瑟伯氏棉TIR-NBS-LRR类蛋白质编码基因的克隆与分析  

Cloning and analysis of genes encoding TIR-NBS-LRR-like proteins in Gossipium thurberi

作　　者：杨晓杰[1,2] 赵付安[1] 唐中杰[1] 李武[1] 赵元明[1] 谢德意[1] 房卫平[1] 

机构地区：[1]河南省农业科学院经济作物研究所,河南郑州450002 [2]棉花生物学国家重点实验室,中国农业科学院棉花研究所,河南安阳455000

出　　处：《江苏农业学报》2013年第6期1228-1235,共8页Jiangsu Journal of Agricultural Sciences

基　　金：棉花生物学国家重点实验室开放课题基金项目(CB2013A03);河南省财政预算项目(棉花抗黄萎病候选基因的克隆与功能验证);国家自然科学基金项目(U1304319)

摘　　要：以对抗黄萎病种质瑟伯氏棉接菌前后高调表达的3个TIR-NBS-LRR类蛋白质点为出发点,根据其对应同源蛋白质B3V7R2(烟草)、B9INW6(棉白杨)和Q19EF1(拟南芥),利用BLAST在线搜索棉花中对应的表达序列标签(EST),分别设计引物,对接菌后的瑟伯氏棉进行RT-PCR扩增,进而以获得的EST序列为母序列,通过电子克隆及RACE技术,克隆了3个编码TIR-NBS-LRR类蛋白质的候选基因。生物信息学分析结果表明,3个候选TIR-NBS-LRR类蛋白质编码基因均具有典型的TIR、NBS及LRR功能域;系统进化分析结果显示,3个候选基因在瑟伯氏棉抗黄萎病机制中可能起重要作用。Based on Proteins B3V7R2 in Nicotiana tabacum, B9INW6 in Populus trichocarpa and Q19EF1 in Arabidopsis thaliana, homologous with the three TIR-NBS-LRR-type protein spots in Gossipium thurberi which were up- regulated after the inoculation with VerticiUium dahliae, the corresponding expressed sequence tags （ESTs） of G. thurberi were found by BLAST, and three novel TIR-NBS-LRR-like candidate genes was cloned from G. thurberi after V. dahliae inoculation by silico cloning and rapid amplification of cDNA ends （RACE）. Bioinformatics analysis ＇re- vealed that all the three candidate genes possessed the typical functional domains including TIR, NBS, and LRR.Phylogenetic analysis implied that the three candidate disease resistant genes might play important roles in the molecular mechanism of Gossipium Thurberi in response to cotton VerticiUium wilt.

关 键 词：瑟伯氏棉 黄萎病 TIR-NBR-LRR蛋白质编码基因 克隆与分析 

分 类 号：S562[农业科学—作物学]