Development of tools for genetic transformation and tissue regeneration in plants

Climate change is negatively affecting agricultural production. Therefore, it is crucial to develop new plants that are tolerant or resistant to abiotic and biotic stresses. Biotechnological tools, such as genetic modification and genome editing, can be used to rapidly create plants with new traits....

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Detalhes bibliográficos
Autor: Yaroshko, Olha
Tipo de documento: tese
Data de publicação:2024
País:España
Recursos:Universidad Miguel Hernández de Elche
Repositório:REDIUMH. Depósito Digital de la UMH
OAI Identifier:oai:dspace.umh.es:11000/35631
Acesso em linha:https://hdl.handle.net/11000/35631
Access Level:Acceso aberto
Palavra-chave:CDU::5 - Ciencias puras y naturales::57 - Biología
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repository_id_str
dc.title.none.fl_str_mv Development of tools for genetic transformation and tissue regeneration in plants
title Development of tools for genetic transformation and tissue regeneration in plants
spellingShingle Development of tools for genetic transformation and tissue regeneration in plants
Yaroshko, Olha
CDU::5 - Ciencias puras y naturales::57 - Biología
title_short Development of tools for genetic transformation and tissue regeneration in plants
title_full Development of tools for genetic transformation and tissue regeneration in plants
title_fullStr Development of tools for genetic transformation and tissue regeneration in plants
title_full_unstemmed Development of tools for genetic transformation and tissue regeneration in plants
title_sort Development of tools for genetic transformation and tissue regeneration in plants
dc.creator.none.fl_str_mv Yaroshko, Olha
author Yaroshko, Olha
author_facet Yaroshko, Olha
author_role author
dc.contributor.none.fl_str_mv Pérez Pérez, José Manuel
Larriba Tornel, Eduardo
Departamentos de la UMH::Biología Aplicada
dc.subject.none.fl_str_mv CDU::5 - Ciencias puras y naturales::57 - Biología
topic CDU::5 - Ciencias puras y naturales::57 - Biología
description Climate change is negatively affecting agricultural production. Therefore, it is crucial to develop new plants that are tolerant or resistant to abiotic and biotic stresses. Biotechnological tools, such as genetic modification and genome editing, can be used to rapidly create plants with new traits. Plant biotechnology utilizes gene editing tools such as zinc finger nucleases, transcription activator-like effector nucleases, and clustered regularly spaced short palindromic repeats (CRISPR)/CRISPR-associated (CRISPR/Cas) proteins. While plant genome editing is a new field, most editing systems are based on established genetic transformation methods. The delivery methods for the editing tools, such as Agrobacterium-mediated delivery, biolistics, and polyethylene glycol-mediated transformation, are the same as those used for classic genetic transformation. To achieve successful editing, intermediate steps such as plant regeneration and transformation must be optimized beforehand. This thesis aims to determine the optimal conditions for Agrobacterium tumefaciens-mediated transformation of Amaranthus caudatus cultivars and establish a regeneration methodology for Solanum lycopersicum and heirloom tomato genotypes. An effective methodology for transforming Amaranthus caudatus cultivars, Karmin and Helios, was established, with results obtained within four days of the experiment’s start due to the speed of the transformation method. The procedure for obtaining and regenerating calli was developed for three commercial tomato varieties and three wild accessions. Optimal conditions for obtaining the maximum number of regenerated shoots and callus formation were evaluated for all the tomatoes studied. Our method allows for obtaining initial results in less than two weeks. The factors that significantly influence the percentage of regeneration and callus formation were assessed.
