棉花尿苷二磷酸糖基转移酶基因GhUGT85O1的克隆及其功能分析  

Cloning and Function Analysis of Uridine Dipfosphate Glycosyltransferase Gene GhUGT85O1 in Cotton(Gossypium hirsutum)

作　　者：黄娟[1,2] 范术丽[2] 宋美珍[2] 庞朝友[2] 魏恒玲[2] 喻树迅[1,2] 

机构地区：[1]西北农林科技大学农学院,杨凌712100 [2]中国农业科学院棉花研究所/棉花生物学国家重点实验室,安阳455000

出　　处：《农业生物技术学报》2015年第6期701-710,共10页Journal of Agricultural Biotechnology

基　　金：国家棉花产业技术体系(No.CARS-18)

摘　　要：在植物糖基转移酶大家族中,多基因家族尿苷二磷酸糖基转移酶(uridine dipfosphate glycosyltransferase,UGT)成员最多,广泛参与植物生长发育调控、逆境响应等生理过程。本研究克隆了陆地棉(Gossypium hirsutum)UGT家族基因Gh UGT85O1(Gen Bank登录号:KP300000),其开放阅读框(open reading frame,ORF)全长1 422 bp,编码473个氨基酸。实时荧光定量PCR(quantitative Real-time PCR,q RT-PCR)结果表明,该基因在衰老叶片中大量表达,并且受脱落酸(abscisic acid,ABA)、茉莉酸(jasmonic,JA)和聚乙二醇(polyethylene glycol,PEG)诱导。该基因在拟南芥(Arabidopsis thaliana)中过表达能够促进拟南芥早花(P<0.01)和提前衰老,衰老时期叶绿素含量显著低于野生型(P<0.01),At ORE1、衰老相关基因12(senescence-associated gene,At SAG12)、At SAG13、WRKY结合蛋白6基因(WRKY DNA-binding protein6,At WRKY6)、NAC转录因子基因(At NAP)和病程相关蛋白1基因(pathogenesis-related protein 1,At PR1)等衰老关键基因的表达量高于野生型(P<0.05或P<0.01)。启动子缺失研究表明,该基因启动子的关键区域在上游-1 077^-1 363 bp内,该区域含有厌氧响应元件(anaerobic response element,ARE)、WRKY转录因子结合位点(WRKY transcription factors binding site,W box)和TGA元件(TGA element,TGA)3个关键调控元件。研究结果初步表明,Gh UGT85O1参与了植物逆境胁迫、开花时间和衰老进程的调控,为在棉花中进一步阐明其功能奠定了基础,为培育生育期短的短季棉提供了优质基因资源。Multigene family of uridine dipfosphate glycosyltransferases （UGTs）, comprising the greatest number of glycosyltransferases, involve in various biological processes, such as regulation of plant growth and development, and response to abiotic stresses. In the present study, one UGT gene, named as GhUGT8501 （GenBank No. KP300000） in cotton （Gossypium hirsutum） was identified. The full length of its open reading frame （ORF） and predicted peptide chain was 1 422 bp and 473 amino acids, respectively. The C-terminal contained a plant specific putative secondary plant glycosyltransferase （PSPG） motif found in UGTs family. Quantitative Real-time PCR （qRT-PCR） result indicated that GhUGT8501 expressed extensively in the mature and old leaves, and was induced by abscisic acid （ABA）, jasmonic acid （JA） and polyethylene glycol （PEG） in cotton. To illustrate its biological function, 35S：：GhUGT8501 vector was constructed and transformed into Arabidopsis thaliana. The over expression of GhUGT8501 in transgenic Arabidopsis thaliana showed significantly early flowering （P〈0.01） and accelerated senescence phenotype compared with that in wild type. The chlorophyll content in transgenic Arabidopsis thaliana was significantly lower than that in wild type at senescence stage（P～0.01）. Meanwhile, the expression levels of AtORE1, senescence-associated genel2 （AtSAG12）, AtSAG13, WRKY DNA-binding protein 6 gene （AtWRKY6）, NAC transcription factor （AtNAP） and pathogenesis-related protein 1 gene （AtPR1） of senescence associated genes were higher than that in wild type （P〈0.05 or P〈0.01）. Promoter deletion analysis result located the essential region of GhUGT8501 promoter on- 1 077--1 363 bp, which contained 3 eis-acting elements of anaerobic response element（ARE）, WRKY transcription factors binding site（W box） and TGA element （TGA）. The result preliminarily suggested the involvement of GhUGT8501 in response to abiotic stresses, regulation of both flowerin

关 键 词：棉花 尿苷二磷酸糖基转移酶基因Gh UGT85O1 表达 开花 衰老 启动子缺失 

分 类 号：Q523[生物学—生物化学]