巴西橡胶NBS类抗病基因同源序列的克隆与分析  

Cloning and Primary Analysis on NBS Type Resistance Gene Analogs in Hevea brasiliensis

作　　者：张影波 庞玉新[2] 莫廷辉[1] 董美超[1] 解辉[1] 蔡秀清[1] 

机构地区：[1]海南大学农学院,海南儋州571737 [2]中国热带农业科学院热带作物品种资源研究所,海南儋州571737

出　　处：《安徽农业科学》2009年第24期11453-11455,共3页Journal of Anhui Agricultural Sciences

基　　金：海南省自然科学基金项目(80659)

摘　　要：[目的]探讨巴西橡胶NBS类抗病基因同源序列的克隆与序列分析。[方法]以高抗炭疽病的巴西橡胶无性系R042为材料,根据抗病基因Prf、L6、N等的P-loop和GLPL保守结构域设计简并引物,从R042扩增具有抗病基因同源序列的片段,对分离获得的抗病基因同源序列进行Blastn、氨基酸同源性分析、序列比较和系统进化分析。[结果]从巴西橡胶高抗炭疽病无性系R042基因组DNA中分离获得1个NBS类抗病基因同源序列,经Blastn比对后发现,该抗病基因同源序列与毛果杨中的一个CC-NBS-LRR抗病蛋白的mRNA的序列同源性为66%,Blastn的E值为2e-24,推测氨基酸序列与已知抗病基因的相似性在26.9%～43.2%。[结论]序列比对与系统进化分析表明,该抗病基因同源序列具有NBS类抗病基因的所有保守序列,且与Xa1的亲缘关系最近。[Objective]The study aimed to discuss the cloning and analysis on NBS type resistance gene analogs （RGA） in Hevea brasiliensis. [ Method ] With H. brasiliensi clone R042 with high resistance to anthraenose as the tested material, the degenerate primer was designed on base of the conserved motif （ P-loop and GLPL） of NBS domain from R-genes such as Prf, L6, N. The segments with RGA was amplified from the clone R042 and the isolated and obtained RGA sequences was made for the analysis of Blastn, amino acid homology, sequence alignment and phylogeny. [ Result ] A RGA sequence was isolated and obtained from the genome DNA of 11. brasiliensi clone R042 with high resistance to anthraenose. Through Blastn aligning, it was found that RGA sequence had the best similarity to the mRNA sequence of CC-NBS-LRR resistance gene in P. tomentosa, with the similarity of 66% and blastn E value of 2e-24 and it was deduced that the similarity of the a mino acid sequence to the known resistance gene ranged from 26.9% to 43.2%. [ Conclusion ] The sequence alignment phylogenetic analyses showed that the RGA sequence had all conserved sequence of NBS class resistance gene and had a closest similarity to the Xal.

关 键 词：巴西橡胶 核苷酸结合位点(NBS) 亮氨酸重复(LRR) 抗病基因同源序列(RGA) 

分 类 号：Q789[生物学—分子生物学]