柑橘類植物非生物脅迫下多胺變化及兩個多胺生物合成基因的克隆與鑒定.pdf

1、華中農(nóng)業(yè)大學(xué)博士學(xué)位論文柑橘類植物非生物脅迫下多胺變化及兩個多胺生物合成基因的克隆與鑒定姓名：王靜申請學(xué)位級別：博士專業(yè)：果樹學(xué)指導(dǎo)教師：劉繼紅200905ABSTRACTCitrusindustryusuallyencountersaseriesofbioticandabioticstressesTherearesomesuperiorrootstocksgermplasmsincitrusrelativespeciesPolyami

2、nesareatypeofplantgrowthregulators，whichhaveimportanteffectsonstressresponseandmorphogenesisItisimportanttoimprovetherootstocks，obtainpotentialgenesthatcallbeusedingenetictransformationandproducestresstolerantgermplasmsI

3、nthisstudytheeffectsofexogenousputrescineoncoldtoleranceofPoneirustrifoliatawereexaminedandthepolyaminecontentsindifferenttolerant(coldandsalinity)cultivarswereinvestigatedusingHPLCTwopolyaminebiosytheticgenes，Sadenosylm

4、ethioninedecarboxylase(SAMDC)andargininedecarboxylase∽Do，wereisolatedandidentifiedbymolecularbiologyandbiochemicaltechniques，suchasSouthernblotRTPCRRealtimeRTPCRNorthemblotandenzymeassayBygenemanipulationandgenetictransf

5、ormationargininedecarboxylasegeneWaStransformedintothemutantofArabidopsisandcitrustoconfirmitsfunctionn坨resultswereasfollows：ADCpathwayWasinvolvedintheputrescinebiosynthesisinPoncirustrifoliataunderlowtemperaturestress，l

6、owdosesofexogenousputrescinealleviatedthestressderiveddamagebutthehighdosesofexogenousputrescineproducedtoxicityOurresultrevealedthatlessthanlmMputrescinecouldbeusedtoprotectcitrusfromfreezingT0“polyaminescontents，especi

7、allyputrescine，wereaccumulatedintolerantcultivaranddecreaSedinsusceptibleoneunderlowtemperatureTotalpolyamineswereincreaSedundercoldacclimationinparticularputrescineandspermineSpermidinewasenhancedinsalttolerantcultivara

8、ndtotalpolyamineswerereduced，whilespermidineandspermineexhibitedthemostprofounddecrementinthesusceptiblecultivarRTPCRincombinationwitllbioinformaticapproachproducedaSAMDCeDNA(CsSAMDC)fromNewhallnavelorange(Citrussinensis

9、Osbeck)whichcontainedanintactopenreadingframeof1095bpThededucedaminoacidssharedllighsequencehomology、析nlSAMDCsinotherplantsandpossessedseveralhighlyconservedaminoacidresiduesormotifsPhylogeneticanalysisindicatedthegenefu

10、nctionWasclosetothoseingrapeandapple，ProkaryoticexpressionofCsSAMDCinEcoligaverisetoproteinofexpectedsize，whichhadbiochemicalactivityviaenzymeanalysis1hegenemaybeallallelicgene、)l，imlittlevariationinthelocusincitrusbySou