马铃薯干旱响应基因StGAMyb-like1克隆、表达及蛋白互作分析
Cloning,Expression and Protein Interaction Analysis of Drought-responsive Gene StGAMyb-like1 in Potato(Solanum tuberosum)
邓玉荣 1魏晶晶 1刘升燕 2杨亮 2王晓凤 2杨江伟 1张宁 1司怀军1
作者信息
- 1. 甘肃农业大学省部共建干旱生境作物学国家重点实验室,兰州 730070;甘肃农业大学生命科学技术学院,兰州 730070
- 2. 甘肃农业大学省部共建干旱生境作物学国家重点实验室,兰州 730070;甘肃农业大学农学院,兰州 730070
- 折叠
摘要
作为MYB转录因子家族中最大的亚家族,R2R3-MYB转录因子广泛参与植物生长发育、物质代谢等过程,并且具有提高植物胁迫耐受性的潜力.前期研究在马铃薯(Solanum tuberosum)中鉴定出1个与拟南芥(Arabidopsis thaliana)AtMYB33 同源的 StGAMyb-like1 基因(GenBank No.XM_015303144.1),该基因是R2R3-MYB的第18亚组GAMYB中的成员,且能够响应干旱胁迫.本研究以马铃薯品种'Desiree'为材料,克隆得到StGAMyb-like1基因,其CDS长为1 572 bp;生物信息学分析发现,该基因启动子区含有多种激素响应元件,其蛋自序列与番茄(Solanum lycopersicum)的相似度最高.组织差异性表达分析发现,StGAMyb-like1基因在叶片中具有更高的表达丰度.对木质素合成途径中的香豆酸-3-羟化酶编码基因StCYP98A3启动子序列进行分析,发现该基因具备与MYB转录因子结合的MBS(MYB binding site)元件.通过酵母单杂交、双荧光素酶报告基因分析和β-D-葡萄糖苷酸酶(β-glucuronidase,GUS)检测,证实StGAMyb-like1可与StCYP98A3基因启动子结合并转录激活该基因表达.本研究可为深入探讨StGAMyb-like1基因功能提供参考依据.
Abstract
R2R3-MYB transcription factors,as the largest subfamily of the MYB transcription factor family,play a crucial role in plant growth and development,material metabolism,and other processes.Additionally,they have the potential to enhance plant stress tolerance.In a previous study,a StGAMyb-like1 gene(GenBank No.XM_015303144),homologous to AtMYB33 in Arabidopsis thaliana,was identified in potato(Solanum tuberosum).The StGAMyb-like1 gene,a member of GAMYB subgroup 18 of R2R3-MYB,is capable of responding to drought stress.In this study,the StGAMyb-like1 gene was cloned from the potato variety'Desiree',and its CDS was 1 572 bp.Bioinformatics analysis revealed that the promoter region of this gene contained multiple hormone response elements,and its protein sequence was similar to that of tomato(Solanum lycopersicum).The expression analysis in different tissues showed that the StGAMyb-like1 gene had higher expression level in leaves.The promoter sequence of the StCYP98A3 gene,which encodes coumarate 3-hydroxylase in the lignin synthesis pathway,was analyzed and found to have MYB binding site(MBS).It was confirmed that StGAMyb-like1 binded to the StCYP98A3 promoter and activated its transcription using yeast one hybrid,dual-luciferase reporter assay and β-glucuronidase(GUS)detection.This study provides a reference basis for further exploring the function of StGAMyb-like1 gene.
关键词
马铃薯/生物信息学/StGAMyb-like1/StCYP98A3Key words
Potato/Bioinformatics/StGAMyb-like1/StCYP98A3引用本文复制引用
基金项目
甘肃省自然科学基金重点项目(22JR5RA832)
甘肃省科技重大专项计划(23ZDNA006)
国家自然科学基金(31960444)
出版年
2024
国家科技期刊平台
NSTL
万方数据