高粱镉胁迫响应基因 sbbHLH168 的表达特征及生物信息学分析  

Expression and Bioinformatics Analysis of Cadmium Stress Response Gene sbbHLH168 in Sorghum bicolor

作　　者：于文慧 郑佳 李杨华 杨明川 王雅利 张子路 龚利娟 康振辉 YU Wen-hui;ZHENG Jia;LI Yang-hua(College of Bioengineering,Sichuan University of Science&Engineering,Yibin,Sichuan 644000;Wuliangye Group Co.,Ltd.,Yibin,Sichuan 644000)

机构地区：[1]四川轻化工大学生物工程学院,四川宜宾644000 [2]五粮液股份有限公司,四川宜宾644000

出　　处：《安徽农业科学》2023年第24期86-92,96,共8页Journal of Anhui Agricultural Sciences

基　　金：四川省科技厅重点项目“利用CRISPR/cas9技术创制五粮型酿酒专用高粱新品种”(2020YFN0023);五粮液产学研合作项目(CXY2021ZR010)。

摘　　要：[目的]以高梁基因sbbHLH168为研究对象,探究其耐镉胁迫发热分子机制,对培育耐镉高粱提供了理论参考。[方法]从高粱品种“BTx623”镉胁迫下转录组数据(RNA-seq)中筛选出一个受镉胁迫后显著上调表达的基因sbbHLH168,使用在线软件对其生物信息进行分析;通过洋葱表皮侵染进行亚细胞定位;构建pGBKT7-sbbHLH168载体验证sbbHLH168基因的转录激活活性;将水培至四叶一心的高粱置于NaCl、PEG、ABA、ACC、GA和JA溶液处理,分别在0、1、12、24和48 h提取RNA,通过RT-qPCR检测sbbHLH168基因的相对表达量。[结果]sbbHLH168基因的开放阅读框长度为567 bp,编码189个氨基酸残基,蛋白分子量为20.55 kD,理论等电点(PI)为8.86,为碱性带正电的蛋白,定位于细胞核,具有较强的转录激活活性。sbbHLH168基因在盐、干旱和植物激素胁迫下表达量整体呈上升趋势,但表达量在不同处理时间存在明显差异。对sbbHLH168基因在不同组织中的表达发现,该基因在高粱的根和叶中均有表达,其中在盐、干旱、ABA、GA和JA胁迫下,sbbHLH168基因在处理12 h时表达量最高。而在ACC处理时,则在处理24 h时表达量最高。[结论]sbbHLH168基因是一个定位在细胞核有转录激活活性的转录因子,属于bHLH家族。[Objective]The study aimed to explore the molecular mechanism of cadmium stress tolerance in Sorghum bicolor gene sbbHLH168,which plays an important role in plant response to biotic and abiotic stresses,it provides a theoretical reference for breeding cadmium-tolerant Sorghum bicolor.[Method]A significantly up-regulated sbbHLH168 was screened from the transcriptome data(RNA-seq)of Sorghum bicolor cultivar‘BTx623’under cadmium stress,and its biological information was analyzed by on-line software subcellular localization of sbbHLH168 gene was carried out by infection of onion epidermis,PGBKT7-sbbHLH168 vector was constructed to verify the transcriptional activation activity of sbbHLH168 gene,and Sorghum bicolor plants cultured in water were treated with NaCl,PEG,ABA,ACC,GA and JA solutions,RNA was extracted at 0,1,12,24 and 48 hours respectively,and the relative expression of sbbHLH168 gene was detected by RT-qPCR.[Result]The open reading frame length of sbbHLH168 gene was 567 bp,encoding 189 amino acid residues.The molecular weight of sbbHLH168 protein was 20.55 kDa,and the theoretical isoelectric point(PI)was 8.86.sbbHLH168 was a basic protein with positive charge,which was located in the nucleus of cells,it has strong transcriptional activation activity.The expression level of sbbHLH168 was increased under salt,drought and plant hormone stress,but the expression level was significantly different at different treatment time.The expression of sbbHLH168 gene in different tissues showed that sbbHLH168 gene was expressed in root and leaf of Sorghum bicolor,and the highest expression level was observed under salt,drought,ABA,GA and JA stress for 12 h.In ACC treatment,the highest expression level was observed at 24 h.[Conclusion]sbbHLH168 gene is a transcription factor with transcriptional activation activity located in the nucleus and belongs to bHLH family.

关 键 词：高粱 sbbHLH168 生物信息学分析 表达特征 转录激活活性 

分 类 号：Q81[生物学—生物工程]