香蕉MaWRKY28基因的克隆及其功能分析  

Cloning and Functional Analysis of MaWRKY28 Gene from Banana(Musa acuminata)

作　　者：孙嘉曼 彭丽云 方辉[1] 陈慧灵 张进忠[2] Sun Jiaman;Peng Liyun;Fang Hui;Chen Huiling;Zhang Jinzhong(Guangxi Crop Genetic Improvement and Biotechnology Key Laboratory,Guangxi Academy of Agricultural Sciences,Nanning,530007;Flowers Research Institute,Guangxi Academy of Agricultural Sciences,Nanning,530007)

机构地区：[1]广西农业科学院,广西作物遗传改良生物技术重点开放实验室,南宁530007 [2]广西农业科学院花卉研究所,南宁530007

出　　处：《分子植物育种》2020年第14期4605-4611,共7页Molecular Plant Breeding

基　　金：国家自然科学基金(31660560);广西自然科学基金项目(2016GXNSFBA380045,2015GXNSFDA139014);广西农业科学院基本科研业务专项(2018YT25,2017YM33)共同资助。

摘　　要：为探究香蕉WRKY28基因响应Foc-TR4侵染的作用及调节机制,本研究从抗病‘桂蕉9号’品种中克隆了香蕉MaWRKY28基因;利用ExPASy、MEGA 6.0等软件对其蛋白序列进行理化性质分析及系统进化树的构建;采用q RT-PCR分析了该基因在抗(‘桂蕉9号’)、感(‘威廉斯’)香蕉品种受Foc-TR4侵染不同时间后的表达量变化;构建过表达MaWRKY28基因载体,转化拟南芥,并检测该基因在转化植株中的表达量。结果表明,从‘桂蕉9号’中克隆了一个WRKY28基因,命名为MaWRKY28 (GenBank登录号为MK941141)。MaWRKY28基因编码区全长999 bp,编码332个氨基酸,属于不稳定的分泌蛋白,具有典型的WRKY结构域;系统进化树分析表明,香蕉与海枣(Phoenix dactylifera)、油棕(Elaeis guineensis)的同源性最高,相似性高达87%;qRT-PCR结果显示,在Foc-TR4侵染的香蕉根系过程中,MaWRKY28基因呈上调表达趋势,且在抗病品种中的表达量整体高于感病品种;筛选获得的7株阳性转化拟南芥植株中,MaWRKY28基因的相对表达量较野生型均显著提高。综上说明,MaWRKY28基因在香蕉抗枯萎病菌侵染过程中参与了抗病相关过程,且能在异源受体拟南芥中成功转化并高效表达。本研究结果为进一步研究香蕉抗病品种的抗性机理提供了帮助。In order to explore the effect of response to Foc-TR4 infection and its regulation mechanism. In this study, the banana WRKY28 gene was cloned from the disease-resistant ’Gujiao9’ variety. The analysis of its protein sequence and the construction of phylogenetic tree were carried out by the software of ExPASy and MEGA 6.0 and so on. The expression of MaWRKY28 under Foc-TR4 infection with ’Guijiao9’ and ’Williams’ at different time points were analyzed by quantitative real-time PCR(qRT-PCR) technique. The overexpression of MaWRKY28 gene vector was constructed and transformed into Arabidopsis thaliana, and the expression of the gene in transformed plants was detected. The results showed that a WRKY28 gene was cloned from the ’Guijiao9’, which named MaWRKY28(GenBank ID: MK941141), and its CDS sequence was 999 bp in full length, encoding 332 amino acids.It belongs to unstable secretory protein and has a typical WRKY domain. Systematic evolution tree analysis showed that banana had the highest homology and similarity as high as 87% between Phoenix dactylifera and Elaeis guineensis. q RT-PCR results showed that the expression of MaWRKY28 gene was up-regulated in the process of banana roots infected by Foc-TR4, and the expression level of resistant varieties was higher than that of susceptible varieties as a whole. The expression of MaWRKY28 in 7 transgenic plants were more abundant than the wild plants.The results showed that MaWRKY28 gene was involved in the process of resistance to Fusarium wilt in bananas,and it can be successfully transformed and highly expressed in Arabidopsis thaliana. The results of this study provide help for further study on the resistance mechanism of banana resistant varieties.

关 键 词：香蕉(Musa acuminata) MaWRKY28 抗病性 转基因 

分 类 号：S668.1[农业科学—果树学]