| 利用SSR、ISSR和RAPD技术构建苜蓿基因组DNA指纹图谱 |
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| 引用本文： | 魏臻武.利用SSR、ISSR和RAPD技术构建苜蓿基因组DNA指纹图谱[J].草业学报,2004,13(3):62-67. |
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| 作者姓名： | 魏臻武 |
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| 作者单位： | 甘肃农业大学草业学院,甘肃,兰州,730070 |
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| 基金项目： | 教育部西部地区高校高级访问学者计划项目
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中-澳合作项目
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国家奶业重大科技专项基金 |
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| 摘 要： | 在建立可靠的苜蓿基因组DNA提取分离和PCR扩增技术体系的基础上,筛选具有稳定多态性位点的RAPD、SSR和ISSR引物,建立苜蓿基因组DNA的SSR、ISSR和RAPD分子标记技术体系,并利用分子标记检测苜蓿品种(系)的DNA分子标记多态性,构建苜蓿品种的DNA指纹图谱.筛选出36个RAPD引物,8个SSR引物和12个ISSR随机引物,通过PCR扩增在55个国内外苜蓿品种(系)中获得了182个RAPD多态性位点和丰富的SSR和ISSR多态性位点,构建了55个苜蓿品种(系)的SSR、ISSR和RAPD指纹图谱.结果表明,不同苜蓿品种(系)的SSR多态性有较大差异;苜蓿品种ISSR指纹图谱多态性丰富,稳定性比RAPD强;可以通过2～3个引物鉴别我国主要苜蓿品种和引进品种,SSR和ISSR指纹图谱可以更好地用于苜蓿品种鉴定和遗传多样性分析.
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| 关 键 词： | 苜蓿 分子标记 DNA指纹图谱 SSR ISSR RAPD |
| 文章编号： | 1004-5759(2004)03-0062-06 |
| 修稿时间： | 2003年6月16日 |
DNA fingerprint of Medicago sativa variety genomes using SSR, ISSR and RAPD |
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| WEI Zhen-wu.DNA fingerprint of Medicago sativa variety genomes using SSR, ISSR and RAPD[J].Acta Prataculturae Sinica,2004,13(3):62-67. |
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| Authors: | WEI Zhen-wu |
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| Abstract: | DNA extraction and the polymerase chain reaction (PCR) were used on DNA genomes of Medicago sativa. DNA polymorphic marker analysis was conducted and DNA fingerprint of 55 domestic and exotic Medicago sativa cultivars was carried out using SSR, ISSR and RAPD. Primers that can reliably find polymorphic bands were screened out. 36 RAPD primers were selected from the many available, and through PCR amplification obtained 182 polymorphic bands. Using 8 pairs of SSR and 12 ISSR screened out primers, through PCR amplification, 120 polymorphic bands were obtained. Results showed large differences in SSR polymorphism (among) the alfalfa varieties. Polymorphism was abundant in the Medicago sativa variety ISSR fingerprints, and stability was greater than for RAPD. 3-6 primers could distinguish between the Medicago sativa varieties. DNA fingerprinting can be applied to variety identification and genetic diversity evaluation of Medicago sativa. |
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| Keywords: | Medicago sativa SSR ISSR RAPD DNA fingerprint |
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