大豆转录因子基因GmbHLH130克隆及在干旱胁迫中的功能分析

Cloning and functional analysis of soybean transcription factor GmbHLH130 gene under drought stress

为了探究大豆GmbHLH130基因在植物干旱胁迫中的调控功能,利用生物信息学分析GmbHLH130与其他物种bHLH家族成员的系统进化关系,检测GmbHLH130基因及其启动子对干旱胁迫的响应,对GmbHLH130蛋白的亚细胞定位和转录激活活性进行分析,最后初步评估GmbHLH130基因过表达拟南芥的耐旱性。结果显示,在进化树中GmbHLH130与拟南芥AtbHLH122进化关系最近;GmbHLH130基因受干旱诱导上调表达;GmbHLH130基因启动子也受干旱诱导激活下游报告基因;烟草叶片瞬时表达分析结果表明GmbHLH130蛋白定位于细胞核,并且具有转录激活活性。此外,GmbHLH130基因过表达拟南芥在干旱处理下的绿色子叶率和根长均显著大于野生型。本研究结果初步证明了大豆GmbHLH130基因在增强植物耐旱方面的功能,为后续探究其参与耐旱性调控的分子机制提供了理论依据。

To explore the regulatory function of soybean GmbHLH130 gene in plant drought stress,bioinformatics was used to analyze the phylogenetic relationship between GmbHLH130 protein and bHLH members of other species.The response of GmbHLH130 and its promoter to drought stress was detected,and the subcellular localization and transcriptional activation of GmbHLH130 protein were analyzed.Finally,the drought tolerance of Arabidopsis which overexpressed GmbHLH130 was preliminarily evaluated.The results showed that GmbHLH130 protein was closest to AtbHLH122 protein in the phylogenetic tree.The expression level of GmbHLH130 gene was up-regulated under drought treatment.The promoter of GmbHLH130 gene was also induced by drought stress to activate downstream reporter gene.Transient expression of GmbHLH130 in tobacco leaves indicated that GmbHLH130 was localized in the nucleus and had transcriptional activation activity.In addition,the green cotyledon percentage and root length of GmbHLH130 overexpressed Arabidopsis were all significantly higher than those of wide type under drought treatment.The results of the study preliminarily proved the function of soybean GmbHLH130 gene in enhancing plant drought tolerance,and provided theoretical basis for further investigation of its molecular mechanism involved in drought tolerance regulation.