爬行衛(wèi)矛與北海道黃楊離體再生及雪花蓮凝集素基因（GNA）的遺傳轉(zhuǎn)化.pdf_第1頁
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1、中國農(nóng)業(yè)大學(xué)博士學(xué)位論文爬行衛(wèi)矛與北海道黃楊離體再生及雪花蓮凝集素基因（GNA）的遺傳轉(zhuǎn)化姓名：尚愛芹申請學(xué)位級別：博士專業(yè)：農(nóng)業(yè)生物環(huán)境與能源工程指導(dǎo)教師：趙梁軍20060601AbstractTheprotocolsfurefficientshootregenerationweresuccessfullydevelopedfromhypocotylexplantsofEuonymusfortuneivaTradicansandEuo

2、nmusjaponicus‘Cuzhi’ThesystemsofgenentictransformationofEuonymusfortuneivarradieans，Euonmusjaponicus‘Cuzhi’andEuonymusfortuneiwereestablishedTheGNAgenewassuccessfullytransferredintothethreematerialsbyAgrobacteriummediate

3、dmethodTheresultswereasfollows：1EstablishmentofdirectshootregenerationsystemofEuonymusfortuneivailradieansHypocotylexplantswerehorizontallyculturedonabasicmediumcomposedofMSmediumsupplementedwith05mg／LBAPand001mg，LNAAfor

4、inductionanddevelopmentofadventitiousshootsRegeneratedshootsproliferatedefficientlyonashootmultiplicationmediumconsistingofMSmediumcontaining10mg／LBAPand01mg／LNAAMicroshootswererootedonamofingmediummadeupofMSmediumenrich

5、edwith05medLIBAand05me：／LIAAAfterhardening，90％ofplantsweresuccessfullyestablishedundergreenhouseconditions，Amongthemanyfactorsknowntoaffecttheregenerationprocesses，severalsweretestedhere：theregenerationabilityofdifferent

6、segmentsofhypocotylexplants，theorientationoftheexplantsinthemedium，typeofmediumandthesourceofcarbon，lingtandtheeffectofAgN03TheHistologicalobservationrevealedthatshootprimordiumoriginatedfromsubepidermalcellsofhypocotyle

7、xplantsanddirectlydevelopedintoadventitiousshootswithoutcallusformation2，EstablishmentofdirectshootregenerationsystemofEuonymusjaponicus‘cuzhi’AdventitiousshootsweresuccessfullyregeneratedfromhypocotyIexplantsofinvitrocu

8、lturedseedlingsofEuonymusjaponicus‘cuzhi’HypoeotylsliceswereculturedonMSandB5basalmediumsupplementedwithvariedconcentrationofdifferentplantgrowth—regulators，eggnaphthaleneaceticacid(NAA)andindole3butyricacid(IBA)incombin

9、ationwith6benzylaminopurine(6BA)andkinetinThestudyshowedthatshootscouldbedirectlyregeneratedfromhypocotylexplantswithouttheinterveningcallusphase；MSmediumwasmoresuitableforadventitiousshootsregenerationTheabilityofhypoco

10、tylssegmentstoproduceshootsvarieddependingupontheirpositionontheseedlingsThehighestregenerationratewasobtainedwithhypocotylsegmentsneartothecotyledonculturedonMSbasalmediumsupplementedwith1，5merlE6BAand005mg／LNAATheregen

11、eratedshootswerereadilyelongatedonthesamemedium叢usedfurmultiplicationandrootedonhalfstrengthMSbasalmediumsupplementedwith10mg／LIBAand100mg／LactivatedcarbonAftertransferredtogreenhouseconditions，96％oftheplanfletsweresucce

12、ssfullyacclimatized3EstablishedthesystemfortransformationofEuonymusfortuneivatradicans，EuonymusfortuneiandEuonmusjaponicus‘Cuzhi’Thefactorsthataffectthetransformationfrequencysuch∞theoptimalconcentrationofantibiotics，the

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