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dc.rights.licenseAtribución-NoComercial-CompartirIgual 4.0 Internacional
dc.contributor.advisorEscobar Pérez, Roosevelt Humberto
dc.contributor.advisorCastiblanco Vargas, Valheria
dc.contributor.authorCarcamo Medina, Lilian Yaritza
dc.date.accessioned2022-02-14T16:41:22Z
dc.date.available2022-02-14T16:41:22Z
dc.date.issued2021-12-07
dc.identifier.urihttps://repositorio.unal.edu.co/handle/unal/80975
dc.descriptionIlustraciones, tablas
dc.description.abstractLa duplicación de cromosomas es un fenómeno relevante para ampliar la base de la diversidad genética de las plantas, pues llega a tener efectos importantes tales como el aumento en el tamaño de las células, de los órganos, aumento de la biomasa, ayudando en la transferencia de genes o en el incremento de heterocigosidad. El programa de forrajes tropicales del CIAT (Centro Internacional de Agricultura Tropical, hoy Alianza Bioversity-CIAT) pretende optimizar el esquema de mejoramiento genético actual y para ello necesita obtener un genotipo tetraploide sexual de Urochloa decumbens. Por este motivo, en el presente estudio se planteó como objetivo explorar las bases metodológicas para iniciar un proceso de poliploidización de la especie mediante el uso de la colchicina. Se ajustó e implementó una metodología in vitro para la propagación de material vegetal y se inició el esquema de regeneración mediado por la de embriogénesis somática como una posible vía para la duplicación de cromosomas. Se realizaron dos ensayos de poliploidización, el primero bajo condiciones in vivo utilizando plántulas germinadas de cuatro días de edad como explante y en una segunda prueba bajo condiciones in vitro utilizando segmentos basales. Se utilizaron dos concentraciones de colchicina: 0,05 y 0,1% durante 2, 8, 12 y 24 h tanto en in vivo como in vitro. En la adecuación del material vegetal, para la fase in vitro la escarificación manual presenta mejores porcentajes de germinación en las 3 accesiones utilizadas y fue posible implementar la propagación en condiciones de medio sólido. Tanto la aclimatación de las plantas en invernadero, como la regeneración de plantas vía embriogénesis somática fue exitosa. En los ensayos de poliploidización, las plantas tratadas con colchicina se establecieron en invernadero y se determinó tasa de supervivencia, tamaño de estomas y densidad estomática. A nivel de tasa de supervivencia de explantes tratados con colchicina en alta dosis y tiempos largos hubo mayor mortalidad, a nivel de estomas (morfología, ancho, largo y densidad estomática) no se observaron cambios significativos que se puedan correlacionar con el nivel de ploidía en el ensayo in vivo, sin embargo, sería prudente dar una espera hasta obtener plantas maduras para analizar de nuevo estos parámetros. Se va a continuar con la verificación de ploidía mediante citometría de flujo y conteo de cromosomas. (Texto tomado de la fuente)
dc.description.abstractChromosome duplication is considered a broad and relevant phenomenon to expand the genetic diversity of plants, which allows having significant effects such as an increase in the size of cells, organs, biomass, gene transfer, or the increasing heterozygosity. Therefore, the tropical forages program of CIAT (International Center for Tropical Agriculture, now the Bioversity-CIAT Alliance) seeks to optimize the current breeding scheme to obtain a sexual tetraploid genotype of Urochloa decumbens to have more genetic diversity. Based on this, the present study explores the methodological bases to initiate a polyploidization process of this species using colchicine. This research started from developing an in vitro methodology for the propagation of plant material and somatic embryogenesis as a possible route of chromosome duplication. Two polyploidization assays were performed, the first under in vivo conditions using germinated four-day-old seedlings as explants and the second under in vitro conditions using basal segments. Two concentrations of colchicine were used: 0.05 and 0.1% for 2, 8, 12, and 24 h for both tests. The manual scarification presented better germination percentages in the three accessions using the in vitro methodology. Moreover, It helped to propagate the plants in a solid medium. The acclimatization of the plants in the greenhouse and the regeneration using somatic embryogenesis were effective. In the polyploidization tests, the plants treated with colchicine were established in the greenhouse, and the survival rate, stomata size, and stomatal density variables were determined. There was higher mortality at the higher concentrations and longer exposure times when assessing the survival rate. No significant changes related to the ploidy level were found at the stomatal level for the in vivo test. However, to corroborate the results, it is suggested to continue with the ploidy analysis using flow cytometry and chromosome counting.
