| dc.contributor.author | Nyaboga, E.N. |
| dc.contributor.author | Njiru, J.M. |
| dc.contributor.author | Tripathi, L. |
| dc.date.accessioned | 2019-12-04T10:58:00Z |
| dc.date.available | 2019-12-04T10:58:00Z |
| dc.date.issued | 2015 |
| dc.identifier.citation | Nyaboga, E.N., Njiru, J.M., & Tripathi, L. (2015). Factors influencing somatic embryogenesis, regeneration and agrobacterium-mediated transformation of cassava (Manihot esculenta Crantz) cultivar TME14. Frontiers in Plant Science, 6, 411. |
| dc.identifier.issn | 1664-462X |
| dc.identifier.uri | https://hdl.handle.net/20.500.12478/893 |
| dc.description.abstract | 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. |
| dc.language.iso | en |
| dc.subject | Cassava |
| dc.subject | Genetics |
| dc.title | Factors influencing somatic embryogenesis, regeneration, and Agrobacterium-mediated transformation of cassava (Manihot esculenta Crantz) cultivar TME14 |
| dc.type | Journal Article |
| dc.description.version | Peer Review |
| cg.contributor.crp | Roots, Tubers and Bananas |
| cg.contributor.affiliation | International Institute of Tropical Agriculture |
| cg.coverage.region | Africa |
| cg.coverage.region | East Africa |
| cg.coverage.country | Kenya |
| cg.isijournal | ISI Journal |
| cg.authorship.types | CGIAR and advanced research institutes |
| cg.iitasubject | Cassava |
| cg.journal | Frontiers in Plant Science |
| cg.howpublished | Formally Published |
| cg.accessibilitystatus | Open Access |
| local.dspaceid | 76392 |
| cg.identifier.doi | https://dx.doi.org/10.3389/fpls.2015.00411 |
