Detección molecular y limpieza de virus en material de siembra de Solanum tuberosum y S. phureja

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dc.contributor.advisorHoyos Sánchez, Rodrigo Alberto
dc.contributor.advisorGutiérrez Sánchez, Pablo Andrés
dc.contributor.authorGarcía Torres, Andrea Stefania
dc.contributor.educationalvalidatorMarìn Montoya, Mauricio Alejandro
dc.contributor.educationalvalidatorCorrea Londoño, Guillermo Antonio
dc.contributor.researchgroupBiotecnología Microbianaspa
dc.contributor.researchgroupBiotecnología Vegetal Unalmed Cibspa
dc.date.accessioned2022-09-02T21:32:26Z
dc.date.available2022-09-02T21:32:26Z
dc.date.issued2022-08-20
dc.descriptionilustraciones, tablasspa
dc.description.abstractEn Colombia, el cultivo de papa (Solanum tuberosum y S. phureja) es uno de los renglones agrícolas más importantes para la zona Andina. Con el fin de aportar nuevos elementos al conocimiento de la situación virológica de la papa en el país, y especialmente en Antioquia, en este proyecto se utilizaron diversas técnicas moleculares (RT-PCR, qPCR, secuenciación Sanger y HTS) para el diagnóstico, identificación, y caracterización genómica de los virus presentes en el material de siembra certificado y no certificado de los cultivares Diacol Capiro y Criolla Colombia, comercializados en Antioquia y en menor proporción en la sabana Cundiboyacense. Los resultados indicaron la presencia de los virus de RNA: virus Y de la papa (Potato virus Y - PVY), virus S de la papa (Potato virus S - PVS), virus X de la papa (Potato virus X - PVX), virus del amarillamiento de las venas de la papa (Potato yellow vein virus - PYVV), virus del enrollamiento de la hoja de papa (Potato leafroll virus - PLRV), virus mop-top de la papa (Potato mop-top virus - PMTV), virus V de la papa (Potato virus V- PVV) y virus B de la papa (Potato virus B - PVB), en ambos cultivares de papa y en ambos tipos de material de siembra. En las evaluaciones directas sobre tubérculos obtenidos en la sabana Cundiboyacense, se detectaron siete (PVY, PVX, PVS, PLRV, PYVV, PMTV y PVB) de los ocho virus en material de siembra certificado de cv. Diacol Capiro, mientras que, en los tubérculos no certificados, se encontraron seis de los virus (PVY, PVX, PVS, PYVV, PLRV y PMTV). Para el caso de Antioquia, las evaluaciones se dividieron en las zonas del oriente y norte del departamento. En el oriente, se detectaron los ocho virus en ambos cultivares; en cuánto al norte, se encontraron siete de los virus (PVY, PVS, PVX, PLRV, PYVV, PMTV y PVB) en el cv. Diacol Capiro. Para el material del cv. Criolla Colombia, no fue posible obtener material certificado en la zona del norte, pero si se detectaron los ocho virus en los tubérculos no certificados. Utilizando la secuenciación masiva de alto rendimiento, se aumentó el número de secuencias disponibles de genomas de los virus PVY, PVV, PVX, PVS, PMTV y PVB, además se realizó el primer reporte de una secuencia de PVV infectando el cv. Diacol Capiro en Colombia y el primer registro del virus D de la necrosis del tabaco (Tobacco necrosis virus D - TNV-D) en el país. Adicionalmente, se realizó un ensayo piloto sobre la distribución viral en brotes de diferentes tubérculos pertenecientes a un mismo lote de semilla, encontrándose que para alcanzar una precisión del 90% en la detección viral, es necesario evaluar de manera independiente al menos tres tubérculos por lote y de dos a tres brotes por tubérculo. Por último, se evaluó el efecto de terapias in vitro para la eliminación viral en plántulas ambos cultivares, evidenciándose una reducción en la proporción de infección en vitroplantas sometidas a quimioterapia por 45 días con respeto a los tratamientos control, de los virus PVY, PVS, PVX, PLRV y PYVV en todas las dosis de ribavirina (50, 75 y 100 ppm); asimismo en las evaluaciones de termoterapia se registraron disminuciones en los virus PVY, PYVV y PMTV en las plántulas sometidas a 35°C, PVV, PVS, PYVV, PMTV y PVX en 36°C, PVS, PVX, PMTV y PYVV en 37°C, y PVS y PLRV en vitroplantas a 38°C por dos semanas. Se espera que los resultados obtenidos en esta tesis se constituyan en un aporte importante para el apoyo de los programas de manejo integrado de las enfermedades virales en papa. (Texto tomado de la fuente)spa
