大豆耐涝bHLH转录因子筛选及生物信息学分析

Selection and Bioinformatics Analysis of bHLH Transcription Factor Response to Submergence Stress in Soybean

为研究大豆bHLH转录因子与涝害响应相关性,为大豆耐涝性研究提供理论基础,本研究对强耐涝性品种齐黄34进行没顶淹水处理,分析转录组测序结果,筛选差异表达的bHLH转录因子并对其进行生物信息学分析,采用荧光定量PCR方法验证主要差异表达bHLH转录因子编码基因的表达情况,并通过生物信息学方法分析基因结构和互作蛋白。结果显示:涝害胁迫下共发现7个差异表达的bHLH转录因子,它们的同源性并不高,属于不同类型的bHLH。主要差异表达bHLH基因GmbHLH25-15(Glyma.15G06680)的表达量变化情况与转录组数据趋势一致,呈现下调表达。预测到10个蛋白可与该蛋白互作,互作蛋白都是铵转运蛋白,其中4个蛋白编码基因的表达量在转录组测序结果中呈现显著性差异。推测在无氧条件下,GmbHLH25-15可能通过调控铵转运蛋白进而调节铵态氮的吸收来供给自身营养,进而抵御涝害胁迫。

In order to study the correlation between bHLH transcription factors and waterlogging responses of soybean,and to provide theoretical basis for the study of waterlogging tolerance of soybean,in this study,Qihuang 34,a soybean variety with strong submergence resistance,was treated with submergence stress.We analyzed the transcriptome sequencing results,selected and took bioinformatics analysis on the differentially expressed bHLH transcription factors.We used fluorescence quantitative PCR to verify the expression of bHLH transcription factor genes,and analyzed the structure and interacting proteins of the gene.The results showed that seven differentially expression bHLH transcription factors were selected,the domain of these protein sequences were not highly homologous,and they belonged to different types of bHLH combining with the transcriptome data.GmbHLH25-15(Glyma.15 G06680)showed the most significant expression changes and the quantitative PCR analysis result was consistent with the trend of transcriptome data,showing down-regulated expression.Furthermore,all the 10 proteins interacting with Glyma.15 G06680 were ammonium transporters.Among them,the coding genes of 4 proteins showed significantly different expression in transcriptome sequencing results.It was speculated that soybean mainly absorbed ammonium nitrogen as its own nutrient supply in the absence of oxygen,and the bHLH transcription factor GmbHLH25-15 might regulate the absorption of ammonium nitrogen by regulating the ammonium transporter,further resist submergence stress.