DOF-binding sites additively contribute to guard cell-specificity of AtMYB60 promoter.docVIP

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DOF-binding sites additively contribute to guard cell-specificity of AtMYB60 promoter.doc

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Cominellietal.BMCPlantBiology2011,11:162 /1471-2229/11/162 RESEARCH ARTICLE OpenAccess DOF-bindingsitesadditivelycontributetoguard cell-specificityofAtMYB60promoter EleonoraCominelli1,4*,MassimoGalbiati1,2,AlessandraAlbertini1,FabioFornara3,5,LucioConti1,2, GeorgeCoupland3andChiaraTonelli1* Abstract Background:WepreviouslydemonstratedthattheArabidopsisthalianaAtMYB60proteinisanR2R3MYB transcriptionfactorrequiredforstomatalopening.AtMYB60isspecificallyexpressedinguardcellsanddown- regulatedatthetranscriptionallevelsbythephytohormoneABA. Results:ToinvestigatethemolecularmechanismsgoverningAtMYB60expression,itspromoterwasdissected throughdeletionandmutagenesisanalyses.BystudyingdifferentversionsofAtMYB60promoter::GUSreporter fusionsintransgenicplantswewereabletodemonstrateamodularorganizationfortheAtMYB60promoter. Particularlywedefined:aminimalpromotersufficienttoconferguardcell-specificactivitytothereportergene;the distinctrolesofdifferentDOF-bindingsitesorganisedinaclusterintheminimalpromoterindeterminingguard cell-specificexpression;thepromoterregionsresponsiblefortheenhancementofactivityinguardcells;a promoterregionresponsibleforthenegativetranscriptionalregulationbyABA.Moreoverfromtheanalysisof singleandmultiplemutantswecouldruleouttheinvolvementofagroupofDOFproteins,knownasCDFs, alreadycharacterisedfortheirinvolvementinfloweringtime,intheregulationofAtMYB60expression. Conclusions:Thesefindingsshedlightontheregulationofgeneexpressioninguardcellsandprovidenew promotermodulesasusefultoolsformanipulatinggeneexpressioninguardcells,bothforphysiologicalstudies andfuturebiotechnologicalapplications. Background stomatalresponsesrequirestheuseofguardcell-specific Land plants uptake carbon dioxide for photosynthesis promoters, or the identification of guard cell-specific andlosewatervapourbytranspirationthroughstomatal mutants, to avoid undesirable side effects on plant pores, present onthesurface ofleaves andstems. The growthandproductivity. opening andclosure of