基于SSR标记的广东含笑等11个含笑属物种亲缘关系研究

    Study on the Genetic Relationship of 11 Michelia Species Based on SSR Markers

    • 摘要:
      目的 含笑属植物是我国南方地区重要的园林绿化和用材树种,但目前种间的遗传关系尚不明确,以开发高效的基因分型手段进行各物种亲缘关系研究,可为含笑属的保护和开发利用提供理论和技术支撑。
      方法 以广东含笑等11个物种为研究材料,筛选高多态性SSR引物,利用基于M13序列的荧光标记方法获取基因分型数据,计算遗传多样性参数,并利用Neighbor-Joining(NJ)方法构建进化树,分析种间的亲缘关系。
      结果 12对SSR引物均具有较高的多态性,扩增成功率为90.9%,可以高效地完成基因分型,12个标记的平均等位基因数为8.17;Shannon多样性指数(I)平均值为1.836;观测杂合度(Ho)、期望杂合度(He)和多态信息含量(PIC)的平均值分别是0.365、0.839和0.796。聚类分析结果显示,分属不同亚属的物种在NJ进化树上可以被明显区分开来,其中后生含笑亚属物种被聚类在两个主要分支上,含笑亚属的3个物种被聚类在一个主要分支上,亲缘关系分析结果与以往的分类学研究结果一致。
      结论 SSR标记基因分型体系可以较好地完成广东含笑等11个物种的基因分型,在未来的杂交亲本评选、亲本选配和杂交子代亲缘关系鉴定等方面均可以发挥重要作用。

       

      Abstract:
      Objective Michelia genus contains important landscaping and timber plants and mainly distributes in South China, however, the genetic relationship among different species is still unclear. Developing efficient genotyping methods and performing genetic relationship researches can provide theoretical and technical support for the utilization and protection of Michelia species.
      Method Using 11 Michelia species as research materials, highly polymorphic SSR primers were screened out. M13-tailed fluorescent label method was used perform the PCR to obtain genotyping data and the genetic diversity parameters were calculated. Furthermore, a Neighbor-Joining(NJ)tree was constructed based on the genetic distance of each species.
      Result All 12 pairs of SSR primers have relatively high polymorphism, and the amplification rate is 90.9%, which can finish genetic typing efficiently. The mean allele number for 12 markers is 8.17 and mean Shannon diversity index(I) is 1.836. The observed heterozygosity(Ho), expected heterozygosity(He)and polymorphic information content(PIC)are 0.365, 0.839 and 0.796, respectively. The cluster analysis results show that species belonging to different subgenus can be clearly distinguished on the NJ evolutionary tree. Among them, seven Subgen. Metamichelia species are clustered on two main divisions, and three species of Subgen. Michelia was clustered on the third branch. The result of genetic relationship analysis in this study is consistent with that of the previous taxonomic study.
      Conclusion SSR marker genotyping system developed in this research can perform the genotyping of 11 Michelia species successfully. In the future, these markers can play an important role in hybrid parent evaluation, parent selection and hybrid offspring genetic relationship identification and other genetic studies.

       

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