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AWARD:$10,000USD|DEADLINE:4/20/15|ACTIVESOLVERS:242|POSTED:2/16/15
Source:InnoCentiveChallengeID:9933678Type:Ideation
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Agreement
Solver Agreements
Detailed Description & Requirements
ThesearetheSolverAgreements
youhavesignedforthisChallenge.
Background
Agrobacteriumtumefaciensmediatedtransformationofsugarbeetsislimitedbythekeyrequirementthatexplants
(typicallyleafcuttings)areabletoproduceregenerabletissueswhenculturedinvitro.Thevastmajorityofplant
tissuesandgenotypesdonotrespondtoinvitrocultureortransformationtechniques,aphenomenonknownas
recalcitrance.Theinabilityofplanttissuestoregenerateinvitrolimitsadvancesinplantscienceandbiotechnology.
Therefore,morerobustmethodstoregeneratewholetransgenicplantsfromtissueculturedmaterialare
desired.
SolverTermsofUse
ChallengeSpecificAgreement
ChallengeSpecificAgreement
Summary
CurrentMethods:
Transgenicsugarbeetplantsaretypicallyproducedusingamethodtotransformshootbasemeristematictissue,
originallydescribedinLindsey&Gallois(1990).Inthismethod,shootbasetissueslicesareisolatedandthen
inoculatedwithAgrobacteriumtumefaciens(hereafterAgrobacterium),whichisabletointegratethedesiredTDNA
intotheplantgenome.SincetheTDNAconfersresistancetoanherbicideorantibiotic,thetransformedcellsare
selectedbyincubatingtheshootbaseslicesinamediumcontainingoneofthesecompounds.Duringtheselection
step,theresistantplanttissuesdeveloptransgenicshoots,whilethenontransgenictissuesdie.However,themethod
hassomedrawbacksinregardtothequalityoftheproducedtransgenicplants(e.g.chimerism)aswellwiththe
overallefficiencyingettingtransgeniclines.
Interestingly,thestomatalguardcellsfromleafderivedprotoplastculturesofsugarbeetshowedanexceptionalability
toregenerateplants.Despitethegreatpotentialofthatdiscovery,thegenerationoftransgenicplantsbyusingthat
systemistediousandtheefficiencyisstilllow(about2%)(Halletal.,1996).Morerecently,alternativeapproaches
usingalsoregenerationsystemsfromsingletransformedcellsaredescribedintheliterature(Kishchenkoetal.,2005
Kagamietal.,2015).Briefly,thesemethodsincludethegenerationofunorganizedcellmasses(callus)whicharise
frompiecesofplantorgans.Inthiscase,Agrobacteriumtransformsindividualcellseitherintheplantorganbeforethe
establishmentofthecalluscultureorinthecallus.Nextthetransgeniccellsareselectedasdescribedabove,and
developmentallyreprogrammedtoregenerateaplantorgan,usuallyashoot.However,thesemethodssufferfromlow
efficiencyinplantregenerationandextremegenotypedependency.
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Challenge Data
(What'sThis?)
Solvers
Eventhoughsomegenotypesshowthepotentialtoregeneratewholeplants,thesuccessrateisoftenlowandvery
variablebetweenindependentexperiments.Thereasonsforthesevariationsarecompletelyunknown.
Weareconfidentthatweareabletoachievehighratesoftransferoftransgenes(throughtheuseofafluorescent
marker).However,duetothelackofregenerationcapabilitythesetransgeniccellsarenotabletodevelopintowhole
plants.
Wehavetestedanumberofmodificationsofdifferentparametersforthosemethodsdescribedintheliterature.Those
modificationsincludehormonetypesandconcentrations,enhancersofcallusproductionandplantregeneration,
diversetissuetypesandgenotypes,variousAgrobacteriumstrains,gellingagents,differentconditionsforgrowingthe
plantsinvitro,etc.However,therateofsuccess(regenerationofaplantinvitrofromeitheranexplantoracallus)is
stillverylowand,moreimportantly,alltheseparametersandmethodsareverygenotypedependent.
Submissions
Solver Map
References:
LindseyK.andGalloisP.,1990.TransformationofSugarbeet(Betavulgaris)byAgrobacteriumtumefaciens.JExp
Bot41,529536.
HallR.D.,RiksenBruinsmaT.,WeyensG.J.,RosquinI.J.,DenysP.N.,EvansI.J.,LathouwersJ.E.,LefbvreM.P.,
DunwellJ.M.,vanTunenA.,KrensF.A.,1996.Ahighefficiencytechniqueforthegenerationoftransgenicsugar
beetsfromstomatalguardcells.NatureBiotech14,11331138
KishchenkoE.M.,KomarnitskiiI.K.,andKuchukN.V.,2005.Productionoftransgenicsugarbeet(BetavulgarisL.)
plantsresistanttophosphinothricin.CellBiolInt29,1519.
