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EXPANSION OF OAT-BASED MICROSATELLITE RESOURCES USING BOTH DIRECT AND INDIRECT STRATEGIES Eric Jackson, J. Anderson, M. Carson, D. Obert, G. Hu, and M. Bonman. USDA ARS Aberdeen, Molecular Genetics Laboratory Robert Campbell – Biological Technician
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EXPANSION OF OAT-BASED MICROSATELLITE RESOURCES USING BOTH DIRECT AND INDIRECT STRATEGIES Eric Jackson, J. Anderson, M. Carson, D. Obert, G. Hu, and M. Bonman USDA ARS Aberdeen, Molecular Genetics Laboratory Robert Campbell – Biological Technician Irene Shackelford – Biological Technician Indirect Use of barley and wheat SSR primer sets 146 barley 210 wheat Direct Development of oat-based SSR enriched libraries AC ATT ATG CATC
Polymorphic 3/6 3/6 3/6 5/6 4/6 3/6 3/6 3/6 3/6 2/6 3/6 2/6 TAM O-405 TAM O-301 MN-Chong MN841810 MN-Rines A.m.-dom Ogle1040 Ogle157 Noble-2 A. m-wt Makuru Kanota Otana Stout SunII
SSR Markers Co-dominant SSR Dominant SSR Dominant deg-SSR
Polymorphic markers: AB_AM_17 Dom. AB_AM_20 Dom. AB_AB_28 Co-dom. AB_AM_29 Dom. AB_AM_31 Dom. AB_AM_32 Dom. AB_AM_35 Co-dom. AB_AM_53 Dom.
Enough candidate SSR markers have been identified, so now the difficult task of validation and mapping will begin. CONCULSIONS 31% that amplified between Ogle and TAM O-301 contained degenerative SSR motifs, while one complete SSR motif was identified. 78.6% sequenced from the AG- and ATG- libraries contained SSR motifs for which working primers could be designed. 25.5% of the markers were polymorphic between Ogle and TAM O-301 (agarose) and 43% of those were polymorphic. Sequencing of the two libraries has yielded approximately 1700 candidate SSR primer sets. 1336 working primers 300 polymorphic in OT 130 co-dominant