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近缘物种筛选法是获取微卫星引物的主要方法之一。利用松属植物EST-SSRs的种间通用性,从马尾松微卫星引物中筛选出红松EST-SSR标记,并与已有的红松基因组SSR标记进行对比分析。采集长白山露水河红松母树林4个种群的248个样本,分析种群遗传多样性。结果表明:马尾松与红松的种间通用率为27.27%,松属内不同亚属物种间的种间通用性偏低;与红松基因组SSRs相比,EST-SSRs的单位点等位基因数(N a)、有效等位基因数(Ne)、观测杂合度(Ho)、期望杂合度(He)等遗传多样性参数均较低;用2组引物分析4个种群的遗传多样性,发现有效等位基因均能反映出不同种群遗传变异的趋势,基因多样性均能反映出遗传同一性的差异,说明两组引物都能准确地反映红松种群的遗传多态性水平,新筛选的EST-SSRs可以用于红松种群遗传多样性研究。
The related species screening method is one of the main methods for obtaining microsatellite primers. According to the interspecific commonality of Pinus sylvestris EST-SSRs, Pinus koraiensis EST-SSR markers were screened out from Pinus massoniana microsatellite primers and compared with the existing Pinus koraiensis SSR markers. 248 samples from 4 populations of Pinus koraiensis forest in Changshu Mountain were collected to analyze the genetic diversity. The results showed that the common rate between Pinus massoniana and Pinus koraiensis was 27.27%, and the interspecific commonality among different subgenus within Pinus was low. Compared with the SSRs of Pinus koraiensis genome, the single locus alleles of EST-SSRs (Ne), observed heterozygosity (Ho), expected heterozygosity (He) and other genetic diversity parameters were all low. The genetic diversity of 4 populations was analyzed by two sets of primers, The results showed that the effective alleles all showed the trend of genetic variation in different populations. The diversity of gene could reflect the difference of genetic identity, which indicated that both primers could accurately reflect the genetic diversity of Korean pine population. EST-SSRs can be used to study the genetic diversity of Korean pine population.