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[1]唐 海 姚银安 张国燕 李 星 丁红霞 吴英青 高永峰**.异源表达胡杨PeBAK1;1基因增强烟草对Pst DC3000的抗性*[J].应用与环境生物学报,2018,24(04):1-14.[doi:10.3724/SP.J.1145.2017.10018]
 TANG Hai,YAO Yinan,ZHANG Guoyan,et al.Heterologous Overexpression of Populus euphratica BAK1;1 Gene Enhanced Tobacco Resistance to Pst DC3000*[J].Chinese Journal of Applied & Environmental Biology,2018,24(04):1-14.[doi:10.3724/SP.J.1145.2017.10018]
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异源表达胡杨PeBAK1;1基因增强烟草对Pst DC3000的抗性*()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
24卷
期数:
2018年04期
页码:
1-14
栏目:
研究论文
出版日期:
2018-08-25

文章信息/Info

Title:
Heterologous Overexpression of Populus euphratica BAK1;1 Gene Enhanced Tobacco Resistance to Pst DC3000*
文章编号:
201710018
作者:
唐 海 姚银安 张国燕 李 星 丁红霞 吴英青 高永峰**
西南科技大学生命科学与工程学院 绵阳 621010
Author(s):
TANG HaiYAO YinanZHANG GuoyanLI XingDING HongxiaWU Yingqing&GAO Yongfeng**
School of Life Science and Engineering, Southwest University of Science and Technology, Mianyang 621010, China
关键词:
PeBAK11抗病相关基因烟草抗病Pst DC3000
Keywords:
PeBAK11pathogenesis-related genestobaccodisease-resistantPst DC3000
DOI:
10.3724/SP.J.1145.2017.10018
摘要:
BAK1(BRI1-ASSOCIATED RECEPTORKINASE1)是一个富亮氨酸重复序列(LRR)的膜受体蛋白激酶,除参与植物油菜素内酯信号的转导外,还可以结合其他的LRR-RLKs蛋白来启动植物的先天免疫反应。为明确胡杨PeBAK1;1基因在烟草中抗Pst DC3000的功能和其在植物抗病中的调控方式。本研究使用以下方法和技术手段对胡杨PeBAK1;1基因的功能进行了研究:首先,利用ClustalX和MEGA5.0软件对胡杨PeBAK1;1蛋白序列以及其他植物同源的BAK1蛋白序列进行多重序列比对,并构建进化树;其次,构建PeBAK1;1基因的过量表达载体pBI121-35S::PeBAK1;1,并利用农杆菌介导法,将其转化野生型烟草,筛选获得PeBAK1;1基因过表达的转基因烟草植株;然后,对野生型和转基因烟草植株进行Pst DC3000的接种,检测其抗病性反应;最后,利用RT-PCR及quantitative Realtime-PCR技术,检测胡杨不同组织中PeBAK1;1基因的表达模式以及转基因烟草中与抗病相关的Marker基因的表达是否受PeBAK1;1的调控。结果表明:胡杨PeBAK1;1蛋白具有植物SERK家族的全部结构特征,系统进化树分析表明胡杨PeBAK1;1与毛果杨PtBAK1的序列同源性最高;组织表达分析表明,PeBAK1;1基因主要在根和叶中表达;野生型烟草植株接种Pst DC3000后表现出明显的感病症状,而PeBAK1;1基因过表达的转基因烟草植株表现出明显的抗病症状;与野生型相比,转基因烟草中抗病防御相关基因PR1、PR3、PR4和PR5的表达量显著升高,且参与植物生长发育与先天防卫反应基因BIR1和BON1的表达量也明显升高。综上所述,异源表达胡杨PeBAK1;1基因在烟草抗Pst DC3000过程中起正调控作用,能够增强植物的抗病能力。
Abstract:
BAK1(BRI1-ASSOCIATED RECEPTORKINASE1)is a leucine-rich repeat (LRR) receptor protein kinase, it plays significant role in brassinosteroid (BR) signaling, and it also combines with other LRR-RLKs protein to initiate the plant immune response.The objective of this study is to investigate the function of the Populus euphratica BAK1;1 gene in transgenic tobacco resistance to Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) , and to discuss the regulation pathway of PeBAK1;1 in resistance to plant pathogen. In this study, the function of the Populus euphratica BAK1;1 gene was analyzed by the following methods and techniques. Firstly, Clustalx and MEGA5.0 softwaers were used to do multiple sequences alignment and construct the phylogenetic tree of BAK1 proteins. Secondly, pBI121-35S::PeBAK1;1 over-expression vector was constructed and then transformed into wild-type tobacco by using Agrobacterium-mediated transformation, and obtained PeBAK1;1 overexpressed transgenic tobacco plants. Then, in order to study the function of the PeBAK1;1 gene in resistance to Pst DC3000, the symptoms and bacterial concentrations of the wild type and transgenic tobacco plants inoculated with Pst DC3000 were tested. Finally, The expression pattern of PeBAK1;1 gene was analysed in root, stem and leaf of wild type Populus euphratica plant by using quantitative real-time PCR analysis. RT-PCR and quantitative RT-PCR technology were used to analyze the expression pattern of PeBAK1;1 gene in Populus euphratica and the expression levels of defence-related genes in transgenic tobacco plants. The main results were as follows: PeBAK1; 1 protein contained all the structural features of the plant SERK family, and the phylogenetic tree showed that PeBAK1;1 is the highest sequence homology with the PtBAK1. The gene expression profile results indicated that the expression level of PeBAK1;1 in root was higher than leaf and stem. The wild type tobacco plants showed an obvious susceptibility to Pst DC3000, while transgenic plants exhibited enhanced resistance to Pst DC3000. Compared with the WT tobacco plants, the expression of pathogenesis-related genes (containing PR1, PR3, PR4 and PR5) BIR1gene (BAK1-Interacting Receptor Kinase 1 gene) and BON1gene (BONZAI1 gene) were upregulated in 35S::PeBAK1;1 transgenic tobacco plants. In conclusion, PeBAK1;1 gene plays a positive regulatory role in 35S::PeBAK1;1 transgenic tobacco against Pst DC3000, can enhance the plant resistance to pathogen.

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备注/Memo

备注/Memo:
收稿日期:2017-10-17 接受日期:2017-12-04
*国家自然基金面上项目(NO.31770644),四川省国际合作项目(No.2017HH0050),四川省教育厅资助科研项目(N0.17ZB0456),四川省青年科技基金项目(No.2014JQ0016),西南科技大学博士基金项目(NO.14zx7157) 资助
**通讯作者(E-mail:gaoyongfeng0263@gmail.com)
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更新日期/Last Update: 2017-12-27