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Resumen
Background: Lettuce is a globally important leafy vegetable and a model plant for biotechnology due to its adaptability to tissue culture and stable genetic transformation. Lettuce is also crucial for functional genomics research in the Asteraceae which includes species of great agronomical importance. The development of transgenic events implies the production of a large number of shoots that must be differentiated between transgenic and non-transgenic [ver mas...]
dc.contributor.authorDarqui, Flavia Soledad
dc.contributor.authorRadonic, Laura Mabel
dc.contributor.authorLopez, Nilda Ester
dc.contributor.authorHopp, Horacio Esteban
dc.contributor.authorLopez Bilbao, Marisa Gisela
dc.date.accessioned2018-03-28T14:36:25Z
dc.date.available2018-03-28T14:36:25Z
dc.date.issued2018-01
dc.identifier.issn0717-3458
dc.identifier.otherhttps://doi.org/10.1016/j.ejbt.2017.10.002
dc.identifier.urihttp://hdl.handle.net/20.500.12123/2147
dc.identifier.urihttps://www.sciencedirect.com/science/article/pii/S0717345817300611
dc.description.abstractBackground: Lettuce is a globally important leafy vegetable and a model plant for biotechnology due to its adaptability to tissue culture and stable genetic transformation. Lettuce is also crucial for functional genomics research in the Asteraceae which includes species of great agronomical importance. The development of transgenic events implies the production of a large number of shoots that must be differentiated between transgenic and non-transgenic through the activity of the selective agent, being kanamycin the most popular. Results: In this work we adjusted the selection conditions of transgenic seedlings to avoid any escapes, finding that threshold concentration of kanamycin was 75 mg/L. To monitor the selection system, we studied the morphological response of transgenic and non-transgenic seedlings in presence of kanamycin to look for a visual morphological marker. Several traits like shoot length, primary root length, number of leaves, fresh weight, and appearance of the aerial part and development of lateral roots were affected in non-transgenic seedlings after 30 d of culture in selective media. However, only lateral root development showed an early, qualitative and reliable association with nptII presence, as corroborated by PCR detection. Applied in successive transgenic progenies, this method of selection combined with morphological follow-up allowed selecting the homozygous presence of nptII gene in 100% of the analyzed plants from T2 to T5. Conclusions: This protocol allows a simplified scaling-up of the production of multiple homozygous transgenic progeny lines in the early generations avoiding expensive and time-consuming molecular assays.eng
dc.formatapplication/pdf
dc.language.isoeng
dc.rightsinfo:eu-repo/semantics/openAccess
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/
dc.sourceElectronic Journal of Biotechnology 31 : 1-9 (January 2018)
dc.subjectLechugas
dc.subjectLettuceseng
dc.subjectPlantas Transgénicas
dc.subjectTransgenic Plantseng
dc.subjectHortalizas de Hoja
dc.subjectLeaf Vegetableseng
dc.titleSimplified methodology for large scale isolation of homozygous transgenic lines of lettuce
dc.typeinfo:eu-repo/semantics/article
dc.typeinfo:ar-repo/semantics/artículo
dc.typeinfo:eu-repo/semantics/publishedVersion
dc.rights.licenseCreative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.description.origenInst. de Biotecnología
dc.gic156665
dc.description.filFil: Darqui, Flavia Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
dc.description.filFil: Radonic, Laura Mabel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
dc.description.filFil: Lopez, Nilda Ester. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
dc.description.filFil: Hopp, Horacio Esteban. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina
dc.description.filFil: López Bilbao, Marisa Gisela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
dc.subtypecientifico


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