Isolation and characterization of thirty-eight microsatellite loci for the Pinus wangii , an endangered species

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MICROSATELLITE LETTERS

Isolation and characterization of thirty-eight microsatellite loci for the Pinus wangii, an endangered species Zhi-Yong Zhang • Binbin Cui • Jian-Feng Mao Xiao-Ming Pang • Ying-Yue Li



Received: 29 October 2014 / Accepted: 1 November 2014 Ó Springer Science+Business Media Dordrecht 2014

Abstract Piuns wangii Hu et Cheng is an endangered five-needle pine species, with scattered populations and scare individuals. To study its genetic diversity and better address conservation programs, 506 novel microsatellite markers were developed by mining the transcriptome database of P. lambertiana. 117 loci were able to amplify a fragment of expected size in P. wangii, of which 38 were polymorphic. The number of alleles of these 38 EST-SSRs in 16 individuals varied from 2 to 6, with an average of 2.6. The observed (Ho) and expected (He) heterozygosity values ranged from 0.000 to 1.000 (averaging 0.532) and 0.117–0.693 (averaging 0.394), respectively. Twenty-three loci showed significant departure from HWE after Bonferroni correction. This was the first report of candidate molecular markers on P. wangii, which will facilitate future analyses of genetic diversity and population structure.

Electronic supplementary material The online version of this article (doi:10.1007/s12686-014-0379-1) contains supplementary material, which is available to authorized users. Z.-Y. Zhang National Engineering Research Center for Floriculture, College of Landscape Architecture, Beijing Forestry University, Beijing 100083, People’s Republic of China B. Cui Department of Biology and Chemistry, Baoding University, Baoding 071000, Hebei, People’s Republic of China J.-F. Mao  X.-M. Pang  Y.-Y. Li (&) National Engineering Laboratory for Tree Breeding, College of Biological Sciences and Technology, Beijing Forestry University, Qinghua East Road No. 35, Beijing 100083, People’s Republic of China e-mail: [email protected]

Keywords Microsatellites  Pinus wangii  Expressed sequence tag (EST)  Conservation genetics Piuns wangii Hu et Cheng, endemic to Malilpo, Maguan, Xichou County, China, has been listed as Endangered species by International Union for the Conservation of Nature in 2012. Overexploitation, especially during the last 5 years, is credited as the greatest cause of population decline (\100 individuals left) prior to its listing (personal conversation). Populations were further threatened by the accelerated degradation of its habitat (with only five remnant natural populations) and low level of natural regeneration (field observation). Up to now, however, no programs of artificial breeding and ex situ conservation have been initiated. In recent years, microsatellites [simple sequence repeats (SSRs)], known as high levels of polymorphism, co-dominance and reproducibility, have become some of the most useful markers for genetic variation analysis. Comparing with traditional methods for SSR markers development, EST-derived SSRs would take less expensive, effort, and time. Another advantage of EST-SSR is relatively

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