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ApplicationTimelineStart
SeedlingsTissueHarvest
DNASequencing
QualityControl
GBSPipelineandGSModels
MakeSelection Plant
GenomicSelectiontoIncreasetheRateofGeneticGainofIntermediateWheatgrass(Thinopyrum intermedium)
JaredCrain1,KevinDorn1,TraciKantarski2,JaneGrimwood3,JeremySchmutz3,LeeDeHaan4,JessePoland11KansasStateUniversity,Manhattan,KS66506.2ColumbiaUniversity,NewYorkCity,NY10027.3HudsonAlphaInstituteforBiotechnology,Huntsville,AL35801.4TheLandInstitute,Salina,KS67401.
Malone Family LandPreservation Foundation
Introduction
MaterialsandMethodsCycle6ofTheLandInstituteIWGbreedingprogramformedthe
trainingpopulation,genotyping-by-sequencing(GBS)wasused
formarkerdiscoveryandgenotyping.Thepopulationincluded:
• 3,658genetsinasinglereplicaterow-columndesign.
• Genetswerephenotyped for46traitsovertwoyears.
Seedyield,spikefertility,seedmass,…,stemstrength.
Foreachtrait,1,880-2,400observationswererecorded.
• 2,974plantsgenotypedusingGBS.
Usingtheavailabledata,bestlinearunbiasedpredictors(BLUPs)
werecalculatedforeachtrait.Heritability(h2)wascalculated
usingthegenomicmarkermatrix.Cross-validationproceeded
byleavingentireparentsoutofthetrainingset,topreventbias
ofcloserelativespredictingperformance,andseveralGS
modelsweretestedtoidentifythebestperformingmodels.
ResultsandDiscussion
Refereneces1. DeHaan,L.R.,S.Wang,S.Larson,T.Kantarski,X.Zhang,andD.
Cattani.2014.CurrenteffortstodevelopperennialwheatanddomesticateThinopyrum intermediumasaperennialgrain.In:C.Batello etal.,editors,Perennialcropsforfoodsecurity:Proc.oftheFAOexpertworkshop.p.72–89.
2. Zhang,X.,Sallam,A.,Kantarski,T.,DeHaan,L.R.,…Anderson,J.A.(2016).EstablishmentandOptimizationofGenomicSelectiontoAcceleratetheDomesticationandImprovementofIntermediateWheatgrass.ThePlantGenome,9(1),1–18.
IntermediateWheatgrass(IWG)isapromisingperennialspecies
thatisbeingdomesticatedasagrainandbiomasscrop.Initial
breedingworkbeganattheRodaleInstitute,Kutztown,PA,inthe
1980’s.In2002,TheLandInstitute,Salina,KS,beganbreedingIWG
usingselectionsmadebytheRodaleInstitute[1].Earlyand
contemporaryworkhasbeenencouraging,withseedyield
increasing77%overtwocyclesofselection,andseedmass
increasing23%[1].Whilethesegainshavebeenimportant,ifthe
currentrateofgainsholds,itisestimatedthatitwouldtake
between24and110yearsofbreedingtoreachtheyieldandseed
sizeofannualwheat[1].Therearenumerouschallengesto
breedingIWGincluding:
• Largegenomesize12.6Gb• Allohexaploid(2n=6x=42)• Outcrossingandheterozygous
Asaperennial,breedingIWGisatimeintensiveprocess,withatypicalbreedingcyclelengthoftwoyears(Figure1).
Recently,genomicselection(GS)hasbeenproposedasawayto
increasetherateofgeneticgaininIWG.WorkbyZhangetal.[2]
showedhighpredictiveabilityofGSforseveralagronomictraits,
includingseedweightandbiomass.Inadditiontohighpredictive
ability,GScouldbeusedtodrasticallyreducethelengthofthe
breedingcycle,allowingforintermating ofselectedprogenyeach
year(Figure2)andresultinginabreedingcyclethatistwiceasfast.
GiventhepotentialofGStoincreasetherateofgeneticgain,we
examinedhowGScouldbeappliedanddevelopedmethodsto
applyGStoTheLandInstituteIWGbreedingprogram.
X
Spring2017:Severalhundredcrossesaremadebetweenselectedparents.
X X
… …Summer2018:F1progenyareevaluatedforyieldandagronomictraits.
Spring2019:Thebestplantsfromsummer2018arecrossed,startinganewcycle.
Figure1. TwoyearphenotypicbreedingcycleforIntermediateWheatgrass.
GBSandGSFall2017
1,000plantsthatareplantedinthefieldtotrain
theGSmodel.
100plantswithhighestpredictedvalue,plantedinthegreenhouseforintermating.
Summer2017:UsingF1seedfromspring2017,4,000seedlingsarestarted.
TheseedlingsaregenotypedandpredictedusingGS.Thebestseedlingsformthecrossingblockandalargeportionoflinesgotofieldevaluation.
X XSpring2018:Crossingoccursinthegreenhouse,whilefieldevaluationsprovidedatatoimprovetheGSmodel.
F1seedthatstartsthenextcycleofselection.
Phenotypicevaluation,withphenotypicdatausedtotrainGSmodelforfollowingcycle.
Fall2018:Anewcycleofselectionbegins.
Figure2.ApplicationofgenomicselectiontoIntermediateWheatgrassbreeding.Thegenomicselectionmodelpredictsplantperformance,withthebestplantsformingtheparentsforthenextcycle.Bysimultaneouslyevaluatingmaterialinthefieldandcrossingplants,thetimetocompletethecyclecaneffectivelybecutinhalf.
Table1. Heritabilityandcross-validationpredictionaccuraciesforphenotypictraitsfromCycle6trainingpopulation.Cross-validationaccuracyisreportedasthecorrelationcoefficientbetweenthephenotypicbestlinearunbiasedpredictorandthepredictedvalue.
Trait h2 r2
FreeThreshing2016 0.61 0.45†
FreeThreshing 2017 0.50 0.91†
Spike yieldingrams2017 0.36 0.89Milligramsperseed2016 0.58 0.86Plantheight2016 0.63 0.80
Heritabilityacrossalltraitsrangedfrom0.28-0.67,andthecross-
validationpredictionaccuracywashighforalltraitsrangingfrom
0.45-0.93(Table1).AcrossmultipleGSmodels,theprediction
valuesamonggenetswerequitestable,withtheGaussianKernel
andrrBLUP havingthemostvariationinprediction.Evenwiththis
variation,foranyindividualtraittheminimumcorrelationbetween
thesetwomodelswasr2 >0.94(Figure3).
Aug.1Aug. 25– Sept.5
Sept. 6- 25Sept.26
Sept.27– Oct. 5Oct. 6-11
(byOct.25)
90DaysfromPlantingtoSelection!
Figure3.Comparisonofdifferentgenomicselection(GS)modelsforseedweight2016.ThediagonalliststhedifferentGSmodelsincludingridgeregression,LASSO,andGaussiankernelusingtherrBLUP andBGLRpackages. Thelowertriangleshowsthescatterplotresultsbetweenthemodels,andtheuppertriangledisplaysthecorrelationcoefficientbetweenthedifferentmodels.
rrBLUP_RR
5 6 7 8 9
1.00*** 1.00***−2 −1 0 1 2 3
−2−1
01
2
0.96***
56
78
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● rrBLUP_GAUS
†Furtherworkisinvestigatingthediscrepancybetween2016and2017modelpredictions.