胡萝卜DcDREB-A6亚族转录因子基因的克隆与非生物胁迫响应分析  

Cloning of DcDREB- A6 Group Transcription Factor Gene from Daucus carota L. and Its Analysis of Expression Profiles Under Abiotic Stress Treatments

作　　者：黄蔚[1] 王枫[1] 徐志胜[1] 李梦瑶[1] 熊爱生[1] 

机构地区：[1]南京农业大学园艺学院/作物遗传与种质创新国家重点实验室/农业部华东地区园艺作物生物学与种质创制重点实验室,南京210095

出　　处：《农业生物技术学报》2014年第10期1213-1222,共10页Journal of Agricultural Biotechnology

基　　金：教育部新世纪优秀人才支持计划(No.NCET-11-0670);国家自然科学基金项目(No.31272175);江苏省杰出青年基金项目(No.BK20130027);江苏高校优势学科建设项目(No.2011PAPD);江苏省双创计划(No.2011JSSC)

摘　　要：干旱响应元件结合蛋白(dehydration responsive element-binding protein,DREB)类转录因子是干旱应答元件的结合蛋白,在植物响应非生物胁迫时发挥重要作用。本研究基于胡萝卜(Daucus carota L.)转录组数据,检索和拼接获得一个胡萝卜DREB类转录因子基因序列,并根据其序列设计引物,采用RTPCR方法从黑田五寸获得该胡萝卜转录因子基因DcDREB-A6(GenBank登录号:KM386646)。序列分析显示,来源于胡萝卜中的DcDREB-A6基因含有993 bp的开放阅读框,编码330个氨基酸。氨基酸组成成分、理化性质、亲水性/疏水性和三级结构分析结果显示,胡萝卜DcDREB-A6转录因子蛋白质分子量为36.68 kD,等电点为6.00,属于亲水性蛋白。进化树分析显示,DcDREB-A6属于APETLA 2/乙烯反应元件结合蛋白(AP2/ERF)家族转录因子中DREB亚族中的A6组。空间结构表明,该转录因子结构域具有典型的植物AP2/ERF家族转录因子的结构特征,有1个α螺旋和3个β折叠。实时定量荧光PCR证实,胡萝卜DcDREB-A6转录因子基因受高温、低温和高盐胁迫诱导,分别在高温、低温处理4和2 h后表达量增加2倍左右;盐胁迫对其影响较大,处理8 h时,基因表达量增加4.6倍;而干旱胁迫下该基因表达变化幅度相对较小。胡萝卜DcDREB-A6转录因子基因能够响应高温、低温、干旱和高盐等非生物胁迫,本研究结果有助于进一步开展胡萝卜DREB类转录因子的逆境调控研究。In higher plant,dehydration responsive element-binding protein （DREB） family transcription factors play an important role under abiotic stress.Transcription factor binds specific DNA sequence to regulate related gene expression.DREB subfamily factors,one of the major subfamilies of the APETLA 2/ethylene responsive element binding protein （AP2/ERF） family transcription factor,were mainly divided into 6 groups,named A1 to A6 group.Carrot （Daucus carota L.）,a biennial vegetable plant,is famous for its taproot and antioxidant compounds.In this study,the gene sequence,which encoded a DREB transcription factor of DcDREB-A6（GenBank accession No.KM386646）,was isolated from carrot based on transcriptome and genome sequence data of carrot.The DcDREB-A6 gene was cloned by RT-PCR using cDNA as template from carrot cultivar Heitianwucun.Sequence analysis indicated that the length of DcDREB-A6 gene was 993 bp,which encoded 330 amino acids.Then,nucleic acid and deduced amino acid sequence,phylogenetic tree,and molecular modeling were further predicted and analyzed.It was predicted that the relative molecular mass of the DcDREB-A6 was 36.68 kD,and the pI was 6.00.The DcDREB-A6 was a hydrophilic protein.DcDREB-A6 from carrot was classified into A6 group of DREB subfamily transcription factor,which belonged to AP2/ERF family transcription factor.The sequence of DcDREB-A6 was highly conserved in the AP2 domain compared to the DREB transcription factors of other plants.The AP2 domain three-dimension structure of DcDREB-A6 of carrot was built with the template structure of lgcc,which was the AP2 domain of the AtERF1 transcription factor.Protein three-dimensional structure prediction and analysis showed that the domain of DcDREB-A6 had typical structural features of AP2 domain,with one α-helix and three [β-sheets.To elucidate the response of the DcDREB-A6 transcription factor gene of carrot to abiotic stresses （high temperature,low temperature,drought and high-salinity stresses）,the gene was chosen for quantitati

关 键 词：胡萝卜 转录因子 干旱响应元件结合蛋白(DREB) 荧光定量PCR 逆境调控 

分 类 号：Q943.2[生物学—植物学]