotide primer 524 ttctgtccat
tctagctcac taaccc 26 525 21 DNA
Artificial oligonucleotide primer 525 aaatcgcgca tctaaattta c
21 526 25 DNA Artificial
oligonucleotide primer 526 acaataactt gttttgcaca ctatt
25 527 24 DNA Artificial oligonucleotide primer
527 gcgcccacat gattttgtac tgat
24 528 25 DNA Artificial oligonucleotide primer 528 tatggtaaat
attaaccatt gtcct 25 529 22 DNA
Artificial oligonucleotide primer 529 cctcgtttta ttccgaaagc cg
22 530 19 DNA Artificial
oligonucleotide primer 530 gggccatgca cctgctact
19 531 23 DNA Artificial oligonucleotide primer
531 ttagattcgg acaccggtct act
23 532 27 DNA Artificial oligonucleotide primer 532 tcttaatttt
tttaggaatg gcatcaa 27 533 26 DNA
Artificial oligonucleotide primer 533 cgctatccct ttgtattttc ttttgc
26 534 27 DNA Artificial
oligonucleotide primer 534 gcgggctaac ttatgttact aaacaat
27 535 26 DNA Artificial oligonucleotide primer
535 cctgtggctg ttatactatg catata
26 536 22 DNA Artificial oligonucleotide primer 536 gggtggcgtt
gacaatgttt ta 22 537 25 DNA
Artificial oligonucleotide primer 537 ttattatcta aacttatatg tgcat
25 538 24 DNA Artificial
oligonucleotide primer 538 gcctcctaag tccgtacaca gcat
24 539 25 DNA Artificial oligonucleotide primer
539 ccctcactcc tagattattt gttgt
25 540 23 DNA Artificial oligonucleotide primer 540 ggatttggtt
tctgtgtgct gtg 23 541 18 DNA
Artificial oligonucleotide primer 541 ggggcacaag ccacactt
18 542 27 DNA Artificial
oligonucleotide primer 542 gcgggtcaga agatgctacc aaactct
27 543 22 DNA Artificial oligonucleotide primer
543 tgggcacttg cagtccaaat ag
22 544 23 DNA Artificial oligonucleotide primer 544 gcgcaaaccg
cacaagttta tgt 23 545 24 DNA
Artificial oligonucleotide primer 545 gcgcgagtta gcgaattatt tgtc
24 546 24 DNA Artificial
oligonucleotide primer 546 tgttgaaatt gaccaaaatt ccca
24 547 26 DNA Artificial oligonucleotide primer
547 gcggaggaat acaagtctct attcaa
26 548 23 DNA Artificial oligonucleotide primer 548 gcgccgctaa
cagacatcca ata 23 549 23 DNA
Artificial oligonucleotide primer 549 tggatttcac ccaagaaaaa agc
23 550 22 DNA Artificial
oligonucleotide primer 550 tcgactaccc gtcgattccg ta
22 551 23 DNA Artificial oligonucleotide primer
551 ggcacaacca tcaccacctt att
23 552 26 DNA Artificial oligonucleotide primer 552 cacaatacct
gtggctgtta tactat 26 553 22 DNA
Artificial oligonucleotide primer 553 atcgaactgt gaccgtccct tc
22 554 24 DNA Artificial
oligonucleotide primer 554 aagggaagag agtcgtgctt cttt
24 555 21 DNA Artificial oligonucleotide primer
555 ccagcaaacc cccaggtaaa g
21 556 22 DNA Artificial oligonucleotide primer 556 gaagaaaacg
cgaataaccc ca 22 557 23 DNA
Artificial oligonucleotide primer 557 gcacgcagac tcaagttcat tca
23 558 22 DNA Artificial
oligonucleotide primer 558 aatagttggg tcgtcgaagg gg
22 559 25 DNA Artificial oligonucleotide primer
559 tccaacttaa cacaagatca gcgaa
25 560 22 DNA Artificial oligonucleotide primer 560 gaaaagtttt
