简介:Precisebaseeditingishighlydesiredinplantfunctionalgenomicresearchandcropmolecularbreeding.Inthisstudy,weconstructedarice-codonoptimizedadeninebaseeditor(ABE)-nCas9toolthatinducedtargetedA·TtoG·Cpointmutationofakeysinglenucleotidepolymorphismsiteinanimportantagriculturalgene.Combinedwiththemodifiedsingle-guideRNAvariant,ourplantABEtoolcanefficientlyachieveadeninebaseeditinginthericegenome.
简介:Homeoboxtranscriptionfactorsparticipateinthegrowthanddevelopmentofplantsbyregulatingcelldifferentiation,morphogenesisandenvironmentalsignalresponse.Torevealthefunctionsofthesetranscriptionfactorsinrice,weconstructedtheRNAivectorsofOsHox9,amemberofhomeoboxfamily,andanalyzedthefunctionofOsHox9usingreversegenetics.TheplantheightandtilleringnumberofRNAitransgenicplantsdecreasedcomparedwiththoseofwild-typeplants.Reversetranscription-polymerasechainreactionanalysisshowedthatOsHox9expressionreducedinthetransgenicplantswithphenotypicvariance,whereasthatinthetransgenicplantswithoutphenotypicvariancewassimilartothatinthewild-typeplants.ThisresultsuggeststhatthephenotypesofthetransgenicplantswerecausedbyRNAieffects.Thetissue-specificityofOsHox9expressionindicatedthatitwasexpressedindifferentorgans,withhighexpressioninstemapicalmeristemandyoungpanicles.SubcellularlocationofOsHox9demonstratedthatitwaslocalizedonthecellmembrane.
简介:EliminationoftheCRISPR/Cas9constructsineditedplantsisaprerequisiteforassessinggeneticstability,conductingphenotypiccharacterization,andapplyingforcommercializationoftheplants.However,removaloftheCRISPR/Cas9transgenesbygeneticsegregationandbybackcrossislaboriousandtimeconsuming.WepreviouslyreportedthedevelopmentofthetransgenekillerCRISPR(TKC)technologythatusesapairofsuicidegenestotriggerself-eliminationofthetransgeneswithoutcompromisinggeneeditingefficiency.TheTKCtechnologyenablesisolationoftransgene-freeCRISPR-editedplantswithinasinglegeneration,greatlyacceleratingcropimprovements.Here,wepresentedtwonewTKCvectorsthatshowgreatefficiencyinbotheditingthetargetgeneandinundergoingself-eliminationofthetransgenes.ThenewvectorsreplacedtheCaMV35SpromoterusedinourpreviousTKCvectorwithtworicepromoterstodriveoneofthesuicidegenes,providingadvantagesoverourpreviousTKCvectorundercertainconditions.Thevectorsreportedhereofferedmoreoptionsandflexibilitytoconductgeneeditingexperimentsinrice.
简介:Thepromoterregionofadroughtandabscisicacid(ABA)induciblegene,osr40c1,wasisolatedfromasalt-tolerantindicaricevarietyPokkali,whichis670bpupstreamoftheputativetranslationstartcodon.Insilicopromoteranalysisofresultedsequenceshowedthatatleast15typesofputativemotifsweredistributedwithinthesequence,includingtwotypesofcommonpromoterelements,TATAandCAATboxes.Additionally,severalputativecis-acingregulatoryelementswhichmaybeinvolvedinregulationofosr40c1expressionunderdifferentconditionswerefoundinthe5′-upstreamregionofosr40c1.TheseareABA-responsiveelement,light-responsiveelements(ATCT-motif,BoxI,G-box,GT1-motif,Gap-boxandSp1),myeloblastosisoncogeneresponseelement(CCAAT-box),auxinresponsiveelement(TGA-element),gibberellin-responsiveelement(GARE-motif)andfungal-elicitorresponsiveelements(BoxEandBox-W1).Aputativeregulatoryelement,requiredforendosperm-specificpatternofgeneexpressiondesignatedasSkn-1motif,wasalsodetectedinthePokkaliosr40c1promoterregion.Inconclusion,thebioinformaticanalysisofosr40c1promoterregionisolatedfromindicaricevarietyPokkaliledtotheidentificationofseveralimportantstress-responsivecisactingregulatoryelements,andtherefore,theisolatedpromotersequencecouldbeemployedinricegenetictransformationtomediateexpressionofabioticstressinducedgenes.
简介:以便揭示在二CCDD染色体种,Oryzaalta和Oryzalatifolia之间的起源和进化关系,在situ杂交(鱼)的荧光被采用从O与C0t-1DNA分析二种的染色体。alta作为一根探针。Karyotype比较地也在O之间被分析。alta和O。latifolia基于他们杂交的类似的乐队模式发信号。在O之间有高相同和靠近的关系。alta和O。然而,在杂交之间的区别表明的latifolia也是清楚的。C0t-1DNA被证明是种类--并且染色体类型特定。C0t-1DNA鱼能是更有效的分析在不同种类之间的genomic关系,这被建议。根据在二allotetraploidy种之间的高度并且中等重复的DNA序列的比较分析,O。alta和O。latifolia,可能的起源和Oryza的allotetraploidy的进化机制被讨论。
简介:RiceisanimportantfoodcropinChina,andthedevelopmentofhybridriceisacrucialwaytoincreasegrainyield.Thecreationofdual-purposenuclear-sterilelinesfortwo-linehybridbreedinghasbecomevitalforcommercialricebreeding.WeconstructedthepC1300-2x35S::Cas9-sgRNAPTGMS2-1expressionvectorforeditingthemalefertilitygenePTGMS2-1intwowidelycompatiblericevarieties,93-11andHuazhan,byusingtheCRISPR/Cas9system.Weobtainedthemarker-freephotoperiod-/thermo-sensitivegenicmale-sterile(P/TGMS)linesinT1generation.Accordingtotheexperimentsinphytotronwithfourtemperatureandphotoperiodtreatments,wefoundthetemperatureisthemainfactorforrestoringthepollenfertilityofptgms2-1mutantsin93-11andHuazhan,andthephotoperiodalsohassomeeffectsonpollenfertilityintwodifferentricebackgrounds.Theapplicationofcultivatingnewmale-sterilelinesbygenomeeditingsystemwillsignificantlyacceleratethericebreedingprocess.
简介:在米饭restorer线C224的条纹疾病抵抗的继承为量的特点加多基因用主要基因的混合效果模型被分析。另外,抵抗与维护者线在C224的七个十字被调查。结果证明C224的条纹抵抗被二主要基因与添加剂优势效果(E-1模型)加多基因与additive-dominance-epistasis效果控制。这二基因有12.47%和24.75%分别地,出现的否定优势效果的添加剂效果。在二主要基因之间有重要epistasis和相互作用效果。当多基因的是2.74%时,二主要基因的可遗传性是92.12%,显示条纹抵抗有主导的主要基因效果。七个十字,五显示了高或中等的抵抗到条纹疾病。