烟草(红花大金元)NBS-LRR类抗病基因同源序列的克隆与分析  

Isolation and Analysis of NBS-LRR Resistance Gene Analogs from Tobacco (Hongda)

作　　者：魏环宇 童文杰[2] 蔺忠龙[3] 莫笑晗[2] 张丽芳[2,6] 何元胜 郑元仙 王继明 许银莲 陈小龙 钟宇 余磊 邓小鹏[2] WEI Huan-yu;TONG Wen-jie;LIN Zhong-long;MO Xiao-han;ZHANG Li-fang;HE Yuan-sheng;ZHENG Yuan-xian;WANG Ji-ming;XU Yin-lian;CHEN Xiao-long;ZHONG Yu;YU Lei;DENG Xiao-peng(College of Agronomy,Yunnan Urban Agricultural Engineering&Technological Research Center,Kunming University,Yunnan Kunming 650214,China;Yunnan Academy of Tobacco Sciences,Yunnan Kunming 650201,China;China National Tobacco Corporation Yunnan Company,Yunnan Kunming 650011,China;Lincang Company of Yunnan Tobacco Company,Yunnan Lincang 677000,China;Tobacco Leaf Purchase Center,China Tobacco Henan Industrial Co.,Ltd.,Henan Zhengzhou 450000,China;College of Biological Resource,Qujing Normal University,Yunnan Qujing 655011,China)

机构地区：[1]昆明学院/云南省都市特色农业工程技术研究中心,云南昆明650214 [2]云南省烟草农业科学研究院,云南昆明650201 [3]中国烟草总公司云南省公司,云南昆明650011 [4]云南省烟草公司临沧市公司,云南临沧677000 [5]河南中烟工业有限责任公司原料采购中心,河南郑州450000 [6]曲靖师范学院生物资源与食品工程学院,云南曲靖655011

出　　处：《西南农业学报》2019年第12期2747-2751,I0001,共6页Southwest China Journal of Agricultural Sciences

基　　金：云南省烟草公司项目(2018530000241016,2016YN 30,2018530000241020,2019530000241028,201953000024101 1);云南省科技厅项目(2018FD080);云南省教育厅项目(2019J0574,2019Y0072)

摘　　要：【目的】探索烟草NBS-LRR类基因在烤烟(红花大金元)抗病中的分子作用机制。【方法】利用生物信息学手段,参考已知植物抗病基因的NBS-LRR类保守结构域,筛选并合成简并引物,扩增烟草中的NBS-LRR类抗病基因。【结果】获得4条具有连续阅读框和NBS结构域的抗病基因同源序列(TRGAH:MN027515、MN027516、MN027517、MN027518);BLAST X分析显示,均与已知植物抗病基因的NBS区域具有高度相似性,氨基酸相似性为64%~98.89%;聚类分析结果表明,4条TRGAHs与已知植物R基因P-loop-GLPL区域氨基酸序列相似性为29%~52%,包含了non-TIR-NBS-LRR和TIR-NBS-LRR两大类抗病基因。【结论】该研究利用分子克隆技术获得烟草抗病基因同源序列,为进一步筛选和构建烟草抗病候选基因及抗病品种的选育提供科学依据。【Objective】Cloning and analysis of the NBS-LRR resistance gene analogs(RGAs)in tobacco provided an opportunity for the cloning and analysis of disease-resistant genes in tobacco.【Method】By means of bioinformatics 2 pairs of degenerate primers were screened and synthesized according to the conserved domains of the NBS-LRR class of known plant disease-resistant genes to amplify the NBS-LRR class of disease-resistant genes in tobacco(Hongda).【Results】Four disease-resistant gene homologous sequences with continuous reading frame and NBS domain were obtained(TRGAH:MN027515,MN027516,MN027517,MN027518).BLAST X analysis found that the four tobacco TRGAHs were highly similar to NBS regions of known plant disease-resistant genes,with amino acid similarity ranging from 64%to 98.89%.Cluster analysis results showed that the similarity between the four TRGAHs and the corresponding regions of the P-loop-GLPL region amino acid sequences of the six R genes of known plants was 29%-52%,including two major classes of disease-resistant genes,namely non-TIR-NBS-LRR and TIR-NBS-LRR.【Conclusion】In this study,molecular cloning technology was used to obtain the homologous sequences of tobacco disease resistance genes,providing references for the cloning of tobacco disease resistance genes by homologous cloning technology,and laying a foundation for the further screening and construction of tobacco disease resistance candidate genes,so as to cultivate new varieties of tobacco disease resistance and transgenic breeding of disease resistance.

关 键 词：烟草红花大金元 NBS-LRR 抗病基因同源序列 

分 类 号：S572[农业科学—烟草工业]