Isolation, via 454 sequencing, characterization and transferability of microsatellites for Grevillea thelemanniana subsp. thelemanniana and cross-species amplification in the Grevillea thelemanniana complex (Proteaceae).

A set of 12 polymorphic microsatellite markers were developed for Grevillea thelemanniana subsp. thelemanniana, a rare and threatened shrub, endemic to southwest Western Australia. Genomic sequences were obtained from next generation (454) sequencing in the target species. Primer pairs for a total of 30 microsatellite loci were designed from these of which 13 were successfully amplified in 25 individuals from Kenwick Nature Reserve, Perth. Twelve loci were polymorphic with observed heterozygosity ranging from 0.12 to 0.6 and the number of alleles per locus ranged from 2 to 8. These markers were trialed in all species within the G. thelemanniana species complex. Amplification success and level of polymorphism varied among loci and taxa, but most (96 %) were successful. These loci will be useful in understanding the genetic variation, molecular ecology and phylogenetic relationships in the G. thelemanniana complex.