大豆GmMYB48基因克隆、生物信息学分析及其参与大豆胞囊线虫病抗性功能分析
Cloning,bioinformatics analysis and functional exploration identification of soybean GmMYB48gene participated in resistance to soybean cyst nema-tode
战宇航 1胡世豪 1曲硕 1宋庚晨 1刘芳 1赵旷宇 2孙浩文 1韩英鹏1
作者信息
- 1. 东北农业大学农学院,哈尔滨 150030;农业农村部东北大豆生物学与遗传育种重点实验室,哈尔滨 150030
- 2. 方正县产品质量综合检验检测中心,黑龙江 方正 150800
- 折叠
摘要
MYB转录因子在植物生长发育过程中执行重要调控功能.以大豆胞囊线虫3号生理小种(SCN 3)胁迫下大豆抗病品种"东农L-10"和感病品种"黑农37"转录组数据为基础,筛选出MYB家族差异表达基因GmMYB48,克隆GmMYB48基因并进行生物信息学、亚细胞定位、基因表达模式分析及过表达大豆毛状根.结果表明,GmMYB48基因编码蛋白为碱性蛋白,与拟南芥存在共线性关系,在植物中分布广泛;靶基因预测分析发现GmMYB48基因参与大豆对盐、干旱、金属盐离子等逆境胁迫反应;启动子元件分析显示,该基因存在多个环境胁迫响应相关WUN-motif、MYB等启动子元件;烟草叶片瞬时表达表明GmMYB48基因定位于细胞核中;GmMYB48基因转录响应大豆胞囊线虫胁迫,在抗病材料根部转录水平较高.通过发根农杆菌介导的遗传转化获得GmMYB48基因过表达大豆毛状根,过表达毛状根较野生型毛状根中单位面积内线虫数目明显减少,从表型鉴定中鉴定GmMYB48基因参与大豆对胞囊线虫3号生理小种的抗性反应.研究初步探究GmMYB48基因在抗大豆胞囊线虫中的功能,为进一步研究抗大豆胞囊线虫病机制奠定基础.
Abstract
MYB transcription factors perform significant regulatory functions in plant growth and development.In this research,using the transcriptome data obtained from the disease-resistant soybean germplasm Dongnong L-10 and the disease-sensitive germplasm Heinong 37,the differentially expressed gene GmMYB48 was screened,which was a member of the MYB family,by using the physiological microspecies of soybean cyst nematode race 3(SCN 3).The GmMYB48 gene was cloned and subjected to bioinformatics,subcellular localization,gene expression pattern analysis and gene overexpression analysis.The results showed that the protein encoded by GmMYB48 transcription factor was a basic protein,which was covalently related to several species and widely distributed in plants;the target gene prediction results showed that the GmMYB48 gene was involved in the response to salt stress,drought stress,metal salt ion stress and other adversity stresses in soybean;the promoter element analysis showed that there were several responses to the gene in response to the environmental stress response related to the WUN-motif,MYB,etc.The gene expression pattern was analyzed and the gene overexpression was performed.The transient expression in tobacco leaves showed that GmMYB48 was subcellularly localized in the nucleus;GmMYB48 responded to soybean cyst nematode,and the relative expression of the gene was higher in the roots of disease-resistant materials than that of susceptible materials,and the overexpressed soybean hairy roots were obtained through Agrobacterium root-mediated genetic transformation.The number of nematodes per unit area in the hairy roots was significantly reduced,and the involvement of GmMYB48 in the resistance response of soybean to cyst nematode race 3 was verified from the phenotypic characterization.In this study,it initially investigated the function of GmMYB48 in resistance to soybean cyst nematode,and laid the foundation for further researches on the mechanism of soybean cyst nematode resistance.
关键词
大豆胞囊线虫/qRT-PCR/生物信息学/亚细胞定位/遗传转化Key words
soybean cyst nematode/qRT-PCR/bioinformatics/subcellular localization/genetic transformation引用本文复制引用
基金项目
黑龙江省重点研发项目(JD22A015)
黑龙江省重点基金项目(ZD2022C002)
国家自然科学基金项目(U22A20473)
国家现代农业岗位体系项目(CARS-04-PS07)
东北农业大学科研项目(NEAU2023QNLJ-003)
出版年
2024
NETL
NSTL
万方数据