摘要
为了了解苦荞抗立枯病的分子机制,以苦荞代表性品种川荞1号为材料,克隆出FtEIN3基因。生物信息学分析表明,FtEIN3基因的CDS序列长度为1623 bp,编码540个氨基酸,FtEIN3蛋白二级结构由α-螺旋(33.52%)、延伸链(6.67%)、β折叠(2.41%)和无规则卷曲(57.41%)组成。系统进化树显示,苦荞FtEIN3蛋白与棉花GhEIN3、榴莲DzEIN3蛋白亲缘关系较近。通过对108份苦荞种质资源EIN3编码区序列比对,检测到5种不同的单倍型,其中Hap3为优异单倍型。qRT-PCR结果表明,FtEIN3基因在苦荞中的表达受立枯丝核菌侵染的诱导。激光共聚焦显微镜观察下,苦荞FtEIN3基因定位在细胞核上。为进一步验证FtEIN3基因的功能,本研究构建了FtEIN3的转基因拟南芥并分析其抗病性。结果显示,相对于野生型拟南芥,过表达FtEIN3基因显著提高了转基因拟南芥株系的抗立枯病能力。以上结果表明FtEIN3基因参与苦荞麦抗立枯病的防御过程,为进一步研究FtEIN3调控苦荞抗立枯病的分子机制奠定了一定的工作基础。
In order to understand the molecular mechanism of tartary buckwheat against bacterial blight,we isolated the transcription factor gene FtEIN3 in Chuanqiao 1,a representative variety of tartary buckwheat.The FtEIN3 contains the CDS sequence length of 1623 bp,encoding 540 amino acids.The secondary structure of FtEIN3 protein was composed ofα-helix(33.52%),extended chain(6.67%),β-fold(2.41%)and irregular coil(57.41%).Phylogenetic tree indicated that FtEIN3 protein was closely related to GhEIN3 and DzEIN3 protein.Five different EIN3 sequence haplotypes were detected in 108 tartary buckwheat germplasm,and out of them Hap3 was an elite haplotype.qRT-PCR revealed that the expression of FtEIN3 gene in tartary buckwheat was induced by Rhizoctonia solani.The FtEIN3 gene in tartary buckwheat was localized in the nucleus under confocal laser microscopy.To further verify the function of FtEIN3 gene,FtEIN3 transgenic Arabidopsis thaliana were constructed and their resistance to blip disease was analyzed.The results showed that overexpression of FtEIN3 gene significantly improved the resistance of transgenic Arabidopsis thaliana to bacterial blight compared with the wild type.These results proved that FtEIN3 gene was involved in the defense process of tartary buckwheat against bacterial blight,and laid a foundation for further research on the molecular mechanism of FtEIN3 regulation of tartary buckwheat resistance to blight.
作者
李光胜
卢翔
刘洋
张凯旋
王海华
唐新科
周美亮
LI Guangsheng;LU Xiang;LIU Yang;ZHANG Kaixuan;WANG Haihua;TANG Xinke;ZHOU Meiliang(College of Life Science and Health,Hunan University of Science and Technology,Xiangtan 411201;Institute of Crop Sciences,Chinese Academy of Agricultural Sciences,Beijing 100081)
出处
《植物遗传资源学报》
CAS
CSCD
北大核心
2023年第5期1389-1400,共12页
Journal of Plant Genetic Resources
基金
国家重点研发计划(2019YFD1001300,2019YFD1001303)。
关键词
苦荞
FtEIN3
克隆
拟南芥
抗立枯病
tartary buckwheat
FtEIN3
cloning
Arabidopsis thaliana
resistance to blight
