Isolation and characterization of 12 microsatellite markers for the vulnerable subshrub Comesperma polygaloides (Polygalaceae)

We isolated 12 new polymorphic microsatellite markers from Comesperma polygaloides, a rare and vulnerable subshrub from southeastern Australia. 454 pyrosequencing was used to identify 81,804 possible primer pairs. Of 96 pairs, 32 amplified a product of expected size in the target species. Subsequently, 12 loci were screened in five populations of C. polygaloides and one population each of C. ericinum and C. calymega. All 12 loci were polymorphic and showed a diploid pattern. Observed and expected heterozygosity were calculated (0.04-0.71 and 0.04-0.76, respectively). Alleles per locus ranged from 2 to 14 (mean = 4.9). Cross species amplification was successful for three primer pairs. These loci will improve understanding of genetic structure and enhance long-term viability of a species under state protection.