海南龙血树WD40转录因子基因DcWD40-1的克隆和表达分析  

Cloning and expression analysis of WD40 transcription factor gene DcWD40-1 from Dracaena cambodiana

作　　者：朱家红[1] 夏栋楠 王颖[1] 郭冬[1] 李辉亮[1] 梅文莉[1] 彭世清[1] ZHU Jiahong;XIA Dongnan;WANG Ying;GUO Dong;LI Huiliang;MEI Wenli;PENG Shiqing(Institute of Tropical Biosciences and Biotechnology,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101,China;Institute of Tropical Agriculture and Forestry,Hainan University,Haikou 570228,China)

机构地区：[1]中国热带农业科学院热带作物生物技术研究所,海口571101 [2]海南大学热带农林学院,海口570228

出　　处：《广西植物》2020年第1期136-142,共7页Guihaia

基　　金：海南省自然科学基金(318MS093);国家自然科学基金(81773845)~~

摘　　要：海南龙血树是国产血竭的主要基源植物,其血竭主要化学成分为类黄酮化合物。为进一步了解DcWD40-1在类黄酮生物合成中的潜在功能和作用机制,该研究根据海南龙血树转录组数据,利用RT-PCR技术在海南龙血树中克隆了一个WD40基因DcWD40-1,该基因全长1550 bp,包含一个1353 bp的开放阅读框,编码450个氨基酸,蛋白质分子量50.77 kD,理论等电点5.71。生物信息学分析显示,DcWD40-1属于WD40蛋白家族成员,具有5个保守的WD40结构域,和其他植物WD40蛋白同源性高,保守性强。利用Genome Walking方法分离了1503 bp的DcWD40-1启动子序列,该区域具有典型真核生物启动子结构特征,并含有多个应答激素和胁迫的响应元件。表达分析显示,血竭诱导剂能够诱导Dc WD40-1的表达,DcWD40-1的变化与血竭形成及类黄酮积累正相关。此外,DcWD40-1也能对茉莉酸甲酯、细胞分裂素、油菜素内酯和UV-B处理做出积极响应。Hainan Dragon trees(Dracaena cambodiana)are the main plant resources of dragon’s blood in China,and the main chemical constituents of the dragon’s blood are flavonoids.The recent studies about dragon’s blood mainly focus on the chemical constituents and pharmacological activities,while the molecular mechanisms of dragon’s blood formation remain unknown.WD40 transcription factor plays an important role in flavonoid accumulation.In this study,a WD40 gene named DcWD40-1 was cloned in Dracaena cambodiana based on transcriptome data and RT-PCR technology.The full-length of c DNA of DcWD40-1 was 1550 bp,containing 1353 bp opening reading frame(ORF),and encoding 450 amino acids with the calculated molecular weight of 50.77 k D and calculated p I 5.71.Bioinformatics analysis showed that DcWD40-1 belonged to a member of WD40 superfamily,had five conserved WD40 domains,and shared high identities to WD40 proteins with other plants.A 1503 bp-length promoter region of DcWD40-1 was isolated by Genome Walking method,which had structural characteristics of typical eukaryotic promoters.The promoter region of DcWD40-1 contained lots of hormone responsible elements,such as abscisic acid-responsive element,auxinresponsive element,salicylic acid-responsive element and jasmonic acid-responsive element;also had many cis acting elements related stress such as light,cold,hot and anaerobic inducer.Expression analysis showed that DcWD40-1 was induced by dragon’s blood inducers,positively related to flavonoids accumulation and formation of dragon’s blood.In addition,DcWD40-1 can also respond positively to jasmonic acid,cytokinin,brassinosteroid and UV-B treatment.These results will lay the foundation for further study of the potential functions and mechanisms of DcWD40-1 in flavonoid biosynthesis in Dracaena cambodiana.

关 键 词：海南龙血树 血竭 类黄酮 WD40转录因子 基因表达 

分 类 号：Q949.45[生物学—植物学]