Publicación: Estandarización de la expresión transitoria de proteínas de virulencia de Phytophthora Palmivora en hojas de palma de aceite
| dc.contributor.advisor | Corzo Herrera, Mariana | spa |
| dc.contributor.advisor | Pachón García, Jorge | spa |
| dc.contributor.author | Sandoval Parra, Karen Ximena | spa |
| dc.date.accessioned | 2019-11-05T19:33:37Z | spa |
| dc.date.available | 2019-11-05T19:33:37Z | spa |
| dc.date.issued | 2018 | spa |
| dc.description | Incluye tablas, figuras, símbolos y abreviaturas. | spa |
| dc.description.abstract | El cultivo de la palma de aceite y su industria se encuentran en constante crecimiento. Sin embargo, su sostenibilidad se ve limitada por factores bióticos como los microorganismos patogénicos. La pudrición del cogollo ha sido una grave enfermedad en la palma de aceite en Colombia y en países vecinos por más de 40 años, causando la destrucción de miles de hectáreas. La búsqueda del patógeno tomó écadas, y en el 2008 se logró identificar a Phytophthora palmivora como el oomicete causal de la enfermedad. Dada la necesidad de encontrar cultivares resistentes a este microorganismo, se planteó el estudio de proteínas efectoras que el patógeno libera al hospedero para iniciar su proceso de infección. Para lo anterior, se normalizaron los parámetros físicos y biológicos del bombardeo de efectores sobre foliolos, estimando su eficiencia por medio de genes reporteros. Se evaluó así la expresión de tres efectores de P. palmivora sobre dos cultivares de palma de aceite mediante dos ensayos histoquímicos indicadores de muerte celular: DAB y NBT. Con este estudio fue posible encontrar la combinación de factores físicos y biológicos que da lugar a una transformación eficaz, según la expresión del gen GUSPlus, para cada uno de los cultivares. El gen GFP probó no ser el adecuado para el seguimiento de la transformación en callo, ya que el callo es autofluorescente. El ensayo DAB señaló, al comparar la respuesta del cultivar OxG frente a cada uno de los tres efectores, que hay diferencias significativas indicando que este se ve más afectado por el E2 y E19. Por otro lado, Elaeis guineensis, no mostró diferencias significativas entre la afectación por cada efector. | spa |
| dc.description.abstract | Oil palm crops and its industry are constantly growing. However, its sustainability is limited through biotic factors like pathogenic microorganisms. The bud rot has been a major oil palm disease in Colombia and nearby countries for over 40 years, where is responsible for the destruction of thousands of hectares. The search for the pathogen took decades, and in 2008 the causing oomycete was identified as Phytophthora palmivora. Given the necessity to find resistant cultivars to this microorganism, the study of effectors, virulence proteins that the pathogen releases to the host in order to start its colonization, was suggested through a standardization of the transient biolistic expression process. To perform the above, the biological and physical parameters of microparticle bombardment of effectors on leaves were optimized, estimating its efficiency through reporter genes. The expression of three effectors on two different oil palm cultivars was evaluated by the means of two histochemical assays that indicate cell death: DAB and NBT. It was possible to achieve the right combination of physical and biological factors that result in an efficient transformation, according to GUSPlus gene expression, for each one of the cultivars. The GFP gene probed not to be a suitable marker gene for transformation in callus, given that these are autofluorescent. DAB assay showed: by comparison of OxG cultivar response against each of the effectors, that there are significant differences indicating that is mostly affected by E2 y E19. On the other hand, Elaeis guineensis, did not displayed significant differences between the affectation for each effector. | eng |
