Plant Biotechnology Laboratory, Department of Botany, North-Eastern Hill University, Shillong 793022, Meghalaya, India.
Gene. 2013 Apr 25;519(1):91-7. doi: 10.1016/j.gene.2013.01.037. Epub 2013 Feb 8.
Molecular genetic fingerprints of seven populations of Vanda coerulea comprising of thirty-two genotypes from Northeast India were developed using PCR based markers. Genetic variability in the wild genotypes of V. coerulea was analyzed using two different single primer amplification reactions (SPAR) methods, viz., random amplified polymorphic DNA (RAPD) and inter-simple sequence repeats (ISSR). A total of 32 genotypes were used to investigate the existing natural genetic diversity at intra-specific level. Two hundred and twenty six (226) amplification products were scored by RAPD and ISSR, both of which collectively showed 58.88% polymorphism with a mean intra-population genetic diversity (Hpop) of 0.119. However, their level of diversity at inter- and intra-population levels was significant, with the percentage of polymorphic loci (Pp) ranging from 17.70% to 45.13%, Shannon's information index (I) from 0.105 to 0.268 and Nei's gene diversity (h) from 0.072 to 0.185 with mean Nei's gene diversity 0.174 and the overall estimate of gene flow being (Nm) 1.165. Analysis of molecular variance (AMOVA) showed 96.07% of variation at intra-population level, whereas 3.93% variation was recorded at inter-population level. Only one major cluster was detected by cluster analysis using the unweighted pair-group method with arithmetic average (UPGMA). Present investigation suggests the efficiency of SPAR methods to estimate the genetic diversity of V. coerulea and can be seen as a starting point for future research on the population and evolutionary genetics of this species.
采用基于 PCR 的标记物，开发了包含来自印度东北部的 32 种基因型的 7 个兜兰种群的分子遗传指纹图谱。使用两种不同的单引物扩增反应（SPAR）方法，即随机扩增多态性 DNA（RAPD）和简单序列重复间（ISSR），对野生兜兰基因型的遗传变异进行了分析。使用 32 种基因型来研究种内水平的现有自然遗传多样性。RAPD 和 ISSR 共记录了 226 个扩增产物，两者均显示出 58.88%的多态性，平均种群内遗传多样性（Hpop）为 0.119。然而，它们在种群间和种群内水平的多样性水平差异显著，多态位点数（Pp）的百分比范围从 17.70%到 45.13%，香农信息指数（I）从 0.105 到 0.268，Nei 基因多样性（h）从 0.072 到 0.185，平均 Nei 基因多样性为 0.174，总体基因流动估计值（Nm）为 1.165。分子方差分析（AMOVA）显示，96.07%的变异发生在种群内水平，而 3.93%的变异发生在种群间水平。聚类分析仅检测到一个主要聚类，使用不带权对组平均法（UPGMA）。本研究表明，SPAR 方法能够有效地估计兜兰的遗传多样性，可以作为该物种种群和进化遗传学未来研究的起点。
