Mejoramiento genético de lechuga a través del knock out del gen LsSPL13 vía CRISPR/Cas9

The transcription factor SPL13 is related to vegetative growth and development. Its silencing in alfalfa produces plants with a longer vegetative phase and an increase in leaf area, characteristics of interest for lettuce production. The objective of this thesis was to implement gene editing via CRI...

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Autor principal: Beracochea, Valeria Cecilia
Otros Autores: López Bilbao, Marisa
Formato: Tesis doctoral acceptedVersion
Lenguaje:Español
Publicado: Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica 2024
Materias:
Lechuga
Edición génica
CRISPR/Cas9
Knock out
SPL13
Biomasa
Floración
Lettuce
Gene editing
Biomass
Flowering.
Ciencias de la vida
Acceso en línea:http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=posgraafa&cl=CL1&d=HWA_7846
https://repositoriouba.sisbi.uba.ar/gsdl/collect/posgraafa/index/assoc/HWA_7846.dir/7846.PDF
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Repositorio Digital de la Universidad de Buenos Aires (UBA) de Universidad de Buenos Aires
id I28-R145-HWA_7846
record_format dspace
spelling I28-R145-HWA_78462025-08-01 The transcription factor SPL13 is related to vegetative growth and development. Its silencing in alfalfa produces plants with a longer vegetative phase and an increase in leaf area, characteristics of interest for lettuce production. The objective of this thesis was to implement gene editing via CRISPR/Cas9 in lettuce (Lactuca sativa), through the knock out of the LsSPL13 gene and to obtain plants with increased biomass and delayed flowering. During this thesis, the lettuce SPL13 gene was identified, 3 guide RNAs were designed and the editing vector was assembled and introduced via Agrobacterium tumefaciens into lettuce. The 20 transgenic events obtained were molecularly characterized, resulting in 3 edited lines. Phenotypic analysis of E1 and E2 lines, up to the T3 generation, showed in comparison with the control plants: increased biomass, greater number of leaves, delayed flowering, larger seeds and greater number of secondary roots, while no differences were observed in the centesimal composition analysis. Furthermore, E1 would be an edited non-transgenic line. In this way, the stated objectives were achieved and the biotechnological interest of the SPL13 gene was demonstrated. Fil: Beracochea, Valeria Cecilia. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Buenos Aires, Argentina López Bilbao, Marisa Rodríguez Talou, Julián Soto, Gabriela C. Beracochea, Valeria Cecilia 2024-04-08 El factor de transcripción SPL13 está relacionado con el crecimiento y desarrollo vegetativo. Su silenciamiento en alfalfa produce plantas con una fase vegetativa más larga y un aumento del área foliar, características de interés para la producción de lechuga. El objetivo de esta tesis fue implementar y poner a punto la tecnología de edición génica vía CRISPR/Cas9 en lechuga (Lactuca sativa), a través del knock out del gen LsSPL13 y obtener plantas con incremento de biomasa y retraso en la floración. Durante esta tesis se identificó el gen SPL13 de lechuga, se diseñaron 3 ARN guías y se ensambló el vector de edición, que se introdujo vía Agrobacterium tumefaciens en lechuga. Se obtuvieron 20 eventos transgénicos y por secuenciación, 3 líneas resultaron editadas. El análisis fenotípico de las líneas E1 y E2, hasta la generación T3, mostró en comparación con las plantas control: aumento de biomasa, mayor número de hojas, retraso de floración, semillas más grandes y mayor cantidad de raíces secundarias, mientras que no se observaron diferencias en el análisis de composición centesimal. Además, E1 sería editada no transgénica. De esta forma, se lograron los objetivos planteados y se demostró el potencial biotecnológico del gen SPL13. application/pdf Gallego, Susana M. Capiati,, Daniela Feingold, Sergio Lechuga Edición génica CRISPR/Cas9 Knock out SPL13 Biomasa Floración Lettuce Gene editing CRISPR/Cas9 Knock out SPL13 Biomass Flowering. spa Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica http://creativecommons.org/licenses/by-nc-nd/2.5/ar/ Ciencias de la vida Doctora de la Universidad de Buenos Aires en Ciencias Biológicas Mejoramiento genético de lechuga a través del knock out del gen LsSPL13 vía CRISPR/Cas9 info:eu-repo/semantics/doctoralThesis info:ar-repo/semantics/tesis doctoral info:eu-repo/semantics/acceptedVersion http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=posgraafa&cl=CL1&d=HWA_7846 https://repositoriouba.sisbi.uba.ar/gsdl/collect/posgraafa/index/assoc/HWA_7846.dir/7846.PDF
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-145
collection Repositorio Digital de la Universidad de Buenos Aires (UBA)
language Español
orig_language_str_mv spa
topic Lechuga
Edición génica
CRISPR/Cas9
Knock out
SPL13
Biomasa
Floración
Lettuce
Gene editing
CRISPR/Cas9
Knock out
SPL13
Biomass
Flowering.
