海岛棉的一个富含亮氨酸重复受体蛋白基因的克隆和功能分析  

Cloning and functional analysis of a leucine-rich-repeat receptor-like protein from island cotton

作　　者：蔡立旺[1] 杨郁文[2] 陈天子[2] 刘廷利[2] 凌溪铁 张保龙[2] 高进[1] 

机构地区：[1]江苏沿海地区农业科学研究所,江苏盐城224002 [2]江苏省农业科学院农业生物技术研究所,江苏南京210014

出　　处：《江苏农业学报》2014年第6期1240-1247,共8页Jiangsu Journal of Agricultural Sciences

基　　金：江苏省科技支撑计划项目(BE2013380);江苏省农业科技自主创新基金项目[CX(13)2029]

摘　　要：通过染色体步移获得了海岛棉的一个受体蛋白基因Gb Vdr1,其开放阅读框为3 387个核苷酸,编码1 128个氨基酸,编码蛋白分子量为1.249×105,等电点为5.86。Gb Vdr1与陆地棉抗病材料常抗棉和感病材料渝棉1号中的同源基因Gh Vdr1和Gh Vdr1-1的相似度高达99.85%。Gbvdr1具有信号肽、跨膜区、LRR重复区、蛋白质降解相关的PEST结构域以及内吞信号。Gb Vdr1在根茎中的表达量高于叶片,其对非落叶型黄萎病菌株BP2的反应较落叶型菌株V991的更为强烈,并且这种差异在茎和根中更为明显。Gb Vdr1定位于细胞膜上。PCR鉴定共获得Gb Vdr1过表达转基因株系23株,对目的基因表达量最高的株系进行黄萎病抗性鉴定,结果表明转基因株系对BP2的抗性显著增强,但是对V991没有抗性。转基因株系接种黄萎病后防卫反应(PR)相关基因PR1、PR5、EDS1以及GST1的表达量较野生型显著增加。对于Gb Vdr1的功能分析结果表明其参与了棉花对BP2的抗性反应。A receptor-like gene Gb Vdr1 was cloned from island cotton by genome walking.It contains an open reading frame of 3 387 bp and encodes 1 128 amino acids.The molecular weight and isoelectric point of Gbvdr1 is 1.249×105and 5.86.Gb Vdr1 shared similarity as high as 99.85% with Gh Vdr1 and homologous Gh Vdr1-1 isolated from resistant cultivar Changkangmian and susceptible cultivar Yumian No.1.Gbvdr1 has signal peptide,transmembrane region,LRR repeat,PEST domain related to protein proteolysis and endocytosis signal.The expression of Gb Vdr1 was higher in root and stem than that in leaves,and was induced more intensively by non-defoliating Verticillium dahlia isolate BP2 than by defoliating isolate V991 in stem and root.Gb Vdr1 was located on the cell membrane.23 Gb Vdr1 overexpressed plants were obtained by PCR identification,and the plant with the highest Gb Vdr1 expression was employed for Verticillium wilt resistance analysis.The results showed Gb Vdr1 could enhance the resistance to BP2 notably but had no function on V991.In addition,the expression of some pathogen related（PR） genes such as PR1,PR5,EDS1 and GST1 were increased more in the transformed plants than that in the wild type inoculated with V.dahlia.The functional analysis of Gbvdr1 indicated it was involved in the resistance reaction to BP2 in cotton.

关 键 词：棉花 黄萎病 受体蛋白基因 抗性基因 诱导表达 

分 类 号：S562.032[农业科学—作物学]