中国水稻科学

• 研究报告 •    下一篇

水稻抗病基因同源序列的克隆及测序分析

杨勤忠1; 杨佩文1; 王 群1; 刘继梅2; 鄢 波2; 李家瑞1; 黄兴奇2   

  1. 1 云南省农业科学院 植物保护研究所,云南 昆明 650205; E-mail:qzyang@public.km.yn.cn; 2 云南省农业科学院 生物技术研究所,云南 昆明 650223
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2001-10-10 发布日期:2001-10-10

Cloning and Sequencing of Disease Resistance Gene Analogues in Rice (Oryza sativa L.)

YANG Qin-zhong 1; YANG Pei-wen 1; WANG Qun 1; Liu Ji-mei 2; YAN Bo 2; LI Jia-rui 1; HUANG Xing-qi 2   

  1. (1Plant Protection Institute; Yunnan Academy of Agricultural Sciences; Kunming 650205; China; E-mail:qzyang@public.km.yn.cn; 2Yunnan Academy of Agricultural Sciences, Kunming 650223, China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2001-10-10 Published:2001-10-10

摘要: 根据已知的NBSLRR类及丝/苏氨酸蛋白激酶类抗病基因结构中氨基酸的保守区域,设计了两组简并引物用于扩增广谱抗稻瘟病品种云系2号中的抗病基因同源序列。结果一共获得11类的NBS-LRR类抗病基因同源片段及16类的丝/苏氨酸蛋白激酶类抗病基因同源片段。所有11类的抗病基因同源系列均含有NBS-LRR类抗病基因的保守序列,如P-loop、Kinase 2、Kinase 3a以及跨膜区域等。所有16类的蛋白激酶的序列均含有丝/苏氨酸蛋白激酶所共有的催化区Ⅵ(共有氨基酸序列:DLKPEN)、Ⅷ(共有氨基酸序列:GT/SXXYXAPE)以及蛋白激酶的其他催化区。两条NBS-LRR类的抗病基因同源片段YR1、YR12分别与抗病基因I2 C-2及Xa1氨基酸同源性很高(>50%)。7条丝/苏氨酸蛋白激酶类的抗病基因同源片段与已克隆的[WTBX][STBX]Xa21、Pto、Lr10[WTBZ][STBZ]等抗病基因的氨基酸序列超过60%的相同以及76%~78%的氨基酸类似。

关键词: 水稻, 核酸结合位点, 丝/苏氨酸蛋白激酶, 稻瘟病, 抗病基因, 同源序列

Abstract: Two sets of degenerate oligonucleotide primers were designed on the basis of nucleotide-binding site (NBS) motifs conserved among the disease resistance genes Xa1, RPM1, RPS2, N, L6, Pib, I2C, Prf, M, and serine/threonine protein kinase (STK) motifs conserved among the disease resistance genes Pto, Fen, Lr10, Xa21, respectively. The primers weresubsequentlyused as PCR primers to amplify resistance gene analogues in a high blast resistant rice cultivar Yunxi 2 (Oryza sativa L.). Sequencing of amplification products indicated that at least 11 classes of NBS resistance gene analogues and 16 classes of STK resistance gene analogues were detected. All of NBS resistance gene analogues contained the conserved motifs of NBS LRR type resistance genes, such as P loop (Kinase 1a), Kinase 2, Kinase 3a and transmembrance domain. All of STK resistance analogues contained the catalytic domains of serine/threonine protein kinase, such as Ⅰ to Ⅴ, Ⅵ (consensus sequence: DLKPEN), Ⅶ, Ⅷ(consensus sequence GT/SXXYXAPE) and Ⅸ. YR12 showed significant amino acid homology (63% identify and 77% similarity) to blight resistance gene Xa1 of rice to Xanthomonas oryzae pv. oryzae. YK19 exhibited 52% deduced amino acid identify and 67% similarity to resistance gene Xa21 of rice to Xanthomonas oryzae pv. oryzae. YK13,YK21,YK25 and YK26 showed significant amino acid homology (61%-64% identify and 76%-78% similarity) to resistance gene Lr10 of wheat to Puccinia recondita. YK1 showed 60% identify and 76% similarity to resistance gene Pto of tomato to Peseudomonas syringae pv. tomato.

Key words: rice, nucleotide binding site, serine/threonine protein kinase, rice blast, resistance gene, analogues