中國水仙轉(zhuǎn)IPT基因的研究.pdf

1、廈門大學(xué)碩士學(xué)位論文中國水仙轉(zhuǎn)IPT基因的研究姓名：曾榮華申請學(xué)位級別：碩士專業(yè)：細(xì)胞生物學(xué)指導(dǎo)教師：沈明山陳亮2001.7.1StudiesonG；eneticTransformationofNarcissustazettaLvarchinens缸RoembyIPTGeneABSTRACTNarcissustazettahasgreatvalueasallimportantflowerplantinChinan坨autolatorys

2、enescenceinhibitionsystemconstructedbyfusingleafsenescencespecificpromoterwitll伊TgeneiSappliedtothetransformationofnarcissusinthisexperiment／PTexpressioninthetransgenicplantCandelayleafsenescencebyincreasingendogerouscyt

3、okininlevelAtthesametime，wehaveinthefirstestablishedgenetictransfonTtationsystemofnarcissusthroughtheguideofGUSreportergeneThemainresultsareasbelow：1TissuecultureprocedureinthenarcissusgenericUansformationWasdevelopedAno

4、fbulbsinitiatedregenerationofbulbilsdirectlywhenculturedinMsmediumsupplementedwith6BA(4mg／L)NAA(05mg／L)thecoefficientofregenerationis112：Attheotherhandabout742％offlowerpedunclesinducedcalliwhenculturedinMS6BA(1mg／L)NAA(5

5、mg／L)，andthecallicarldifferentiategreatmanybulbilsinMS6一BA(4mg／L)NAA(05meJL)，thecoefficientofdifierentiationis1292EstablishedemcienttransgenieteehnologyofnarcissusThechimericPCMV35SGUSandPSAOl2儼Tgenesweretrailsforrnedint

6、onarcissusmediatedbyAgrobacteriumtumefaciensAbout95％ofbulbsindicatedexpressionofGUSreportergeneailerCO—cultivating3dayswithagrobacteriumSOdid85％offlowerpedunclesOtherwisetherewere15％oftransformedbulbsandallofflowerpedunc

7、lesthatCOuldregeneratebulbils3Intervalscreeningwasadoptedtoscreentransformednarcissustissue293％oftransformedtissueObtainedtheresistancetoHygand20％oftheHygtissueindicatedexpressionofGUSgeneTransformedtissueregeneratedbulb

8、ilsinMS6BA(4mg／L)NAA(05mg／L)，thecoefficientofregenerationis34TherehadexpressionofGUSgeneintransgenicplantleaf4PCRanalysisshowedthat14％oftheHygrtissuehadpositiveband；PCRSouthemindicatedthatitwasthefragmentofPSAGl2Southern