ncomms-tomato


Чтобы посмотреть этот PDF файл с форматированием и разметкой, скачайте файл и откройте на своем компьютере.
ARTICLE Received6Mar2015 Accepted14Sep2015 Published26Oct2015 Multi-levelengineeringfacilitatestheproduction ofphenylpropanoidcompoundsintomato YangZhang *,EugenioButelli *,SalehAlseekh ,TakayukiTohge ,GhanasyamRallapalli ,JieLuo PrashantG.Kawar ,LionelHill ,AngeloSantino ,AlisdairR.Fernie &CathieMartin DOI:10.1038/ncomms9635 NATURECOMMUNICATIONS |6:8635|DOI:10.1038/ncomms9635|www.nature.com/naturecommunications MacmillanPublishersLimited.Allrightsreserved. heuseoftranscriptionfactors(TFs)toactivateorsuppress Shikimate pathway Pentose phosphate pathway Figure1|Schematicrepresentationofthephenylpropanoidpathwayin ARTICLE NATURECOMMUNICATIONS|DOI:10.1038/ncomms9635 NATURECOMMUNICATIONS |6:8635|DOI:10.1038/ncomms9635|www.nature.com/naturecommunications MacmillanPublishersLimited.Allrightsreserved. Fig.1b).SinceSlMYB12isnotexpressedintomatoesh ,esh offeredarobustcontrolonwhichtoestablishenrichmentinChIP experiments(Fig.3b).ChIP-qPCRthatscannedbindingof SlMYB12acrossthepromotersof inpeel ofWTtomatoes,showedthatSlMYB12bindstothesameDNA motifsasAtMYB12(Fig.3c).Consistentwiththisresult,the 01234 5.005.0 NATURECOMMUNICATIONS|DOI:10.1038/ncomms9635 ARTICLE NATURECOMMUNICATIONS |6:8635|DOI:10.1038/ncomms9635|www.nature.com/naturecommunications MacmillanPublishersLimited.Allrightsreserved. expressionof PeelFlesh PeelFleshPeelFlesh PeelFlesh PeelFlesh SIENOSIENO SICHS1SIF3HSIFLSproSICHS1 abc Figure3|AtMYB12andSlMYB12binddirectlytothepromoterregionsof SlDAHPS )BindingofAtMYB12totheregionsof SlDAHPS wasanalysedbyChIP-qPCR.Pericarp(peelplusesh)samplesofWTand AtMYB12 fruitwereharvestedat3daysafterbreaker.The numbersonthehorizontalaxisbelowthebarscorrespondtotheleftandrightborderoftheampliedregionrelativetothetranscriptionstartsitean dbars indicatemeansands.e.m.s( 3);asterisksindicatesignicantdifferencescomparedwiththenegativecontrol( value 0.05,Students lMYB12 expressionispredominantlyinthepeelofWTtomatofruitandisassociatedwithelevatedexpressionofgenesinvolvedinbothprimaryand ARTICLE NATURECOMMUNICATIONS|DOI:10.1038/ncomms9635 NATURECOMMUNICATIONS |6:8635|DOI:10.1038/ncomms9635|www.nature.com/naturecommunications MacmillanPublishersLimited.Allrightsreserved. aminoacids(exceptphenylalanine)weresignicantlyhigherthan inWTfruit(SupplementaryFigs2and3).Greatercarbonux throughglycolysis,thepentosephosphatepathwayandtheTCA cyclein AtMYB12 tomatoes,wouldprovideadditionalATP, 123456 00.511.5 00.0040.008 00.020.04 00.0050.010.0150.02 00.00050.0010.0015 0123 00.0020.0040.006 Figure4|AtMYB12changestheuxofcarbonintomatofruit. NATURECOMMUNICATIONS|DOI:10.1038/ncomms9635 ARTICLE NATURECOMMUNICATIONS |6:8635|DOI:10.1038/ncomms9635|www.nature.com/naturecommunications MacmillanPublishersLimited.Allrightsreserved. Arabidopsis 2.00.08.0 abc Figure5|Co-expressionof AtMYB12 withothertranscriptionfactorsintomatofruitenhancesphenylpropanoidproduction. )PhenotypesofWT, AtMYB12 Del/Ros1 andIndigo( Del/Ros1 AtMYB12 )tomatofruit.Picturesweretakenatsevendaysafterbreaker.