atgttctgag tg 22 561 25 DNA
Artificial oligonucleotide primer 561 ccacacacac tgacacagta aagca
25 562 17 DNA Artificial
oligonucleotide primer 562 ccaaggtctc tcagagg
17 563 22 DNA Artificial oligonucleotide primer
563 acggaggcac ttcccattgt ta
22 564 26 DNA Artificial oligonucleotide primer 564 caaaacgaca
tgacaaggaa acttca 26 565 28 DNA
Artificial oligonucleotide primer 565 tcctccataa aaattaaaaa cattggaa
28 566 22 DNA Artificial
oligonucleotide primer 566 gcccccagac acaacataat ca
22 567 22 DNA Artificial oligonucleotide primer
567 cgagagaagc agaagcagag gc
22 568 22 DNA Artificial oligonucleotide primer 568 gagaaaggtg
acgatggacg ct 22 569 36 DNA
Artificial oligonucleotide primer 569 catggtttct cttatcttat agacattgtt
gccaag 36 570 36 DNA Artificial
oligonucleotide primer 570 caattcatgg tttctcttat agacattgtt gccaag
36 571 28 DNA Artificial oligonucleotide primer
571 cactgtccct gctcctgttt caagtatc
28 572 31 DNA Artificial oligonucleotide primer 572 caattcttgt
gggttgaagc cttgttctga c 31 573 30 DNA
Artificial oligonucleotide primer 573 ggaatcaact tcttcgtgag tgggttgttc
30 574 28 DNA Artificial
oligonucleotide primer 574 tggtggctat ggaaatctca tgtgtgga
28 575 33 DNA Artificial oligonucleotide primer
575 ctctcattta ccaaactcca acatttgatc acc
33 576 29 DNA Artificial oligonucleotide primer 576 cacactggta
gatgggaagc aagaatagg 29 577 32 DNA
Artificial oligonucleotide primer 577 gaagaagatt ccacccagat catcatcagt
ag 32 578 26 DNA Artificial
oligonucleotide primer 578 gcttgtgcag ctccaatcgg tgtaac
26 579 27 DNA Artificial oligonucleotide primer
579 gtgcaatcca agacatctgg ttcggac
27 580 31 DNA Artificial oligonucleotide primer 580 cagctaaacc
ttacaaggat gattggtcaa g 31 581 28 DNA
Artificial oligonucleotide primer 581 ccctggactg aagttgccat aatgtatc
28 582 23 DNA Artificial
oligonucleotide primer 582 gctggttggg agaaagcact tcc
23 583 30 DNA Artificial oligonucleotide primer
583 gaatctaaca ttacgctctg ctggagtatc
30 584 28 DNA Artificial oligonucleotide primer 584 gagggactat
gtgaaatgga gaggagtg 28 585 31 DNA
Artificial oligonucleotide primer 585 gtatgctaaa agaggagact tgactggtga g
31 586 32 DNA Artificial
oligonucleotide primer 586 gatgaaggaa ccaacacttg cataacaatt tg
32 587 27 DNA Artificial oligonucleotide primer
587 tgtcagcact cctcactcat ttgccga
27 588 29 DNA Artificial oligonucleotide primer 588 cacttaacag
gagtgctcct gatcaccag 29 589 36 DNA
Artificial oligonucleotide primer 589 gagaacaagg acaaatcaat aggtgagacg
aagaaa 36 590 28 DNA Artificial
oligonucleotide primer 590 ctcatctgct cagaaccttc agtcagtc
28 591 33 DNA Artificial oligonucleotide primer
591 cggatcatgt ctagtacatt agagatgctt gtg
33 592 32 DNA Artificial oligonucleotide primer 592 ctacacttct
aatgcctatt taggtgtgct tg 32 593 32 DNA
Artificial oligonucleotide primer 593 gtcatatcta gggagatttc taaccagttg
tc 32 594 27 DNA Artificial
