大豆胞囊线虫病抗性候选基因GmPEBP4-1克隆及表达分析  

Cloning and expression analysis of soybean cyst nematode resistance candidate gene GmPEBP4-1

作　　者：郭杨 陈立新[2] 姜海鹏 战宇航 GUO Yang;CHEN Lixin;JIANG Haipeng;ZHAN Yuhang(Heilongjiang Academy of Agricultural Sciences Postdoctoral Programme,Harbin 150086,China;Horticultural Sub-Academy,Heilongjiang Academy of Agricultural Sciences,Harbin 150069,China;School of Agriculture,Northeast Agricultural University,Harbin 150030,China)

机构地区：[1]黑龙江省农业科学院博士后科研工作站,哈尔滨150086 [2]黑龙江省农业科学院园艺分院,哈尔滨150069 [3]东北农业大学农学院,哈尔滨150030

出　　处：《东北农业大学学报》2020年第10期10-19,共10页Journal of Northeast Agricultural University

基　　金：黑龙江省杰出青年基金项目(JC2018007);国家自然科学基金面上项目(31671717)。

摘　　要：研究利用抗病材料东农L-10转录组测序结果,选取差异表达的磷脂酰乙醇胺结合蛋白(PEBP)基因命名为GmPEBP4-1。通过基因克隆、生物信息学和表达模式分析研究GmPEBP4-1基因对SCN响应及生理功能的影响。结果表明,GmPEBP4-1基因CDS全长519 bp,编码172个氨基酸,蛋白分子质量为19.34 ku,是一种亲水性蛋白;氨基酸组分中精氨酸(Arg)、脯氨酸(Pro)和缬氨酸(Val)占比最高;等电点为9.16,初步预测为酸性蛋白;蛋白保守结构域分析发现含有保守PEBP结构域;蛋白质二级结构中无规则卷曲所占比例最高(54.07%),其次为延伸链(25.58%)、α-螺旋(16.28%)、β-转角(4.07%);系统进化树分析表明,GmPEBP4-1蛋白与相思子PEBP蛋白亲缘性最近;启动子元件分析表明,存在茉莉酸甲酯(MEJA)、TCT-rich repeats,P-box和ABRE等应激、胁迫相关顺式作用元件,推测该基因参与逆境胁迫反应;利用实时荧光定量PCR分析发现,在抗病材料中,大豆胞囊线虫4号生理小种胁迫各时间段GmPEBP4-1基因表达量均显著高于对照,而感病材料中GmPEBP4-1基因表达量仅在胁迫7 d时显著高于对照。综上表明,GmPEBP4-1基因响应大豆胞囊线虫4号生理小种胁迫过程,且在感病材料和抗病材料中响应模式存在差异。In this study,the differential expression of GmPEBP4-1 gene under soybean cyst nematode(SCN)stress was selected based on the results of transcriptome sequencing of diseaseresistant material Dongnong L-10.The response and physiological function of GmPEBP4-1 gene to SCN were studied by gene cloning,bioinformatics analysis and expression pattern analysis.The results showed that the CDS of GmPEBP4-1 gene was 519 bp,encoding 172 amino acids.The coded protein had a molecular mass of 19.34 ku.It was a hydrophilic protein.Among the amino acid components,arginine(Arg),proline(Pro)and valine(Val)accounted for the highest proportion.The isoelectric point of the protein encoded by the gene was 9.16,which was preliminarily predicted to be an acidic protein.Protein conserved domain analysis showed that there was a conserved PEBP domain,and the proportion of irregular crimp in the secondary structure of protein was the highest(54.07%),followed by extended chain(25.58%),α-helix(16.28%)andβ-rotation(4.07%).Phylogenetic tree analysis showed that GmPEBP4-1 protein was closely related to the PEBP protein of Abrus precatorius.Promoter element analysis showed that there were stress-related cis-acting elements,such as methyl jasmonate(MEJA),TCT-rich repeats,P-box and ABRE,which suggested that the gene was involved in stress response.Real-time quantitative PCR analysis showed that in the resistant materials,the expression of GmPEBP4-1 gene was significantly higher than that of the control in different periods of stress of soybean cyst nematode race 4,while the expression of GmPEBP4-1 gene in susceptible materials was significantly higher than that of the control only after 7 days of stress.In summary,GmPEBP4-1 gene responded to the stress process of soybean cyst nematode race 4,and there were differences in response patterns between the susceptible materials and the resistant materials.

关 键 词：大豆胞囊线虫 GmPEBP4-1 基因克隆 生物信息学 表达分析 

分 类 号：Q78[生物学—分子生物学] S565.03[农业科学—作物学]