Reduced enzymatic browning in potato tubers by specific editing of a Polyphenol oxidase gene via Ribonucleoprotein complexes delivery of the CRISPR/Cas9 System.
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...
| Autores principales: | , , , , , , , , , |
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| Formato: | info:ar-repo/semantics/artículo |
| Lenguaje: | Inglés |
| Publicado: |
Frontiers Media, Switzerland
2020
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| Materias: | |
| Acceso en línea: | 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 |
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| author | 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_browse | Andersson, Mariette Decima Oneto, Cecilia Andrea Feingold, Sergio Enrique Fält, Ann-Sofie Gonzalez, Matías Nicolás Hofvander, Per Massa, Gabriela Alejandra Olsson, Niklas Storani, Leonardo Turesson, Helle |
| 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_sort | Gonzalez, Matías Nicolás |
| collection | INTA Digital |
| 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. |
| format | info:ar-repo/semantics/artículo |
| id | INTA6872 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2020 |
| publishDateRange | 2020 |
| publishDateSort | 2020 |
| publisher | Frontiers Media, Switzerland |
| publisherStr | Frontiers Media, Switzerland |
| record_format | dspace |
| spelling | INTA68722020-03-04T11:04:31Z 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 Massa, Gabriela Alejandra Andersson, Mariette Turesson, Helle Olsson, Niklas Fält, Ann-Sofie Storani, Leonardo Decima Oneto, Cecilia Andrea Hofvander, Per Feingold, Sergio Enrique Papa Solanum Tuberosum Pardeamiento Enzimático Catecol Oxdasa Genomas Potatoes Enzymic Browning Catechol Oxidase Genomes Polifenol Oxidase 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. 2020-03-03T16:45:34Z 2020-03-03T16:45:34Z 2020-01-09 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 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 eng 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) application/pdf Frontiers Media, Switzerland Frontiers in plant science 10 : article 1649. (09 January 2020) |
| spellingShingle | Papa Solanum Tuberosum Pardeamiento Enzimático Catecol Oxdasa Genomas Potatoes Enzymic Browning Catechol Oxidase Genomes Polifenol Oxidase 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 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. |
| 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_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_sort | reduced enzymatic browning in potato tubers by specific editing of a polyphenol oxidase gene via ribonucleoprotein complexes delivery of the crispr cas9 system |
| topic | Papa Solanum Tuberosum Pardeamiento Enzimático Catecol Oxdasa Genomas Potatoes Enzymic Browning Catechol Oxidase Genomes Polifenol Oxidase |
| 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 |
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