辣椒抗黄瓜花叶病毒同源序列的克隆与分析  

Analysis and Cloning of Cucumber Mosaic Virus Resistance Gene Analogs in Pepper(Capsicum annuum L.)

作　　者：杨学玲[1,2] 王述彬[2] 刘金兵[2] 潘宝贵[2] 万红建[1] 陈劲枫[1] 

机构地区：[1]南京农业大学园艺学院,江苏南京210095 [2]江苏省农业科学院蔬菜研究所,江苏南京210014

出　　处：《江苏农业学报》2009年第4期838-842,共5页Jiangsu Journal of Agricultural Sciences

基　　金：国家"863"计划(2008AA102150);国家"十一五"科技支撑项目(2006BAD01A7-04-03;2008ADB1B04);江苏省农业科技自主创新资金项目(CX08610);江苏省科技基础设施建设计划(BM200808)

摘　　要：根据已知抗病基因的核苷酸结合位点（NBS）保守结构域设计简并引物，以高抗黄瓜花叶病毒的辣椒Yv2007001为试材，获得10条抗病基因同源序列（RGA）。经序列分析，这些RGAs皆具有开放阅读框和NBS保守结构域（P-loop、Kinase-2、Kinase-3a、RNBS-C和GLPL），属于nonTIR—NBS-LRR类抗病基因同源序列。其中RGA46与番茄花叶病毒抗性基因Tm-2^2的氨基酸同源性为78％，可能与辣椒抗黄瓜花叶病毒（CMV）相关。其它序列与已知抗病基因Prf、RPP13、I2C-1、Mi-1．2、L6、M氨基酸相似性为21％～70％。所得RGA序列的Ka／Ks值为0．0110—0．3051，均显著小于1，表明“纯化选择”在辣椒NBS区域进化中起主要作用。Ten resistance gene analogs （RGAs） were cloned from a pepper variety （ Yv2007001 ） with high resistance to cucumber mosaic virus（ CMV ） , using a PCR approach with degenerate primers designed from conserved motifs found in a number of plant resistance genes. Sequence analysis showed that all RGAs belong to nonTIR-NBS-LRR subgroup, containing ORF and five conserved motifs including P-loop, kinase-2a, kinase-3a, RNBS-C and GLPL. RGA46 showed 78% homology at the deduced amino acid with Tin-2^2（ ToMV）, suggesting that RGA46 has correlation with resistance to CMV. The highest and the lowest homology, between the RGAs and known resistance genes （Prf, RPP13,I2C-1, Mi-1. 2, L6 and M） were found 70% and 21% respectively. The ratios of non-synonymous （Ka） to synonymous nueleotide substitution （Ks） of the NBS domains of RGAs were 0. 011 0 - 0. 305 1, less than which showed that purifying selection plays an important role in evolutionary process of resistance genes NBS region of pepper.

关 键 词：辣椒 黄瓜花叶病毒 抗病基因同源序列 核苷酸结合位点-富含亮氨酸重复 

分 类 号：Q785[生物学—分子生物学]