publishDate 2024
dc.date.none.fl_str_mv 2024
2025
2025
dc.type.none.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
dc.identifier.none.fl_str_mv https://hdl.handle.net/11000/35631
url https://hdl.handle.net/11000/35631
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
rights_invalid_str_mv Attribution-NonCommercial-NoDerivatives 4.0 Internacional
http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.format.none.fl_str_mv application/pdf
144
application/pdf
dc.publisher.none.fl_str_mv Universidad Miguel Hernández de Elche
publisher.none.fl_str_mv Universidad Miguel Hernández de Elche
dc.source.none.fl_str_mv reponame:REDIUMH. Depósito Digital de la UMH
instname:Universidad Miguel Hernández de Elche
instname_str Universidad Miguel Hernández de Elche
reponame_str REDIUMH. Depósito Digital de la UMH
collection REDIUMH. Depósito Digital de la UMH
repository.name.fl_str_mv
repository.mail.fl_str_mv
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spelling Development of tools for genetic transformation and tissue regeneration in plantsYaroshko, OlhaCDU::5 - Ciencias puras y naturales::57 - BiologíaClimate change is negatively affecting agricultural production. Therefore, it is crucial to develop new plants that are tolerant or resistant to abiotic and biotic stresses. Biotechnological tools, such as genetic modification and genome editing, can be used to rapidly create plants with new traits. Plant biotechnology utilizes gene editing tools such as zinc finger nucleases, transcription activator-like effector nucleases, and clustered regularly spaced short palindromic repeats (CRISPR)/CRISPR-associated (CRISPR/Cas) proteins. While plant genome editing is a new field, most editing systems are based on established genetic transformation methods. The delivery methods for the editing tools, such as Agrobacterium-mediated delivery, biolistics, and polyethylene glycol-mediated transformation, are the same as those used for classic genetic transformation. To achieve successful editing, intermediate steps such as plant regeneration and transformation must be optimized beforehand. This thesis aims to determine the optimal conditions for Agrobacterium tumefaciens-mediated transformation of Amaranthus caudatus cultivars and establish a regeneration methodology for Solanum lycopersicum and heirloom tomato genotypes. An effective methodology for transforming Amaranthus caudatus cultivars, Karmin and Helios, was established, with results obtained within four days of the experiment’s start due to the speed of the transformation method. The procedure for obtaining and regenerating calli was developed for three commercial tomato varieties and three wild accessions. Optimal conditions for obtaining the maximum number of regenerated shoots and callus formation were evaluated for all the tomatoes studied. Our method allows for obtaining initial results in less than two weeks. The factors that significantly influence the percentage of regeneration and callus formation were assessed.El cambio climático está afectando negativamente a la producción agrícola. Por eso es crucial desarrollar nuevas plantas tolerantes o resistentes al estrés abiótico y biótico. Las herramientas biotecnológicas, como la modificación genética y la edición del genoma, pueden utilizarse para crear rápidamente plantas con nuevos rasgos. La biotecnología vegetal utiliza herramientas de edición genética como las nucleasas de dedos de zinc, las nucleasas efectoras similares a activadores de la transcripción y las proteínas asociadas a repeticiones palindrómicas cortas agrupadas regularmente espaciadas (CRISPR)/Cas. Aunque la edición del genoma vegetal es un campo nuevo, la mayoría de los sistemas de edición se basan en métodos establecidos de transformación genética. Los métodos de administración de las herramientas de edición, como la administración mediada por Agrobacterium spp., la biolística y la transformación mediada por polietilenglicol, son los mismos que los utilizados para la transformación genética clásica. Para que la edición genética tenga éxito, es necesario optimizar previamente los pasos intermedios, como la regeneración de la planta y la transformación. Esta tesis pretende determinar las condiciones óptimas para la transformación mediada por Agrobacterium tumefaciens de cultivares de Amaranthus caudatus y establecer una metodología de regeneración para genotipos de Solanum lycopersicum. Se estableció una metodología eficaz para transformar los cultivares de Amaranthus caudatus, Karmin y Helios, con resultados obtenidos a los cuatro días del inicio del experimento debido a la rapidez del método de transformación. En esta tesis se optimizó el procedimiento de obtención y regeneración de callos para tres variedades comerciales de tomate y tres entradas silvestres. Se evaluaron las condiciones óptimas para obtener el máximo número de brotes regenerados y la formación de callo para todos los tomates estudiados. Nuestro método permite obtener resultados iniciales en menos de dos semanas. Se evaluaron también los factores que influyen significativamente en el porcentaje de regeneración y formación de callo.Universidad Miguel Hernández de ElchePérez Pérez, José ManuelLarriba Tornel, EduardoDepartamentos de la UMH::Biología Aplicada202520252024info:eu-repo/semantics/doctoralThesisapplication/pdf144application/pdfhttps://hdl.handle.net/11000/35631reponame:REDIUMH. Depósito Digital de la UMHinstname:Universidad Miguel Hernández de ElcheInglésinfo:eu-repo/semantics/openAccessAttribution-NonCommercial-NoDerivatives 4.0 Internacionalhttp://creativecommons.org/licenses/by-nc-nd/4.0/oai:dspace.umh.es:11000/356312026-05-27T13:36:21Z
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