Reduced enzymatic browning in potato tubers by specific editing of a Polyphenol oxidase gene via Ribonucleoprotein complexes delivery of the CRISPR/Cas9 System.

Autores
Gonzalez, Matías Nicolás; Massa, Gabriela Alejandra; Andersson, Mariette; Turesson, Helle; Olsson, Niklas; Fält, Ann-Sofie; Storani, Leonardo; Decima Oneto, Cecilia Andrea; Hofvander, Per; Feingold, Sergio Enrique
Año de publicación
2020
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Polyphenol Oxidases (PPOs) catalyze the conversion of phenolic substrates to quinones, leading to the formation of dark-colored precipitates in fruits and vegetables. This process, known as enzymatic browning, is the cause of undesirable changes in organoleptic properties and the loss of nutritional quality in plant-derived products. In potato (Solanum tubersoum L.), PPOs are encoded by a multi-gene family with different expression patterns. Here, we have studied the application of the CRISPR/Cas9 system to induce mutations in the StPPO2 gene in the tetraploid cultivar Desiree. We hypothesized that the specific editing of this target gene would result in a lower PPO activity in the tuber with the consequent reduction of the enzymatic browning. Ribonucleoprotein complexes (RNPs), formed by two sgRNAs and Cas9 nuclease, were transfected to potato protoplasts. Up to 68% of regenerated plants contained mutations in at least one allele of the target gene, while 24% of edited lines carried mutations in all four alleles. No off-target mutations were identified in other analyzed StPPO genes. Mutations induced in the four alleles of StPPO2 gene, led to lines with a reduction of up to 69% in tuber PPO activity and a reduction of 73% in enzymatic browning, compared to the control. Our results demonstrate that the CRISPR/Cas9 system can be applied to develop potato varieties with reduced enzymatic browning in tubers, by the specific editing of a single member of the StPPO gene family.
EEA Balcarce
Fil: González, Matías Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina
Fil: Massa, Gabriela Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agraria; Argentina.
Fil: Andersson, Mariette. Swedish University of Agricultural Sciences, Department of Plant Breeding; Suecia
Fil: Turesson, Helle. Swedish University of Agricultural Sciences, Department of Plant Breeding; Suecia
Fil: Olsson, Niklas. Swedish University of Agricultural Sciences, Department of Plant Breeding; Suecia
Fil: Fält, Ann-Sofie. Swedish University of Agricultural Sciences, Department of Plant Breeding; Suecia
Fil: Storani, Leonardo. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina
Fil: Decima Oneto, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina
Fil: Hofvander, Per. Swedish University of Agricultural Sciences, Department of Plant Breeding; Suecia
Fil: Feingold, Sergio. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina.
Fuente
Frontiers in plant science 10 : article 1649. (09 January 2020)
Materia
Papa
Solanum Tuberosum
Pardeamiento Enzimático
Catecol Oxdasa
Genomas
Potatoes
Enzymic Browning
Catechol Oxidase
Genomes
Polifenol Oxidase
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/6872
Acceder
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oai_identifier_str oai:localhost:20.500.12123/6872
network_acronym_str INTADig
repository_id_str l
network_name_str INTA Digital (INTA)
spelling Reduced enzymatic browning in potato tubers by specific editing of a Polyphenol oxidase gene via Ribonucleoprotein complexes delivery of the CRISPR/Cas9 System.Gonzalez, Matías NicolásMassa, Gabriela AlejandraAndersson, MarietteTuresson, HelleOlsson, NiklasFält, Ann-SofieStorani, LeonardoDecima Oneto, Cecilia AndreaHofvander, PerFeingold, Sergio EnriquePapaSolanum TuberosumPardeamiento EnzimáticoCatecol OxdasaGenomasPotatoesEnzymic BrowningCatechol OxidaseGenomesPolifenol OxidasePolyphenol Oxidases (PPOs) catalyze the conversion of phenolic substrates to quinones, leading to the formation of dark-colored precipitates in fruits and vegetables. This process, known as enzymatic browning, is the cause of undesirable changes in organoleptic properties and the loss of nutritional quality