農(nóng)桿菌介導(dǎo)的白三葉遺傳轉(zhuǎn)化體系的建立及優(yōu)化.pdf

1、首都師范大學(xué)碩士學(xué)位論文農(nóng)桿菌介導(dǎo)的白三葉遺傳轉(zhuǎn)化體系的建立及優(yōu)化姓名：赫衛(wèi)清申請(qǐng)學(xué)位級(jí)別：碩士專(zhuān)業(yè)：生物學(xué)植物學(xué)指導(dǎo)教師：洪劍明唐益雄2003.5.1——一夔苧塑堇點(diǎn)堂塑!蘭燕堡塞AbstractManylegumeplantsmayproducethebeneficialisoflavones，whicharevitaltotheformationofnitrogen。fixingroot。nodulesandthediseaser

2、esistanceresponseofplantsWhiteclover(trifoliumrepens)，ahighqualitylegumeforage，howeverhasliRleisoflavonesToimprovetheisoflavonecontentsofsuchforage，atlavailablestrategyistointroducegene(s)encodingthekeyenzyme(s)inthebios

3、ynthesispathwayofinsoflavonesAspartoftheprojecttoimproveisoflavonecontentsoflegumeforagebyintroducingisoflavonesynthase(1FS)gene，inthiswork，wehaveestablishedtheAgrobacterium—mediatedtransformationsystemofwhitecloverAgoba

4、cteriumtumefaciensstrainA311carryingtheplanttranfervectorpBll21，whichcontainstheneomycinphosphotransferasellgene(nptlI)and0glucuronidasereportergene(GUS)bothunderthecontroloftheCaMV35Spromoterwasusedintheestablishmentoft

5、hegenetictranformationofwhitecloverThefactorsaffectingthetransformationefficiencyandregernerationefficiencyofwhitecloverhavebeeninvestigatedandoptimizedTotransformwhitecloverthecotyledonwithpetiolefrom45daysoldseedlingsw

6、erepreculturedfor2days，andthenCOcultivatedwiththeAtumefaciensstrainA311for45daysThecotyledonwereCOcultivated，andthenincubatedontheregenerationmedium(blBlmgL6一BAO1mgL’NAASmgL—AgN03)containing40ragL‘kanamyeinand400mgL～carb

7、ennieillinfor6weeks，thetransformedbudsappearedontheexplantsobivouslytheregenerationpercentageoftransformedbudswasabout226％Thetransformedbudsgrewupintosubculturerootingmedium(I／2MBOimg，LNAA)containing30mg，∥kanamycinand400

8、mgL～carhennicillinthepercentageoftransformationisabout65％。SotheestablishedtransformationsystemofwhiteclovercallbeimpliedtotransfertheIFSgeneKeywords：whiteclover(捫如癢Ⅻrepens)，cotyledonarypetiole，GUSgene，Agobacteriumtumefae