基于EST-SSR标记的南方型黑杨主栽无性系鉴定及亲缘关系分析  

Genetic Identification and Relationship Analysis of Main Southern-type Poplar Clones with EST-SSR

作　　者：袁佳秋 田野 洑香香 YUAN Jia-Qiu;TIAN Ye;FU Xiang-Xiang(Co-Innovation Center for the Sustainable Forestry in Southern China,College of forestry,Nanjing forestry university,Nanjing 210037)

机构地区：[1]南方现代林业协同创新中心南京林业大学林学院

出　　处：《植物研究》2019年第5期770-778,共9页Bulletin of Botanical Research

基　　金：国家科技支撑计划课题(2015BAD09B02);国家自然科学基金项目(31370618);国家重点研发计划课题(2016YFD0600402)~~

摘　　要：以南方型黑杨引进后主栽区重点推广栽植的12个无性系为研究对象,利用EST-SSR分子标记进行多态性分析,并建立无性系鉴定的指纹图谱,为优良黑杨无性系的推广提供依据。用EST-trimmer对NCBI数据库中的EST序列进行分析、MISA软件找出SSR位点后运用primer3在线设计获得的30对EST-SSR引物,对12个黑杨无性系进行PCR扩增。经过筛选获得18对多态性引物,对12个黑杨无性系进行扩增获得88个等位基因,多态率为70.5%,平均观察等位基因数为2.1463,平均Shannon’s多样性指数为0.5927。通过5对引物组合(EU147、EU43、EU11、EU164和EU81)可以将12个黑杨无性系进行区分,并在此基础上构建指纹图谱。亲缘关系分析发现无性系间的遗传相似系数为0.6020~0.9040,平均值为0.7691;无性系间的遗传距离较近,与其均来源于美洲黑杨和欧美杨的相似遗传背景有关。总之,EST-SSR标记可以对南方型黑杨主栽无性系进行有效鉴定。Twelve main southern-type poplar clones were selected to assess genetic polymorphism based on EST-SSR markers and to establish fingerprint for clone identification in the main planting region. The results could provide information and tools for the effective and protective extension of superior clones in this area. Thirty pairs of EST-SSR primers were obtained with stepwise approaches including EST sequences analysis from NCBI database by EST-trimmer, SSR loci screening by MISA, and primers design using primer 3. The obtained EST-SSR primers were then applied to PCR amplification for the 12 poplar clones. Eighteen pairs of polymorphic primers were selected and 88 alleles were obtained by sequence amplification of 12 poplar clones. The polymorphism ratio of the locus was 70.5%, the average number of the observed alleles was 2.1463, and the average shannon’s diversity index was 0.529 7. The 12 clones could be effectively identified based on a combination of 5 pairs of SSR primers, which were EU147, EU43, EU11, EU164, and EU81. The genetic fingerprint map of the 12 clones was also constructed successfully using the same SSR primers. The genetic similarity coefficient of the 12 clones ranged from 0.602 0-0.904 0, with a mean value of 0.769 1. All the clones showed close genetic distance, which revealed their genetic background of the same origination. In a word, EST-SSR marker is an effective tool for the identification of the southern type poplar clones.

关 键 词：南方型杨树 EST-SSR标记 无性系鉴定 多态性 

分 类 号：S722.3[农业科学—林木遗传育种]