dc.description.abstractIn Colombia, potato (Solanum tuberosum y S. phureja) is one of the most important staple food crops in the Andean region. This work was designed to update our current knowledge of the potato virome in Colombia using a wide range of molecular tools (RT-PCR, qPCR, Sanger and HTS) for the diagnostics, identification and genomic characterization of the viruses present in certified and uncertified planting material used by potato farmers in the province of Antioquia, and some regions of Cundinamarca and Boyacá. The results presented here, reveal a high prevalence of the RNA viruses Potato virus Y (PVY), Potato virus S (PVS), Potato virus X (PVX), Potato yellow vein virus (PYVV), Potato leafroll virus (PLRV), Potato mop-top virus (PMTV), Potato virus V (PVV) and Potato virus B (PVB) in planting material from both cultivars. Direct testing on seed-tubers produced in the highlands of Boyacá and Cundinamarca revealed widespread infection with PVY, PVX, PVS, PLRV, PYVV, PMTV, and PVB in certified cv. Diacol Capiro seeds, and PVY, PVX, PVS, PYVV, PLRV, and PMTV, in uncertified ones. In eastern Antioquia, this investigation detected all eight viruses infecting both cultivars. As for northern Antioquia, seven viruses (PVY, PVS, PVX, PLRV, PYVV, PMTV y PVB) were found in cv. Diacol Capiro. All eight viruses under study were identified in uncertified Criolla Colombia seeds, unfortunately, certified seeds for this cultivar were not available in northern Antioquia at the time of study. HTS allowed either the partial or complete genome characterization of new PVY, PVV, PVX, PVS, PMTV and PVB isolates, in addition to the first PVV isolate infecting S. tuberosum in Colombia, and the first report for Tobacco necrosis virus D - TNV-D. A pilot study on the distribution of viruses across sprouts within the same tuber was performed revealing that to achieve 90% accuracy it is necessary to test a minimum of two or three sprouts from three different tubers per lot. Finally, this work also tested the effect of chemotherapy with ribavirin at 50, 75, and 100 ppm for 45 days; and thermotherapy at 35, 36, 37 and 38°C for 15 days, in reducing the proportion of vitroplants infected with PVY, PVV, PVS, PVX, PYVV, PLRV and PMTV in potato cultivars Diacol Capiro and Criolla Colombia. Chemotherapy proved to be effective in reducing the proportion of PVY, PVS, PVX, PLRV y PYVV at all ribavirin concentrations (50, 75 and 100 ppm). Additionally, thermotherapy reduced the proportion of vitroplants infected with PVY, PYVV and PMTV at 35°C, PVV, PVS, PYVV, PMTV and PVX at 36°C, PVS, PVX, PMTV and PYVV at 37°C, and PVS and PLRV at 38°C. These results are expected to be an important contribution in support of viral disease management programs of potato in Colombia.eng
dc.description.curricularareaÁrea curricular Biotecnologíaspa
dc.description.degreelevelMaestríaspa
dc.description.degreenameMagíster en Ciencias - Biotecnologíaspa
dc.description.researchareaFitopatologíaspa