oligonucleotide primer 594 taggcagcgt gacaaactga gcatagg
27 595 30 DNA Artificial oligonucleotide primer
595 gggttgatgt ccgatgggta aatgaagttg
30 596 36 DNA Artificial oligonucleotide primer 596 gcactaatac
tttagttgac ttttgaggtg gttgag 36 597 37 DNA
Artificial oligonucleotide primer 597 gctatgtggt agaagtatat gaaaaggtag
atgacag 37 598 37 DNA Artificial
oligonucleotide primer 598 gaatatacta gcttgatgcc tatttgtttc taaaccc
37 599 33 DNA Artificial oligonucleotide primer
599 agcagtcata caatgctctt tattgtggtg aag
33 600 33 DNA Artificial oligonucleotide primer 600 gctcatagcc
tgcttcttaa tcttgttatt ctg 33 601 35 DNA
Artificial oligonucleotide primer 601 ctatttgttt caggagtttc acaaccatct
caagt 35 602 30 DNA Artificial
oligonucleotide primer 602 catcttgacc agtcaccaat ctgagtacag
30 603 30 DNA Artificial oligonucleotide primer
603 ggcttcagtg agaaaggtgg atcaaatgga
30 604 32 DNA Artificial oligonucleotide primer 604 caacgtgtaa
aatcaagaga ttgagcttct gg 32 605 30 DNA
Artificial oligonucleotide primer 605 tcggtgtgtt caactatgac tttggttctg
30 606 36 DNA Artificial
oligonucleotide primer 606 cccccaacaa aactaaaaat agaaccctca acaacc
36 607 35 DNA Artificial oligonucleotide primer
607 ggtccaagaa cattatgatc ttgaacaatc ttcac
35 608 30 DNA Artificial oligonucleotide primer 608 tggctagagc
atcaacacat ctataccttc 30 609 33 DNA
Artificial oligonucleotide primer 609 cattgggcat gattcttgaa tagccttttt
acc 33 610 30 DNA Artificial
oligonucleotide primer 610 gagggctatg ttttcttctc cagatgtgag
30 611 26 DNA Artificial oligonucleotide primer
611 aaggtcggct tggtggttaa aggcag
26 612 33 DNA Artificial oligonucleotide primer 612 gccttttatg
cacatttttc ctggggatct aac 33 613 37 DNA
Artificial oligonucleotide primer 613 ctcaatgtca tgggatcaat ttggaaattc
aatgacc 37 614 37 DNA Artificial
oligonucleotide primer 614 gattacatta attaccgcta tgactatatc ttgggac
37 615 32 DNA Artificial oligonucleotide primer
615 gctaccttct ccattgcttc tatgtattgg tc
32 616 32 DNA Artificial oligonucleotide primer 616 gggcatcaca
tacatgaaaa caactacact tg 32 617 27 DNA
Artificial oligonucleotide primer 617 caacagctct cttccaccac aatcctg
27 618 33 DNA Artificial
oligonucleotide primer 618 gagattggaa attgtagctc tctttacttg ctg
33 619 35 DNA Artificial oligonucleotide primer
619 ctttgaggac ttatttggtt gttataggca tttgg
35 620 30 DNA Artificial oligonucleotide primer 620 gtgtttccct
ccatttttgc caaaagacag 30 621 31 DNA
Artificial oligonucleotide primer 621 tatacacact aagaattcgc tcgctgtaca a
31 622 28 DNA Artificial
oligonucleotide primer 622 ggtctagact ttcactcaga caaggaac
28 623 38 DNA Artificial oligonucleotide primer
623 caaaatacta cagacctaat ttgtaactaa ttgctccc
38 624 29 DNA Artificial oligonucleotide primer 624 gggaagaaga
agaacactcg gtacagtag 29 625 37 DNA
Artificial oligonucleotide primer 625 aagctaggaa atccacactc aaattatcga
cttgtgt 37 626 27 DNA Artificial
