三叶青种质资源遗传多样性的SSR荧光标记分析

目的研究三叶青64个种质的遗传多样性和亲缘关系。方法利用SSR荧光标记对其进行PCR扩增,利用POPGENE32软件分析三叶青64个种质的遗传多样性和亲缘关系,利用UPGMA法构建其亲缘关系树状图,利用NTSYS软件构建其主成分分析二维图和三维散点图。结果从14对引物中筛选出8对条带清晰、重复性好的引物,对64份供试材料的基因组DNA进行扩增。8个SSR标记的观察等位基因数(Na)变化范围为3～13,均值为7.875 0;有效等位基因数(Ne)变化范围为1.424 9～6.087 4,均值3.605 2;Shannon信息指数(I)变化范围为0.689 5～2.082 4,均值1.424 0;观测杂合度(Ho)变化范围为0.206 3～0.734 4,均值0.524 7;期望杂合度(He)变化范围为0.300 6～0.842 4,均值0.658 4;Nei’s基因多样性(H)0.298 2～0.835 7,均值0.653 2;多态信息含量(PIC)变化范围为0.288 0～0.817 5,均值0.614 5,遗传相似系数变化范围为0.115 4～0.954 5,遗传距离变化范围为0.000 0～3.218 1,说明64份三叶青种质亲缘关系较远,遗传分化程度较大。在遗传距离1.018 9处,64份三叶青种质可分为5大组。结论地理差异与种质遗传差异无必然联系。三叶青种质资源具有丰富的遗传多样性,SSR荧光标记分析结果可为三叶青种质资源的利用和品种选育提供一定的参考。

Genetic diversity analysis of Tetrastigma hemsleyanum germplasm resources based on fluorescently labeled SSR markers

Objective To understand the genetic diversity and genetic relationship of 64 samples of Tetrastigma hemsleyanum germplasm resources. Methods Fluorescently labeled SSR were used for PCR amplification, POPGENE32 software was used to analyze genetic diversity and genetic relationship of 64 samples of T. hemsleyanum germplasm resources, UPGMA method was used to construct their genetic tree map, NTSYS software was used to construct their two-dimensional principal component analysis map and three-dimensional scatter map. Result Eight pairs of primers with clear and reproducible bands were screened from 14 pairs of primers and used for genomic DNA amplification of 64 sample materials. The observed number of alleles (Na) ranged from 3 to 13, and the mean value was 7.875 0; The effective number of alleles (Ne) ranged from 1.424 9 to 6.087 4, and the mean value was 3.605 2; The Shannon information index (I) ranged from 0.689 5 to 2.082 4, and the mean value was 1.424 0; The observed heterozyghosity (Ho) ranged from 0.206 3 to 0.734 4, and the mean value was 0.524 7; The expected heterozygosity (He) ranged from 0.300 6 to 0.842 4, and the mean value was 0.658 4; The Nei''s gene diversity (H) ranged from 0.298 2 to 0.835 7, and the mean value was 0.653 2; The polymorphism information content (PIC) ranged from 0.288 0 to 0.817 5, and the mean value was 0.614 5; The genetic similarity coefficient ranged from 0.115 4 to 0.954 5; The genetic distance ranged from 0.000 0 to 3.218 1. It was indicated that the genetic relationship of 64 T. hemsleyanum germplasm resources was far, and the degree of genetic differentiation was high. At the genetic distance of 1.018 9, 64 T. hemsleyanum germplasm resources could be divided into five groups. Conclusion There was no necessary connection between geographical difference and genetic difference of germplasm. T. hemsleyanum germplasm resources are rich in genetic diversity. The results of fluorescently labeled SSR analysis can provide some references for the utilization and variety breeding of T. hemsleyanum germplasm resources.