麦族WKS基因的重组和遗传转化  

MOLECULAR DESIGN AND GENETIC TRANSFORMATION OF WKS GENES FROM TRITICEAE SPECIES

作　　者：李坤[1] 吕波[1] 姜辉[1] 吴佳洁[1] 付道林[1] 

机构地区：[1]作物生物学国家重点实验室,山东省作物生物学重点实验室,山东农业大学,山东泰安271018

出　　处：《山东农业大学学报（自然科学版）》2012年第4期483-490,共8页Journal of Shandong Agricultural University：Natural Science Edition

基　　金：国家自然基金重大国际合作项目(31110103917);山东省自然科学杰出青年基金(JQ201107);国家转基因专项任务(2011ZX08009-003-001-007)

摘　　要：小麦条锈病是由专性寄生真菌Puccinia striiformis f.sp.tritici引起的重要病害,在世界范围内广泛流行。小麦条锈病抗性基因Wheat Kinase-START 1(WKS1),是近年克隆的高温抗条锈病基因。本研究从二粒小麦、长穗偃麦草、百萨薄冰草和顶芒山羊草中克隆了6个WKS同源基因(WKS1或WKS2)的cDNA片段;通过基因切换创建了WKS1和WKS2之间的重组基因8个;并构建了WKS同源和重组基因相应的植物表达载体。为验证基因功能,分别将WKS同源和重组基因导入了高感锈病的短柄草株系,获得了转基因稳定整合和表达的阳性植株。本项研究为创建新型WKS基因和揭示WKS1基因的作用机理奠定了基础。Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most devastating disea- ses worldwide. The high temperature sensitive gene, Wheat Kinase - START 1 （ WKS1 ）, has been cloned re- cently. In this study, six WKS cDNA homologues （ WKS1 or WKS2） lmere clonde from Aegilops comosa, Thinopy- rum elongatum, Th. bessarabicum, and Triticum turgidum ssp. dicoccoides. Eight recombinant genes were cre- ated by motif swapping between Kinase and START. All native and derived WKS genes were used to construct Ubi： WKS expression vectors. To validate the gene function, they were transformed into a rust - susceptible ac- cession of Brachypodium distachyon. Stable transgenic events and cDNA transcription of investigated WKS genes were confirmed in To generation. The study lays a foundation for introducing novel WKS genes and revealing mo- lecular mechanism of WKS1 function.

关 键 词：小麦 短柄草 基因切换 条锈病 WKS1 

分 类 号：Q784[生物学—分子生物学]