Waxy基因的RNA沉默使转基因小麦种子中直链淀粉含量下降

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1、维普资讯http://www.cqvip.com遗传学报ActaGeneticaSinica，August2005，32(8)：846～854SSN0379—4172AmyloseintheSccdsofTransgenicWheat(TriticumaestivumL．)LIJia．Rui’ZHAOWei’'。LIQuan-Zi’YEXing-Guo，ANBao-Yan’．，，，LIXiang‘．ZHANGXian-Sheng。’(1．TheKeyLaboratoryofCropBiology，CollegeofLifeSciences，ShandongAgricultur

2、alUniversity，Taian271018，Chma2．MinistryKeyLabforCropGeneticsandBreeding，InstituteofCropBreedingandCultivation，ChmeseAcademyofAgriculturalSciences．Beijing100081，China；3．Sc『entechDocumen妇tion占InformationCenter，ChineseAcademyofAgricuItura，Sciences，Beijing100081，China)Abstract：Inthisstudy，thele

3、velofamylosewasreducedinwheatseedsbyRNAistrategy．Becausethesyn·thesisofamyloseiscatalyzedbythegranule-boundstarchsynthaseI(GBSSIorWAXYprotein)，theWaxygeneofwheatwasisolatedfrOmwheatseedsbyusingRT-PCR．SouthernanalysisconfirmedthattherewerethreeWaxygenesinwheatgenome．Northernhybridizationshow

4、edthatWaxymRNAaccumulatedinseedsfol_lowingpollination．ByRNAistrategy，the683bpsenseandantisensefragmentsinreverseorientationseparatedbya15ObpintronwereclonedintopCAMBIA3300justdownstreamofthemaizeubilpromoter．ByAgrobacteri-um-mediatedwheattransformationmethod，fourtransgenicplants(CultivarYan

5、gmai10)wereidentifiedbyPCR．RT-PCRandleafpaintingassay．Thelevelofamyloseintheendospermweresignificantlyreducedintransgenicseedsascheckedbyiodinestainingandanalysisofamylosecontent．TheresultsindicatedthatRNAsilencingofWaxygeneresultedinlowlevelofamyloseintheseedsoftransgenicwheat．Keywords：whe

6、at；Waxy；RNAinterference；Agrobacterium-mediatedtransformation；amyloseWaxy基因的RNA沉默使转基因小麦种子中直链淀粉含量下降李加瑞’，赵伟’一，李全梓’，叶兴国2，安宝燕’，李祥’，张宪省’’①(1．山东农业大学生命科学学院，山东省作物生物学重点实验室，泰安271018；2．中国农业科学院作物育种栽培研究所．农业部作物遗传育种重点实验室，北京100081；3．中国农业科学院文献信息中心，北京100081)摘要：通过RNAi策略转化小麦，以降低小麦种子中直链淀粉的含量。小麦中直链淀粉合成的关键酶是颗粒结收稿日

7、期：2005—04—18；修回日期：2005—05—18基金项目：国家转基因专项(编号：JO0一A一006)和国家863项目(编号：2001AA212131)[SuppodedbytheNationalProjectofPlantTrans—formation(No．J00一A-006)andChineseNationalProgramsforHighTechnologyResearchandDevelopment(No．2001AA212131)]作者简介：李加瑞(1978一)，男，博士，研究方向：植