梨属植物SSR分子标记核心引物的筛选及其聚类分析  

Core primers screening for SSR markers and cluster analysis in the plants of Pyrus spp.

作　　者：赵瑞娟 薛华柏[1] 王磊[1] 温晶晶 杨健[1] 王龙[1] 王苏珂[1] 苏艳丽[1] 李秀根[1] 

机构地区：[1]中国农业科学院郑州果树研究所,郑州450009

出　　处：《果树学报》2017年第10期1229-1238,共10页Journal of Fruit Science

基　　金：中国农业科学院科技创新工程(CAAS-ASTIP);中央级科研院所基本科研业务费专项(1610192016612;1610192017709);现代农业(梨)产业技术体系(CARS-29)

摘　　要：【目的】筛选适合梨种质资源遗传多样性研究、品种鉴定分析的SSR核心引物。【方法】利用来源于不同系统、遗传背景差异较大的12份梨种质资源对已报道的800对梨和苹果的SSR引物和本课题组开发的534对SSR引物进行初步筛选,再利用48份不同地理来源的梨种质资源对初筛引物进行复筛,筛选一套适合梨种质资源遗传多样性研究、系统发育和品种鉴定分析的SSR核心引物,并通过遗传多样性分析和聚类分析验证引物的有效性。【结果】初筛出SSR引物131对,进一步复筛筛选出25对清晰稳定、多态性高的核心引物。25对核心引物对48份梨种质资源进行遗传多样性分析,共得到387个等位基因,平均每个位点等位基因数15.48个。各位点杂合度变幅为0.44~0.92,均值0.76;多态性信息含量变幅为0.70~0.92,均值0.85;基因多样性变幅为0.73~0.92,均值为0.87,说明引物的多态性高,能够有效揭示48份梨种质资源的遗传多样性。48份梨种质资源的聚类分析结果显示,西洋梨和东方梨分别聚成了2个类群,东方梨类群中的栽培种和野生种分别聚成了2个亚群;栽培种中的秋子梨品种聚在了一起,白梨和砂梨聚在了一起。【结论】筛选出的25对梨SSR引物,带型清晰、稳定,多态性信息含量均在0.70以上,可作为核心引物用于梨种质资源鉴定和遗传多样性分析等研究。[ Objective] China is one of the origin centers of Pyrus plants with abundant deposits of Pyrus germplasm resources. To date, experts have carried out a lot of research on pear cultivars by SSR markers, such as genetic diversity, population structure analysis and construction of fingerprint database, etc. Screenings of primers were necessary for these researches. However, the screening of SSR core primers on genetic diversity analysis, construction of fingerprint database and identification of pear cuhivars had rare- ly reported in the literature yet. SSR core primers should have the features of high efficiency, good repeat- ability and high polymorphism. The purpose of our study is to screen out suitable SSR core primers, which can be accurately used for evaluation of germplasm genetic diversity and genetic relationship of pear cultivats. [Methods] 1 334 pairs of SSR primers were initially screened using 12 pear （Pyrus. L） germplasm resources with distant genetic relationship. 1 334 pairs of SSR primers that consisted of 800 pairs of SSR primers for apples and pears searched from the articles and 534 pairs of SSR primers developed from pear transcriptome were screened. The markers with clear PCR amplified results, unique amplified loci, good stability and repeatability were Chosen. Then 48 pear germplasm resources with different geographical ori- gins were chosen to screen the initial selected primers. Polyacrylamide gel electrophoresis and ABI 3730XL DNA analyzer were used to describe the amplified results, and the secondly selected primers were screened by fluorescence SSR markers detection technique with capillary electrophoresis. Finally, a set of SSR core primers were identified, which could be suitable for genetic diversity analysis, construction of fingerprint database and identification of pear materials. Moreover, Genetic diversity and cluster analysis of 48 pear materials were performed to verify the effectiveness of these core primers. The genetic diversity of 47 pear cultivars and 1 wild germplas

关 键 词：梨 SSR核心引物 遗传多样性 聚类分析 

分 类 号：S661.2[农业科学—果树学]