Comparative Analysis Reveals Dynamic Changes in miRNAs and Their Targets and Expression during Somatic Embryogenesis in Longan Dimocarpus longan Lour.).docVIP

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Comparative Analysis Reveals Dynamic Changes in miRNAs and Their Targets and Expression during Somatic Embryogenesis in Longan Dimocarpus longan Lour.).doc

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Comparative Analysis Reveals Dynamic Changes in miRNAs and Their Targets and Expression during Somatic Embryogenesis in Longan Dimocarpus longan Lour.)

ComparativeAnalysisRevealsDynamicChangesin miRNAsandTheirTargetsandExpressionduring SomaticEmbryogenesisinLongan(Dimocarpuslongan Lour.) YulingLin,ZhongxiongLai* InstituteofHorticulturalBiotechnology,FujianAgricultureandForestry,Fuzhou,Fujian,China Abstract Somaticembryogenesis(SE),whichresembleszygoticembryogenesis,isanessentialcomponentoftheprocessofplantcell differentiationandembryodevelopment.AlthoughmicroRNAs(miRNAs)areimportantregulatorsofmanyplantdevelop- mental processes, their roles in SE have not been thoroughly investigated. In this study, we used deep-sequencing, computational, and qPCR methods to identify, profile, and describe conserved and novel miRNAs involved in longan (Dimocarpuslongan)SE.Atotalof643conservedand29novelmiRNAs(includingstarstrands)frommorethan169miRNA families were identified in longan embryogenic tissue using Solexa sequencing. By combining computational and degradome sequencing approaches, we were able to predict 2063 targets of 272 miRNAs and verify 862 targets of 181 miRNAs. Target annotation revealed that the putative targets were involved in a broad variety of biological processes, includingplantmetabolism,signaltransduction,andstimulusresponse.Analysisofstage-andtissue-specificexpressionsof 20 conserved and 4 novel miRNAs indicated their possible roles in longan SE. These miRNAs were dlo-miR156 family members and dlo-miR166c* associated with early embryonic culture developmental stages; dlo-miR26, dlo-miR160a , and families dlo-miR159, dlo-miR390, and dlo-miR398b related to heart-shaped and torpedo- shaped embryo formation; dlo- miR4a, dlo-miR24, dlo-miR167a, dlo-miR168a*, dlo-miR397a, dlo-miR398b.1, dlo-miR398b.2, dlo-miR808 and dlo-miR5077 involved in cotyledonary embryonic development; and dlo-miR17 and dlo-miR2089*-1 that have regulatory roles during longanSE.Inaddition,dlo-miR167a,dlo-miR808,anddlo-miR5077mayberequiredformatureembryoformation.Thisstudy is thefirst reportedinvestigationof longanSEinvolving large-scal