馬鈴薯A病毒外殼蛋白（PVA CP）基因的遺傳轉(zhuǎn)化研究.pdf

1、Y785563國內(nèi)圖書分類號：$532國際圖書分類號：學(xué)位論文密級：馬鈴薯A病毒外殼蛋白(PVACP)基因的遺傳轉(zhuǎn)化研究于風(fēng)麗學(xué)善：砧≥峭j導(dǎo)師姓名：盧翠華研究員申請學(xué)位級別：農(nóng)學(xué)碩士專業(yè)名稱：作物遺傳育種論文提交日期：2005年4月27日答辯日期：2005年6月8目所在單位：東北農(nóng)業(yè)大學(xué)學(xué)位授予單位：東北農(nóng)業(yè)大學(xué)學(xué)位授予日期：答辯委員會主席：障伊里教授2005年6月StudyonqransferingofCoatProteinGene

2、ofPotatoVirusA(PVAcP)intoPotato(SoIanumtuberosumL)AbstractAregenerationsystemavailableforpotatotransformationwasestablishedconsideringbyseverialfactors，suchasthepotatogenetype，theexplanttype，theconcentrationofdifferentph

3、ytohormonesandbactericidesthetimeofcocultureInthisstudyPVACPgenewastransferredintoCVSDongnon9303andDadiNO1，maintwolocalcultivars，withAgrobacteriummediated—methodTheexplantsincludedthestemsegmentsofplantletsinvitro，discso

4、fmicrotubersandminitubersThetransgenicplantsweretestedbyPCRandPCR—Southem，andthePVACPgenewasconfirmedtobeintegratedinthegenomeofpotatoThemainresultswefeshowedasfollowing：1Establishmentoftheregenerationsystemforportotrans

5、formationThestemoptimalmediumforcallusculturewasMS01mg／L24D25mgmBAandthebestmediumfordifferentiationwasMS5mg，LGAs2SmUtBAAndtheoplimatregenerationmediumfortuberdiscwasMS35mg／LZT088mg／LIAAUnderthesarneconditions，wecompared

6、thestemregenerationfrequencyoftheDongnon9303，Atlantic，KexinNo4andKexinNo13，andfoundthatthehighestoneisAtlantic，whichis7500％ThedifferentiationfrequencyofDongnon9303washigherthanthatofDadiNo12Establishmentofhighfrequencytr

7、ansformationsystemofpotato21Theoptimaltimeofpreculturewas2daysandtheoptimaltimeofco—culturewas3daysforstemexplantTheoptimaltimeofcoculturewas2daysfortuberexplant22Differentexplanttypesneededdifferentrespomdencefortransfo

8、rmationForthestemsexplant，theoptimalconcentrationofAgrobacteriumwasOD60005，theoptimaltimeofinfectionwas10minutesThefrequencyofcallusresistanttoKanamycinwashighestunderthiscondition，reaching7850％Forthetuberdiscs，theoptima

9、lconcentrationofAgrobacteriumwasOD600O5，theoptimaltimeofinfectionwas5minutesThefrequencyofshootresistanttoKanamycinwashi【ghestunderthiscondition，reaching5340％23Inthecomparativeexperimentofseveralbactericidesthebestonewas

10、CefotaximesodiummadeinUSA，anditdidn’tinhibittheexplantregenerationItsconcentrationwasl50mg／Lforthetuberdiscsand200m∥Lforthestemsegments24KanamycinwasusedastheselectiveagentDifferentexplanttypesneededdifferentselectivecon