14858.棘孢木霉刺激植物響應(yīng)蛋白基因epl1功能研究

1、UniversitvCode：l0225RegiSterCode：S13772DissertationfortheDegreeofMasterFunctionalStudyofElicitingPlantResponseProteinGene印Z仔om乃Zc辦D沈廠聊臼a印e陀擾Z砌Candidate：Supervisor：AssociateSupervisor：AcademicDegreeAppliedfor：Speciali夠：Da

2、teofOralExamination：Universi坶：GulijimilaMijitiZhiyingWangZhihuaLiuMasterForestProtectionJune2013NortheastForest巧UniVersityAbstractAbstractFungiarethemostcommonplantpathogensand凡ngaldiseaseisaccountingfor80％ofplantdisease

3、BiologicalcontrolofplantfungalphytopathogenissafetohumanandotherliVingbeings，enViromentallysound，andsociallyacceptablecontr01BasedontheoverwhelminglypositiVefeaturesofbiologicalcontrol，itistheprimecandidateinthesearchfor

4、reducingdependencyonchemicalpesticidesInordertoinvestigatethefunctionofelicitingplantresponseprotein1gene(互p，，)fombiocontrolfungus乃七辦D沈，聊臼臼印P比，，“mACCC30536，theseriesofprimersweredesignedbasedonthetranscriptomicssequenceo

5、f印，』andthetotalRNAandgenomeDNAextractedfrom丁口spP廠P，f“聊ACCC30536asthetemplateUsingPCRmethod，t11efulllengthcDNAandDNAsequenceof印，，genefinallywereobtained，withaccessionnumbersofHM572232andJF970203，respectivelyThecDNAsequenc

6、eof—Ep，Jwas690bpinlength，encoding139aminoacidsBlastPsearchindicatedthatEpllaminoacidsequencesharedthehighestsimilari母of92％withEpllofr口驢Dv護(hù)i沈(CAL80754)Funhemlore，signalPpredictionshowedthattheEpl1aminoacidsequencewascleaV

7、edbysignalpeptidasebetweenpos“ions18andl9(VSA／／DT)PfamproteinfamilypredictionshowedthatEpllpmteinbelongedtocerato—plataninfamilyToobtainhomologousgenesequencesof上≯，J『fromthegenomedatabasesoffourspecies療記辦D(紀(jì)r聊甜，meaminoac

8、idssequencesofEpl1from丁口印P旭朋“朋ACCC30536wasusedtoasqueries，resultingthat3elicitingplantresponseproteinsequenceswithhighsimilaritywereobtainedfromthe丁v護(hù)P珊Gv298，r口護(hù)Dvf，f沈ATCC74058andZ辦日陀砌行“聊CBS22695，respectively，and5similar

9、itysequences仔omr礎(chǔ)汐P，P刀“朋CBS43397Epllaminoacidsequencesharedthehighestsimilarit)，of91％withEPLlTas，locatedonscaffold7of丁口點(diǎn)pPrP，，“聊CBS43397RealtimeRTPCRwasconductedtoanalyzetheexpressionof印，，in丹fc向D比，m口儺pP肥，，“聊aRertreatedby

10、eightdifferentcultureconditionsTheresultsindicatedthatt11e五≯，Jgeneisdif詫rentiallyregulatedbytreatmentsofdifferentcultureconditionTheexpressionoftheJ巨p，，genewasdownregulatedduringtheMM(O5％glucose)，NstarVation，1％PDp甜，淞坊vf西

11、口門口P6D，彪口門口motpowdercultureandshowedlowestexpressionat4，72，4hand1297，1082，3191timeslowerthanpretreatmentGene點(diǎn)p，，ismainlyupregulated吼derCS切Ⅳation，1％Stempowder，1％leaVespowder，1％4拋r玎a，婦口，陀朋口細(xì)ceUwall，5％A口，陀，門口幻fementationliq

12、uorcultureconditionandshowedpeakexpressionlevelat72，12，8，72，72h，respectivelyThepeakexpressionwashigher715，37356，4500，7557，14992timesthanbeforetreatment，respectiVelyTheclonedEpfJgenewasinsertedintoProkaryoticexpressionVec