developing next generation grapevine rootstocks with long ... · -score for hr (resistance) x...

FORFURTHERINFORMATION

Figure3:SNPmarkerslinkedtoMJR1andRDV2wereiden2fiedonchromosome18and14,respec2vely,inVcinereaC2-50.

REFERENCESHarley M. Smith, Brady P. Smith, Norma B. Morales, Sam Moskwa, Peter R. Clingeleffer, Mark R. Thomas (2018a) SNP markers tightly linked to root knot nematode resistance in grapevine (Vitis cinerea) identified by a genotyping-by-sequencing approach followed by Sequenom MassARRAY validation. PLoS ONE 13(2): e0193121. Harley M. Smith, Catherine W. Clarke, Brady P. Smith, Bernadette M. Carmody, Mark T. Thomas, Peter R. Clingeleffer, Kevin S. Powell (2018b) Genetic identification of SNP markers linked to a new grape phylloxera resistant locus in Vitis cinerea for marker-assisted selection. BMC Plant Biology 18, 360.

Developing next generation grapevine rootstocks with long-term resistance to phylloxera and root knot nematode

BiosecurityThreatstoAustralianViAcultureA.PhylloxeraB.Rootknotnematode

Rootstocksareakeymanagementtoolforincreasingvineyardperformancebysafeguardingwinegrapevarie2esfromsoilbornepests.However, the current set of rootstocks that provide resistance to phylloxera and root knot nematode were derived from a limitednumber of breeding lines. As a result, these rootstocks likely inherited similar resistance mechanism(s) to phylloxera and root knotnematode.Thisisamajorconcern,asabreakdowninphylloxeraand/orrootknotnematoderesistancewouldseverelylimitrootstockop2ons forreplan2ng.TheCSIRORootstockBreedingTeamisu2lizingnextgenera2ontechnologiescombinedwithrapidphenotypingmethodstodevelopeliterootstockswithnewpedigreesforlong-termresistancetophylloxeraandrootknotnematode.

HarleyM.Smith,JakeD.Dunlevy,NormaB.Morales,AmyQ.SmithandPeterR.ClingelefferCSIROAgriculture&Food,WaiteCampus,UrrbraeSA5064harley.smith@csiro.auCSIROAGRICULTURE&FOOD

HarleyM.SmithE harley.smith@csiro.auw h_ps://people.csiro.au/S/H/Harley-Smith

TheRootstockBreedingTeamisfundedbyCSIROandWineAustralia

NextGeneraAonMappingPestResistance

IdenAficaAonofMarkersLinkedtoMJR1andRDV2

A.F1MappingpopulaAon

B.ScreeningF1Individuals

C.GBS-SNPPipelineVi/scinereaC2-50xV.viniferaRiesling

F1ProgenyGenotypes

M.Javanica‘pt1103P’S

GenomicDNA

359016828SequenceReads

1,361,275SNPs

18124SNPs

3974SNPsV.cinereaC2-502973SNPsV.viniferaRiesling

Smithetal.,2018a

MJR1 & RDV2V. cinerea

MJR2(V. champinii)

RDV1V. cinerea x V. ripariaX

MJR1 & RDV1/RDV2V. cinerea x (V. cinerea x V. riparia)

MJR1/MJR2 & RDV1/RDV2V. champinii x (V. cinerea x [V. cinerea x V. riparia])

Nursery & Field trial EvaluationRooting, graft compatibility, vigour, yield, berry & wine quality

LibraryPreparaAonIlluminaHiSeq2000

TASSELSNPCallingPipeline

VCFtoolsDP>10MAF>0.2MD=1GQ>98

Smithetal.,2018b

Figure 2: Flow chart for next genera2onmapping. (A) AV cinerea x Riesling F1mapping popula2on of 90individuals was established and (B) screened withM. Javanica ‘pt 1103P’. Note: this popula2on was alsoscreenedwithG1andG4phylloxera(datanotshown).(C)Agenotyping-by-sequencingpipelinewasusedtoiden2fysegrega2ngSNPs.

D.GeneAcMapConstrucAon

Figure1:Biosecurityofrootpests.(1)PhylloxeraisthemajorbiosecuritythreattoAustralianVi2culture.(B) The emergence of aggressive root knot nematodes that effec2vely feed on 1103 Paulsen and othersimilarrootstockmaterialisanotherbiosecuritythreat.

FirstGeneraAonDurableResistantRootstocks

RapidPhenotypingF1MappingPopulaAonsGlasshouseScreening(1-2years)

InvitroScreening(4-6months)

Establishinglasshouse(2months)PropagaAonofmaterial(2months)Screeningmaterial(2-24months)-OctobertoApril-Limitedspace(100genotypesx3reps)-Scoreforgallsonly

Establishinvitro(1-2months)Propagatedroots(2wks)Screeningmaterial(2-4wks)-Allyearround-Unlimitedspace-Scoreforgalls(suscepAble)-ScoreforHR(resistance)

X

Figure5:Marker-assistedselec2onoffirstgenera2onrootstocks.ThisbreedingschemewillallowustocombineRDV1,RDV2,MJR1andMJR2fordurableresistancetophylloxeraandrootknotnematode.

Figure4:Rapidphenotypingforrootknotnematoderesistance.U2liza2onofglasshousescreeningcantakeupto2yearstocompleteforeachgenotypeduetolimita2onsofspaceandrestric2onofscreeningduringwarmermonthsoftheyear.Furthermore,onlythesuscep2blephenotypecanbescored.Invitroscreeningcanbeperformedallyearroundandthesuscep2ble and resistant phenotypes can be scoredwith isolated rootswithin 2-4weeks. This procedure is extremelyefficientand isdependentuponmaintaininghighnumbersofnematodes in vitro. For in vitro screeningfigure.ControlrootsinAandDhavenocellnecrosis(HR,hypersensi2veresponse)orgall/eggmassdevelopment.Inrootknotnematodegenotypes,cellnecrosisinthe(B)root2pand(C)cortexcellsofroots.Suscep2blegenotypesaremarkedby(E)galland(F)eggmassdevelopment.