乙烯受體基因LeETR1和LeETR2反義轉(zhuǎn)化番茄純合體的鑒定及其表型的初步研究.pdf

1、浙江大學(xué)碩士學(xué)位論文乙烯受體基因LeETR1和LeETR2反義轉(zhuǎn)化番茄純合體的鑒定及其表型的初步研究姓名：向慶寧申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：蔬菜學(xué)指導(dǎo)教師：張明方應(yīng)鐵進(jìn)2003.5.1SummaryEthyleneisasimple，readilydiffusiblegaseoushormoneanditregulatesmultipledevelopmentalprocessesSuchethylenedependentprocesses

2、havelongbeenpresumedtobedependentonthesynthesisoractivationofethylenereceptorsAlthoughtheregulationofethylenebiosynthesisisespeciallywellcharacterized，muchlessisknownabouttheregulationofethyleneperceptionBasedontheresult

3、sthatwereinvestigatedonthemutantsETRl，amemberofethylenereceptorgenes，ETRl，ERSl，ETR2，EIN4，ERS2ethylenereceptorgenesmightplayanegativeroleinregulatingresponsetOethyleneHoweverSOfaronlytheresultsobtainedbyWhitelawetal(2002)

4、concerningLeETRlgenewerepartlyconsistentwiththenegativemodelandpartlyagainstitObviouslytheactionmodelsfortheseethylenereceptorsarefarfrombeingclearlyunderstoodThepurposeofthisstudyistoverifythepossiblefunctionsoftomatoet

5、hylenereceptorgenesLeETRlandLeETR2inregulatingethyleneresponseByintroducingantisenseLeETRlorLeETR2genesintotomato，thefollowingaspectswereinvestigatedMethods：(1)ThesegregationofforeigntargetgeneintheT1byhistochmicalGUSass

6、ays；(2)Identificationofpurelinefromtransgenictomatoes口1)throughexaminingGUSexpressioninpollensinconjunctionwithPCRanalysisofmarkergene∞fⅡ)；(3)ThetranscriptlevelsofLeETRlorLeETR2inanti—sensetransgenicplants；(4)Thephenotyp

7、esofthetransgenicplantsintomatoduringwholelifecycleunderethylenetreatedandnon—treatedconditionsResultsshowedthat：(1)Ingeneral，thesegregationratiosoftargetgeneinsomeT1linesarcconformedto3：1howeversomearenotpossiblyduetoge

8、nesilenceormissingintheselfpollinatedprogeny；(2)Twohomozygousplantswereidentifiedfrom10putativetransgenicplantsTheirfrequenciesofGUSexpressioninpollenswere97755％0800％and96556％0600％respectivelyTovedfytheseresults，theselfp

9、ollinatedprogeniesofthesetwoputativehomozygoteswereexaminedinT2byhistochenmicalGUSstainingofvegetativetissuesandPCRanalysisusingspecificprimersTheseresultsdemonstratedthatmerelyexaminingthesegregationofreportergeneinpoll

10、ensisareliablemethodthatcouldavoidthelaborioustestingprocessofthepurelinefromtransformedplants；(3)ThetranscriptlevelsofLeETRlandLeETR2werecheckedbyNorthernblottinganalysisExpressionoftargetgeneswassuppressedinLeETRlorLeE