Acta Agrestia Sinica

›› 2011, Vol. 19 ›› Issue (2): 299-305.DOI: 10.11733/j.issn.1007-0435.2011.02.022

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ISSR Maker Analysis of Stylosanthes spp.Strains by Space Mutation

BAI Chang-jun, YAN Lin-ling   

  1. 1. Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan Province, 571737, China;
    2. Ministry of Agriculture Key Laboratory for Utilization of Tropical Crops Germplasm Resources, Danzhou, Hainan Province, 571737, China
  • Received:2009-11-03 Revised:2010-12-28 Online:2011-04-15 Published:2011-04-15

空间诱变柱花草的ISSR分析

白昌军, 严琳玲   

  1. 1. 中国热带农业科学院作物品种资源研究所, 海南, 儋州, 571737;
    2. 农业部热带作物种质资源利用重点开放实验室, 海南, 儋州, 571737
  • 作者简介:白昌军(1967- ),男,甘肃民勤人,研究员,博士,从事热带牧草种质资源保存、鉴定评价、新品种选育等方面研究,E-mail:baichangjun2000@yahoo.com.cn
  • 基金资助:
    国家科技支撑计划(2008BADB3B08);物种资源保护项目;国家重点研究发展计划(973)项目(2007CB108903)资助

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Abstract: High quality DNA was extracted from the young leaves of Stylosanthes spp.by improved CTAB method;The ISSR PCR system suitable for Stylosanthes was developed after testing influenced factors.A total of 133 bands were amplified by 12 ISSR primers in 86 Stylosanthes spp.accessions,and 109 bands(81.95%) were polymorphism.The clustering dendrogram from the analysis with NTSYS software shows that: the genetic similarity coefficient of these accessions was 0.750～0.983,indicating the close genetic relationship of these materials.When similarity coefficient is set at Dice’s coefficient of 0.81,the 86 Stylosanthes spp.accessions could be divided into five groups.The results showed that only No.85 strain was closely related to the original strain Reyan 2.This fact suggested that the most of Stylosanthes spp.strains induced by space mutation would have some difference from the original referred ones in DNA level.

Key words: Stylosanthes spp., Space Mutation, ISSR-PCR reaction system

摘要： 采用改良CTAB法从空间诱变柱花草(Stylosanthes spp.)嫩叶中提取得到高质量的DNA,并对柱花草ISSR反应体系中的影响因子进行优化,建立柱花草ISSR反应的最佳体系。在此基础上,用12条引物对86份柱花草进行遗传多样性分析,结果表明:共获得133条谱带,其中多态性条带数为109条,占总条带数的81.95%,品种中间的遗传相似系数在0.75~0.98,表明其遗传关系比较近。当相似系数在0.81水平时,可将供试86份材料分为5大类,其中只有品系85号与原对照品种(热研2号)聚在一起,说明大部分空间诱变柱花草株系在DNA水平上与原对照品种存在差异。

关键词: 柱花草, 空间诱变, ISSR-PCR

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