烟草NBS-LRR类抗病基因同源序列的克隆及分析  

Cloning and Analysis of the Homologous Sequence of NBS-LRR Resistance Genes in Tobacco

作　　者：魏环宇 邓小鹏[3] 晋艳[3] 蔺忠龙[4] 郑元仙[5] 何元胜 陈小龙 魏薇 黄飞燕 余磊 童文杰[3] Wei Huanyu;Deng Xiaopeng;Jin Yan;Lin Zhonglong;Zheng Yuanxian;He Yuansheng;Chen Xiaolong;Wei Wei;Huang Feiyan;Yu Lei;Dong Wenjie(Yunnan Urban Agricutural Engineering&Technological Research Center,Cllege of Agronomy,Kunming University,Kunming,650214;College of Plant Protection,Yunnan Agricultural University,Kunming,650201;Yunnan Academy of Tobacco Agricultural Sciences,Kunming,650201;China National Tobacco Corporation Yunnan Company,Kunming,650011;Lincang Cormpany of Yunnan Tobacco Company,Lincang,677000;China Tobacco Henan Industrial Co,Lid,Zhengzhou,450000)

机构地区：[1]昆明学院农学院,云南省都市特色农业工程技术研究中心,昆明650214 [2]云南农业大学植物保护学院,昆明650201 [3]云南省烟草农业科学研究院,昆明650201 [4]中国烟草总公司云南省公司,昆明650011 [5]云南省烟草公司临沧市公司,临沧677000 [6]河南中烟工业有限责任公司,郑州450000

出　　处：《分子植物育种》2020年第11期3524-3529,共6页Molecular Plant Breeding

基　　金：中国烟草总公司云南省公司资助项目(2018530000241020,2018530000241016,2019530000241028,201953-0000241011)资助。

摘　　要：通过同源克隆法获得烟草抗病基因同源序列(RGAs),为烟草抗病基因的筛选和相关研究提供帮助。基于抗病基因的NBS-LRR保守结构域,筛选并合成了3对简并引物,扩增烟草中的NBS-LRR类抗病基因。获得了4条与抗病基因具有高度同源性的NBS-LRR类烟草抗病基因同源序列(TRGA,登录号:MK634316,MK634317,MK634318,MK634319),目的片段大小均在500 bp左右;BLAST X分析表明,获得的4个烟草RGAs与已知RGAs具有高度相似性,氨基酸相似性为97%~100%;聚类分析将其分成2大类,包括TIRNBS-LRR和non-TIR-NBS-LRR两类抗病基因,其编码氨基酸序列含有NBS保守区的典型特征基序。通过简并引物进行同源扩增是分离烟草RGAs的有效方法,可为进一步抗病基因筛选及抗病分子育种奠定基础。The resistance gene analogs(RGAs)were measured by homologous cloning method,which could be helpful for the detection and characterization of disease resistance genes in tobacco.Based on the conserved structural domain of disease-resistant genes,three pairs of degenerate primers were screened and synthesized to amplify the disease-resistant genes of NBS-LRR in tobacco.Four NBS-LRR tobacco disease resistance gene homologous sequences(TRGA,registration number:MK634316,MK634317,MK634318,MK634319)with high homology to disease resistance genes were obtained.BLAST X analysis showed that the 4 tobacco RGAs were highly similar to the known RGAs,with amino acid similarity ranging from 97%to 100%.Cluster analysis divided them into two categories,including TIR-NBS-LRR and non-TIR-NBS-LRR disease-resistant genes,whose coded amino acid sequences contained typical motifs of the conserved region of NBS.Homologous amplification with degenerate primers is an effective method to isolate tobacco RGAs,which can lay a foundation for further screening of disease-resistant genes and disease-resistant molecular breeding.

关 键 词：烟草(Nicotiana tabacum L.) NBS-LRR 抗病基因同源序列 

分 类 号：Q943.2[生物学—植物学] S435.72[农业科学—农业昆虫与害虫防治]