菊花转基因体系的优化与瞬时转化体系的建立

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1、万方数据oPT邛ⅢZATIoNoFTRANSGENICSYSTEMANDESTABLIS丑姗NToFTRANS皿NTTRANSFoRMm’IoNSⅣSTEM矾CHRYs．ANTHEⅣn月讧Chrysanthemum(ChrysanthemummorifoliumRamat．)isoneofthe‘‘Chinesetoptenfamousflowers’’whichisonghatedinChina．Ithasoccupiedabout30％oftheOohalcutflowermarketasthetopof“fourmostfamouscutfl

2、owersallaroundtheworld'’．(Chrysanthemumnankingense)isoneofthediploidwildspeciesinchrysanthemum，withthemostsimplegeneticbackgroundinvolvedintheoriginofcdtiva：tcdchrysanthemum．Duetothelargechromosomenumberofcultivatedchrysanthemum,researchershavemetwithmanydifficultiesontheanaly

3、sisofmoleculargeneticsandmolecularbiologyinit．W砬eCh．nankingensecallbeallexcellentmodelplanttosolvethisproblemforit’Ssimplegeneticbackground．Transgenictechnologyhasbecomeallimportantmeansofimprovingcropvarietiesandgenefunctionstudy．Geneticengineeringinchrysanthemumhasaratherlat

4、estart,manyfactorsCanaffecttheefficiencyresultoftransgenic，forexample：thekindofvectorsorpromotor．Onchrysanthemum,successfultransgenicorstableexogenousgeneexpressionintransgenicplantsarerarelyreported．Onthestudyofgeneticengineeringinchrysanthemum，thereisanurgentneedtoovercomedi

5、fficultiessuchasthelowefficiencyofgenetictransformationandlowexpressionlevelofthetargetgenenotasweexpected．ResearchontheinfluenceoftransformationmethodsandpromoterforchrysanthemumtransgenicCancontributetofindingwayoutofthesedifficulties．Themainresultsaresummarizedasfollows：1．E

6、xpressionanalysisofGUSin35S：GUS／2x35S：GUStransgenicchrysanthemum．Inthisstudy,thesequenceof35Spromoterand2x35SpromoterwereclonedfrompCAMBIAl301vectorbyusingtechnologyofprimerdesignandP咀The35Spromoterand2x35SpromoterwereinsertedinpORERIVectorandpORER2Vector,andobtainedthefollowf

7、ourrecombinantvectors：pORER1--35S：GUS,pORER2--33S：GUS,pORER1-2x35S：GUSandpORER2—2x35S：GUS．ThefourconstructedgenetictransformationIll万方数据菊花转基因体系的优化与瞬时转化体系的建立vectorswereintroducedintothecutchrysanthemum‘Jinba’bytheagrobaeterium-mediatedgenetictransformationsystem．Wehavetransform

8、ed4090piecesofleaf-disc,andreceived22overexpressedtransgenicl