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

doi:10.11733/j.issn.1007-0435.2007.06.010

›› 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 Fang^1,2, YUAN Qing-hua¹, SU De-rong², GAO Jian-ming³

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. 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

Abstract

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

CLC Number:

S432.21
Q943

MENG Fang, YUAN Qing-hua, SU De-rong, GAO Jian-ming. Study on ISSR Markers Linked to Common Leaf Spot Disease Resistance Gene of Tetraploid Alfalfa[J]. , 2007, 15(6): 561-565.

孟芳, 袁庆华, 苏德荣, 高建明. 紫花苜蓿抗褐斑病ISSR分子标记研究[J]. , 2007, 15(6): 561-565.

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

紫花苜蓿抗褐斑病ISSR分子标记研究

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