Genome editing of soybean inoculant using CRISPR/Cas9 system: enhancing agricultural sustainability

Autores
Serantes, Maria Laura; Stritzler, Margarita; Brambilla, Silvina Maricel; Soto, Gabriela Cynthia; Ayub, Nicolás Daniel
Año de publicación
2024
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
In modern agriculture, inoculants, consisting of various bacteria and fungi, are essential for promoting crop growth and sustainability while reducing reliance on agrochemicals. Despite their benefits, there have been no recent significant advances in enhancing their efficacy. Notably, Bradyrhizobium japonicum E109 and Bradyrhizobium japonicum SEMIA 5079, the most globally utilized, were isolated decades ago, underscoring the need for genetic enhancement. Recently, efforts have focused on selecting spontaneous mutations in genes associated with denitrification and oxidative stress. While this technology has shown promise in reducing nitrous oxide emissions and enhancing root colonization, selecting mutants remains costly and challenging without clear phenotypic markers. The CRISPR/Cas9 system offers a potential solution, although validation is limited to model strains such as Sinorhizobium meliloti 1021. Using the CRISPR/Cas9 system and three sgRNAs, we have edited the genes 16S, napA and glxA from strains E109 and SEMIA5079, generating mutants with streptomycin tolerance, reduced nitrate reductase activity, and increased catalase activity, respectively. Finally, we discuss how the CRISPR/Cas9 system can contribute to bridging the gap between crop and inoculant improvement, and its possible role in producing a new generation of climate-smart inoculants.
Instituto de Biotecnología
Fil: Serantes, Maria Laura. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Serantes, Maria Laura. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina
Fil: Stritzler, Margarita. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Stritzler, Margarita. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina
Fil: Brambilla, Silvina Maricel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Brambilla, Silvina Maricel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina
Fil: Soto, Gabriela Cinthia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Soto, Gabriela Cinthia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina
Fil: Ayub, Nicolás Daniel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Ayub, Nicolás Daniel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina
Fuente
Plant Cell, Tissue and Organ Culture (PCTOC) 157 : 35 (Published online: 22 April 2024)
Materia
Inoculation
Rhizobiaceae
Bacteria
Soybeans
Gene Editing
CRISPR
Inoculación
Soja
Edición de Genes
Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas
Inoculants
Inoculantes
Nivel de accesibilidad
acceso restringido
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:20.500.12123/19060
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oai_identifier_str oai:localhost:20.500.12123/19060
network_acronym_str INTADig
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network_name_str INTA Digital (INTA)
spelling Genome editing of soybean inoculant using CRISPR/Cas9 system: enhancing agricultural sustainabilitySerantes, Maria LauraStritzler, MargaritaBrambilla, Silvina MaricelSoto, Gabriela CynthiaAyub, Nicolás DanielInoculationRhizobiaceaeBacteriaSoybeansGene EditingCRISPRInoculaciónSojaEdición de GenesRepeticiones Palindrómicas Cortas Agrupadas y Regularmente InterespaciadasInoculantsInoculantesIn modern agriculture, inoculants, consisting of various bacteria and fungi, are essential for promoting crop growth and sustainability while reducing reliance on agrochemicals. Despite their benefits, there have been no recent significant advances in enhancing their efficacy. Notably, Bradyrhizobium japonicum E109 and Bradyrhizobium japonicum SEMIA 5079, the most globally utilized, were isolated decades ago, underscoring the need for genetic enhancement. Recently, efforts have focused on selecting spontaneous mutations in genes associated with denitrification and oxidative stress. While this technology has shown promise in reducing nitrous oxide emissions and enhancing root colonization, selecting mutants remains costly and challenging without clear phenotypic markers. The CRISPR/Cas9 system offers a potential solution, although validation is limited to model strains such as Sinorhizobium meliloti 1021. Using the CRISPR/Cas9 system and three sgRNAs, we have edited the genes 16S, napA and glxA from strains E109 and SEMIA5079, generating mutants with streptomycin tolerance, reduced nitrate reductase activity, and increased catalase activity, respectively. Finally, we discuss how the CRISPR/Cas9 system can contribute to bridging the gap between crop and inoculant improvement, and its possible role in producing a new generation of climate-smart inoculants.Instituto de BiotecnologíaFil: Serantes, Maria Laura. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Serantes, Maria Laura. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; ArgentinaFil: Stritzler, Margarita. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Stritzler, Margarita. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; ArgentinaFil: Brambilla, Silvina Maricel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Brambilla, Silvina Maricel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; ArgentinaFil: Soto, Gabriela Cinthia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Soto, Gabriela Cinthia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; ArgentinaFil: Ayub, Nicolás Daniel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Ayub, Nicolás Daniel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; ArgentinaSpringer2024-08-22T10:19:47Z2024-08-22T10:19:47Z2024-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/19060https://link.springer.com/article/10.1007/s11240-024-02764-yhttps://doi.org/10.1007/s11240-024-02764-yPlant Cell, Tissue and Organ Culture (PCTOC) 157 : 35 (Published online: 22 April 2024)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/restrictedAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-09-29T13:46:46Zoai:localhost:20.500.12123/19060instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-29 13:46:46.56INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Genome editing of soybean inoculant using CRISPR/Cas9 system: enhancing agricultural sustainability
