二穗短柄草BdDREB-30基因的表达及其启动子的功能分析  

The Expression BdDREB-30 Gene from Brachypodium distachyon and Functional Analysis of Its Promoter

作　　者：黄钢 谭生龙 孙婧 陈利红 HUANG Gang;TAN Sheng-Long;SUN Jing;CHEN Li-Hong(Institute for Systems Biology,Jianghan University,Wuhan 430056,China;School of Information and Communication Engineering,Hubei University of Economics,Wuhan 430205,China)

机构地区：[1]江汉大学系统生物学研究院,武汉430056 [2]湖北经济学院信息与通信工程学院,武汉430205

出　　处：《农业生物技术学报》2022年第7期1257-1267,共11页Journal of Agricultural Biotechnology

基　　金：湖北省教育厅科学研究计划指导性项目(B2019115)。

摘　　要：脱水应答元件结合蛋白(dehydration responsive element binding protein,DREB)转录因子在植物生长发育和响应逆境胁迫中发挥着重要作用。研究二穗短柄草(Brachypodium distachyon)DREB转录因子对禾本科作物DREB类转录因子抗逆分子机制的阐明具有一定的促进作用。本课题组前期针对二穗短柄草DREB转录因子芯片数据的研究发现BdDREB-30(GenBank No.XM_003561067.4)响应多种逆境胁迫。为进一步了解其功能,本研究采用qPCR技术研究了BdDREB-30基因在不同组织和7种非生物胁迫条件下的表达模式,并采用GUS染色法研究了其启动子的功能。结果表明:BdDREB-30基因在茎中的表达量最高,在干旱和HO条件下其表达量显著高于对照。与其他物种同源蛋白的多序列比对结果表明,BdDREB-30蛋白质含DREB/CBF亚家族典型的AP2/ERF结构域和A-1亚类的特征序列。进化分析显示其与大麦(Hordeum vulgare)和黑麦(Secale cereale)等麦类作物的同源蛋白亲缘关系相对较近。启动子顺式作用元件分析结果显示,其启动子含有众多光响应元件,以及一些植物激素应答元件和一些胁迫应答元件。构建BdDREB-30启动子的植物表达载体并转入烟草(Nicotiana tabacum),在低温、干旱和盐胁迫处理条件下对转基因烟草叶片进行GUS染色。结果表明BdDREB-30启动子受干旱诱导较明显,是一种干旱诱导型启动子。本研究为二穗短柄草BdDREB-30基因响应逆境分子机制的研究提供了理论依据。Dehydration responsive element binding protein(DREB)transcription factors play an important role in plant development and response to various stresses.The study on its DREB transcription factors of Brachypodium distachyon would promote the elucidation of molecular mechanism of the stress resistance of DREB transcription factors in gramineous crops.The early research work on the transcription profiles from the microarray study of DREB transcription factors showed that BdDREB-30(GenBank No.XM_003561067.4)responds to various stresses.In order to further explore its function,the expression patterns of BdDREB-30 gene in different tissues and under 7 abiotic stresses were studied by qPCR,and the function of its promoter were studied by GUS staining.The results showed that the expression of BdDREB-30 was the highest in stems and its expression under drought and HOstresses was significantly higher than that of the control.The results of multi-sequence alignment with other proteins showed that BdDREB-30 protein contained the typical AP2/ERF domain of DREB/CBF subfamily and the characteristic sequence of A-1subclass.Evolutionary analysis showed that BdDREB-30 was relatively close to the homologous of wheat crops such as barley(Hordeum vulgare)and rye(Secale cereale).The analysis results of cis-acting elements of promoter showed that the promoter of BdDREB-30 gene contained many light response elements,plant hormone response elements and stress response elements.In order to further study the expression characteristics of the promoter,it was constructed into plant expression vector and transferred into tobacco(Nicotiana tabacum).The plant leaves of transgenic tobacco under low temperature,drought and salt stresses were stained with GUS.GUS staining showed that the promoter of BdDREB-30 gene was strongly induced under drought stress,indicated that the promoter of BdDREB-30 gene was a drought inducible promoter.This study provides a theoretical basis for the study of the molecular mechanism of BdDREB-30 gene in response to str

关 键 词：二穗短柄草 脱水应答元件结合蛋白(DREB) 启动子 干旱诱导 GUS染色 

分 类 号：S51[农业科学—作物学] Q781[生物学—分子生物学]