辣椒NBS类抗病基因同源序列的克隆与分析

Cloning and Analysis of NBS Class Resistant Gene Analogues in Capsicum annuum

辣椒RGA的分离将为进一步从辣椒中分离功能性抗病基因和抗病机理分析打下基础,为辣椒相关抗病基因的克隆提供依据。根据已知抗病基因保守氨基酸结构域设计简并引物,以高抗辣椒品种88为试材,通过PCR扩增抗病基因同源序列。结果表明:从辣椒基因组DNA中分离出1条辣椒抗病基因同源序列WD1。对其编码的氨基酸序列进行分析表明:该序列同时含有P-环(GGVGKTT)、kinase-2(VLDD)、kinase-3(GSRII)及HD(即GLPLAL)保守域结构,分离的辣椒抗病基因同源序列(RGA)与已报道的辣椒抗病基因同源序列A5和A13都有99%的同源性。

This RGA isolated from the pepper used in present study would provide the base for the further cloning of disease-resistance genes and the analysis of resistance mechanism in pepper,and provide theoretical basis for the cloning of resistance gene-related in pepper.According to the conserved amino acid domains that exist in well-known disease-resistance genes（R-genes）,the corresponding degenerated primer had been synthesized,which is used pepper strain 88 with high resistance as materials.Resistance gene analogues has been isolated by PCR（polymerase chain reaction） amplification.1 NBS（nucleotide binding site） types of resistance gene analogues（WD1）from the pepper were obtained by PCR amplification.The amino acid sequences translated from WD1 contain conserved domains,such as P-loop,kinase-2（VLDD）,kinase-3（GSRII） and GLPL.This RGA isolated from pepper respectively have 99% homology with the reported resistance gene analogues A5 and A13.