水稻矮化多分蘖突变体的表型研究与基因克隆-phenotypic study and gene cloning of dwarf multi - tillering mutant in rice.docxVIP

ABSTRACTRiceisoneofthemostimportantgraincropsintheworld,andthenitisanimportantgoaltobreedhighyieldingricevarieties.Theshootbranchingisankeyfactorondetermininggrainyieldofrice.Strigolactones(SLs)isagroupofnovelplanthormones,playinganimportantroleincontrollingtheshootbranchingofplant.Inthisstudy,mutants2-69asmaterialwasstudiedinfollowingaspects:analyzationofmutantphenotype,determinationofmutationgeneZEBRA2bythewayofmap-basedcloning,analyzationofgeneexpressionpattern,subcellularLocalizationanalysisoftheZEBRAproteinandhowZEBRA2effecttheSLsbiosynthesis.Theresultsareasfollows:Themutants2-69presentshightilleringanddwarfphenotypeandleafvariegationphynotype;Fine-mappingshowedthatthemutationsiteoccurredinfirstexonoftheZEBRA2gene.Achangeofanucleotide(fromGtoA)thatconverts77thaminoacidfromglycinetoarginine.Besides,wehavedetectedthreeallelicmutants,nameds1-62,s1-115,s1-167Arespectively.ZEBRA2encodesacarotenoidisomerase(CRTISO),akeyenzymecatalyzingconversionofcis-lycopenetoall-trans-lycopeneincarotenoidbiosynthesis；BothRT-PCRandZEBRA2-promoter-β-glucuronidase(GUS)transgenicriceindicatesthatZEBRA2mainlyexpressedinmesophyllcells.SubcellularLocalizationanalysisshowsthatZEBRAproteinwaslocalizationinchloroplasts;GR24treatmentofriceseedlingindicatesthats2-69isaSLssensitivemutant.Themeasurementofepi-5DSshowsthatthestrigolactonecontentwasdecreasedinthemutants2-69.Alltogether,theSLsbiosynthesiswasimpairedinmutants2-69;CarotenoidcomponentscontentweremeasuredbyHPLCwhichwascarriedoutusingtheleafofthewide-typeands2-69mutant,showingthatthecarotenoidcomponentscontent,measuredinthisstudy,werechanged.Theleveloftheβ-carotenoid(precursorofSLssynthesis)ofs2-69mutantwasdecreased,indicatingthatZEBEA2isinvolvedineffectingtheSLsbiosynthesis.s2-69mutantexhibitsthecharacteristicleafvariegationphenotypeanddecreasedleveloflutein,thexanthophyllthatisessentialforefficientchltripletquenching.WeconcludethatZEBRA2isinvolvedinphotoprotectioninrice.KeyWords:Rice;Strigolactones;CRTISO目录第一章前言………………………………………………………………