Identification and characterization of microsatellite markers in Pinus kesiya var. langbianensis (Pinaceae)1

This is an open access article distributed under the terms of the Creative Commons Attribution License (CC-BY-NC-SA 4.0), which permits unrestricted noncommercial use and redistribution provided that the original author and source are credited and the new work is distributed under the same license as the original. Pinus kesiya Royle ex Gordon var. langbianensis (A. Chev.) Gaussen (Pinaceae) is an important forest tree species in Yunnan Province, China. It has been recorded at altitudes from 600–1800 m in the southern, semihumid climate zone of Yunnan (Editorial Committee of Flora of China, 1978; Wu, 1986) and accounts for 11% of the forest area and 1.0 × 10 8 m 3 of the forest volume (Jiang et al., 2007). The wood is extensively used in building, furniture, and the fiber industry. Pinus kesiya var. langbianensis is also highly valued for its high resin content, Output of gum turpentine from P. kesiya var. langbianensis accounted for more than 90% of the total output in Yunnan (Yin et al., 2005). However, germplasm resources of P. kesiya var. langbianensis have decreased in recent years as a result of overexploitation (Zhao et al., 2016). Information on genetic diversity and spatial structure in P. kesiya var. langbianensis is important for its future conservation and can be used to help guide local forest management (Sanchez et al., 2014). No specific conservation strategy is available for this species, in part due to the limited understanding of genetic diversity and structure of the natural populations. As a primary forest tree species in southern Yunnan Province, resource conservation of P. kesiya var. langbianensis will benefit the entire ecological system in the region (Li et al., 2015). Therefore, in this study we developed novel microsatellite markers for P. kesiya var. lang-bianensis by applying next-generation sequencing to investigate the genetic diversity and population structure of this species at the molecular level. METHODS AND RESULTS Needle samples of 60 individuals from four P. kesiya var. langbianensis populations located in Yunnan Province, China, and 59 individuals from four related species (P. massoniana D. Don, P. densata Mast, P. tabuliformis Carrière, P. yunnanensis Franch.) were collected (Appendix 1). All needle samples were dried and preserved in silica gel. Total genomic DNA was isolated from dried needle samples using the cetyltrimethylammonium bromide (CTAB) method (Doyle and Doyle, 1990). Paired-end libraries were constructed on four individuals sampled from Puer City Institute of Forestry Sciences and sequenced by a …