枇杷幼果在低溫脅迫下基因的差異表達.pdf

1、福建農林大學碩士學位論文枇杷幼果在低溫脅迫下基因的差異表達姓名：吳海波申請學位級別：碩士專業(yè)：果樹學指導教師：鄭國華20100401fl繾農林大學碩：E學位論文枇杷幼果在低溫脅迫下基因的差異表達AbstractInordertoanalyzethemolecularmechanismofcoldtoleranceinloquatwithearly—maturingandsusceptibleto—frostdamageZaoZhongN

2、o6Ioquatfruitastheresearchobject，DDRTPCRtechniquewasusedtOstudythegeneexpressionduringthecourseofcoldstressinloquatTheSDSmethodwasimprovedtofindoutakindoffastandefficientextractionmethodoftotalRNAinloquatfruit，andtheRNAe

3、xtractedusingthismethodwassuitableforDDRTPCRanalysisByoptimizingthetest，thebestDDRTPCRanalysissystemforloquatfruitduringthecourseofcoldstresswasestablished，In20Itlsystem，eachcomponentwasidentifiedas：TemplatecDNA1Obtl，Pri

4、mer05州，dNTPs50rt0，10x3C5EulliLf2p1，Taqenzyme1U，annealingtemperature40。COnthebasisofthissystem，themRNAchangesofloquatfruitwasanalyzedundertheconditionof0“Ctreatedfor6hNineteendifferentiallyexpressedgenesegmentsinducedbyco

5、ldstresswereobtainedandfourofthemwasconfirmedaspositiveclonesbyReverseNorthemdotblotting(ItwasnamedCSIGE1～4__C艘6tms’nQ岬GHlHin(0RE醛UD№即iml～4)ThefourpositivefragmentsweresequencedandhavebeensubmittedtoGenBankCSIGE!4followe

6、dbytheaccessionnumberofGT617703，GT617704，GT617705，GT617706Theresultofsequencesimilarityanalysisshowedthat：(!)CSIGE1sequencehashighlyhomologoustOHomosapienschromosome8cloneRpl1420F14mapp21pl1，whichWasacoldresistancegeneTh

7、isshowsthatplantandanimalgenomeshavecertaincommoncodingsequence，maybetheyhavesomecommongeneexpressionmechanism；：(g)CSIGE2hasdifferentdegreesofhomologywithsomegenes，suchasalcoholdehydrogenase(ADH)inricinuscommunis，NADPdep

8、endentoxidoreductasesinarabidopsisthalianasalicylicacidstress—relatedgenesinsoybean，CSIGE3hassomehomologoustoESTsstressedbyadversityinvitisviniferaandcucumbentheregulationoftheirexpressionplaysomerolesinresponsetocoldstr