dc.description.sponsorshipEsta tesis fue financiada por el Fondo de Ciencia, Tecnología e Innovación del Sistema General de Regalías del Departamento de Antioquia (Colombia) (Convenio No. 4600007658–779), a través del Proyecto: "Desarrollo de una plataforma molecular y bioinformática para el diagnóstico de virus en cultivos y material de siembra de papa (Solanum tuberosum y S. phureja) en Antioquia" (Código: 1101-805-62787), ejecutado por la Universidad Nacional de Colombia sede Medellín, Universidad CES y Fedepapa. El proyecto fue supervisado por la Secretaria de Agricultura y Desarrollo Rural de Antioquia y por el Ministerio de Ciencia, Tecnología e Innovación de Colombia. Las muestras vegetales fueron colectadas bajo el Permiso marco de la Universidad Nacional de Colombia y el permiso RGE152-27 del Ministerio del Medio Ambiente y Desarrollo Sostenible (Resolución 0208, 9/03/2020).spa
dc.format.extentxxiii, 325 páginasspa
dc.format.mimetypeapplication/pdfspa
dc.identifier.instnameUniversidad Nacional de Colombiaspa
dc.identifier.reponameRepositorio Institucional Universidad Nacional de Colombiaspa
dc.identifier.repourlhttps://repositorio.unal.edu.co/spa
dc.identifier.urihttps://repositorio.unal.edu.co/handle/unal/82244
dc.language.isospaspa
dc.publisherUniversidad Nacional de Colombiaspa
dc.publisher.branchUniversidad Nacional de Colombia - Sede Medellínspa
dc.publisher.departmentEscuela de biocienciasspa
dc.publisher.facultyFacultad de Cienciasspa
dc.publisher.placeMedellín, Colombiaspa
dc.publisher.programMedellín - Ciencias - Maestría en Ciencias - Biotecnologíaspa
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dc.rights.accessrightsinfo:eu-repo/semantics/openAccessspa
dc.rights.licenseAtribución-NoComercial-SinDerivadas 4.0 Internacionalspa
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/spa
dc.subject.ddc570 - Biología::575 - Partes específicas de y sistemas fisiológicos en plantasspa
dc.subject.ddc630 - Agricultura y tecnologías relacionadas::632 - Lesiones, enfermedades, plagas vegetalesspa
dc.subject.ddc630 - Agricultura y tecnologías relacionadas::633 - Cultivos de campo y de plantaciónspa
dc.subject.lembVirología agrícola
dc.subject.lembPapa - Enfermedades y plagas
dc.subject.lembPapa - Cultivo
dc.subject.lembCultivo in vitro
dc.subject.proposalCertificación de semillaspa
dc.subject.proposalqPCReng
dc.subject.proposalSecuenciación de alto rendimientospa
dc.subject.proposalSolanaceaeother
dc.subject.proposalVirología vegetalspa
dc.subject.proposalHigh-throughput sequencingeng
dc.subject.proposalPlant virologyeng
dc.subject.proposalSeed certificationeng
dc.subject.proposalPapa (Solanum tuberosum y S. phureja)spa
dc.titleDetección molecular y limpieza de virus en material de siembra de Solanum tuberosum y S. phurejaspa
dc.title.translatedMolecular detection and characterization of viruses and development of in vitro virus removal methods in Solanum tuberosum and S. phurejaeng
dc.typeTrabajo de grado - Maestríaspa
dc.type.coarhttp://purl.org/coar/resource_type/c_bdccspa
dc.type.coarversionhttp://purl.org/coar/version/c_ab4af688f83e57aaspa
dc.type.contentTextspa
dc.type.driverinfo:eu-repo/semantics/masterThesisspa
dc.type.redcolhttp://purl.org/redcol/resource_type/TMspa
dc.type.versioninfo:eu-repo/semantics/acceptedVersionspa
dcterms.audience.professionaldevelopmentEstudiantesspa
dcterms.audience.professionaldevelopmentInvestigadoresspa
dcterms.audience.professionaldevelopmentMaestrosspa
dcterms.audience.professionaldevelopmentPúblico generalspa
oaire.accessrightshttp://purl.org/coar/access_right/c_abf2spa
oaire.awardtitlecódigo 1101-805-62787 "Desarrollo de una plataforma molecular y bioinformática para el diagnóstico de virus en cultivos y material de siembra de papa (Solanum tuberosum y S. phureja) en Antioquia"spa
oaire.fundernameFondo de Ciencia, Tecnología e Innovación del Sistema General de Regalías del Departamento de Antioquia (Colombia) (Convenio No. 4600007658–779)spa

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