白粉菌诱导的小麦类萌发素蛋白的克隆、定位及表达分析  

Cloning and Chromosome Mapping of a Germin-Like Protein Gene in Wheat and Its Expression in Response to Infection with Wheat Powdery Mildew

作　　者：王俊美[1,2] 孙燕飞[1] 刘红彦[1,2] 康振生[1,3] 徐红明[2] 

机构地区：[1]西北农林科技大学植保资源与病虫害治理教育部重点开放实验室,陕西杨凌712100 [2]河南省农业科学院植物保护研究所,郑州450002 [3]西北农林科技大学陕西省农业分子生物学重点实验室,陕西杨凌712100

出　　处：《中国农业科学》2009年第9期3104-3111,共8页Scientia Agricultura Sinica

基　　金：河南省杰出青年基金项目(04120001400);西北农林科技大学植保资源与病虫害治理教育部重点开放实验室资助项目;国家"十一五"支撑计划(2006BAD08A05);现代农业产业技术体系建设专项资金资助;长江学者和创新团队发展计划资助项目(200558);高等学校学科创新引智计划资助项目(B07049)

摘　　要：【目的】克隆受白粉菌诱导的小麦类萌发素蛋白基因,分析其在感、抗病单株中的表达模式,探讨其在小麦抗白粉病过程中的作用机制。【方法】根据基因芯片结果,结合电子克隆与RT-PCR方法,克隆到一个受白粉菌诱导的小麦类萌发素蛋白基因。通过中国春缺体-四体系对获得的基因片段进行定位。运用荧光定量PCR方法分析了该基因片段在抗病植株、感病植株白粉菌诱导的时空表达模式。【结果】获得了一个具有完整开放阅读框的小麦类萌发素蛋白基因片段,命名为TaGLP5(GenBank登录号为FJ594470)。系统进化分析表明该基因片段与已知的来自禾本科的萌发素蛋白分属不同的进化分支,很可能是新的小麦萌发素蛋白成员。通过中国春缺体-四体系将该基因片段定位在小麦的5A染色体上。定量PCR分析结果表明,该基因的表达受白粉菌诱导,且在接菌后的24h以前抗病中的表达量高于同期感病株。【结论】本实验获得的类萌发素蛋白是一个新的成员。该类萌发素蛋白在感、抗植物受白粉菌诱导上调表达,但是表达量、表达时间上存在差异。推测该基因参与了小麦对白粉菌的防御反应。Objective The aim of this study is to clone a wheat germin-like protein gene induced by Bgt（Blumeria graminis f.sp.tritici）,analyse the expression pattern in susceptible and resistant plants and elucidate the molecular mechanism of the gene in resistance to powdery mildew.Method Gene chip hybridization,in silico cloning and RT-PCR methods were used to clone a wheat germin-like gene induced by Bgt,named as TaGLP5（GenBank accession number FJ594470）.A set of nulli-tetrasomic lines of Chinese Spring were employed to locate the gene. Its expression pattern at transcription level was investigated by quantitative PCR in resistant and susceptible progenies. [Result] A wheat germin-like protein, TaGLP5 with a complete ORF was cloned. Phylogenetic assay indicated that this gene differed from the known germin-like proteins from gramineous family. It was located on chromosome 5A. Quantitative PCR showed that the gene was induced by Bgt and higher expression in resistant plants than in susceptible ones before 24 h of inoculation. These will provide an important reference for further study on the gene. [ Conclusion ] A new wheat germin-like protein was obtained in this study. The expression patterns results revealed that TaGLP5 was up-regulated in both resistant and susceptible plants. However, the expression in quantity and time was different. It was speculated that TaGLP5 was involved in wheat defense response to Bgt.

关 键 词：白粉菌 类萌发素蛋白 克隆 表达模式 染色体定位 

分 类 号：S512.1[农业科学—作物学]