rolC基因轉(zhuǎn)化‘美人指’葡萄的研究.pdf

1、南京農(nóng)業(yè)大學碩士學位論文rolC基因轉(zhuǎn)化‘美人指’葡萄的研究姓名：楊麗娜申請學位級別：碩士專業(yè)：果樹學指導教師：陶建敏20080601Abs仃actAneffectivesystemregenerationsystemof‘ManicureFinger’孕apevinevinewasdevelopedinthisstudyAgrobacteriummediatedtransformationandpollenrobepathwayWeI

2、eadoptedtointroducetherolCgeneinto‘ManicureFinger’grapevinetoimprovetheabilityofabloomThedetailedresultswerelisted蠲follows：1Plantregenerationwereinducedfromleafpetioleandsteinexplantsof‘ManicureFinger’grapevinewithdiffer

3、encemediasupplemented、城mdifferentcombinafio璐。of6BATDZandIBAthroughorganogenesisTheresultsindicatedthatthebasicmediaMS，B5andNN69allinduceadventitiousbudsfromleafpetioleandstemexplantsThehighestregenerationrateofadventitio

4、usbudswas600％，whenexplantsWereincubatedonMSmedium誡也6BA30mg／LandIBA01mg／LTheregenerationrateofadventitiousbudswas400％whenexplantsw豇eincubatedonMSTDZI0mg／LmA02mg／L2Transgenicplantsweregeneratedfromleaves，petioleandstemexpl

5、antsof‘ManicureFinger’grapevinebyAgrobacteriummediatedtransformationwithroICgene，andthefollowingfactorswereinvestigated：Agrobacteriumdippingtime，coculturetime，concentrationofKanamycinandconcentrationofCefotaxineTheoptima

6、lconditionsincludedthatinfected4minbyAgrobacterium，cocultured3d，Km30mg／L，andCef250mg／L3TwelveKmresistantshootsof‘ManicureFinger’grapevinewereobtainedbyAgrobacterium—mediatedtransformationHistocheraicalGUSassayontheseresi

7、stantshootsshowedthattwoshootsWCItGUSpositiveplants；oneofthemwasconfirmedthatrolCgenew越integratedintothehostgenomeof‘MaJlicltlrcFmger’grapevinebybothPCRandRTPCRanalysis4Agrobacterium—mediatedtransformationwasadoptedtoint

8、roducetherolCgeneintotheinflorescenceof‘ManicureFinger’grapevineviathepollen—tubepathway40planfletswereobtainedfrom‘ManicureFingcr’seedsByGUShistochemistryassay、PCRamplifiationandRTPCRanalysis，fiveplantsshowedGUSpositive

9、witha125％positiverate，andtwoshootsshowedPCRandRTPCRpositive，whichinitiallydemonstratedthattherolCgenewasintegratedint0thegenomeof‘ManicureFinger’grapevineKeywords：‘ManicureFinger’grapevine。rolCgerm，Agrobacteriummediated，