Development of microsatellite markers for the mycoheterotrophic species Burmannia nepalensis (Miers) Hook.f. based on RAD sequencing.

Mycoheterotrophic plants can derive carbon from fungi rather than from photosynthesis. Habitat destruction and sensitivity to environmental perturbation may result in the loss of biodiversity including genetic variation of mycoheterotrophic plants. Burmannia nepalensis (Miers) Hook.f. (Burmanniaceae) is a mycoheterotrophic plant with a wide distribution across southern China and southern and eastern Asia. As part of our endeavor to reveal population genetic patterns of mycoheterotrophic plants, fifteen microsatellite loci were developed by RAD (restriction site-associated DNA) sequencing in 89 individuals from four populations of B. nepalensis. A total of 49 alleles were amplified. The number of alleles per locus ranged from two to six with an average of 3.3. The observed and expected heterozygosity per population varied from 0.000 to 1.000 and from 0.000 to 0.722, respectively. A transferability test showed that only one to five loci could be cross-amplified successfully in four other congeneric species of Burmannia. These markers can be used to reveal population genetic diversity in B. nepalensis, and will help to elucidate the evolutionary history and to enhance efforts for conservation of mycoheterotrophic plants.