草地学报 ›› 2021, Vol. 29 ›› Issue (9): 2081-2090.DOI: 10.11733/j.issn.1007-0435.2021.09.027

• 技术研发 • 上一篇    下一篇

菊苣EST-SSR分子标记开发及通用性分析

梁小玉1, 季杨1, 胡远彬1, 易军1, 白史且3, 张靓1, 张新全2   

  1. 1. 四川省畜牧科学研究院, 四川 成都 610066;
    2. 四川农业大学动物科技学院, 四川 成都 611130;
    3. 四川省林业和草原局, 四川 成都 610082
  • 收稿日期:2021-03-24 修回日期:2021-05-10 出版日期:2021-09-15 发布日期:2021-10-14
  • 通讯作者: 张新全,E-mail:zhangxq@sicau.edu.cn
  • 作者简介:梁小玉(1976-),女,四川合江人,博士,研究员,主要从事牧草资源评价、育种等研究,E-mail:liangxiaoyucao@163.com
  • 基金资助:
    四川省科技计划项目(2021YFYZ0013-4);四川省现代畜牧业发展类项目(SASA2021CZYX003)资助

Exploitation and Universal Analysis of EST-SSR Molecular Markers in Cichorium intybus

LIANG Xiao-yu1, JI Yang1, HU Yuan-bin1, YI Jun1, BAI Shi-qie3, ZHANG Jing1, ZHANG Xin-quan2   

  1. 1. Sichuan Academy of Animal Sciences, Chengdu, Sichuan Province 610066, China;
    2. College of Animal Science and Technology, Sichuan Agricultural University, Chengdu, Sichuan Province 611130, China;
    3. Sichuan Forestry and Grassland Administration, Chengdu, Sichuan Province 610082, China
  • Received:2021-03-24 Revised:2021-05-10 Online:2021-09-15 Published:2021-10-14

摘要: 为了开发菊苣(Cichorium intybus)简单重复序列(Simple sequence repeat,SSR)分子标记,采用菊苣表达序列标签(Expressed sequence tags,EST)序列设计EST-SSR引物,并验证引物的有效性和通用性。结果表明:菊苣EST序列中共搜索到648个SSR位点,146种重复基序,SSR出现频率为1.61%;三核苷酸重复类型最多(46.45%),TC/GA基序出现频率最高(14.35%);152对EST-SSR引物在菊苣中的有效扩增率和多态率分别为78.29%和57.24%。其中37对引物在31份莴苣亚族(Lactucinae Less.)材料中的扩增转移率为86.05%,遗传多样性分析显示不同种间存在明显的遗传分化,聚类结果与材料的地理来源有较高相关性。综上所述,基于菊苣EST序列开发的SSR引物可用于菊苣及其近缘属种遗传多样性分析及种间鉴定,为菊苣族(Lactuceae Cass.)种质资源评价、系统进化研究及分子标记辅助育种奠定了重要基础。

关键词: 菊苣, EST-SSR分子标记, 信息分析, 遗传多样性, 通用性

Abstract: To develop the molecular markers of the simple sequence repeat (SSR), the EST-SSR primer was designed based on the expressed sequence tags (ESTs) of chicory, and the applicability of EST-SSR primers were validated. The results showed that a total of 648 SSRs loci were identified in chicory ESTs, with 146 types of motifs and a 1.61% occurrence frequency of SSR. The trinucleotide had the most abundant of repeat types (46.45%), and TC/GA had the highest frequency of occurrence (14.35%).The effective amplification rate and polymorphism rate of the 152 designed primers in chicory was 78.29% and 57.24%, respectively. The amplification transferability rate of the 37 primers in 31 Lactucinae Less. was 86.05%. The genetic diversity analysis showed obvious genetic differentiation among different species, and the clustering results were highly correlated with the geographical origin of materials. The above results indicated that the development of EST-SSR markers could be used for the genetic diversity and genetic variation analysis in chicory and related species, which established an important foundation for germplasm resources evaluation, system evolution research and marker-assisted breeding of (Lactuceae Cass.).

Key words: Chicory, EST-SSR, Information analysis, Genetic diversity, Universality

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