Microsatellite markers for Corybas (Orchidaceae) species in New Zealand.

Premise of the Study: Microsatellite markers were developed for New Zealand species of Corybas (Orchidaceae) to investigate population genetics and species delimitation.

Methods and Results: From sequencing a total genomic DNA library (using Illumina MiSeq), we developed 22 microsatellite markers for C. obscurus . The di- and trinucleotide repeat loci were initially trialed on individuals representing seven Corybas taxa ( C . "rimutaka," C. confusus , C. hypogaeus , C. macranthus , C. obscurus , C. trilobus , and C. walliae ) and had one to eight alleles per locus. Twelve polymorphic markers were further tested on six Corybas populations from three of the seven taxa ( C. obscurus , C . "rimutaka," and C. trilobus ). Observed and expected heterozygosities ranged from 0-1 and 0-0.859, respectively. The utility of these 12 loci was further validated in five related Corybas species ( C. hypogaeus , C. obscurus , C. vitreus , C. walliae , and C . "rimutaka"; 38 individuals) representing populations from across the North and South Islands. The average value for genetic diversity among populations ( F ST ) of 0.439 shows differentiation among species.

Conclusions: These markers will be useful for future studies aimed at delimiting species boundaries and examining the genetic diversity of the New Zealand Corybas species.