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1、棉花GhMYB0基因的克隆、表达分析及功能鉴定1,2221222王诺菡于霁雯吴嫚马启峰李兴丽裴文锋李海晶221,2,黄双领张金发喻树迅12西北农林科技大学农学院,陕西杨凌712100;中国农业科学院棉花研究所/棉花生物学国家重点实验室,河南安阳455000摘要:MYB类转录因子是植物转录因子最大的家族之一,参与控制植物腺毛细胞的模式和形态建成。本研究利用雷蒙德氏棉(Gossypiumraimondii)D5基因组数据库以AtMYB0(GL1,NM_113708)蛋白为参比序列获得同源基因GrMYB0,从徐州142中克
2、隆了陆地棉的GhMYB0,其开放阅读框长度为843bp,编码280个氨基酸。经过保守结构域分析和亚细胞定位确定GhMYB0为R2R3-MYB转录因子。qRT-PCR的结果表明,GhMYB0在徐州142开花当天开始高调表达,开花后20d表达量达高峰;在所有的组织器官中,花中表达量最高,其次为胚珠。转基因功能分析结果表明,在野生型拟南芥(Columbia)中过表达GhMYB0,使其叶片表皮毛与野生型相比明显减少;该基因在拟南芥突变体gl-1中过表达,能恢复表皮毛缺失型突变体的表型,说明该基因可能对拟南芥表皮毛的形态建成发挥一
3、定作用,本试验为研究R2R3-MYB转录因子在棉纤维起始和伸长过程中的调控作用提供有力证据。关键词:棉花;MYB转录因子;植物表达载体构建;遗传转化Cloning,Expression,andFunctionalAnalysisofGhMYB0GenefromCotton(GossypiumhirsumtumL.)1,221222WANGNuo-Han,YUJi-Wen,WUMan,MAQi-Feng,LIXing-Li,PEIWen-Feng,LIHai-Jing,HUANG221,2,Shuang-Ling,ZH
4、ANGJin-Fa,andYUShu-Xun12CollegeofAgronomy,NorthwestA&FUniversity,Yangling712100,China;StateKeyLaboratoryofCottonBiology/CottonResearchInstitute,ChineseAcademyAgriculturalSciences,Anyang455000,ChinaAbstract:MYBtranscriptionfactor,oneofthemostimportantproteinfamili
5、esinplants,isinvolvedintheregulationofsecondarymetabolism,morphogenesisofplant,respondingtoenvironmentstressandplanthormone.Inthisstudy,weusedD5genomicbankofGossypiumraimondiiasthereferencetoAtMYB0(GL1,NM_113708)protein,andclonedthefull-lengthcDNAofanewMYBtranscr
6、iptionfactorgeneGhMYB0fromcotton(GossypiumhirsutumL.).TheopenreadingframeofGhMYB0is843bpinlength,whichencodes280aminoacidresidues.GhMYB0wasconfirmedasR2R3-MYBtranscriptionfactorviaconservedstructureanalysisandsubcellularlocalization.TheqRT-PCRresultindicatedthatG
7、hMYB0washighlyexpressedattheblossomday,itsexpresssionamountreachedthepeakafter20days,withthemostamountinflower,theninovules.TransgenicfuntionanalysisindicatedthatGhMYB0over-expressedinArabidopsislines,showingfewertrichomesinleafepidermthaninthatofthewildtype,whic
8、hsuggeststhatthegenecanrestorethenotrichomephenotypeofgl-1mutantofArabidopsis.Inaddition,thetransgeniclineshadshorterplantheights,longervegetativegrowthtime,an