oligonucleotide primer 626 gacctgaagc aaaacgccac catttcc
27 627 26 DNA Artificial oligonucleotide primer
627 cgttggagac gacgccgttt gattac
26 628 22 DNA Artificial oligonucleotide primer 628 cagtcgacac
gtcttctact cc 22 629 21 DNA
Artificial oligonucleotide primer 629 tggaaggcat gtcggaactt g
21 630 30 DNA Artificial
oligonucleotide primer 630 cctttttgcc ctcacttcat gccttctatg
30 631 27 DNA Artificial oligonucleotide primer
631 gcataaccca agagctggac tgacaag
27 632 28 DNA Artificial oligonucleotide primer 632 caagctctgc
tgccaggtta agtgtttc 28 633 32 DNA
Artificial oligonucleotide primer 633 gactagaaca aattggggct agtgtgtttg
ag 32 634 26 DNA Artificial
oligonucleotide primer 634 tggacttgcg ggactatgcc ttagag
26 635 28 DNA Artificial oligonucleotide primer
635 gcaggaacac gttcgtaacc atcaactg
28 636 32 DNA Artificial oligonucleotide primer 636 gtccctttct
gaaccactta aagagtcaac ag 32 637 30 DNA
Artificial oligonucleotide primer 637 ggcatagtga gttgaatacc aggaggaatc
30 638 27 DNA Artificial
oligonucleotide primer 638 attgaagggt gggcgttacc aggttac
27 639 32 DNA Artificial oligonucleotide primer
639 gagttaatgg ggctatgcta ttggctattc ac
32 640 36 DNA Artificial oligonucleotide primer 640 gtcactatat
ggagtcaagg taattattgt gttcac 36 641 38 DNA
Artificial oligonucleotide primer 641 ggtctagagt ttgaatatta gtaatgactt
gtattgac 38 642 35 DNA Artificial
oligonucleotide primer 642 gcatattgtg ccatagagag agaaaatgta gtaag
35 643 34 DNA Artificial oligonucleotide primer
643 cgctactgca agttatcagt caagagatta ttcc
34 644 30 DNA Artificial oligonucleotide primer 644 gccgtgtaag
cgtgtttacc aatctagttg 30 645 32 DNA
Artificial oligonucleotide primer 645 caaaactgag cgaaacttgt gttgggagaa
ag 32 646 31 DNA Artificial
oligonucleotide primer 646 gtggggactc tttattcgaa gtttgctgaa c
31 647 36 DNA Artificial oligonucleotide primer
647 ctcatgtaac caactctcta tgaagtttga gatcca
36 648 33 DNA Artificial oligonucleotide primer 648 ctctaatcgg
atttggtgtt tcacttcggt aag 33 649 29 DNA
Artificial oligonucleotide primer 649 gatggctgtc attgctacag aggagtatc
29 650 38 DNA Artificial
oligonucleotide primer 650 gtgactccaa aggaaagaga aatgtttctt aaatcatc
38 651 32 DNA Artificial oligonucleotide primer
651 ggatagcaag tcaatttcat gccttgtgat ag
32 652 34 DNA Artificial oligonucleotide primer 652 gcaggacatg
aagatgtact tagtgaatgt gaag 34 653 27 DNA
Artificial oligonucleotide primer 653 actggaagag ggtgcttagg gaatctg
27 654 30 DNA Artificial
oligonucleotide primer 654 gagaatctag tctaccacca taccacgaac
30 655 35 DNA Artificial oligonucleotide primer
655 gggactctct ttatattgga aggtataact cagtg
35 656 33 DNA Artificial oligonucleotide primer 656 gttggaccct
tgtaattaga cccgaaacaa atg 33 657 30 DNA
Artificial oligonucleotide primer 657 atctgtccaa cgatctctcc atgttcattc
30 658 33 DNA Artificial
oligonucleotide primer 658 cgaataagaa gttgggtatc acttacacgt tgg
