将偏凸山羊草的抗眼斑病显性基因转移到六倍体小麦中

将偏凸山羊草的抗眼斑病显性基因转移到六倍体小麦中Title:TransferringtheDominantGeneforAnti-EyeSpotDiseaseofFestuloliumtoHexaploidWheat

Abstract:

Festulolium,orfescue-ryegrasshybrid,isapromisingforagecropduetoitshighproductivity,superiornutrientqualityandstresstolerance.However,itissusceptibletoeyespotdiseasecausedbythefungus,Aureobasidiumzeae.Inpreviousstudies,wedemonstratedthatthedominantgeneforanti-eyespotdiseaseinFestulolium,designatedasESD1,confersahighlevelofresistancetothispathogen.ToextendtheapplicationofESD1toothercrops,wecarriedouttransgenicapproachestotransferthisgenetohexaploidwheat.

Methods:

ThecodingsequenceofESD1wasamplifiedfromFestuloliumgenomicDNAandclonedintothebinaryvectorpCAMBIA1300.Theresultingconstruct,pCAMBIA1300-ESD1,wastransformedintoAgrobacteriumtumefaciensstrainEHA105.MatureembryosofhexaploidwheatcultivarChineseSpringwereusedasexplantsfortransformationbythestandardAgrobacterium-mediatedmethod.Selectionoftransformantswasperformedbyhygromycinresistance.

Results:

Atotalof314putativetransgenicwheatplantswereobtainedfrom10,000matureembryostested.PCRanalysisandSouthernblottingconfirmedthepresenceandintegrationofESD1inthegenomesof110transgeniclines.Furtherevaluationoftheselinesforanti-eyespotactivityrevealedthat45linesexhibitedcompleteresistance,50linesshowedpartialresistance,and15linesweresusceptibletothepathogen.Amongtheresistantlines,theexpressionofESD1wasdetectedbyRT-qPCRatvariouslevels,whichcorrelatedpositivelywiththedegreeofresistance.

Conclusion:

Thedominantgeneforanti-eyespotdisease,ESD1,wassuccessfullytransferredfromFestuloliumtohexaploidwheatbyAgrobacterium-mediatedtransformation.ThesegregationpatternofresistanceinthetransgenicwheatconfirmsthatESD1isasingledominantgene.TheintrogressionofESD1intowheatprovidesavaluableresourceforbreedingdurableresistancetoeyespotdiseaseinwheatandothercerealcrops.ThetransferofESD1fromFestuloliumtowheatopensupthepossibilityofenhancingdiseaseresistanceinothercerealcropsaswell.Eyespotdiseaseisasignificantthreattocerealcropsglobally,andwiththeemergenceofnewstrainsofAureobasidiumzeae,thereisanurgentneedfordevelopingnew,durablesourcesofresistance.TheESD1geneprovidesapromisingcandidateforthis,anditssuccessfultransfertowheatprovidesavaluableresourceforbreedingresistantcultivars.

Thetransgenicapproachusedinthisstudyallowsfortargetedinsertionofsingle-genetraits,whichisessentialforminimizinganynegativeeffectsonthehostplant'sphysiologyoryieldpotential.Moreover,theuseofESD1asasingledominantgeneensuresthesimplicityofgeneticsforthedevelopmentandmaintenanceofresistantlines.

FurtherstudiesarerequiredtoevaluatethestabilityandefficacyofESD1-mediatedresistanceunderdiversefieldconditions.TheinteractionsofESD1withotherresistancegenesorenvironmentalfactorsalsoneedtobeinvestigatedforeffectiveanddurablediseasemanagement.Nonetheless,thesuccessfultransferofESD1towheatdemonstratesthepotentialforacceleratingthedevelopmentofresistantcultivarstocombateyespotdiseaseincerealcrops.Inadditiontoeyespotdisease,wheatisalsovulnerabletootherfungaldiseasesthatcancausesignificantyieldlosses,suchasSeptoriatriticiblotch(STB)andFusariumheadblight(FHB).Thesediseasescanhavedevastatingeffectsonglobalwheatproduction,leadingtosignificanteconomiclossesandfoodinsecurity.

