辣椒NBS-LRR类型抗病基因同源序列的克隆及分析  

Cloning and analysis of NBS-LRR type gene analogs in pepper(Capsicum annuum L.)

作　　者：赵敬[1,2] 万红建[2] 杨悦俭[2] 侯喜林[1] 

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室,江苏南京210095 [2]浙江省农业科学院蔬菜研究所,浙江杭州310021

出　　处：《江苏农业学报》2012年第3期611-616,共6页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(31071800);浙江省公益技术研究农业项目(2011C22007);浙江省重大科技专项(2009C02006-1)

摘　　要：为研究辣椒NBS-LRR类抗病基因的结构和功能特点,运用前人根据NBS-LRR类抗病基因保守结构域(P-Loop和GLPL)设计的简并引物,以辣椒PBC631B的基因组DNA为模板进行PCR扩增,共获得17个具有完整开放阅读框的辣椒抗病基因同源序列(CaRGA01～CaRGA17)。多重序列比对显示这些序列都含有NBS-LRR类基因的6个保守结构域(P-loop、RNBS-A-nonTIR或RNBS-A-TIR、Kinase-2、RNBS-B、RNBS-C和GLPL)。与已知6个抗病基因(Gpa2、L6、M、N、Prf和RPM1)构建系统进化树,发现这17个序列被分为TIR-NBS-LRR和nonTIR-NBS-LRR2组,进一步划分为4个亚组(CaRGAⅠ～CaRGAⅣ)。其中,CaRGA01和CaRGA13分别与来自番茄Solanum lycop-ersicum抗细菌性斑点病基因Bs4和番茄Solanum sp.VFNT抗线虫基因Mi-1.4相似性最高,分别为90%和91%,推测这2个序列可能是相关抗病基因的一部分或与相关抗病基因属于同一家族。这些结果将为研究辣椒抗病相关基因提供理论依据。To study the structure and function of nucleotide-binding site leucine-rich repeat （NBS-LRR） type of disease-resistance genes in pepper, two pairs of degenerate primers designed based on conserved domains （P-Loop and GL- PL） in NBS-LRR resistance genes in plants, were used to isolate NBS-LRR sequences from pepper （ Capsicum annuum L. ） cultivar PBC631B. Seventeen resistance gene analogues （RGAs） with uninterrupted open reading frames （ORFs） were obtained from pepper （designated CaRGAO1-CaRGA17）. Multiple sequence alignments revealed six conserved motifs （P-loop, RNBS-A-nonTIR/RNBS-A-TIR, Kinase-2, RNBS-B, RNBS-C and GLPL）. Phylogenetic tree analysis between these CaRGAs and six known resistance genes （RPM1, Gpa2, L6, M, N and Prf） showed that 17 RGAs were grouped into non TIR-NBS-LRR and TIR-NBS-LRR families and further seperated into four subgroups （CaRGA I-CaRGA IV）. Amongthem, CaRGA01 and CaRGA13 showed high sequence similarities （90% and 91%, respectively） with bacterial spot disease resistance protein gene Bs4 and Mi-1.4 dis- ease resistance gene from Solanum lycopersicum and Sola- num sp. VFNT, respectively, which suggested that these two genes might be parts of the related resistance genes in pepper. The results might be conducive to clone resistance genes from pepper in future.

关 键 词：NBS-LRR 抗病基因同源序列 辣椒 克隆 

分 类 号：Q78[生物学—分子生物学]