33 659 27 DNA Artificial oligonucleotide primer
659 gatggggttc tagactggga tctggat
27 660 32 DNA Artificial oligonucleotide primer 660 cttttcctac
aggattgtca ggcttatcgt ca 32 661 27 DNA
Artificial oligonucleotide primer 661 gaagagtaac agagtctacg caccgac
27 662 37 DNA Artificial
oligonucleotide primer 662 gtcaacgaac atactatgca tgatgatttc tgattag
37 663 28 DNA Artificial oligonucleotide primer
663 catatgcaga cagcaggcta aggaactc
28 664 38 DNA Artificial oligonucleotide primer 664 gcagcaatat
aaccaggatt cagaattaat ctagttag 38 665 30 DNA
Artificial oligonucleotide primer 665 cgatggggtt gacttagaaa tggcatatac
30 666 28 DNA Artificial
oligonucleotide primer 666 gcaccaattc ctcaagcata ctccaaac
28 667 30 DNA Artificial oligonucleotide primer
667 ctgacaaggt gttttggtag ggagagattc
30 668 32 DNA Artificial oligonucleotide primer 668 gcatcctccg
ttactccaat cagagtttcc at 32 669 30 DNA
Artificial oligonucleotide primer 669 ctgacaaggt gttttggtag ggagagattc
30 670 32 DNA Artificial
oligonucleotide primer 670 gcatcctccg ttactccaat cagagtttcc at
32 671 32 DNA Artificial oligonucleotide primer
671 cacactatca acacctattg gtgaccattg ta
32 672 33 DNA Artificial oligonucleotide primer 672 ggagggtgct
tatgtaaatg atgtaaagac cat 33 673 35 DNA
Artificial oligonucleotide primer 673 gatgaccatt ttgattccct catgctatta
gtacc 35 674 33 DNA Artificial
oligonucleotide primer 674 ccaataagtt agcagcatgt ggatcacagt gta
33 675 30 DNA Artificial oligonucleotide primer
675 gtctctcgga gttgcttcaa ttgctcatac
30 676 36 DNA Artificial oligonucleotide primer 676 gagagtgtgg
aattgtaagt cattgattga aaactc 36 677 30 DNA
Artificial oligonucleotide primer 677 gtctctcgga gttgcttcaa ttgctcatac
30 678 36 DNA Artificial
oligonucleotide primer 678 gagagtgtgg aattgtaagt cattgattga aaactc
36 679 26 DNA Artificial oligonucleotide primer
679 gcagaaggag cattgaggct ttccag
26 680 32 DNA Artificial oligonucleotide primer 680 gaaaaggttt
gttatgcttc gtactctgtc tc 32 681 33 DNA
Artificial oligonucleotide primer 681 catgaagctc caccatttgc tagtacatga
aac 33 682 33 DNA Artificial
oligonucleotide primer 682 ccagagttac caaaccatct gtgagaaata tcc
33 683 33 DNA Artificial oligonucleotide primer
683 catgaagctc caccatttgc tagtacatga aac
33 684 33 DNA Artificial oligonucleotide primer 684 ccagagttac
caaaccatct gtgagaaata tcc 33 685 28 DNA
Artificial oligonucleotide primer 685 caaacgctcc caacagcttc agaatctc
28 686 28 DNA Artificial
oligonucleotide primer 686 ttgaaggttg taagagtctc ggtcgtcg
28 687 30 DNA Artificial oligonucleotide primer
687 cccattcttc atgtactcat acaccaagag
30 688 33 DNA Artificial oligonucleotide primer 688 gcagccacca
ataatttctc atttgacaac aag 33 689 31 DNA
Artificial oligonucleotide primer 689 ctctgtatat gayatatgtg ctcagtgcct c
31 690 32 DNA Artificial
oligonucleotide primer 690 gacttaccaa atgagtttga ccaggtttta cc
32 691 30 DNA Artificial oligonucleotide primer
