玉米莖尖分生組織叢生芽高頻誘導和再生體系建立的中期報告

玉米莖尖分生組織叢生芽高頻誘導和再生體系建立的中期報告摘要：玉米莖尖作為植物多能干細胞的重要來源之一，已被廣泛應(yīng)用于植物遺傳轉(zhuǎn)化和基因編輯等領(lǐng)域。本研究通過優(yōu)化不同影響因素，建立了玉米莖尖分生組織的叢生芽高頻誘導和再生體系。結(jié)果顯示，誘導培養(yǎng)基組合優(yōu)化、激素濃度合理調(diào)配以及過渡到再生培養(yǎng)基的適當時間，可以明顯提高叢生芽誘導的有效率和質(zhì)量，進而提高再生體的發(fā)生率和生長速度。通過對誘導和再生體系中關(guān)鍵激素的研究，發(fā)現(xiàn)ABA可以顯著促進叢生芽的分化和發(fā)生，而GA、KT和NAA等激素則對芽的生長和分枝起到了重要作用。此外，本研究還探討了不同基因型對誘導和再生的響應(yīng)差異，為后續(xù)利用玉米莖尖進行遺傳轉(zhuǎn)化和基因編輯提供了重要的實驗基礎(chǔ)。關(guān)鍵詞：玉米莖尖；分生組織；叢生芽；高頻誘導；再生體系IntroductionCornisoneofthemostimportantcropsintheworld,providingfood,feed,fuel,andindustrialproductsforhumanbeings.Asoneofthemostwidelycultivatedcerealcrops,ithasbeensubjectedtoextensivegeneticimprovementbytraditionalbreedingandbiotechnologyapproaches.Inrecentyears,theapplicationofgeneticengineeringhasgreatlyexpandedthepossibilitiesforcropimprovementbyintroducingnewtraitsthatarenoteasilyaccessiblethroughtraditionalbreedingmethods,suchasdiseaseresistance,herbicidetolerance,andstresstolerance.MaterialsandmethodsPlantmaterialsCorncultivarsHX151andJY31wereusedastestmaterialsinthisstudy.Theseedsweresoakedinsterilizedwaterfor12handthensurfacesterilizedwith70%ethanolfor1min,followedby6%NaClOfor15min.ThesterilizedseedswererinsedwithsterilizedwaterforthreetimesandculturedonsolidifiedMSmediumat25±2°Cwithaphotoperiodof16hlight/8hdark.InductionofcallusandshootorganogenesisThestemapicalmeristemsof7-day-oldseedlingswereusedforcallusinductionandshootorganogenesis.Thestemapiceswereexcisedandculturedoninductionmedium(IM)containingMSsalts,3%(w/v)sucrose,2mg/L2,4-D,and0.2mg/Lkinetin(KT)forcallusinduction.After14daysofculture,thecalliweretransferredtoshootorganogenesismedium(SOM)containingMSsalts,3%(w/v)sucrose,0.5mg/Lzeatin(ZT),and0.2mg/Lindole-3-aceticacid(IAA)forshootinduction.ResultsanddiscussionOptimizationofcallusinductionandshootorganogenesisTheinductionfrequencyandthequalityofregeneratedshootsarekeyfactorsfortheestablishmentofanefficientregenerationsystem.Inthisstudy,weoptimizedseveralfactorsaffectingthecallusinductionandshootorganogenesis,includingthecompositionoftheinductionandregenerationmedia,theconcentrationandcombinationofplantgrowthregulators,andthedurationofthecultureperiod.EffectsofdifferentplantgrowthregulatorsoncallusinductionandshootorganogenesisToinvestigatetheeffectsofdifferentplantgrowthregulatorsoncallusinductionandshootorganogenesis,weused2,4-D,KT,ZT,IAA,gibberellin(GA),andabscisicacid(ABA)atvariousconcentrationsandcombinations.Ourresultsshowedthat2,4-DandKTwerethemosteffectiveininducingcallusformation,buttheywerealsoinhibitorytoshootformation(Figure1).EffectsofdifferentconcentrationsofABAoncallusinductionandshootorganogenesisToinvestigatetheeffectsofABAoncallusinductionandshootorganogenesis,weculturedstemapicesonIMsupplementedwithdifferentconcentrationsofABA.OurresultsshowedthatABAsignificantlypromotedcallusinductionandshootorganogenesisinbothHX151andJY31,withthehighestinductionfrequencyat5mg/LABA(Figure2).ConclusionInthisstudy,weestablishedanefficientsystemforinducingandregeneratingshootsfrommaizestemapicalmeristems.Ourresultsshowedthattheoptimizationofmediumcomposition,plantgrowthregulatorcombinations,andculturedurationsignificantlyimprovedthefrequencyandqualityofshootorganogenesis.ABAwasfoundtobeakeyregulatorinpromotingcallusinductionandshootorganogenesis,whileGA,KT,andNAA