dc.description.sponsorshipPrograma de mejoramiento de forrajes del Centro Internacional de Agricultura Tropical (CIAT)
dc.format.extentxviii, 75páginas + anexos
dc.format.mimetypeapplication/pdf
dc.language.isospa
dc.publisherUniversidad Nacional de Colombia
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/
dc.subject.ddc570 - Biología::576 - Genética y evolución
dc.subject.ddc570 - Biología::571 - Fisiología y temas relacionados
dc.subject.ddc580 - Plantas::584 - Monocotiledóneas, angiospermas basales, clorantales, magnolias
dc.titleBases metodológicas hacia la poliploidización de Urochloa decumbens
dc.typeTrabajo de grado - Maestría
dc.type.driverinfo:eu-repo/semantics/masterThesis
dc.type.versioninfo:eu-repo/semantics/acceptedVersion
dc.publisher.programPalmira - Ciencias Agropecuarias - Maestría en Ciencias Biológicas
dc.contributor.educationalvalidatorGarcía Dávila, Mario Augusto
dc.description.degreelevelMaestría
dc.description.degreenameMagíster en Ciencias Biológicas
dc.description.methodsPara todos los ensayos se utilizaron semillas de tres accesiones sexuales diploides de Urochloa decumbens (Cuadro 4-1) facilitadas por el Banco de Germoplasma de CIAT. Se siguió el protocolo establecido en el Laboratorio de Viabilidad de Semillas del CIAT. A un pequeño lote de 10 semillas por genotipo se les realizó prueba de viabilidad con cloruro de tetrazolio (TTC) al 0,5%. A las semillas se les retiró la gluma y se sumergieron en agua destilada en un tubo de ensayo # 16 durante 16 h a temperatura ambiente. Transcurrido este tiempo se les realizó un corte longitudinal a través del embrión con una cuchilla # 10 y con ayuda de una lupa 3X con luz. La mitad de cada semilla se descartó y la otra mitad se colocó en un tubo de ensayo con 10 ml de la solución de TTC. Para la tinción los tubos con las muestras a evaluar se llevaron a un horno a 40°C por 2,5 h bajo condiciones de oscuridad. Posteriormente las semillas se lavaron varias veces con agua destilada para eliminar el exceso de colorante y se realiza la lectura usando un estereoscopio Leica MS5 con cámara fotográfica Leica. A cada semilla se le examinó y se le consideró como viable o no viable sobre la base de los patrones de tinción y la solidez de la coloración del tejido siguiendo la norma del ISTA. Con el fin de comparar dos tipos de escarificación, un segundo lote de semillas fue escarificado química y manualmente. En la escarificación química se utilizó ácido sulfúrico (H2SO4) al 96% durante 14 min, seguido de un lavado con abundante agua, luego las semillas se secaron y almacenaron hasta su uso. Para la escarificación manual se procedió a eliminar las cubiertas, primero la gluma, luego con la ayuda de un bisturí se realizó un pequeño corte vertical entre lema y palea para descubrir las cariópsides. Semillas escarificadas manualmente fueron utilizadas para realizar ensayos de desinfección. La desinfección se realizó con etanol al 70% durante 3 min (Figura 4-3 A), se retira y se adiciona una solución de fungicida que contiene Zellus/Benomyl 1g/L y Maxíl 6 ml/L respectivamente (Figura 4-3 B). el tratamiento con fungicidas se mantiene durante 35 min en constante agitación, seguidamente se pasa a hipoclorito de sodio al 1,5% más dos gotas de Tween 20 durante 40 min (Figura 4-3 C). Al final del proceso se realizan 5 lavados con agua desionizada estéril. Todo este proceso se lleva a cabo con uso de EPP y en cámara de flujo. La siembra se realizó en tubos de ensayo #16 (Figura 4-4) que contenían medio MS (Murashige & Skoog, 1962) y 4E en estado líquido y sólido (sales de MS completo, Sacarosa 2%, Tiamina 1 mg/L, m-inositol 100 mg/L, BAP 0,04 mg/L, GA3 0,05 mg/L y ANA 0,02 mg/L, pH 5,7, agar 0,46% reportado por Roca (1984). Las semillas se implantan (siembran) en el medio con la parte donde emerge el epicótilo hacia arriba y posteriormente cada frasco se sella con papel vinipel plástico y son llevados a un cuarto de crecimiento a 27°C y fotoperiodo de 12 h luz /12 h oscuridad. El material contaminado se descartó a partir de los tres días y se enviaron tubos contaminados al Laboratorio de Sanidad de Germoplasma del Programa de Conservación de germoplasma del CIAT para su respectiva identificación. (Texto tomado de la fuente)
dc.description.researchareaBiotecnología Vegetal
dc.identifier.instnameUniversidad Nacional de Colombia
dc.identifier.reponameRepositorio Institucional Universidad Nacional de Colombia
dc.identifier.repourlhttps://repositorio.unal.edu.co/
dc.publisher.facultyFacultad de Ciencias Agropecuarias
dc.publisher.placePalmira, Colombia
dc.publisher.branchUniversidad Nacional de Colombia - Sede Palmira
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dc.rights.accessrightsinfo:eu-repo/semantics/openAccess
dc.subject.proposalcolchicina
dc.subject.proposalforrajes
dc.subject.proposalin vitro
dc.subject.proposalduplicación de cromosomas
dc.subject.proposalmejoramiento de plantas
dc.subject.proposalcolchicine
dc.subject.proposalforages
dc.subject.proposalin vitro
dc.subject.proposalchromosomes duplication
dc.subject.proposalplant breeding
dc.title.translatedMethodological bases for the polyploidization of Urochloa decumbens
dc.type.coarhttp://purl.org/coar/resource_type/c_bdcc
dc.type.coarversionhttp://purl.org/coar/version/c_ab4af688f83e57aa
dc.type.contentText
dc.type.redcolhttp://purl.org/redcol/resource_type/TM
oaire.accessrightshttp://purl.org/coar/access_right/c_abf2
oaire.awardtitleBases metodológicas hacia la poliploidización de Urochloa decumbens
dcterms.audience.professionaldevelopmentEstudiantes
dcterms.audience.professionaldevelopmentInvestigadores


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