developing next generation grapevine rootstocks with long ... · -score for hr (resistance) x...
TRANSCRIPT
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,[email protected]&FOOD
HarleyM.SmithE [email protected] 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.