水稻系统获得抗性基因ossrt2的克隆及功能研究-cloning and functional study of the resistance gene os srt 2 obtained by rice system.docxVIP

AbstractPeoplehavefoundanumberofRgeneparticularlybygeneticengineering.Diseaseresistance(R)genesprotectplantsfromfungal,bacterial,viral,andnematodepathogensasacomplexgeneticdefensesystem.NumerousRgeneshavebeenclonedandcharacterizedfromdicotyledons.Diseaseresistance(R)genesinplantsencodeproductsthatspecificallyrecogniseincompatiblepathogensandtriggeracascadeofeventsleadingtodiseaseresistanceinthehostplant.BiosynthesisofSAinresponsetopathogensisbelievedtobecontrolledbyPAD4,EDS5andSID2.Pathogen-inducedexpressionofPAD4,EDS5andSID2wassuppressedbyAtSRT2,suggestingthatAtSRT2playsanimportantroleinregulatingSAsynthesis.SinceSir2wasfirstisolatedinyeast,itshomologshavebeenclonedinmammaliancellssuccessively.Nevertheless,thefeaturesofSir2proteinsarenottotallycomprehendedinplants.InordertodetecttheroleSir2playedintheresistanceprogressofrice,wedetectedwherethegeneislocated,therelationshipofexpressionlevelsofOsSRT2andthedisease-resistancerelatedlevelsofSAintransgenicplants，howstrongthetransgenicplantsresisttoXoopathogen.Thedetailsareasthefollowing:First,OsSRT2geneisobtained,electrophoreticshowsasignof938bp,identicalwiththeforecastedsizeofthegene,andnophenomenaoftowingandmulti-banddispersions.Aftersequencing,itwassubmittedtoNCBIBlastwebsite.Thereisasequencehomologyof99%withtheregisteredSRT2geneonNCBI,withwhichthereis100%proteinhomology.LogintheExpansywebsiteandenterproteinsequenceofOsSRT2,itwasdeducedthatOsSRT2geneencodeaprotein,ofwhichmolecularweightMw=34.56kD,isoelectricpointpI=8.76,whichiscoorderationwiththeproteinexpressioninprotokaryon.Thus,synonymousmutationswasoccurredinthenucleotideofSRT2.InordertofindthefuctionallocationofOsSRT2andlaythefoundationofspeculationoftheactionmode,genegunwasusedtotransforme35S-OsSRT2-GFPfusionexpressionvectorintoonionepidermalcells.Then,subcellularlocalizationof35S-OsSRT2-GFPwasdetectedbyfluorescencemicroscopy.TheresultshowedthatOsSRT2instantaneouslocatedinthenucleus.Viaagrobacterium-mediatedconversiontechnology,theywereintroducedintotherice.9T1ge