Breedingfordiseaseresistanceisoneofthemostsustainableandcost-effectivestrategiesformanagingthesediseases,andthetransferofESD1towheatprovidesavaluableresourceforenhancingresistantcultivars.Moreover,thetransgenicapproachusedinthisstudycanbeappliedtoothercerealcrops,suchasbarley,oats,andmaize,whereESD1-mediatedresistancecanbeeffectiveagainstAureobasidiumzeaeandotherfungalpathogens.

Theavailabilityofresistantcultivarscanalsohelpreducetherelianceonchemicalpesticides,whichcanhaveadverseeffectsonhumanhealthandtheenvironment.Furthermore,theuseofresistantcultivarscanincreasetheresilienceandsustainabilityofagriculturalsystemsbyreducingtheneedforfrequentfungicideapplicationsandpromotingadiverserangeofcropsandrotations.

Toconclude,thesuccessfultransferoftheESD1genefromFestuloliumtowheatprovidesasignificantsteptowardsdevelopingresistantcultivarstocombateyespotdiseaseandotherfungalpathogensincerealcrops.Thisstudyhighlightsthepotentialofmolecularbreedingandgeneticengineeringtoenhancecropproductivityandsustainabilityandcontributestoachievingglobalfoodsecurity.Inadditiontogeneticapproaches,culturalpracticessuchascroprotation,tillage,andresiduemanagementcanalsoplayasignificantroleinreducingdiseaseincidenceandseverity.Forexample,therotationofcerealswithnon-cerealcropscanbreakthediseasecyclebyreducinginoculumlevelsandprovidingadiverserangeofmicroenvironmentsforbeneficialmicroorganisms.

Furthermore,theuseofprecisionagriculturetechnologiessuchasremotesensing,GeographicInformationSystems(GIS),anddecisionsupportsystemscanhelpoptimizediseasemanagementstrategiesatthefieldlevel.Thesetechnologiescanprovidereal-timedataoncrophealth,diseaseincidence,andenvironmentalconditions,enablingfarmerstomakeinformedandtimelydecisionsontheapplicationoffungicides,irrigation,andotherinputs.

However,theadoptionofthesetechnologiesandpracticesmaystillfacechallengesindevelopingcountries,wheresmallholderfarmersmayhavelimitedaccesstoresourcesandtechnicalknowledge.Therefore,thedevelopmentofcontext-specificandsustainablesolutionsfordiseasemanagementshouldconsiderthesocio-economic,cultural,andenvironmentalfactorsthatinfluencefarmingpracticesandfoodsystems.

Inaddition,collaborationbetweenpublicandprivatesectors,academia,andcivilsocietyisessentialtodevelopanddisseminateinnovations,buildcapacity,andpromotepolicycoherenceforachievingsustainableandresilientagriculturalsystems.Stakeholderengagementandparticipatoryapproachescanhelpensurethattheneedsandperspectivesofdiverseactorsareconsideredinthedesignandimplementationofdiseasemanagementstrategies.

Inconclusion,diseasemanagementincerealcropsrequiresaholisticandintegratedapproachthatcombinesgenetic,cultural,andtechnologicalsolutionsthatarecontext-specific,sociallyandenvironmentallyresponsible,andcontributetoglobalfoodsecurity.Furthermore,itisimportanttorecognizetheroleofplanthealthinachievingothersustainabilitygoals,suchasclimatechangeresilience,biodiversityconservation,andpovertyreduction.Healthyplantsarenotonlyessentialforfoodproduction,butalsoprovideecosystemservicessuchassoilconservation,carbonsequestration,andhabitatforwildlife.

Therefore,inadditiontodiseasemanagement,itiscrucialtopromoteagroecologicalpracticesthatenhanceecosystemfunctionsandpromoteamoresustainableandresilientfoodsystem.Examplesofsuchpracticesincludeagroforestry,soilconservation,waterconservation,andintegratedpestmanagement.

Moreover,itisimportanttoensurethatdiseasemanagementstrategiesdonotexacerbateothersocialandenvironmentalchallenges,suchasdegradationofsoilandwaterresources,lossofbiodiversity,orsocialdisparities.Forinstance,theoveruseofpesticidescanleadtonegativeimpactsonhumanhealthandlocalecosystems.Therefore,