Development of AFLP and STS markers for identifying wheat-rye translocations possessing 2RL

Wheat-rye translocation line (2BS/2RL) has been developed for resistance to biotype L of Hessian fly and agronomically useful traits. AFLP analysis us ing 64 primer pairs was conducted in order to identify 2RL-specific polymor phisms between `Coker 797' (non-2RL), near-isogenic line (NIL) carrying 2RL , and `Hamlet'. Nine primer combinations identified twelve reproducible pol ymorphic fragments in the NIL carrying 2RL. These twelve fragments were clo ned and sequenced with an aim towards converting AFLP markers into sequence tagged sites (STS). A comparison of the 12 sequences with non-redundant ac cessions in the NCBI database using the BLAST search option indicated that one fragment of approximately 200 bp in length (amplified using primer comb ination E+AAC / M+CTA) was highly homologous with the rye-specific repetiti ve sequence R173-1 and Wis-2-1A, a retrotransposon-like element in wheat. T wo STS primers (SJ07 and SJ09) out of twelve STS primer sets enabled the de tection of polymorphisms between Coker 797 and NIL carrying 2RL. In order t o verify whether the polymorphism detected by primers SJ07 and SJ09 was in fact the result of the presence of 2RL, additional plant material was exami ned. Amplified products of about 260 bp fragment with the SJ07 primer set w ere generated in rye cvs. `Chilbohomil' and `Jochunhomil', triticale experi mental line Suwon 15, and wheat experimental line K-14 (1AL/1RS & 2BS/2RL), as well as NIL carrying 2RL and Hamlet, but not in Coker 797 (non-2RL), `K eumgangmil' (non-translocation wheat), KS92WGRC17 (PI592729 /;/ 6BS/6BL-6RL ), KS92WGRC19 (P1592731 /;/ 4BS/4BL-6RL), `TAM200' (1AL/1RS), and `Siouxlan d' (1BL/1RS). Our data suggest that primer set SJ07 amplifies a `2RL-specif ic' fragment of diagnostic value.