Isolation and characterization of simple sequence repeat markers for the tetraploid forage grass Dactylis glomerata

With 1 table Abstract To isolate simple sequence repeat (SSR) markers from Dactylis glomerata , four SSR‐enriched genomic libraries for different repeat motifs from genomic DNA of a Japanese orchardgrass variety, ‘Akimidori II’, were constructed. A total of 969 unique SSR‐containing clones were identified and used for primer design. A BLAST search revealed that 63 of the sequences showed significant homology with sequences of other plant species including 18 that showed significant gene homology. After screening of a panel consisting of eight orchardgrass individuals, 606 SSR markers showed polymorphism, 70 generated monomorphic bands and the other primers either generated multiple bands or did not provide any amplification product. In the 606 polymorphic SSR markers, most had a large number of alleles, ranging from 1 to 13 with an average of 4.5 alleles per SSR primer pair. The number of alleles per individual ranged from 1 to 4. These SSR markers developed in this study will have broad utility for genotyping applications in this species, including gene targeting, quantitative trait locus mapping, variety or species identification and marker‐assisted selection.