基于SSR分子标记的东莞广宁红花油茶遗传多样性及群体结构研究

    Study on Molecular Genetic Diversity and Population Structure of Camellia semiserrata in Dongguan Based on SSR Molecular Markers

    • 摘要:
      目的 基于简单重复序列(Simple Sequence Repeat,SSR)分子标记技术对东莞广宁红花油茶(Camellia semiserrata)群体开展遗传多样性和群体结构研究,为广宁红花油茶遗传驯化适应机制解析及良种分子选育提供一定的理论基础。
      方法 对东莞樟木头林场近533 hm2广宁红花油茶林中的200份随机种质样本,利用聚合酶链式反应(PCR)扩增和荧光毛细管电泳检测技术,进行SSR多态性引物筛选及SSR位点分型工作,基于分型结果统计位点的等位基因数量,计算多态性信息含量(PIC值)、Shannon’s信息指数、Nei’s遗传多样性指数、观测和期望杂合度、近交系数(Fis)、遗传分化系数(Fst)等群体遗传学参数,开展群体结构分析和非加权组平均法(UPGMA)聚类分析等,以评价该种质样本群体的遗传多样性水平,并解析其群体结构。
      结果 共筛选出20对SSR多态性引物,并扩增出187个等位基因,SSR位点的平均等位基因数为9.4,平均有效等位基因数(Ne)为2.832,平均PIC值为0.5854;平均Shannon’s信息指数为1.028,平均Nei’s遗传多样性指数为0.6230,广宁红花油茶样本群体的观测杂合度和期望杂合度平均值分别为0.4690和0.6248;平均Fis为0.127,平均Fst为0.12;群体遗传结构分析将样本群体分为6个类群,其中混合个体比例为41%;UPGMA聚类分析结果显示群体的分组和遗传背景较复杂。
      结论 筛选得到的20对SSR多态性引物适合广宁红花油茶群体的遗传多样性和群体结构研究;东莞广宁红花油茶群体的遗传多样性水平和杂合度较高,属中等程度的遗传分化,群体结构和遗传背景较复杂。

       

      Abstract:
      Objective Based on Simple Sequence Repeat (SSR) molecular marker technology, this study conducted investigations into the genetic diversity and population structure of Camellia semiserrata populations in Dongguan, aiming to provide a theoretical basis for elucidating the genetic domestication and adaptive mechanisms of C. semiserrata and facilitating the molecular breeding of elite varieties.
      Method A total of 200 randomly selected germplasm samples were collected from nearly 533 hm2 C. semiserrata forest in Zhangmutou Forest Farm, Dongguan. Using polymerase chain reaction (PCR) amplification and fluorescent capillary electrophoresis detection technology, SSR polymorphic primer screening and SSR locus genotyping were performed. Based on the genotyping results, the number of alleles per locus was counted, and population genetic parameters including Polymorphism Information Content (PIC), Shannon's Diversity Index, Nei's genetic diversity index, observed heterozygosity, expected heterozygosity, inbreeding coefficient (Fis), and genetic differentiation coefficient (Fst) were calculated. Additionally, population structure analysis and unweighted pair group method with arithmetic mean (UPGMA) cluster analysis were conducted to evaluate the genetic diversity level of the germplasm samples population and elucidate its population structure.
      Result A total of 20 pairs of polymorphic SSR primers were screened out, and 187 alleles were amplified. For the SSR loci, the average number of alleles was 9.4, the average effective number of alleles (Ne) was 2.832, and the average PIC was 0.5854. The average Shannon's Diversity Index was 1.028, and the average Nei's genetic diversity index was 0.6230. For the C. semiserrata germplasm samples population, the average observed heterozygosity and expected heterozygosity were 0.4690 and 0.6248, respectively. The average Fis was 0.127, and the average Fst was 0.12. Population genetic structure analysis divided the germplasm samples population into 6 clusters, with admixed individuals accounting for 41%. UPGMA cluster analysis revealed that the population exhibited complex grouping patterns and genetic backgrounds.
      Conclusion The 20 pairs of polymorphic SSR primers screened in this study are suitable for investigating the genetic diversity and population structure of C. semiserrata populations. The C. semiserrata population in Dongguan exhibits high levels of genetic diversity and heterozygosity, accompanied by a moderate degree of genetic differentiation, as well as a complex population structure and genetic background.