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    Factors influencing somatic embryogenesis, regeneration, and Agrobacterium-mediated transformation of cassava (Manihot esculenta Crantz) cultivar TME14

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    Date
    2015
    Author
    Nyaboga, E.N.
    Njiru, J.M.
    Tripathi, L.
    Type
    Journal Article
    Metadata
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    Abstract/Description
    Routineproductionoflargenumbersoftransgenicplantsisrequiredtofullyexploitadvancesincassavabiotechnologyandsupportdevelopmentofiprovedgermplasmfordeploymenttofarmers.Thisarticledescribesanimproved,high-efficiencytransformationprotocolforrecalcitrantcassavacultivarTME14preferredinAfrica.Factorsthatfavorproductionoffriableembryogeniccalli(FEC)werefoundtobeuseofDKWmedium,crushingoforganizedembryogenicstructures(OES)through1–2mmsizedmetalwiremesh,washingofcrushedOEStissuesandshortexposureoftyrosinetosomaticembryos;andtransformationefficiencywasenhancedbyuseoflowAgrobacteriumdensityduringco-cultivation,co-centrifugationofFECwithAgrobacterium,germinationofparamomycinresistantsomaticembryosonmediumcontainingBAPwithgradualincreaseinconcentrationandvariationsofthefrequenyofsubcultureofcotyledonary-stageembryosonshootelongationmedium.Byapplyingtheoptimizedparameters,FECwereproducedforcassavacultivarTME14andtransformedusingAgrobacteriumstrainLBA4404harboringthebinaryvectorpCAMBIA2301.About70–80independenttransgeniclinespermlsettledcellvolume(SCV)ofFECwereregeneratedonselectivemedium.HistochemicalGUSassaysconfirmedtheexpressionofgusAgeneintransformedcalli,somaticembryosandtransgenicplants.ThepresenceandintegrationofthegusAgenewereconfirmedbyPCRandSouthernblotanalysis,respectively.RT-PCRanalysisoftransgenicplantsconfirmedtheexpressionofgusAgene.Thisprotocoldemonstratessignificantlyenhancedtransformationefficiencyoverexistingcassavatransformationprotocolsandcouldbecomeapwerfultoolforfunctionalgenomicsandtransferringnewtraitsintocassava.
    https://dx.doi.org/10.3389/fpls.2015.00411
    Multi standard citation
    Permanent link to this item
    https://hdl.handle.net/20.500.12478/893
    Digital Object Identifier (DOI)
    https://dx.doi.org/10.3389/fpls.2015.00411
    IITA Subjects
    Cassava
    Agrovoc Terms
    Cassava; Genetics
    Regions
    Africa; East Africa
    Countries
    Kenya
    Journals
    Frontiers in Plant Science
    Collections
    • Journal and Journal Articles4835
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