in plant-derived products. In potato (Solanum tubersoum L.), PPOs are encoded by a multi-gene family with different expression patterns. Here, we have studied the application of the CRISPR/Cas9 system to induce mutations in the StPPO2 gene in the tetraploid cultivar Desiree. We hypothesized that the specific editing of this target gene would result in a lower PPO activity in the tuber with the consequent reduction of the enzymatic browning. Ribonucleoprotein complexes (RNPs), formed by two sgRNAs and Cas9 nuclease, were transfected to potato protoplasts. Up to 68% of regenerated plants contained mutations in at least one allele of the target gene, while 24% of edited lines carried mutations in all four alleles. No off-target mutations were identified in other analyzed StPPO genes. Mutations induced in the four alleles of StPPO2 gene, led to lines with a reduction of up to 69% in tuber PPO activity and a reduction of 73% in enzymatic browning, compared to the control. Our results demonstrate that the CRISPR/Cas9 system can be applied to develop potato varieties with reduced enzymatic browning in tubers, by the specific editing of a single member of the StPPO gene family.EEA BalcarceFil: González, Matías Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Massa, Gabriela Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agraria; Argentina.Fil: Andersson, Mariette. Swedish University of Agricultural Sciences, Department of Plant Breeding; SueciaFil: Turesson, Helle. Swedish University of Agricultural Sciences, Department of Plant Breeding; SueciaFil: Olsson, Niklas. Swedish University of Agricultural Sciences, Department of Plant Breeding; SueciaFil: Fält, Ann-Sofie. Swedish University of Agricultural Sciences, Department of Plant Breeding; SueciaFil: Storani, Leonardo. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Decima Oneto, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Hofvander, Per. Swedish University of Agricultural Sciences, Department of Plant Breeding; SueciaFil: Feingold, Sergio. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina.Frontiers Media, Switzerland2020-03-03T16:45:34Z2020-03-03T16:45:34Z2020-01-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/6872https://www.frontiersin.org/articles/10.3389/fpls.2019.01649/full1664-462Xhttps://doi.org/10.3389/fpls.2019.01649Frontiers in plant science 10 : article 1649. (09 January 2020)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-09-04T09:48:22Zoai:localhost:20.500.12123/6872instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-04 09:48:23.302INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Reduced enzymatic browning in potato tubers by specific editing of a Polyphenol oxidase gene via Ribonucleoprotein complexes delivery of the CRISPR/Cas9 System.
title Reduced enzymatic browning in potato tubers by specific editing of a Polyphenol oxidase gene via Ribonucleoprotein complexes delivery of the CRISPR/Cas9 System.
spellingShingle Reduced enzymatic browning in potato tubers by specific editing of a Polyphenol oxidase gene via Ribonucleoprotein complexes delivery of the CRISPR/Cas9 System.
Gonzalez, Matías Nicolás
Papa
Solanum Tuberosum
Pardeamiento Enzimático
Catecol Oxdasa
Genomas
Potatoes
Enzymic Browning
Catechol Oxidase
Genomes
Polifenol Oxidase
title_short Reduced enzymatic browning in potato tubers by specific editing of a Polyphenol oxidase gene via Ribonucleoprotein complexes delivery of the CRISPR/Cas9 System.
title_full Reduced enzymatic browning in potato tubers by specific editing of a Polyphenol oxidase gene via Ribonucleoprotein complexes delivery of the CRISPR/Cas9 System.
title_fullStr Reduced enzymatic browning in potato tubers by specific editing of a Polyphenol oxidase gene via Ribonucleoprotein complexes delivery of the CRISPR/Cas9 System.
title_full_unstemmed Reduced enzymatic browning in potato tubers by specific editing of a Polyphenol oxidase gene via Ribonucleoprotein complexes delivery of the CRISPR/Cas9 System.
title_sort Reduced enzymatic browning in potato tubers by specific editing of a Polyphenol oxidase gene via Ribonucleoprotein complexes delivery of the CRISPR/Cas9 System.
