Experimental evidence for the ancestry of allotetraploid Trifolium repens and creation of synthetic forms with value for plant breeding.docVIP

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Experimental evidence for the ancestry of allotetraploid Trifolium repens and creation of synthetic forms with value for plant breeding.doc

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Experimental evidence for the ancestry of allotetraploid Trifolium repens and creation of synthetic forms with value for plant breeding

Williamsetal.BMCPlantBiology2012,12:55 /1471-2229/12/55 RESEARCH ARTICLE OpenAccess Experimentalevidencefortheancestryof allotetraploidTrifoliumrepensandcreationof syntheticformswithvalueforplantbreeding WarrenMWilliams1,2*,NicholasWEllison1,HelalAAnsari1,IsabelleMVerry1andSWajidHussain1 Abstract Background:Whiteclover(Trifoliumrepens)isaubiquitousweedofthetemperateworldthatthroughuseof improvedcultivarshasalsobecomethemostimportantlegumeofgrazedpasturesworld-wide.Ithaslongbeen suspectedtobeallotetraploid,butthediploidancestralspecieshaveremainedelusive.Putativediploidancestors wereindicatedbyDNAsequencephylogenytobeT.pallescensandT.occidentale.Here,weusefurtherDNA evidenceaswellasacombinationofmolecularcytogenetics(FISHandGISH)andexperimentalhybridizationtotest thehypothesisthatwhitecloveroriginatedasahybridbetweenT.pallescensandT.occidentale. Results:T.pallescensplantswereidentifiedwithchloroplasttrnLintronDNAsequencesidenticaltothoseofwhite clover.Similarly,T.occidentaleplantswithnuclearITSsequencesidenticaltowhitecloverwerealsoidentified. ReciprocalGISHexperiments,alternatelyusinglabeledgenomicDNAprobesfromeachoftheputativeancestral speciesonthesamewhiteclovercells,showedthathalfofthechromosomeshybridizedwitheachprobe.F1 hybridsweregeneratedbyembryorescueandtheseshowedstronginterspecificchromosomepairingand producedasignificantfrequencyofunreducedgametes,indicatingthelikelymodeofpolyploidization.TheF1 hybridsareinter-fertilewithwhitecloverandfunctionassyntheticwhiteclovers,avaluablenewresourceforthe re-incorporationofancestralgenomesintomodernwhitecloverforfutureplantbreeding. Conclusions:EvidencefromDNAsequenceanalyses,molecularcytogenetics,interspecifichybridizationand breedingexperimentssupportsthehypothesisthatadiploidalpinespecies(T.pallescens)hybridizedwithadiploid coastalspecies(T.occidentale)togeneratetetraploidT.repens.Thecomingtogetherofthesetwonarrowlyadapted species(onealpineandtheothermaritime),alongwithallotetraploidy,hasledtoatransgressivehybri