1、中國農業(yè)大學碩士學位論文棉纖維次生壁增厚啟始相關基因的cDNA克隆及分析姓名：秦治翔申請學位級別：碩士專業(yè)：作物栽培與耕作學指導教師：楊佑明翟志席2002.5.1AbstractUptonowgenesrelatedtoeoRonfiberdevelopmentareclonedfromelongatingfiberDozensofgeneshavebeenisolated，butnoneisthemaingenedetermining

2、thedevelopmentandformationofqualitytraitofeoRonfiberBecausefiberstrengthisthechiefquailtytraitanddeterminedduringsecondarywallthickening，it’SimportanttostudyonthesecondarywallthickeningofcottonfibeLCotton(Gossypiumhfrsut

3、umLevXuZhoul42)fiberoffieldgrownplantcollectedon14DPA，18DPA，24DPA，30DPA。whichwerenttmberedX1，X一2，X3，X4，respectivelyandthatofovulesculturedfrom2DEAto14D隊collectedaftersubcultureonthemediumcontained100umelL‘ABAor0umelL1ABA

4、forfivehours，whichwerenumberedP14andP14CK，respectivelywereusedforeachexperimentInordertoisolatecDNAsbombeginningtoexpressduringsecondarywallthickeningandexpressinginducedbyABA，DDRTPCRwasusedtoanalyzedifferenceofgeneexpre

5、ssionofthefiberdescribed∞aboveTheresultsaleshownasfollows：1ThehotphenolmethodwasadoptedtoextractRNAandhighqualityRNAWasobtained2DDRTPCRwasusedtodisplaygeneexpressionofcoRonfiberdescribedasaboveDifferenceofgeneexpressiono

6、fonsetofsecondarywallthickeningandthatofexpressionofgenesinducedbyABAwereanalyzedTwentydifferentialeDNAfragmentswererecoveredAmongthese，thirteenwereobtainedfromP143ThefirstcycleofReverseNorthernhybridizationWasprocessedw

7、iththeprobeRNAofP14andninepositiveeDNAfragmentswereobtained4ThesecondWasdonewiththeprobeRNAofPt4、P14CK、Xl、X2、X3andX一4respectivelyFivepositiveeDNAfragmentswereobtainedOneofthesenumbered14hadpositivesignalwithRNAofF14，X2，X

8、3，X4ItWaSshownthat14expressedatonsetofsecondarywallthickeningandWaSinducedbyABAOthersnumberedlit。112，113，114，respectivelyhadpositivesignalwithILNAofbothP14，Pt4CKandXl、X2、X3X4buttheintensityamongthemWaSdifferent111eresult

9、sshowedthattheexpressionof11WasraisedbyABAwhilethatofll2。113andlhWaSdecreaSedbyABA5ThefiveESTswerenewincottonanalyzedbyBlaStx1dentityof14withproteinkinaSeinArabidopsisthaBanaWaS5】％ItWaSsupposedtobeproteinkinaseIdentityof

10、JIf112，113，114withcytoehromeP450monooxygenaSeinNicotianatabacum，glyoxalaseI，phenylconmaranbenzylicetherreductasehomologTP5，TH6andlsofiavonereductaseWas92％、85／、44％and65％respectivelyThefiveESTsweresupposedtoberelatedtoonse