利用轉(zhuǎn)錄后基因沉默（PTGS）防治作物黃瓜花葉病毒病.pdf

1、分類號(hào)UDC密級(jí)單位代碼湖南農(nóng)業(yè)大學(xué)碩士學(xué)位論文Y93231910537利用轉(zhuǎn)錄后基因沉默(PTGS)防治作物黃瓜花葉病毒病StudyoncontI：olplantcucumbermosaicvirusthroughpost—transcriptionalgenesilencingapproach研究生姓名指導(dǎo)教師專業(yè)領(lǐng)域研究方向盤查哩型要盟基旦丞堡嗑副班童雖植物瘟翌堂擅塑痘垂堂提交論文日期2QQ生旦!目論文答辯日期2QQ生旦8目答辯委

2、員會(huì)主席：受亙豆煎論文評(píng)閱人夏童丞￡耋援2戴良墓(教援≥學(xué)位授予日期2QQ生旦目二00六年六月StudyoncontrolplantCucumbermosaicvirusthroughpost—transcriptionalgenesilencingapproachAbstract：RNA2fragment、MPgenefragmentofCMVisolateP3613，andCPgenefragmentofCMVisolateANwe

3、reamplifiedbyRTPCR，respectiveltyThreefragmentswereclonedintovectorpBluescriptsic(一)TofacilitatetheconstructionofinvertedrepeatexpressionvectorofCMVgenome，thecorrespondingrestrictionenzymesitesofCPgene(Bspl20I)，RNA2fragme

4、nt(KpnIandBspl20I)andMPgene(PsyIandBspl20I)wereintroducedbyPCR，respectivelyTwoinverted—repeatexpressionvectors，usingCPgeneasaspace，ofCMVRNA2andMP，namelyRNA2一CPRNA2andMPCP—MP，werebuiltupunderthecontrolofT7promotorbyconven

5、tionalmolecularcloningtechniqueFurthermore，thetworecombinantplasmidswereconfirmedbyPCR、restrictionenzymedigestionandsequencingThetwoexpecteddsRNAfragmentswereexpressedthroughinvitrotranscriptionThetwoplasmidsweretransfor

6、medintoEcoliHTl15，whichisRNaseIIIde矗cient，respectivelyMoreover，theexpressioncrudeoflargeamountdsRNAwereusedfortreatingplantstopreventvirusinfectionTwotypesofexperimentsweredesigned：theCMVwereinoculatedbeforedsRNAtreatmen

7、torviceversaTheexistenceofCMVintobaccowereconfirmedbyRT—PCRofCPgeneNoviruswasdetectedinbothtreatmentsTheresultsshowedthatthebacterialcrudecaninduceplantresistancetocorrespondingvirusOurresultsleadawayforplantviruscontrol

8、usingdsRNAthroughPTGSapproachNowwearetreatingthevirusinfectedplantsingreenhousetofindthebestconditionforusingthistechniqueinfieldinnearfutureKeywords：CMV(Cucumbermosaicvirus)；PTGS(posttranscriptionalgenesilencing)：dsRNA；