KagamiH.,KurataM.,MatsuhiraH.,TaguchiK.,MikamiT.,TamagakeH.,andKuboT.,2015.SugarBeet(Beta
vulgarisL.).KanWang(ed.),AgrobateriumProtocols:Volume1,MethodsinMolecularBiology,vol.1223,335347.
TheChallenge
Takingalloftheseintoaccount,theSeekerislookingforimprovedregenerationmethodsfromexplantsculturedin
vitrodefiningregenerationastheprocesstoobtainaplantthroughorganogenesisorsomaticembryogenesis.The
methodsoranswersshouldimprovetheregenerationofstabletransformedcellsinrecalcitrantplantspecies(Sugar
beets)ortissues.Thesemethodsshouldberobustandreliable,meaningthattheresultscanbereproducedand
producetransgenicplantswithadesiredefficiencyexceeding10%(calculatedastheamountofindependent
transgeniceventsobtainedperprimaryexplant,whichmeansthatin10outof100isolatedexplants,the
Agrobacteriumhassuccessfullyinfectedaplantcellthatwilldevelopintoatransgenicplant).Inordertoallowthe
furtheruseofthesetransgenicevents,itisnecessarytotransferthetransgeneintothenextgeneration.Itisimportant
tonotethattheregeneratedplantsmustbefertile.
Weareparticularlyinterestedinsolutionswhichcouldinvolvetheearlydetectionofthetissueorgenotypeamenable
fortransformationandregeneration,thereprogrammingofanonregenerabletissuetoaregenerableone,the
enrichmentofatissueinregenerablecells,etc.
Additional Information
Theproposedmethodshouldmeetthefollowingrequirements:
1.
RegenerationoffertilesugarbeetplantsfromasinglecelloriginwhichisamenabletoAgrobacterium
tumefaciensmediatedtransformationmethods
https://www.innocentive.com/ar/workspace/challengeDetail?challenge=9933678
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ProjectRoomChallengeDetails
2.
Efficiencyexceeding10%fortheregenerationofplants(calculatedasthepercentageofexplantswhich
canbeculturedinvitrosuccessfullytoafullplantaftertransformation).
3.
Themethodshouldbeapplicabletomultiplegenotypesinthebestcaseitwouldbegenotype
independent.
Challenge Attachments
Therearenoattachmentsforthis
Challenge.
Project Criteria
Thesubmittedproposalshouldincludethefollowing:
1.
AdetaileddescriptionoftheproposedplantregenerationprotocolrelevanttotheSolution.
2.
Ifapplicable,adetailedplanorguidelineontestsorexperimentswhicharenecessarytovalidatethe
solution.
3.
Referencetoliterature(commercial,patent,orpeerreviewed)tosupportyourclaims.
4.
Examplesofwherethesolutionisbeingconsideredtobeutilizedorhasbeenutilizedbefore.
5.
Lengthofproposalnotlimitedaslongasitincludesexecutivesummarynotexceeding500words.
Aprotocolforplantregenerationisarecipeinvolvingdifferentexplants,mediacomponentsandcultureconditions.
Wewouldliketoavoidproposalsthatonlyincludethetestingofjustdifferentphytohormonesconcentrations(e.g.use
0.25mg/lofthephytohormone6BenzyladenineforcallusinductionasdescribedinKagamietal.,2015),media
componentcombinations,genotypesandtissuetypes.ThepreferenceforthisChallengeistolookfor(an)outofthe
boxanswerstotheproblemofsugarbeettransformation,meaning,ideastodiminishrecalcitrance.
Inthebestcase,theSolverhasfoundinanotherrecalcitrantplantspeciessomewaystoimprovetheshoot
regenerationfromtransformedcellsandhasgoodargumentstobelievethatthisdiscoverycouldbealsoapplicableto
sugarbeet.
SubmittedproposalsalongwithallrelevantsupportingdatashouldincludetheinformationdescribedintheDetailed
DescriptionoftheChallenge.
SubmittedproposalsshouldnotincludeanypersonalidentifyinginformationtheSolversdonotwanttomakepublic,
oranyinformationtheSolversmayconsiderastheirIntellectualPropertytheydonotwanttoshare.
ThisisanIdeationChallenge,whichhasthefollowinguniquefeatures:
Thereisaguaranteedaward.Theawardswillbepaidtothebestsubmission(s)assolelydeterminedbythe
Seeker.Thetotalpayoutwillbe$10,000,withatleastoneawardbeingnosmallerthan$5,000andnoaward
beingsmallerthan$2,500.
TheSolversarenotrequiredtotransferexclusiveintellectualpropertyrightstotheSeeker.Rather,by
submittingaproposal,theSolversgrantstotheSeekeraroyaltyfree,perpetual,andnonexclusivelicense
touseanyinformationincludedinthisproposal.
AftertheChallengedeadline,theSeekerwillcompletethereviewprocessandmakeadecisionwithregardstothe
WinningSolution(s).AllSolversthatsubmittedaproposalwillbenotifiedonthestatusoftheirsubmissionshowever,
nodetailedevaluationofindividualsubmissionswillbeprovided.
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