Development and characterization of 20 polymorphic microsatellite markers from RAPD product in Populus euphratica
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TECHNICAL NOTE
Development and characterization of 20 polymorphic microsatellite markers from RAPD product in Populus euphratica Chunhua Wang1 • Ling-Xiao Liu2 • Yun-Guo Liu3
Received: 17 April 2015 / Accepted: 4 May 2015 Ó Springer Science+Business Media Dordrecht 2015
Abstract Twenty polymorphic mirosatellite markers have been developed and characterized from RAPD product in Populus euphratica for protection of its natural resources. Polymorphisms of these microsatellite markers were evaluated in a natural population of 30 individuals collected from Xinjiang Region in China. The number of alleles ranged from 3 to 12. Observed and expected heterozygosities varied from 0.53 to 0.97, and from 0.56 to 0.94, respectively. All the microsatellite loci conformed to Hardy–Weinberg equilibrium after Bonferroni correction. They have showed sufficient level of polymorphisms to estimate the population structure, genetic diversity and species conservation. It is an efficient method to isolate microsatellite markers from RAPD product in P. euphratica. Keywords Populus euphratica Microsatellite markers Population structure Genetic conservation Populus euphratica, which is the only arborescent species naturally distributed in the semi-arid areas in central Asia and the Middle East, is well known for its tolerance to soil salinity (Gu et al. 2004). However, natural stands of P. & Ling-Xiao Liu [email protected] & Yun-Guo Liu [email protected] 1
College of Earth and Environment Science, Anhui University of Science and Technology, Huainan 232001, China
2
Linyi Academy of Agricultural Sciences, Linyi 276012, China
3
College of Life Sciences and Technology, Xinjiang ¨ ru¨mqi 830046, China University, U
euphratica have decreased in the past decade and some populations have disappeared due to habitat changes in its natural distributions. Therefore, it is very important to protect the natural resources of P. euphratica. In the long term, a good knowledge of the genetic diversity, population structure and genetic differentiation of P. euphratica is required in order to establish adequate management plans for both conservation and utilization of this important tree species. Microsatellites, or simple sequence repeats (SSRs), are of great utility in species conservation and management because of their high polymorphism, abundance, co-dominance and small length of sequences. In the present paper, a total of 20 polymorphic microsatellite markers were isolated from P. euphratica for the purpose of protecting its natural populations. Randomly amplified polymorphism DNA (RAPD) method is a PCR-based DNA fingerprinting technique (Williams et al. 1990). RAPDs are known to enrich for microsatellites (Ender et al. 1996). Genomic DNA was amplified with RAPD oligonucleotide primers. RAPD reactions were carried out in a 20-ll reaction mixture that included 20 pmol of RAPD primer, 100 lM of dNTPs, 10 mM Tris-HCl (pH 8.3), 50 mM KCl, 2.0 mM MgCl2, 1 unit of Taq polymerase (TaKaRa Corp.), and about 150 ng of template DNA. Amplification w
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