Development of polymorphic microsatellite markers for the Tertiary relict tree species Taiwania cryptomerioides (Cupressaceae) in East Asia.

Taiwania cryptomerioides Hayata is an endangered relict tree species which is endemic to mainland China, Taiwan, Myanmar, and northern Vietnam. It is an economically important tree species and has been used for reforestation in mountain areas of mainland China and Taiwan. In order to investigate its genetic diversity for conservation and restoration, we developed and characterized 15 nuclear microsatellite markers based on next-generation sequencing data. A total of 100 microsatellite primer pairs were initially designed and tested based on the restriction-site associated DNA sequencing data. 60 of 100 loci (60%) were successfully amplified, of which 42 loci exhibited polymorphism. Fifteen polymorphic microsatellite loci with clear peaks were selected for further analyses in four T. cryptomerioides populations sampled from China (Hubei, Fujian, Guizhou, and Yunnan). The number of alleles per locus ranged from 2 to 24, and the levels of observed and expected heterozygosity ranged from 0.000 to 0.950 and from 0.000 to 0.860, respectively. This set of microsatellite markers will be useful for future population genetic studies of T. cryptomerioides in East Asia.