梁山慈竹DfMYB3基因克隆及启动子分析  

Cloning and Promotor Analysis of DfMYB3 from Dendrocalamus farinosus

作　　者：杜恬恬 李会萍 王博雅 欧倩 黄艳 曹颖 胡尚连 DU Tian-Tian;LI Hui-Ping;WANG Bo-Ya;OU Qian;HUANG Yan;CAO Ying;HU Shang-Lian(Lab of Plant Cell Engineering Southwest University of Science and Technology,Mianyang 621010;Engineering Research Center for Biomass Resource Utilizaiton and Modification of Sichuan Province,Mianyang 621010)

机构地区：[1]西南科技大学植物细胞工程实验室,绵阳621010 [2]四川省生物质资源利用与改性工程技术研究中心,绵阳621010

出　　处：《植物研究》2021年第5期729-737,共9页Bulletin of Botanical Research

基　　金：四川省应用基础研究项目(18YYJC0920);四川省“十三五”重点攻关项目(2016NYZ0038);四川省重点研发项目(2019YFN0005);环境友好能源材料国家重点实验室项目(19fksy0113)。

摘　　要：基于梁山慈竹转录组数据库,以梁山慈竹叶片为材料,克隆得到一个MYB转录因子,命名为DfMYB3,其开放阅读框长度为1287 bp,编码428个氨基酸,GenBank注册号为KY963358。保守结构域分析显示,DfMYB3有典型的SANT结构域,具有DNA-Binding结构域。系统进化树分析发现,DfMYB3与甘蔗、拟南芥、毛白杨等物种的R2R3-MYB转录因子聚集在一枝上。亚细胞定位结果显示,DfMYB3蛋白在细胞核以及细胞膜中均有表达,在细胞核上更为显著。通过染色体步移法克隆得到DfMYB3基因5′侧翼2000 bp左右的序列。PlantCARE在线软件分析表明,该序列具有典型的启动子特征,并含有GA、ABA、MeJA等激素以及干旱等胁迫响应元件。100μmol·L^(-1)GA、100μmol·L^(-1)ABA处理梁山慈竹后,显著上调了DfMYB3基因的表达,表明DfMYB3基因能对GA及ABA处理产生应答。为探究DfMYB3启动子的功能,构建了DfMYB3启动子融合GUS基因的表达载体,并遗传转化烟草。在转基因烟草叶片及茎杆中都检测到了GUS信号,在叶脉处最为显著。Based on the transcriptome database of Dendrocalamus farinosus,a MYB transcription factor named DfMYB3 was cloned from the leave of D.farinosus.Its open reading frame length was 1287 bp,encoding 428 amino acids,and GenBank accession Number was KY963358.The conserved domain analysis showed that DfMYB3 had the typical SANT domains and DNA-Binding domains.Phylogenetic tree analysis showed that DfMYB3 clustered with R2 R3-MYB transcription factors in Saccharum,Arabidopsis and Populus.Subcellular localization showed that DfMYB3 protein was expressed in both the nucleus and cell membrane,and was more significant in the nucleus.The sequence of 5′flanking of DfMYB3 gene about 2000 bp was cloned by chromosome walking method.Analysis of PlantCARE online software showed that the sequence had typical promoter characteristics,and contained hormones responsive elements such as GA,ABA,MeJA as well as stress responsive elements such as drought.The expression level of DfMYB3 gene was significantly up-regulated after 100μmol·L^(-1) GA and 100μmol·L^(-1) ABA treatment,indicating that DfMYB3 gene was involved in response to GA and ABA treatments.In order to explore the function of DfMYB3 promoter,the expression vector of GUS gene fused with DfMYB3 promoter was constructed and genetically transformed into tobacco.The results showed that GUS signal was detected in both leaf and stem of transgenic tobacco,especially in the veins.

关 键 词：梁山慈竹 MYB转录因子 启动子 GA、ABA处理 GUS染色 

分 类 号：S795.9[农业科学—林木遗传育种]