简介：Precisebaseeditingishighlydesiredinplantfunctionalgenomicresearchandcropmolecularbreeding.Inthisstudy,weconstructedarice-codonoptimizedadeninebaseeditor(ABE)-nCas9toolthatinducedtargetedA·TtoG·Cpointmutationofakeysinglenucleotidepolymorphismsiteinanimportantagriculturalgene.Combinedwiththemodifiedsingle-guideRNAvariant,ourplantABEtoolcanefficientlyachieveadeninebaseeditinginthericegenome.

简介：ThetransgenicriceKMD1,expressingasyntheticCry1AbgenefromBacillusthuringiensis,showedeffectiveresistancetoSignificantdeclineswererevealedinfoodconsumptionandgrowthoftheolderRLFnymphsfedonthecut-leavesoftransgenicKMD1plants.TheincreaserateoffoodconsumptionbylarvaefedonKMD1wasdrasticallylowerthanthoseonXiushui11.FoodconsumptionwasvariedwithdifferentinstarswhenthelarvaefedontheBtrice.Thoseoffourth-andfifth-instarlarvaeweredifferentcomparedtothethird-instar,lowerthanthoseonthenon-transgenicricebutstillincreasedalittlewhenthefeedingtimeprolonged.ItisindicatedthatyoungerRLFlarvaearemoresensitivetoBtricethanolderones.Also,about81%,78%and68%ofthethird-,fourth-andfifth-instarRLFlarvaediedwithin72hoursbioassayperiodonKMD1leaves,respectively.TheseresultsdemonstratedthatBt-transgeneinKMD1riceconferssubstantialprotectionagainstinfestationswitholderRLFlarvae.

简介：Homeoboxtranscriptionfactorsparticipateinthegrowthanddevelopmentofplantsbyregulatingcelldifferentiation,morphogenesisandenvironmentalsignalresponse.Torevealthefunctionsofthesetranscriptionfactorsinrice,weconstructedtheRNAivectorsofOsHox9,amemberofhomeoboxfamily,andanalyzedthefunctionofOsHox9usingreversegenetics.TheplantheightandtilleringnumberofRNAitransgenicplantsdecreasedcomparedwiththoseofwild-typeplants.Reversetranscription-polymerasechainreactionanalysisshowedthatOsHox9expressionreducedinthetransgenicplantswithphenotypicvariance,whereasthatinthetransgenicplantswithoutphenotypicvariancewassimilartothatinthewild-typeplants.ThisresultsuggeststhatthephenotypesofthetransgenicplantswerecausedbyRNAieffects.Thetissue-specificityofOsHox9expressionindicatedthatitwasexpressedindifferentorgans,withhighexpressioninstemapicalmeristemandyoungpanicles.SubcellularlocationofOsHox9demonstratedthatitwaslocalizedonthecellmembrane.

简介：Smallubiquitin-likemodifier(SUMO)-conjugatingenzymesareinvolvedinpost-translationalregulatoryprocessesineukaryotes,includingtheconjugationofSUMOpeptidestoproteinsubstrate(SUMOylation).SUMOylationplaysanimportantroleinimprovingplanttolerancetoabioticstresssuchassalt,drought,heatandcold.Herein,wereportedtheisolationofOsSCE1(LOC_Os10g39120)geneencodingaSUMO-conjugatingenzymefromrice(Oryzasativacv.Nipponbare)anditsfunctionalvalidationinresponsetodroughtstress.TheE2enzyme,OsSCE1,isoneofthreekeyenzymesinvolvedintheconjugationofSUMOtoitstargetproteins.ActivatedSUMOistransferredtothecysteineofanE2enzymeandthentothetargetlysineresidueofthesubstrate,withorwithoutthehelpofanE3SUMOligase.ExpressionofOsSCE1wasstronglyinducedbypolyethyleneglycol6000(PEG6000)treatment,whichsuggestedOsSCE1maybeinvolvedinthedroughtstressresponse.OverexpressionofOsSCE1(OsSCE1-OX)inNipponbarereducedthetolerancetodroughtstress.Conversely,thedroughttolerancewasslightlyimprovedbytheknockdownofOsSCE1(OsSCE1-KD).TheseresultswerefurthersupportedbymeasurementofprolinecontentinOsSCE1-OXandOsSCE1-KDtransgeniclinesunderinduceddroughtstress,whichshowedOsSCE1-KDtransgeniclinesaccumulatedhigherprolinecontentthanthewildtype,whereasOsSCE1-OXlinehadlowerprolinecontentthanthewildtype.ThesefindingssuggestedOsSCE1mayplayaroleasanegativeregulatorinresponsetodroughtstressinrice.

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：Twonewlybredhybridricecombinations,superhigh-yieldingLiangyoupeijiu(Pei'ai64S×9311)andPei'ai64S/E32(Pei'ai64S×E32)wereusedtoinvestigatethephotosyntheticcharacteristicsunderhightemperatureincomparisonwithhybridriceShangyou63.Hightemperaturecausedadecreasedphotosyntheticefficiencyandaggravatedphotoinhibition.TheoptimumtemperatureforphotosyntheticelectrontransportationandphotosyntheticCO2fixationwereabout28℃and35-40℃respectively.Linearelectrontransportationismoresensitivetohightemperaturethanthephotochemicalprocess.Themechanismofhightemperatureadaptationwaspossiblyasfollows:superhigh-yieldingricehasquicklyincreasingcarotenoid,whichactedasamorefavorableantioxidantsystemtoreducetheactiveoxygenproductionandavoiddamagetothephotosynthesissystem;superhigh-yieldingricehasahigherefficiencyofxanthophyllscycletodissipateexcessheatenergy;superhigh-yieldingricehasamorestablephotosyntheticfunction,higherphotosyntheticefficiencyandmoreheatstableproteincontent.

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.