番茄核苷酸结合位点(NBS)-亮氨酸富集重复(LRR)型抗病基因同源序列的分离与分析  

Isolation and Analysis of Nucleotide Binding Site(NBS)-leucine-richrepeat(LRR) Type Resistance Gene Analogs in Tomato(Solanum lycopersicum L.)

作　　者：倪小勋[1] 杨文才[1] 

机构地区：[1]设施蔬菜生长发育调控北京市重点实验室,中国农业大学农学与生物技术学院蔬菜学系,北京100193

出　　处：《农业生物技术学报》2013年第10期1145-1151,共7页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.30972003);高等学校博士学科点专项科研基金(No.20110008110019);北京市果类蔬菜产业技术体系

摘　　要：携带核苷酸结合位点(nucleotide binding site,NBS)和亮氨酸富集重复(leucine-rich-repeat,LRR)结构的NBS-LRR类抗性基因是迄今从植物中克隆到的最大一类抗病基因,在植物抵抗病原菌入侵和扩展过程中起着重要作用。为了分析番茄(Solanum lycopersicum L.)基因组内NBS-LRR类抗病基因同源序列的序列特征和结构域的保守性,本研究根据其保守结构域设计简并引物,以6份番茄材料OH88119、Ha7981、Ha7998、PI114490、PI128216和TR的基因组DNA为模板,通过PCR技术分离番茄抗病基因同源序列,经测序后分析其进化关系。共分离到了分布于12条染色体上的157条抗病基因同源序列,推导的氨基酸序列都具有P-Loop、Kinase2、Kinase3和GLPL几个保守区域。系统进化分析可将这些抗病基因同源序列分为两类:Toll interleukin-1受体域(TIR)-NBS-LRR类型和卷曲螺旋结构(CC)-NBS-LRR类型。研究结果为从番茄中分离抗性基因和进行抗性标记辅助选择育种提供参考。The nucleotide binding site （NBS） - leucine-rich-repeat （LRR） gene family is the largest group of resistance genes cloned from plant to date. They play important roles during pathogen infection and expansion in plants. To characterize the sequences and their conserved domains of NBS-LRR type of resistance genes in tomato（Solanum lycopersicum L.）, degenerate primers were designed using sequences of conserved domains and used to amplify resistance gene analogs （RGAs） from six tomato lines OH88119, Ha7981, Ha7998, PI114490, PI128216, and TR. The PCR products were sequenced and the sequences were subjected to evolutionary analysis. A total of 157 RGAs scattering on 12 chromosomes were isolated. The deduced amino acid sequences of all RGAs contained domains such as P-loop, Kinase2, Kinase3 and GLPL, which were conserved among NBS-LRR resistance genes. Phylogenetic analysis divided these RGAs into two classes, toll interleukin-1 receptor domain （TIR）-NBS-LRR and coiled-coil （CC）-NBS-LRR. These results will provide some information for isolation of disease resistance genes and marker-assisted selection in tomato.

关 键 词：番茄 抗病基因类似物(RGAs) 核苷酸结合位点-亮氨酸富集重复(NBS—LRR) 

分 类 号：S565.1[农业科学—作物学]