不同秋眠性苜蓿SRAP体系优化及遗传多样性分析

草业学报 ›› 2011, Vol. 20 ›› Issue (2): 201-209.

不同秋眠性苜蓿SRAP体系优化及遗传多样性分析

何庆元^1,2,吴萍¹,张晓红¹,王松华^1*,李正鹏¹,汪竹香¹

1.安徽科技学院生命科学学院, 安徽凤阳 233100;
2.南京农业大学大豆研究所国家大豆改良中心作物遗传与种质创新国家重点实验室, 江苏南京 210095

收稿日期:2010-03-29 出版日期:2011-02-25 发布日期:2011-04-20
通讯作者: E-mail:shwang70@yahoo.com.cn
作者简介:何庆元(1978-),男,安徽宿松人,助理研究员,在读博士。E-mail:heqingyuan1@163.com
基金资助:
国家“973”计划基金项目(2007CB108901),安徽省教育厅重大基金项目(ZD200910)和安徽科技学院校级重点学科(AKXK20102-1)资助。

A study on optimization and primer screening of a SRAP reaction system and genetic diversity of different fall dormancy alfalfas

HE Qing-yuan^1,2, WU Ping¹, ZHANG Xiao-hong¹, WANG Song-hua¹, LI Zheng-peng¹, WANG Zhu-xiang¹

1.Life Science College of Anhui Science and Technology University, Fengyang 233100, China;
2.Soybean Research Institute of Nanjing Agricultural University, National Center for Soybean Improvement, National Key Laboratory for Crop Genetics and Germplasm Enhancement, Nanjing 210095, China

Received:2010-03-29 Online:2011-02-25 Published:2011-04-20

摘要/Abstract

摘要： 用改良CTAB(十六烷基三甲基溴化铵)法提取苜蓿基因组DNA,用L₁₆(4⁵)正交设计,研究了模板DNA、dNTPs、Mg²⁺、Taq酶和引物的浓度对扩增迁移率重现性的影响,结果表明,在20 μL体系中含有50 ng DNA、1.4 mmol/L Mg²⁺、1.2 U Taq酶、0.4 mmol/L dNTPs和0.25 μmol/L浓度的引物最为稳定,重现率达到100%。利用最佳体系从100对引物中筛选出8对在34个苜蓿上扩增效果好的引物,共扩增出87个SRAP标记,有84个多态性位点。聚类分析结果表明,来自安徽和江苏两地的野生南苜蓿和其他栽培苜蓿的遗传差异最大,单独聚为一类;其他32个苜蓿品种在相似系数0.68附近聚为3个亚群,秋眠型苜蓿品种主要分布在第2个亚群中,半秋眠和非秋眠型苜蓿品种分布散乱,并不表现出成族分布,表明秋眠性与苜蓿的亲缘关系不完全一致。

Abstract: Genomic DNA was extracted by advanced CTAB methods from Medicago sativa. The L₁₆(4⁵) orthogonal diagram was applied to study the effects of mobility reproducible in different concentrations of template DNA, dNTPs, Mg²⁺, Taq polymerase, and primer on M. sativa. A suitable SRAP reaction system had 50 ng DNA,1.4 mmol/L Mg²⁺,1.2 U Taq polymerase, 0.4 mmol/L dNTPs and 0.25 μmol/L in a 20 μL reaction system. Better primers were screened out from 100 pairs of primers using optimizated systems, that could amplify clear fragments. A total of 87 amplicons were resolved by 8 pairs of well-chosen primers, of which 84 were polymorphic. These varieties had high genetic polymorphisms. A dendrogram from the analysis clustered two Medicago hispida cultivars from Anhui and Jiangsu into one group. Thirty two other cultivars were classified into sub-groups with a similarity coefficient of 0.68. Fall dormancy alfalfa cultivars were mostly distributed in the second sub-group. Semi-fall dormancy and non-fall dormancy alfalfa cultivars did not fall in the same sub-group. There was no absolute distinction of fall dormancy of alfalfa.

中图分类号:

何庆元,吴萍,张晓红,王松华,李正鹏,汪竹香. 不同秋眠性苜蓿SRAP体系优化及遗传多样性分析[J]. 草业学报, 2011, 20(2): 201-209.

HE Qing-yuan, WU Ping, ZHANG Xiao-hong, WANG Song-hua, LI Zheng-peng, WANG Zhu-xiang. A study on optimization and primer screening of a SRAP reaction system and genetic diversity of different fall dormancy alfalfas[J]. Acta Prataculturae Sinica, 2011, 20(2): 201-209.

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