转化山葡萄ice基因培育抗寒葡萄新种质的分析

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1、ABSTRACTTHESTUDYoFBREEDINGNEWCOLD．RESISTANTGRAPEGERMPLASMTRANSFoRMED舰GENEFRoMAMURGRAPEGrapesbelongtotheVitaeeaeVitis，whichareveryhighvalueoffruittreecultivation．However,grapefreezingdamagebringsgreatlosstotheproduction,SOcultivatethecold-resistantvarietiesimminently．Thegrapeisaperennialf

2、ruittreeswithacomplexgeneticbackground，longlifecycle，andSOon,thatconventionalbreedingmetho&havebeenextremelylimitedinbreeding，andtheapplicationofgeneticengineeringhasopenedupanewwayfortheimprovementofgrapebreeding．1CEasimilaractivatorofgenesencodingbHLHtranscriptionfactorMYC,underthelow-

3、temperature-induced，CanmakeaseriesofcoldinducedgenesCOR，aswellasinworktheexpressionofothergenesinplantcoldadaptation，thusincreasingcold-resistanceoftransgenicplants．Inthisstudy,rootstockvarieties‘1202’(Mourvedre*Rupestris)andcultivatedvarieties’ManicureFinger’grape(VitisviniferaL．’Manicu

4、reFinger’)asmaterials，establishing‘1202’graperootstockadventitiousbudregenerationsystem；Agrobacterium—mediatedtransformationwasadoptedtointroducetheICEgeneinto‘ManicureFinger’grapevine，andtheICEgeneintothe‘1202’thatisdifficulttransformationofgraperootstockWasusedbypollen-tubepathwaymetho

5、d，inordertoobtaincold-resistanttransgenicgrapevineandcultivatethenewgermplasmofgrape．neexperimentalresultsofthestudyareasfollows：1．ICEgeneWasclonedfromtheVamurensis，‘ManicureFinger’grapetransformationWasmediatedbytheoptimizationofAgrobacterium·mediatedtransformationsystem，receivedatotalo

6、f5resistantlines，afterGUSdetection,testedbyPCIL5linescontainedthetargetband；furtherdetectedbyRT-PCR,inwhich3linesthroughRT-PCRdetection．Preliminaryconfirmedthat，ICEgenehasbeenintegratedintothe’ManicureFinger’grapegenome．2．Bymoleculardetectionof3linesoftransgenicManicureFingerGrapewereinl

7、owtemperaturetocoldresistance，choosingthemostcoldlineofmalondialdehyde(Mna)，thecontentofsolublesugar,proline，superoxidedismutase(SOD)，peroxidase(POD)andcatalase(CAT)wasmeasured．Theresultsshowedthatthetransgenicline3Canstand一8"Clowtemperatureandrecoverynormalgrowthunderroo