大豆遺傳轉(zhuǎn)化系統(tǒng)的建立與農(nóng)桿菌介導(dǎo)Bt基因轉(zhuǎn)移的研究.pdf

1、東北農(nóng)業(yè)大學(xué)碩士學(xué)位論文大豆遺傳轉(zhuǎn)化系統(tǒng)的建立與農(nóng)桿菌介導(dǎo)Bt基因轉(zhuǎn)移的研究姓名：孔凡江申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：農(nóng)業(yè)昆蟲與害蟲防治指導(dǎo)教師：趙奎軍2002.11.1碩士學(xué)位論文摘要2002年11月EstablishmentofSoybeanGeneticTransfonllationSystemandStudyonBtGeneTransferWithAgrobacterium—mediatedMethodAbstractInthisre

2、search，tenvarietiesofsoybeanwerechosenandthecotyledonarynodesofthesevarietieswereusedtobethetargetexplantsofBtgeBetransformationThefactorsaffectingtheAgrobacteriummediatedtransformationandtheshootregenerationwerestudiedT

3、heresistantplantswereobtainedfromthetwostepsselectionFourpositiveplantswasobtainedSouthernanalysisshowthatBtgenewasintegratedintosoybean’sgenomeThemainresultsofthisresearchasfollows：1Therewerelargedifferencesofregenerati

4、onfrequencybetweensoybeangenotypesRegenerationfrequencywasbetween06504％in10soybeangenotypesthatwereusedinthisresearchTheresultSinthisresearchhasshownt11atregenerationfrequencywashighofHeinong35Hefeng35andHefeng25，whichwe

5、re6504％6402and6082％respect；elySothe3genetypeswerechosentobetheresearchmaterialsinthisstudy2TheconcentrationofBAandIBAwasdesignedasanalternationchangingatthedi行erentiationstagcoftheadventitiousshootfromcotyledonarynodeThe

6、suitableproportionbetweenBAandIBAfordifferentgenotypeswassceenedForHeinong35，whentheconcentraonofBAwas150mg／LandtheconeentrationofIBAwas020mg／L，thedjfrerentiationfrequenceisthehighestwith7857％ForHefeng25，whentheconcentra

7、tionofBAwas130mg／L，andtheconcentraionofIBAwas025mg／L，thedifierentiationfrequencyisthehighest4857％Withthesuitableproportion，theshootsofeachexplantswere7to10，notover10，whichisbenefitforelongationForHefen235，whentheconcentr

8、ationofBAwas120mg／L，andtheconcentrationofIBAwas025m“L，thedifferentiationfrequencyisthehighestwith7407％Theshootsofeachexplantswere4to93Thefrequencyofrootregeneraionwasremarkablyincreasedbytheadditionof1BAattherootregenera

9、tionstageThebasicmediumforrootregenefationwasl／2MSB，andtheconcentrationofsugarwas2％WhentheconcentrationofIBAwas20mg／LtherootregenerationfrequencyforHeinon935andHefeng25werehigherwith7188％and7241％respectivelyWhentheconcen

10、trationofIBAwas15mg／L，therootregenerationfrequencyforHefeng35werehigherwith6897％4TheselectfongenesofBtwereNPTII，andtheselectiveregeantwasKanThelateselectionwasusedtoovercomethedifficultyofadventitiousshootdifferentiation

11、andtherootregenerationWhentheshootswere03lcm，andbeforebeingtransferredtorootregenerationmedium，theshootswerescreenedwithKan，andthesuitableconcentrationofKanforHeinon935，Hefen935andHefen925waslOOmg／L5TheCefwasusedasantibi