Acta Agrestia Sinica

›› 2007, Vol. 15 ›› Issue (6): 561-565.DOI: 10.11733/j.issn.1007-0435.2007.06.010

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Study on ISSR Markers Linked to Common Leaf Spot Disease Resistance Gene of Tetraploid Alfalfa

MENG Fang1,2, YUAN Qing-hua1, SU De-rong2, GAO Jian-ming3   

  1. 1. Institute of Animal Sciences, CAAS, Beijing 100094, China;
    2. College of Resources and Environment, Beijing Forestry University, Beijing 100083, China;
    3. Tianjin Agricultural College, Tianjin 300384, China
  • Received:2007-06-27 Revised:2007-08-31 Online:2007-12-15 Published:2007-12-15

紫花苜蓿抗褐斑病ISSR分子标记研究

孟芳1,2, 袁庆华1, 苏德荣2, 高建明3   

  1. 1. 中国农业科学院北京畜牧兽医研究所, 北京, 100094;
    2. 北京林业大学资源与环境学院, 北京, 100083;
    3. 天津农学院, 天津, 300384
  • 通讯作者: 袁庆华,E-mail:yuanqinghua@hotmail.com
  • 作者简介:孟芳(1983- ),女,汉族,安徽人,硕士研究生,研究方向为牧草资源,E-mail:mengfang1021@126.com
  • 基金资助:
    国家自然科学基金(30471230);国家奶业项目(2006BAD04A04-03-07)资助

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Abstract: In order to establish breeding technique system of marker-assisted selection,inter-simple sequence repeat(ISSR) was employed and accompanied with bulked-segregant analysis(BSA) to detect molecular markers linked to the resistant gene of Common leaf spot in the F1 of alfalfa(Medicago sativa L.).The results show that 32 of 93 ISSR primers could amplify clear and steady bands,and there is 6 ISSR primers in the F1 of moderately resistant could amplify polymorphic bands with length of 430-900 bp between resistant and susceptible DNA pools composed of 12 highly resistant plants and 12 highly susceptible plants.The 6 ISSR markers were further verified by 12 plants apiece from resistant and susceptible DNA pools,25 plants apiece from 3 combinations of highly resistant×highly resistant,and 25 plants apiece from 2 combinations of highly susceptible×highly susceptible.In F1 progeny,9-R920、20-R750、21-R430、818-R680 were found to be significantly associated with resistance and 866-S800 significantly associated with susceptibility.

Key words: Alfalfa, Common leaf spot, ISSR markers

摘要： 运用ISSR分子标记技术,采用集群分离分析法,对四倍体紫花苜蓿(Medicago sativa)F1代进行抗褐斑病基因连锁的分子标记筛选,进而建立苜蓿褐斑病抗性遗传资源筛选和分子标记辅助选择(MAS)育种技术体系。结果表明:在93个随机引物中,有32个能够产生清晰稳定的扩增条带,其中6个在中抗杂交F1代高抗、高感各12个单株所组成的抗、感DNA池间产生差异性条带;在抗、感DNA池的各12个单株中,挑选出高抗×高抗杂交F1代3个组合中各25个单株,高感×高感杂交F1代2个组合各25个单株进行验证,结果9-R920、20-R750、21-R430、818-R680均与F1代苜蓿褐斑病抗病基因紧密连锁,866-S800与F1代苜蓿褐斑病感病基因紧密连锁。

关键词: 苜蓿, 褐斑病, ISSR标记

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