| dc.description.degreelevel | Pregrado | spa |
| dc.description.degreename | Biología | spa |
| dc.description.notes | Trabajo de grado presentado como requisito parcial para optar al título de Bióloga. | spa |
| dc.description.tableofcontents | Planteamiento del problema. -- Justificación. -- Objetivos. -- Marco teórico. -- Palma de aceite y su cultivo. -- Pudrición del cogollo. -- Generalidades de phytophthora palmivora. -- Interacciones moleculares planta-patógeno. -- Efectores y efectorómica. -- Transformación genética en plantas. -- Expresión transitoria en palma de aceite. – Metodología. -- Material vegetal. -- Figura 7. Arreglo final del callo en cajas de petri previo al bombardeo. -- Constructos plasmídicos. -- Preparación ADN-microcarriers. -- Ensayos de transformación mediante bombardeo por biobalística. -- Figura 8. Pistola de biobalística empleada en los ensayos. --Figura 9. Secuencia de montaje set de biobalística. -- Ensayos con genes marcadores. -- Detección de β-glucuronidasa mediante tinción con buffer GUS (x-GLUC). -- Ensayo de microscopía fluorescente con GFP. -- Ensayos histoquímicos indicadores de posible muerte celular (HR). -- Prueba DAB (3,3’-diaminobenzidina). -- Prueba NBT (nitro blue tetrazolio). -- Resultados y discusión. -- Eficiencia y seguimiento de transformación. -- Ensayo GUS. -- Ensayo GFP. -- Ensayos histoquímicos para detectar posible HR. -- Tinción DAB. -- Prueba NBT. -- Conclusiones. -- Apreciaciones finales. -- Recomendaciones. -- Productos. -- Bibliografía. | spa |
| dc.format.extent | 101 páginas | spa |
| dc.format.mimetype | application/pdf | spa |
| dc.identifier.citation | Sandoval Parra, Karen X. (2018). Estandarización de la expresión transitoria de proteínas de virulencia de Phytophthora Palmivora en hojas de palma de aceite [Trabajo de grado, Universidad de los Llanos]. Repositorio digital Universidad de los Llanos. | spa |
| dc.identifier.instname | Universidad de los Llanos | |
| dc.identifier.reponame | Repositorio digital Universidad de los Llanos | |
| dc.identifier.uri | https://repositorio.unillanos.edu.co/handle/001/1465 | spa |
| dc.language.iso | spa | spa |
| dc.publisher | Universidad de los Llanos | spa |
| dc.publisher.faculty | Facultad de Ciencias Básicas e Ingeniería | spa |
| dc.publisher.place | Villavicencio | spa |
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| dc.rights | Derechos Reservados - Universidad de los Llanos, 2018 | spa |
| dc.rights.accessrights | info:eu-repo/semantics/openAccess | spa |
| dc.rights.coar | http://purl.org/coar/access_right/c_abf2 | spa |
| dc.rights.creativecommons | Atribución-NoComercial 4.0 Internacional (CC BY-NC 4.0) | spa |
| dc.rights.uri | https://creativecommons.org/licenses/by-nc/4.0/ | spa |
| dc.subject.proposal | Biobalística | spa |
| dc.subject.proposal | Palma de aceite | spa |
| dc.subject.proposal | Pudrición del Cogollo | spa |
| dc.subject.proposal | ERO (ROS) | spa |
| dc.subject.proposal | Efectores | spa |
| dc.subject.proposal | Phytophthora palmivora | eng |
| dc.title | Estandarización de la expresión transitoria de proteínas de virulencia de Phytophthora Palmivora en hojas de palma de aceite | spa |
| dc.type | Trabajo de grado - Pregrado | spa |
| dc.type.coar | http://purl.org/coar/resource_type/c_7a1f | spa |
| dc.type.coarversion | http://purl.org/coar/version/c_970fb48d4fbd8a85 | spa |
| dc.type.content | Text | spa |
| dc.type.driver | info:eu-repo/semantics/bachelorThesis | spa |
| dc.type.redcol | https://purl.org/redcol/resource_type/TP | spa |
| dc.type.version | info:eu-repo/semantics/publishedVersion | spa |
| dspace.entity.type | Publication |
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