691 aggattcaac ctctagccat gatgatgttg
30 692 28 DNA Artificial oligonucleotide primer 692 ccaagctctt
tccgtgtgta tcaatctg 28 693 12 DNA
Artificial oligonucleotide probe 693 gtttcagata ac
12 694 12 DNA Artificial
oligonucleotide probe 694 gaatgacttt ga
12 695 13 DNA Artificial oligonucleotide probe
695 tcattctttc atg
13 696 12 DNA Artificial oligonucleotide probe 696 tctccagaaa ca
12 697 10 DNA Artificial
oligonucleotide probe 697 gtgatccgtg
10 698 13 DNA Artificial oligonucleotide probe
698 atcttttcag gtt
13 699 13 DNA Artificial oligonucleotide probe 699 ggattacata cta
13 700 13 DNA Artificial
oligonucleotide probe 700 tttgaagctt tat
13 701 13 DNA Artificial oligonucleotide probe
701 aagaatcttc cta
13 702 15 DNA Artificial oligonucleotide probe 702 aatctaaaat ttagt
15 703 14 DNA Artificial
oligonucleotide probe 703 actaaattta tacc
14 704 11 DNA Artificial oligonucleotide probe
704 attaggggca g
11 705 12 DNA Artificial oligonucleotide probe 705 actatagttc gc
12 706 12 DNA Artificial
oligonucleotide probe 706 aacctttctg tc
12 707 12 DNA Artificial oligonucleotide probe
707 ttgaggattt ag
12 708 12 DNA Artificial oligonucleotide probe 708 tctcaacttg ga
12 709 12 DNA Artificial
oligonucleotide probe 709 ttcagtcaaa cc
12 710 13 DNA Artificial oligonucleotide probe
710 ttcttttgtg aca
13 711 12 DNA Artificial oligonucleotide probe 711 tgtgacaacc ga
12 712 14 DNA Artificial
oligonucleotide probe 712 aatctttttt aaag
14 713 13 DNA Artificial oligonucleotide probe
713 tcatctgtga taa
13 714 12 DNA Artificial oligonucleotide probe 714 aagagaaggc ta
12 715 13 DNA Artificial
oligonucleotide probe 715 atataagtaa ggg
13 716 10 DNA Artificial oligonucleotide probe
716 gcatgtcgac
10 717 13 DNA Artificial oligonucleotide probe 717 ttggaagtta tac
13 718 11 DNA Artificial
oligonucleotide probe 718 ggcatgtgag t
11 719 14 DNA Artificial oligonucleotide probe
719 attaacagta aagt
14 720 15 DNA Artificial oligonucleotide probe 720 tatctatgta tatta
15 721 11 DNA Artificial
oligonucleotide probe 721 tgggaatgat g
11 722 15 DNA Artificial oligonucleotide probe
722 taaataaata agatg
15 723 13 DNA Artificial oligonucleotide probe 723 aaaaaaaatg agg
13 724 12 DNA Artificial
oligonucleotide probe 724 tcaatgttgg at
12 725 13 DNA Artificial oligonucleotide probe
725 ttgtgaccaa tat
13 726 11 DNA Artificial oligonucleotide probe 726 atcaagccca a
11 727 11 DNA Artificial
oligonucleotide probe 727 agaagctcgt g
11 728 12 DNA Artificial oligonucleotide probe
728 gaaaaagaaa gg
12 729 14 DNA Artificial oligonucleotide probe 729 acatataata gtag
14 730 14 DNA Artificial
oligonucleotide probe 730 ttttgtatct gtat
14 731 12 DNA Artificial oligonucleotide probe
731 ttaacttgcc ag
12 732 14 DNA Artificial oligonucleotide probe 732 aatgataatt tagt
14 733 13 DNA Artificial
oligonucleotide probe 733 gaatgaattt ttc
13 734 11 DNA Artificial oligonucleotide probe
