Construcción de promotores trampa basados en efectores TAL de Xanthomonas axonopodis pv. manihotis

dc.contributorDíaz Tatis, Paula Alejandra
dc.contributorLópez Carrascal, Camilo Ernesto
dc.contributorManihot Biotec
dc.creatorSánchez Ferro, Juan Sebastian
dc.date.accessioned2020-07-24T23:36:55Z
dc.date.available2020-07-24T23:36:55Z
dc.date.created2020-07-24T23:36:55Z
dc.date.issued2020-02-07
dc.identifierSánchez-Ferro, J. (2020). Construcción de promotores trampa basados en efectores TAL de Xanthomonas axonopodis pv. manihotis. Tesis de Maestría en Ciencias - Biología UNAL. Universidad Nacional de Colombia - Sede Bogotá.
dc.identifierhttps://repositorio.unal.edu.co/handle/unal/77853
dc.description.abstractCassava bacterial blight caused by Xanthomonas axonopodis pv. manihotis (Xam), is the main bacterial disease that affects Cassava crops. Xam virulence has been attributed to TALEs (Transcription Activator-Like Effectors), that bind to promoter elements of the host target genes to induce their expression. The TALome characterization of diverse Colombian strains of Xam have led to the identification of TAL14, TAL20, and TAL22 as the most recurrent TAL effectors in the pathogen populations in the country. With the purpose to develop a biotechnological tool for producing broad-spectrum resistance against Xam, this study generated a genetic construction of a trap promotor containing EBEs (Effector Binding Elements) for TALEs 14, 20 and 22. The EBE sequences were inserted in the minimal Bs3 promoter (pBs3min) using site-directed mutagenesis, to construct the TriEBE promoter EBETAL14Xam and EBETAL22Xam were cloned flanking EBETAL20Xam. The reporter gene GUSplus and 35s terminator were inserted flanking the promoters, the resulting vectors were used to transform Agrobacterium tumefaciens and perform the functional evaluation in Nicotiana tabacum. Finally, the promoters activation was evidenced because of TAL14 and TAL20 presence in co-infiltrated leaves. These results suggest that trap promoters developed in the present research can be activated by any Xam strain with TAL14 or TAL20 in its TALome, therefore representing a novel recognition cassette for most strains of the pathogen in Colombia.
dc.description.abstractLa bacteriosis vascular de la yuca, causada por Xanthomonas axonopodis pv. manihotis (Xam), es la principal enfermedad bacteriana que afecta al cultivo de yuca. La virulencia de Xam ha sido atribuida principalmente a los TALEs (Transcription Activator-Like Effectors), los cuales se unen a elementos en el promotor de genes blanco del hospedero para inducir su expresión. La caracterización del TALoma en diversas cepas colombianas de Xam ha llevado a la identificación de los efectores TAL14, TAL20 y TAL22 como los más frecuentes en las poblaciones del patógeno. Con el fin de desarrollar una herramienta para producir resistencia de amplio espectro a Xam, en este trabajo se generó una construcción genética de un promotor trampa conteniendo los EBEs (Effector Binding Elements) para los TALEs 14, 20 y 22. Los EBEs fueron insertados por mutagénesis dirigida en el promotor mínimo de Bs3 (pBs3min) y para la construcción de un promotor TriEBE se clonaron EBETAL14Xam y EBETAL22Xam alrededor del EBETAL20Xam. Las secuencias del gen GUSplus y el terminador 35s fueron clonadas flanqueando los promotores. La expresión transitoria de los constructos en Nicotiana tabacum mediada por Agrobacterium tumefaciens evidenció la activación de los promotores debido a la presencia de los TALEs 14 y 20 en hojas coinfiltradas con los promotores junto los efectores. Estos resultados indican que los promotores trampa generados en este estudio tienen la capacidad de ser activados por cualquier cepa de Xam que presente TAL14 o TAL20 en su TALoma, planteándose como un cassette de reconocimiento para la mayoría de las cepas del patógeno en el país.
dc.languagespa
dc.publisherBogotá - Ciencias - Maestría en Ciencias - Biología
dc.publisherUniversidad Nacional de Colombia - Sede Bogotá
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dc.rightsAtribución-NoComercial 4.0 Internacional
dc.rightsAcceso abierto
dc.rightshttp://creativecommons.org/licenses/by-nc/4.0/
dc.rightsinfo:eu-repo/semantics/openAccess
dc.rightsDerechos reservados - Universidad Nacional de Colombia
dc.titleConstrucción de promotores trampa basados en efectores TAL de Xanthomonas axonopodis pv. manihotis
dc.typeDocumento de trabajo


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