大豆转录因子GmWIN1-3介导逆境胁迫响应的功能分析  

Function analysis of soybean transcription factor GmWIN1-3 mediating plant response to abiotic stress

作　　者：薛金爱 蔡桂萍 周雅莉 陈莹 刘宝玲 张莉 李润植 XUE Jin'ai;CAI Guiping;ZHOU Yali;CHEN Ying;LIU Baoling;ZHANG Li;LI Runzhi(Institute of Molecular Agriculture&Bioenergy,College of Agriculture,Shanxi Agricultural University,Taigu,Shanxi 030801,China)

机构地区：[1]山西农业大学农学院/分子农业与生物能源研究所,山西太谷030801

出　　处：《植物生理学报》2022年第9期1703-1714,共12页Plant Physiology Journal

基　　金：国家自然科学基金(31201266和31401430);山西省重点研发计划项目(201803D221005-9和201803D221019-1);山西省自然科学基金项目(201901D111221和20210302124170);山西农业大学生物育种工程项目(YZGC101);山西省晋中市科技重点研发计划项目(Y172007-5)。

摘　　要：为挖掘提高大豆逆境胁迫抗性的优异基因,本文聚焦大豆(Glycine max)WIN1转录因子基因克隆和生物学功能分析。基于转录组学分析,RT-PCR克隆到GmWIN1-3基因的编码序列(720 bp)。亚细胞定位分析显示该转录因子定位于细胞核。PlantCARE软件预测GmWIN1-3启动子中含有LTR和TC-rich repeats等胁迫顺式作用元件。基因表达谱分析显示,在低温和高盐胁迫下,GmWIN1-3呈上调表达,而在干旱胁迫下呈下调表达。在低温胁迫下,过表达GmWIN1-3显著促进大豆毛状根油脂积累和减低MDA含量。油脂合成途径相关酶基因GmBCCP1-4和GmDGAT2-3在GmWIN1-3转基因大豆毛状根中表达显著上调,然而GmDGAT2-1和Gm LPAT5-2则呈下调表达。异源过表达GmWIN1-3显著提高烟叶油脂积累和烟草幼苗低温胁迫耐性。研究表明转录因子GmWIN1-3可通过上调GmDGAT2-3等酶基因表达和总脂质积累,进而提高大豆幼苗抵御逆境胁迫的能力。To explore excellent target genes for improving soybean stress resistance,this study focused on the cloning and biological function characterization of WIN1 transcription factor from soybean(Glycine max).The conding sequence(720 bp)of Gm WIN1-3 gene was isolated by RT-PCR based on the transcriptome data.Subcellular localization anaylsis indicated that GmWIN1-3 was located in the nucleus.Gm WIN1-3 promoter contained multiple stress cis-acting elements such as LTR and TC-rich repeats,predicted by PlantCARE.Further analysis of the expression profiles of Gm WIN1-3 gene under three kinds of stress.Gene expression profiling reveaked that Gm WIN1-3 was up-regulated under low temperature and salt stress,respectively while it was down-regulated by drought stress.Under low temperature stress,overexpression of Gm WIN1-3 significantly promoted oil accumulation and reduced MDA content in soybean hairy roots.Notably,expression analysis of genes related to the lipid synthesis pathway showed that the expressions of Gm BCCP1-4 and Gm DGAT2-3 enzyme genes were significantly up-regulated in the Gm WIN1-3 transgenic soybean hairy roots,whereas GmDGAT2–1 and Gm LPAT5-2 were down-regulated in the transgenics compared to the controls.Furthermore,heterologous overexpression of Gm WIN1-3 significantly improved oil accumulation in tobacco leaves and tobacco seedling resistance to low temperature stress.Collectively,these data indicate that Gm WIN1-3 may upregulate the expression of DGAT2-3 and Gm BCCP1-4 gene,and consequently promote the synthesis and accumulation of total lipids in the cell,thereby enhancing soybean seedling resistance to abiotic stresses.

关 键 词：大豆 WAX INDUCER1(WIN1)转录因子 功能分析 逆境胁迫 大豆毛状根 

分 类 号：S565.1[农业科学—作物学]