苦瓜抗白粉病SCAR分子标记的开发  

Development of SCAR from SRAP molecular markers closely linked to powdery mildew-resistant genes in bitter melon(Momordica charantia L.)

作　　者：周生茂[1,2] 尚小红[1] 梁任繁[1] 郭元元[1] 文俊丽[1] 黄皓[1] 陈振东[1] 班美玲 黄如葵[1] 

机构地区：[1]广西农业科学院蔬菜研究所,南宁530007 [2]广西作物遗传改良生物技术重点开放实验室,南宁530007 [3]广西环境保护科学研究院,南宁530022

出　　处：《南方农业学报》2013年第10期1595-1601,共7页Journal of Southern Agriculture

基　　金：国家自然科学基金项目(31240060);广西科学研究与技术开发计划项目(桂科攻10100005/6);广西自然科学基金项目(2012GXNSFAA053061);广西农业科学院基本科研业务专项项目(桂农科2012YT05);广西农业科学院公益性维持费项目(桂农科2012GW04)

摘　　要：【目的】获得与苦瓜抗白粉病基因紧密连锁的分子标记,为加快苦瓜抗白粉病新品种的选育奠定基础。【方法】以高抗白粉病野生苦瓜MC18为父本、高感白粉病苦瓜栽培种MC1-2为母本创建F2代分离群体;经单株抗病性鉴定后,以BSA法构建F2代单株的高抗和高感白粉病苦瓜DNA近等基因池;利用SRAP技术筛选多态扩增片段,对仅在抗白粉病近等基因池和父本中出现的差异片段进行回收、测序、比对和转化成SCAR标记,并利用已知抗病性的单株DNA分析标记与苦瓜抗白粉病的相关性。【结果】从1188对SRAP引物组合中筛选到稳定阳性差异条带的引物组合5对,其中ME20EM5引物对扩增的差异条带长度为332 bp,与葡萄抗体蛋白基因(抗性基因)的DNA序列有较高相似性,并将其成功转化成与苦瓜白粉病抗性相关、大小为320 bp的SCAR标记。【结论】开发的SCAR-ME20EM5分子标记可用于苦瓜抗白粉病分子标记辅助选择。[ Objective ]Molecular markers closely linked to powdery mildew resistant genes in bitter melon （Momordi- ca cbarantia L.） were obtained to provide basis on accelerating the development of new powdery mildew resistant cultivars. [Method]The plants from F2 generations were created through the hybridization of highly powdery mildew-sensitive and cultivated bitter melon MC1-2 as female parent with strongly powdery mildew- resistant and wild bitter melon MC18 as male parent. After identification powdery mildew resistance abilities with their parents, the near-isogenic DNA pools from F2 population plants were classified on the basis of bulked segregation analysis （BSA） method. Polymorphic amplified fragments were screened from near-isogenic DNA pools by sequence-related amplified polymorphism （SRAP） technique. The differential fragments only belonging to powdery mildew-resistant near-isogenic pool and paternal plant were re- trieved, sequenced, blasted, and changed into SCAR marker. The correlations of sequence characterized amplified region （SCAR） molecular marker with powdery mildew-resistant abilities were analyzed based on DNA of the known disease-re- sistant or-sensitive plants. [Result]Five pairs of SRAP primers in 1188 had stable and positive differential fragments, of which ME20EM5 primer combination not only got a differential fragment with 332 bp sharing comparatively high simi- larity with gene encoded grape antibody protein （disease-resistant gene）, but also changed into 320 bp SCAR molecu- lar marker significantly correlated with powdery mildew resistance. [ Conclusion ]SCAR molecular marker screened could be used to develop powdery mildew-resistant bitter melon using marker-assisted selection.

关 键 词：苦瓜 白粉病 集团分离分析法 SRAP SCAR 

分 类 号：S642.5[农业科学—蔬菜学]