734 acactgctta c
11 735 13 DNA Artificial oligonucleotide probe 735 acaactaata agg
13 736 14 DNA Artificial
oligonucleotide probe 736 ttctgataaa aaaa
14 737 11 DNA Artificial oligonucleotide probe
737 tgtaatgcgt g
11 738 13 DNA Artificial oligonucleotide probe 738 gacaatctaa aaa
13 739 15 DNA Artificial
oligonucleotide probe 739 taataataat tgtgt
15 740 14 DNA Artificial oligonucleotide probe
740 ttccttcttt tttt
14 741 11 DNA Artificial oligonucleotide probe 741 ggaacgttac c
11 742 14 DNA Artificial
oligonucleotide probe 742 aattttttag tatg
14 743 13 DNA Artificial oligonucleotide probe
743 ttatagacac ttg
13 744 14 DNA Artificial oligonucleotide probe 744 ataaaccata tatg
14 745 14 DNA Artificial
oligonucleotide probe 745 ttaattacct taag
14 746 11 DNA Artificial oligonucleotide probe
746 atgccatttt g
11 747 14 DNA Artificial oligonucleotide probe 747 taatatctta tgca
14 748 12 DNA Artificial
oligonucleotide probe 748 agtgcgatga aa
12 749 12 DNA Artificial oligonucleotide probe
749 gaggagatgt ag
12 750 14 DNA Artificial oligonucleotide probe 750 taaaattgtt ggtt
14 751 14 DNA Artificial
oligonucleotide probe 751 tgaagaaaaa tatg
14 752 11 DNA Artificial oligonucleotide probe
752 ctgtccacta a
11 753 11 DNA Artificial oligonucleotide probe 753 agaggcagtg a
11 754 12 DNA Artificial
oligonucleotide probe 754 gtttctgata ac
12 755 13 DNA Artificial oligonucleotide probe
755 gaatgatttt gac
13 756 13 DNA Artificial oligonucleotide probe 756 tcattcattc atg
13 757 10 DNA Artificial
oligonucleotide probe 757 gtttcgggag
10 758 12 DNA Artificial oligonucleotide probe
758 atcacgaatc ac
12 759 13 DNA Artificial oligonucleotide probe 759 tatggagtaa ttg
13 760 14 DNA Artificial
oligonucleotide probe 760 tggattttat acta
14 761 14 DNA Artificial oligonucleotide probe
761 tttgaatctt tatc
14 762 14 DNA Artificial oligonucleotide probe 762 aaagaatatt ccta
14 763 14 DNA Artificial
oligonucleotide probe 763 atctaacatt tagt
14 764 13 DNA Artificial oligonucleotide probe
764 ggtatacatt tag
13 765 11 DNA Artificial oligonucleotide probe 765 attagggggc a
11 766 12 DNA Artificial
oligonucleotide probe 766 actatacttc gc
12 767 11 DNA Artificial oligonucleotide probe
767 accttgctgt c
11 768 12 DNA Artificial oligonucleotide probe 768 ttgagcattt ag
12 769 13 DNA Artificial
oligonucleotide probe 769 tctcaatttg gaa
13 770 12 DNA Artificial oligonucleotide probe
770 ttcagttaaa cc
12 771 12 DNA Artificial oligonucleotide probe 771 ttctttggtg ac
12 772 11 DNA Artificial
oligonucleotide probe 772 gtgacacaac c
11 773 14 DNA Artificial oligonucleotide probe
773 aatcttcttt aaag
14 774 13 DNA Artificial oligonucleotide probe 774 tcatgtgtga taa
13 775 12 DNA Artificial
oligonucleotide probe 775 aagagatggc ta
12 776 14 DNA Artificial oligonucleotide probe
776 aatataaata aggg
14 777 10 DNA Artificial oligonucleotide probe 777 gcatctcgac
