基于SSR标记的100份葡萄资源亲缘关系和群体遗传结构分析  

Genetic relationship and population genetic structure analysis of 100 accessions of grape germplasm resources based on SSR markers

作　　者：赵旗峰 黄丽萍[1] 王敏[1] 刘晓婷 张晓萍 马小河 ZHAO Qifeng;HUANG Liping;WANG Min;LIU Xiaoting;ZHANG Xiaoping;MA Xiaohe(Pomology Institute of Shanxi Agricultural University,Taigu 030815,Shanxi,China)

机构地区：[1]山西农业大学果树研究所,山西太谷030815

出　　处：《果树学报》2021年第8期1217-1230,共14页Journal of Fruit Science

基　　金：农业农村部保种项目(111821301354052002);科技部资源平台专项(NHGRC2020-NH12-2);国家现代农业产业技术体系专项(CARS-29-yz-5);院优势课题组自选项目(YCX2018D2YS05)。

摘　　要：【目的】通过表型性状鉴定分析,从山西太谷国家葡萄种质资源圃内保存的资源中,筛选出不同葡萄种质资源进行SSR分析,揭示这些不同来源的葡萄种质资源之间的亲缘关系和群体遗传结构,为葡萄种质资源的科学管理和分子标记辅助育种提供参考。【方法】以100份葡萄种质为材料,依据基因组DNA测序挖掘出的SSR引物数据,筛选出33对多态性核心引物,根据标准分子质量记录扩增DNA多态性条带,获得数据矩阵。利用每对SSR引物在100份葡萄种质资源中的等位基因组成,计算33对SSR引物的位点多态性信息含量(PIC)、100份葡萄种质资源个体间的Nei’s遗传距离,并利用MEGA 7.0软件构建NJ邻接聚类树,推断葡萄原始集合与核心集合的遗传结构。【结果】100个样本中的33对SSR标记共检测到423个等位基因,每对引物扩增条带数4~26条,平均扩增条带数为12.82条。葡萄种质资源之间的Nei’s遗传距离为0.4~0.8,占资源总量的97.80%。通过遗传结构分析并预测其最佳分组数K为2,即葡萄核心集合主要分为2个亚群,分别为欧亚种群的鲜食葡萄和杂种群,其中杂种群涵盖了欧美杂种、美洲种、中国野生种及欧亚种的酿酒品种。来自国外的美洲种的砧木和来自山西的野葡萄聚在一起,同时与欧美杂种和酿酒品种也聚为一类。可能是由于人为选种和选用目的的不同造成各品种之间遗传背景的混杂。这一结论还需进一步研究证明。【结论】33对引物将100份葡萄分为两个集群,第一集群主要为葡萄亚种鲜食葡萄品种,第二集群主要包含了欧亚种酿酒品种、欧美杂种、美洲种、中国野生种4个亚种群。【Objective】100 accessions of grape resources were selected from the National Grape Germplasm Resource Center in Taigu,Shanxi through phenotypic character identification for SSR analysis in order to reveal their genetic relationship and population genetic structure and provide references for the scientific management of grape germplasm resources and molecular marker-assisted breeding.【Methods】The modified CTAB was used to extract the genomic DNA of the tested grape germplasm.Based on the SSR primers extracted from the grape genome sequencing,33 pairs of core polymorphic primers were screened out,and the DNA was amplified according to standard molecular weight records using capillary electrophoresis technology.According to the polymorphic bands,the data matrix was obtained.The number of alleles Na(Na),the number of effective alleles(Ne),the Shannon’s information index(I),the expected heterozygosity(He),and the observed heterozygosity were calculated using Gen Al Ex6.503 software.At the same time,Gen Al Ex6.503 software were used to count the allelic composition of each pair of SSR primers in 100 accessions of grape germplasm resources,including the number of rare alleles(≤1%),the number of common alleles(1%–20%),and the most common allele(20%).The site polymorphism information content(PIC)of 33 pairs of SSR primers and the Nei’s Genetic distance between 100 accessions of grape germplasm resources were calculated using Power Marker software.The NJ adjacent cluster trees of 100 grape germplasm resources based on the Nei’s genetic distance were constructed using MEGA7.0 software.The STRUCTURE software was used to infer the genetic structure of the original grape set and the core set based on the Bayesian clustering analysis method.The population number(K)was set to 1-10,and each K value was simulated 10 times,and the number of iterations(length of burn-in period)was set.The Markov Chain Monte Carlo(MCMC)at the beginning was 100000 times,and the MCMC after no-count iterations was 1000000 times.Finally

关 键 词：葡萄 SSR分子标记 亲缘关系 遗传结构 

分 类 号：S663.1[农业科学—果树学]