Ciencias de la vida
spellingShingle Lechuga
Edición génica
CRISPR/Cas9
Knock out
SPL13
Biomasa
Floración
Lettuce
Gene editing
CRISPR/Cas9
Knock out
SPL13
Biomass
Flowering.
Ciencias de la vida
Beracochea, Valeria Cecilia
Mejoramiento genético de lechuga a través del knock out del gen LsSPL13 vía CRISPR/Cas9
topic_facet Lechuga
Edición génica
CRISPR/Cas9
Knock out
SPL13
Biomasa
Floración
Lettuce
Gene editing
CRISPR/Cas9
Knock out
SPL13
Biomass
Flowering.
Ciencias de la vida
description The transcription factor SPL13 is related to vegetative growth and development. Its silencing in alfalfa produces plants with a longer vegetative phase and an increase in leaf area, characteristics of interest for lettuce production. The objective of this thesis was to implement gene editing via CRISPR/Cas9 in lettuce (Lactuca sativa), through the knock out of the LsSPL13 gene and to obtain plants with increased biomass and delayed flowering. During this thesis, the lettuce SPL13 gene was identified, 3 guide RNAs were designed and the editing vector was assembled and introduced via Agrobacterium tumefaciens into lettuce. The 20 transgenic events obtained were molecularly characterized, resulting in 3 edited lines. Phenotypic analysis of E1 and E2 lines, up to the T3 generation, showed in comparison with the control plants: increased biomass, greater number of leaves, delayed flowering, larger seeds and greater number of secondary roots, while no differences were observed in the centesimal composition analysis. Furthermore, E1 would be an edited non-transgenic line. In this way, the stated objectives were achieved and the biotechnological interest of the SPL13 gene was demonstrated.
author2 López Bilbao, Marisa
author_facet López Bilbao, Marisa
Beracochea, Valeria Cecilia
format Tesis doctoral
Tesis doctoral
acceptedVersion
author Beracochea, Valeria Cecilia
author_sort Beracochea, Valeria Cecilia
title Mejoramiento genético de lechuga a través del knock out del gen LsSPL13 vía CRISPR/Cas9
title_short Mejoramiento genético de lechuga a través del knock out del gen LsSPL13 vía CRISPR/Cas9
title_full Mejoramiento genético de lechuga a través del knock out del gen LsSPL13 vía CRISPR/Cas9
title_fullStr Mejoramiento genético de lechuga a través del knock out del gen LsSPL13 vía CRISPR/Cas9
title_full_unstemmed Mejoramiento genético de lechuga a través del knock out del gen LsSPL13 vía CRISPR/Cas9
title_sort mejoramiento genético de lechuga a través del knock out del gen lsspl13 vía crispr/cas9
publisher Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica
publishDate 2024
url http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=posgraafa&cl=CL1&d=HWA_7846
https://repositoriouba.sisbi.uba.ar/gsdl/collect/posgraafa/index/assoc/HWA_7846.dir/7846.PDF
work_keys_str_mv AT beracocheavaleriacecilia mejoramientogeneticodelechugaatravesdelknockoutdelgenlsspl13viacrisprcas9
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