10 778 13 DNA Artificial
oligonucleotide probe 778 ttggaagata tac
13 779 10 DNA Artificial oligonucleotide probe
779 ggcttgcgag
10 780 15 DNA Artificial oligonucleotide probe 780 tattaacatt aaagt
15 781 16 DNA Artificial
oligonucleotide probe 781 tatctatata tattaa
16 782 11 DNA Artificial oligonucleotide probe
782 atcatcccca a
11 783 14 DNA Artificial oligonucleotide probe 783 aaataagtaa gatg
14 784 13 DNA Artificial
oligonucleotide probe 784 aaaaaatatg agg
13 785 13 DNA Artificial oligonucleotide probe
785 tcaatgatgg ata
13 786 13 DNA Artificial oligonucleotide probe 786 atattgatca caa
13 787 11 DNA Artificial
oligonucleotide probe 787 tgggtttgat c
11 788 10 DNA Artificial oligonucleotide probe
788 gaaggtcgtg
10 789 13 DNA Artificial oligonucleotide probe 789 gaaaaaaaaa gga
13 790 13 DNA Artificial
oligonucleotide probe 790 acatatagta gta
13 791 12 DNA Artificial oligonucleotide probe
791 tttgtggctg ta
12 792 12 DNA Artificial oligonucleotide probe 792 ttaactagcc ag
12 793 13 DNA Artificial
oligonucleotide probe 793 aatgatcatt tag
13 794 13 DNA Artificial oligonucleotide probe
794 aatgaacttt ttc
13 795 12 DNA Artificial oligonucleotide probe 795 tttttgctag ag
12 796 13 DNA Artificial
oligonucleotide probe 796 tacaactaag gta
13 797 13 DNA Artificial oligonucleotide probe
797 tctgatgaaa aaa
13 798 11 DNA Artificial oligonucleotide probe 798 tgtaacgcgt g
11 799 13 DNA Artificial
oligonucleotide probe 799 gacaatttaa aaa
13 800 13 DNA Artificial oligonucleotide probe
800 aataatgatt gtg
13 801 13 DNA Artificial oligonucleotide probe 801 tccttccttt ttt
13 802 12 DNA Artificial
oligonucleotide probe 802 tggaacatta cc
12 803 14 DNA Artificial oligonucleotide probe
803 aatttttgag tatg
14 804 12 DNA Artificial oligonucleotide probe 804 tataggcact tg
12 805 14 DNA Artificial
oligonucleotide probe 805 ataaacctta tatg
14 806 14 DNA Artificial oligonucleotide probe
806 ttaattatct taag
14 807 12 DNA Artificial oligonucleotide probe 807 atgccctttt gt
12 808 13 DNA Artificial
oligonucleotide probe 808 aatatcgtat gca
13 809 12 DNA Artificial oligonucleotide probe
809 agtgcactga aa
12 810 12 DNA Artificial oligonucleotide probe 810 agaggagatg ta
12 811 14 DNA Artificial
oligonucleotide probe 811 taaaattatt ggtt
14 812 13 DNA Artificial oligonucleotide probe
812 gaacaaaaag atg
13 813 12 DNA Artificial oligonucleotide probe 813 actgtcaact aa
12 814 11 DNA Artificial
oligonucleotide probe 814 agagccagtg a
11 815 13 DNA Artificial oligonucleotide probe
815 tcatctctga taa
13 816 14 DNA Artificial oligonucleotide probe 816 ttggaatata tact
14 817 14 DNA Artificial
oligonucleotide probe 817 agatgttaga gtta
14 818 12 DNA Artificial oligonucleotide probe
818 tgtaacgcat gt
12 819 13 DNA Artificial oligonucleotide probe 819 attttttggt atg
13
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