( )RT-qPCRdataindicatedthat ARTICLE NATURECOMMUNICATIONS|DOI:10.1038/ncomms9635 NATURECOMMUNICATIONS |6:8635|DOI:10.1038/ncomms9635|www.nature.com/naturecommunications MacmillanPublishersLimited.Allrightsreserved. inavanone3-hydroxylaseactivity(F3H)(Fig.6a) tomake AtMYB12are .Comparedwith tomatofruithadonly30%avonolsintheirfruit (SupplementaryFig.6).Wechosethe linewhichhadthe highestgenistincontentinfruit(SupplementaryFig.5)and crossedittoan AtMYB12are tomatolineandselected differentgenotypesintheF2generation(Fig.6aand SupplementaryFig.7).Infruitexpressing only,the StSyMYB12StSy StSyMYB12StSy IFSIFS IFSIFS Phenylalanine 4-Coumarate CoA Naringenin chalcone Naringenin Anthocyanins Flavonols Dihydroflavonol Resveratrol PAL Total isoflavones Total flavones Total flavonol Total resveratrol Res-Glc isoform DWmg g DW DW DW abc Figure6|Co-expressionof AtMYB12 withotherstructuralgenesintomatofruitenhancesnovelphenylpropanoidproduction. )Schematic representationofresveratrolandgenisteinbiosynthesis.Bothpathwaysarisefromavonoidbiosynthesis(inblack)bytheadditionof VvStSy respectively.Aftersynthesis,bothresveratrolandgenisteinareglycosylatedtoformpiceidandgenistin.Thenaturallyoccurring mutantisdecientin F3Hactivity.( )ThecontentsofCGA,totalavonolsandtotalresveratrolcompoundsindifferentgenotypes.Fruitwereharvested10daysafterbreaker. Errorbarsshows.e.m.( 3).( NATURECOMMUNICATIONS|DOI:10.1038/ncomms9635 ARTICLE NATURECOMMUNICATIONS |6:8635|DOI:10.1038/ncomms9635|www.nature.com/naturecommunications MacmillanPublishersLimited.Allrightsreserved. possibilitythatthelargeeffectsofAtMYB12areduesolelytothe increasedsinkstrengthofthephenylpropanoidpathwaygiven thatinhibitorstudiesorRNAiofPALindevelopingfruitwould ARTICLE NATURECOMMUNICATIONS|DOI:10.1038/ncomms9635 NATURECOMMUNICATIONS |6:8635|DOI:10.1038/ncomms9635|www.nature.com/naturecommunications MacmillanPublishersLimited.Allrightsreserved. aftercentrifugationat2,000 at4 Cfor10minandresuspendedagainwith1ml Hondabuffer,repeatingthecentrifugationandresuspensionatleastthreetimes, tovisualizethepeaksinthetomatogenome. Feedingexperimentswith C-and C-labelledglucose Pericarpdiscsof NATURECOMMUNICATIONS|DOI:10.1038/ncomms9635 ARTICLE NATURECOMMUNICATIONS |6:8635|DOI:10.1038/ncomms9635|www.nature.com/naturecommunications MacmillanPublishersLimited.Allrightsreserved. Aminoacidmeasurements Fruitwereharvestedsevendaysafterbreakerand at4 Cfor10min,and centrifugationat11,000 at4 Cfor10min,wasevaporatedwithoutheating. ARTICLE NATURECOMMUNICATIONS|DOI:10.1038/ncomms9635 NATURECOMMUNICATIONS |6:8635|DOI:10.1038/ncomms9635|www.nature.com/naturecommunications MacmillanPublishersLimited.Allrightsreserved. 36.Zhang,Y.,Butelli,E.&Martin,C.Engineeringanthocyaninbiosynthesisin Curr.Opin.Plant.Biol. 8190(2014). 37.Bilal,I.,Chowdhury,A.,Davidson,J.&Whitehead,S.Phytoestrogensand preventionofbreastcancer:thecontentiousdebate. WorldJ.Clin.Oncol. 705712(2014). 38.Rallapalli,G. ThisworkislicensedunderaCreativeCommonsAttribution4.0 InternationalLicense.Theimagesorotherthirdpartymaterialinthis articleareincludedinthearticlesCreativeCommonslicense,unlessindicatedotherwise inthecreditline;ifthematerialisnotincludedundertheCreativeCommonslicense, userswillneedtoobtainpermissionfromthelicenseholdertoreproducethematerial. Toviewacopyofthislicense,visithttp://creativecommons.org/licenses/by/4.0/ NATURECOMMUNICATIONS|DOI:10.1038/ncomms9635 ARTICLE NATURECOMMUNICATIONS |6:8635|DOI:10.1038/ncomms9635|www.nature.com/naturecommunications MacmillanPublishersLimited.Allrightsreserved.

Приложенные файлы

  • pdf 14430241
    Размер файла: 2 MB Загрузок: 0

Добавить комментарий