二穗短柄草根部木栓质调控基因BdMYB92的克隆及功能研究  

Cloning and Functional Research of BdMYB92,Which Regulates the Suberin Biosynthesis in Roots of Brachypodium distachyon

作　　者：呼宁 何兆峰 朱雨遥 贺佳佳 李翀照 汪勇[1] 王中华[1] HU Ning;HEZhaofeng;ZHU Yuyao;HE Jiajia;LI Chongzhao;WANG Yong;WANG Zhonghua(College of Agronomy,Northwest A&F University,Yangling,Shaanxi 712100,China)

机构地区：[1]西北农林科技大学农学院,陕西杨陵712100

出　　处：《西北植物学报》2023年第3期359-365,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：陕西省自然科学基金(2019JZ-19)。

摘　　要：为了探究二穗短柄草(Brachypodium distachyon)根部木栓质合成的调控机制,该研究以二穗短柄草Bd21为试验材料,利用生物信息学分析方法克隆了二穗短柄草根部木栓质合成的调控转录因子基因BdMYB92(GenBank登录号为OP497966);采用荧光定量PCR方法,分析BdMYB92基因在二穗短柄草不同组织中的表达模式以及对6种非生物胁迫处理(空气中干旱、20%PEG-6000模拟干旱、4℃中冷处理、200 mmol/L NaCl、100μmol/L ABA和机械损伤)的响应表达特征;利用双荧光素酶和酵母单杂交验证BdMYB92蛋白和BdFAR4基因启动子的互作关系。结果表明:(1)二穗短柄草BdMYB92基因cDNA全长为1343 bp,开放阅读框为990 bp,编码329个氨基酸,蛋白分子量为36.4 kD,理论等电点为5.54。(2)亚细胞定位结果显示,BdMYB92定位在细胞核。(3)荧光定量PCR分析表明,BdMYB92在二穗短柄草根中的表达量显著高于叶鞘、节、穗、节间等其他组织;6种非生物胁迫处理均能诱导BdMYB92的上调表达,且对干旱胁迫的响应最为迅速,表明BdMYB92参与二穗短柄草响应逆境的过程。(4)双荧光素酶和酵母单杂交试验结果表明,BdMYB92与木栓质合成基因BdFAR4启动子区存在相互作用,且能够直接调控木栓质合成基因BdFAR4的转录表达。研究推测,BdMYB92可能与二穗短柄草中其他的木栓质合成基因存在相互作用,进而调控木栓质在根部的沉积。In order to explore the regulation mechanism of suberin biosynthesis in Brachypodium distachyon,we cloned the transcription factor gene BdMYB92 regulating suberin synthesis in the roots of B.distachyon by bioinformatics analysis using Bd21 as the experimental material(GenBank accession number OP497966).The expression pattern of BdMYB92 gene in different tissues of B.distachyon and its response to six abiotic stress treatments(drought in the air,simulated drought in the 20%PEG-6000,4℃cold treatment,200 mmol/L NaCl,100μmol/L ABA and mechanical damage)were analyzed by quantitative PCR.The interaction between BdMYB92 protein and the promoter of BdFAR4 gene was verified by dual-luciferase report assays and yeast one-hybrid.The results showed:(1)the full-length cDNA of BdMYB92 was 1343 bp,with an open reading frame was 990 bp,encoding 329 amino acids.The molecular weight of the protein was 36.4 kD,and the theoretical isoelectric point was 5.54.(2)The subcellular localization results confirmed that BdMYB92 was located in the nucleus.(3)Quantitative PCR analysis showed that the expression of BdMYB92 was significantly higher in the root than in other tissues such as sheats,nodes,spikelets and internodes.All six abiotic stress treatments could induce the up-regulated expression of BdMYB92,and the response to drought stress was the fastest,indicating that BdMYB92 was involved in the response to stresses in B.distachyon.(4)The results of dual-luciferase report assays and yeast one-hybrid showed that BdMYB92 interacted with the promoter region of the suberin synthesis gene BdFAR4 and could directly regulate the transcription expression of the suberin synthesis gene BdFAR4.It is speculated that BdMYB92 may interact with other suberin synthesis genes in B.distachyon,thereby regulating the deposition of suberin in roots.

关 键 词：二穗短柄草 木栓质 BdMYB92 亚细胞定位 表达模式分析 非生物胁迫 相互作用 

分 类 号：Q785[生物学—分子生物学] Q786Q789