Position: Home > Articles > Cloning and Analysis of NBS-LRR Type Disease Resistance Gene Analogs in Erianthus arundinaceum
Chinese Journal of Tropical Crops
2009,30
(2)
75-80
斑茅NBS-LRR类抗病基因同源序列的克隆与分析
作 者：
阙友雄;许莉萍;林剑伟;张木清;陈如凯
单 位：
福建农林大学农业部甘蔗遗传改良重点开放实验室
关键词：
班茅;保守区域;抗病基因同源序列;简并引物
摘 要：
根据已知NBS-LRR抗病基因[含有核苷酸结合位点(NBS)和富亮氨酸重复(LRR)的胞内受体蛋白基因]NBS结构域蛋白质的保守序列,设计简并引物,对斑茅基因组进行体外扩增,获得了对应区段的DNA片段,回收、克隆这些特异片段,测序分析,共获得8个片段序列。序列分析发现其中7个编号分别为RGA-Q1、RGA-Q2、RGA-Q3、RGA-Q4、RGA-Q5、RGA-Q6和RGA-Q7的片段推导的氨基酸序列均具有典型的NBS结构域,即P-loop(GGVGKTT)、Kinase-2a(VLDDVW)、Kinase-3a(GSR/KILVTTR)及疏水结构域HD(hydrophobic domain)。它们在NCBI上的登录号为EU685828、EU685829、EU685830、EU685831、EU685832、EU685833和EU685834。这些抗病基因同源片段(RGA)与已经克隆的N、L6、RPS2和Mi等11个抗病基因在氨基酸水平上的同源性为2.3%～39.8%,可进一步用作斑茅抗病候选基因的分子筛选及遗传图谱的构建。
译 名：
Cloning and Analysis of NBS-LRR Type Disease Resistance Gene Analogs in Erianthus arundinaceum
作 者：
Que Youxiong, Xu Liping, Lin Jianwei, Zhang Muqing, Chen Rukai Ministry of Agriculture Key Lab for Sugarcane Genetic Improvement, Agriculture and Forestry University, Fuzhou, Fujian 350002, China
关键词：
Erianthus arundinaceum; conserved motif; disease resistance gene analogs (RGA); degenerate oligonucleotide primers
摘 要：
Most of known plant disease resistance genes are featured with a nucleotide-binding site (NBS) and leucine-rich repeats (LRR). Degenerate oligonucleotide primers were designed according to the conserved NBS motifs among some disease resistance genes. These primers were used for amplification of disease resistance gene analog (RGA) from the genome (ic DNA) of Erianthus arundinaceum Retz. by polymerase chain reaction (PCR). After cloning and sequencing, 8 sequences in Erianthus arundinaceum Retz. were obtained, of which 7 were NBS-LRR type RGAs (RGA-Q1, RGA-Q2, RGA-Q3, RGA-Q4, RGA-Q5, RGA-Q6, RGA-Q7), the accession numbers of these RGAs were EU685828, EU685829, EU685830, EU685831, EU685832, EU685833 and EU685834, respectively. The deduced amino acid sequences of these DNA fragments contained the conserved motifs of NBS-LRR type RGAs, such as P-loop (Kinase 1a), Kinase 2a, Kinase 3a and hydrophobic domain. The sequences of each RGA were compared with those of eleven known resistance genes (N, L6, RPS2, Mi, etc.), and the percentage of amino-acid identity ranged from 2.3%～39.8%. These RGAs may further be used as molecular markers for screening of candidate disease resistance genes and genetic map construction in Erianthus arundinaceum Retz. The result indicated that PCR-based homologous cloning approach could be a potentially useful strategy in cloning of disease resistance genes and related genomic research.