dc.creator.none.fl_str_mv Gonzalez, Matías Nicolás
Massa, Gabriela Alejandra
Andersson, Mariette
Turesson, Helle
Olsson, Niklas
Fält, Ann-Sofie
Storani, Leonardo
Decima Oneto, Cecilia Andrea
Hofvander, Per
Feingold, Sergio Enrique
author Gonzalez, Matías Nicolás
author_facet Gonzalez, Matías Nicolás
Massa, Gabriela Alejandra
Andersson, Mariette
Turesson, Helle
Olsson, Niklas
Fält, Ann-Sofie
Storani, Leonardo
Decima Oneto, Cecilia Andrea
Hofvander, Per
Feingold, Sergio Enrique
author_role author
author2 Massa, Gabriela Alejandra
Andersson, Mariette
Turesson, Helle
Olsson, Niklas
Fält, Ann-Sofie
Storani, Leonardo
Decima Oneto, Cecilia Andrea
Hofvander, Per
Feingold, Sergio Enrique
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Papa
Solanum Tuberosum
Pardeamiento Enzimático
Catecol Oxdasa
Genomas
Potatoes
Enzymic Browning
Catechol Oxidase
Genomes
Polifenol Oxidase
topic Papa
Solanum Tuberosum
Pardeamiento Enzimático
Catecol Oxdasa
Genomas
Potatoes
Enzymic Browning
Catechol Oxidase
Genomes
Polifenol Oxidase
dc.description.none.fl_txt_mv Polyphenol Oxidases (PPOs) catalyze the conversion of phenolic substrates to quinones, leading to the formation of dark-colored precipitates in fruits and vegetables. This process, known as enzymatic browning, is the cause of undesirable changes in organoleptic properties and the loss of nutritional quality in plant-derived products. In potato (Solanum tubersoum L.), PPOs are encoded by a multi-gene family with different expression patterns. Here, we have studied the application of the CRISPR/Cas9 system to induce mutations in the StPPO2 gene in the tetraploid cultivar Desiree. We hypothesized that the specific editing of this target gene would result in a lower PPO activity in the tuber with the consequent reduction of the enzymatic browning. Ribonucleoprotein complexes (RNPs), formed by two sgRNAs and Cas9 nuclease, were transfected to potato protoplasts. Up to 68% of regenerated plants contained mutations in at least one allele of the target gene, while 24% of edited lines carried mutations in all four alleles. No off-target mutations were identified in other analyzed StPPO genes. Mutations induced in the four alleles of StPPO2 gene, led to lines with a reduction of up to 69% in tuber PPO activity and a reduction of 73% in enzymatic browning, compared to the control. Our results demonstrate that the CRISPR/Cas9 system can be applied to develop potato varieties with reduced enzymatic browning in tubers, by the specific editing of a single member of the StPPO gene family.
EEA Balcarce
Fil: González, Matías Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina
Fil: Massa, Gabriela Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agraria; Argentina.
Fil: Andersson, Mariette. Swedish University of Agricultural Sciences, Department of Plant Breeding; Suecia
Fil: Turesson, Helle. Swedish University of Agricultural Sciences, Department of Plant Breeding; Suecia
Fil: Olsson, Niklas. Swedish University of Agricultural Sciences, Department of Plant Breeding; Suecia
Fil: Fält, Ann-Sofie. Swedish University of Agricultural Sciences, Department of Plant Breeding; Suecia
Fil: Storani, Leonardo. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina
Fil: Decima Oneto, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina
Fil: Hofvander, Per. Swedish University of Agricultural Sciences, Department of Plant Breeding; Suecia
Fil: Feingold, Sergio. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina.
description Polyphenol Oxidases (PPOs) catalyze the conversion of phenolic substrates to quinones, leading to the formation of dark-colored precipitates in fruits and vegetables. This process, known as enzymatic browning, is the cause of undesirable changes in organoleptic properties and the loss of nutritional quality in plant-derived products. In potato (Solanum tubersoum L.), PPOs are encoded by a multi-gene family with different expression patterns. Here, we have studied the application of the CRISPR/Cas9 system to induce mutations in the StPPO2 gene in the tetraploid cultivar Desiree. We hypothesized that the specific editing of this target gene would result in a lower PPO activity in the tuber with the consequent reduction of the enzymatic browning. Ribonucleoprotein complexes (RNPs), formed by two sgRNAs and Cas9 nuclease, were transfected to potato protoplasts. Up to 68% of regenerated plants contained mutations in at least one allele of the target gene, while 24% of edited lines carried mutations in all four alleles. No off-target mutations were identified in other analyzed StPPO genes. Mutations induced in the four alleles of StPPO2 gene, led to lines with a reduction of up to 69% in tuber PPO activity and a reduction of 73% in enzymatic browning, compared to the control. Our results demonstrate that the CRISPR/Cas9 system can be applied to develop potato varieties with reduced enzymatic browning in tubers, by the specific editing of a single member of the StPPO gene family.
publishDate 2020
dc.date.none.fl_str_mv 2020-03-03T16:45:34Z
2020-03-03T16:45:34Z
2020-01-09
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12123/6872
https://www.frontiersin.org/articles/10.3389/fpls.2019.01649/full
1664-462X
https://doi.org/10.3389/fpls.2019.01649
url http://hdl.handle.net/20.500.12123/6872
https://www.frontiersin.org/articles/10.3389/fpls.2019.01649/full
https://doi.org/10.3389/fpls.2019.01649
identifier_str_mv 1664-462X
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Frontiers Media, Switzerland
publisher.none.fl_str_mv Frontiers Media, Switzerland
dc.source.none.fl_str_mv Frontiers in plant science 10 : article 1649. (09 January 2020)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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