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草业学报  2015 

燕麦隐性核不育cSSR标记的开发及其验证

DOI: 10.11686/cyxb2014232, PP. 146-154

Keywords: 燕麦,cSSR,SSR标记

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Abstract:

构建燕麦隐性核不育转录组数据库,开发cSSR标记,丰富燕麦分子标记数据库,并对隐性核不育相关cSSR标记进行验证,为充分利用燕麦种质资源及深入研究隐性核不育奠定基础。对燕麦雄性不育近等基因系CAMS1进行转录组测序,利用软件对得到的EST序列进行SSR位点查找和分析,开发cSSR引物。并用CAMS1×品燕2号F2代群体对所开发的雄性不育相关的cSSR引物进行验证。通过转录组测序获得的EST序列中6409条存在SSR位点,736条序列中有2个及2个以上SSR位点;在所有SSR位点中三核苷酸、二核苷酸重复序列最多,分别为4340(56.66%)和1768(24.30%)个。三核苷酸重复中,CCG/CGG(26.68%)、AGG/CTT(21.29%)、AGC/CTG(18.48%)出现的频率较多,二核苷酸重复中,以AG/CT(60.63%)和AC/GT(26.24%)出现的频率较高;根据开发的cSSR设计了13344对cSSR引物,选择与雄性不育相关且能设计引物的21条序列合成45对SSR引物。PCR检测表明,32对引物(71%)有扩增产物,其中11对cSSR引物在亲本间的扩增产物具有多态性,7对引物在可育和不育性状池间存在差异。本研究对燕麦雄性不育近等基因系CAMS1进行了转录组测序,根据得到的EST序列开发了13344对cSSR引物,并利用CAMS1×品燕2号F2代群体进行有效性检测,32对引物有扩增产物,7个标记可用于燕麦雄性不育材料的分子检测。

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