title Genome editing of soybean inoculant using CRISPR/Cas9 system: enhancing agricultural sustainability
spellingShingle Genome editing of soybean inoculant using CRISPR/Cas9 system: enhancing agricultural sustainability
Serantes, Maria Laura
Inoculation
Rhizobiaceae
Bacteria
Soybeans
Gene Editing
CRISPR
Inoculación
Soja
Edición de Genes
Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas
Inoculants
Inoculantes
title_short Genome editing of soybean inoculant using CRISPR/Cas9 system: enhancing agricultural sustainability
title_full Genome editing of soybean inoculant using CRISPR/Cas9 system: enhancing agricultural sustainability
title_fullStr Genome editing of soybean inoculant using CRISPR/Cas9 system: enhancing agricultural sustainability
title_full_unstemmed Genome editing of soybean inoculant using CRISPR/Cas9 system: enhancing agricultural sustainability
title_sort Genome editing of soybean inoculant using CRISPR/Cas9 system: enhancing agricultural sustainability
dc.creator.none.fl_str_mv Serantes, Maria Laura
Stritzler, Margarita
Brambilla, Silvina Maricel
Soto, Gabriela Cynthia
Ayub, Nicolás Daniel
author Serantes, Maria Laura
author_facet Serantes, Maria Laura
Stritzler, Margarita
Brambilla, Silvina Maricel
Soto, Gabriela Cynthia
Ayub, Nicolás Daniel
author_role author
author2 Stritzler, Margarita
Brambilla, Silvina Maricel
Soto, Gabriela Cynthia
Ayub, Nicolás Daniel
author2_role author
author
author
author
dc.subject.none.fl_str_mv Inoculation
Rhizobiaceae
Bacteria
Soybeans
Gene Editing
CRISPR
Inoculación
Soja
Edición de Genes
Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas
Inoculants
Inoculantes
topic Inoculation
Rhizobiaceae
Bacteria
Soybeans
Gene Editing
CRISPR
Inoculación
Soja
Edición de Genes
Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas
Inoculants
Inoculantes
dc.description.none.fl_txt_mv In modern agriculture, inoculants, consisting of various bacteria and fungi, are essential for promoting crop growth and sustainability while reducing reliance on agrochemicals. Despite their benefits, there have been no recent significant advances in enhancing their efficacy. Notably, Bradyrhizobium japonicum E109 and Bradyrhizobium japonicum SEMIA 5079, the most globally utilized, were isolated decades ago, underscoring the need for genetic enhancement. Recently, efforts have focused on selecting spontaneous mutations in genes associated with denitrification and oxidative stress. While this technology has shown promise in reducing nitrous oxide emissions and enhancing root colonization, selecting mutants remains costly and challenging without clear phenotypic markers. The CRISPR/Cas9 system offers a potential solution, although validation is limited to model strains such as Sinorhizobium meliloti 1021. Using the CRISPR/Cas9 system and three sgRNAs, we have edited the genes 16S, napA and glxA from strains E109 and SEMIA5079, generating mutants with streptomycin tolerance, reduced nitrate reductase activity, and increased catalase activity, respectively. Finally, we discuss how the CRISPR/Cas9 system can contribute to bridging the gap between crop and inoculant improvement, and its possible role in producing a new generation of climate-smart inoculants.
Instituto de Biotecnología
Fil: Serantes, Maria Laura. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Serantes, Maria Laura. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina
Fil: Stritzler, Margarita. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Stritzler, Margarita. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina
Fil: Brambilla, Silvina Maricel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Brambilla, Silvina Maricel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina
Fil: Soto, Gabriela Cinthia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Soto, Gabriela Cinthia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina
Fil: Ayub, Nicolás Daniel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Ayub, Nicolás Daniel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina
description In modern agriculture, inoculants, consisting of various bacteria and fungi, are essential for promoting crop growth and sustainability while reducing reliance on agrochemicals. Despite their benefits, there have been no recent significant advances in enhancing their efficacy. Notably, Bradyrhizobium japonicum E109 and Bradyrhizobium japonicum SEMIA 5079, the most globally utilized, were isolated decades ago, underscoring the need for genetic enhancement. Recently, efforts have focused on selecting spontaneous mutations in genes associated with denitrification and oxidative stress. While this technology has shown promise in reducing nitrous oxide emissions and enhancing root colonization, selecting mutants remains costly and challenging without clear phenotypic markers. The CRISPR/Cas9 system offers a potential solution, although validation is limited to model strains such as Sinorhizobium meliloti 1021. Using the CRISPR/Cas9 system and three sgRNAs, we have edited the genes 16S, napA and glxA from strains E109 and SEMIA5079, generating mutants with streptomycin tolerance, reduced nitrate reductase activity, and increased catalase activity, respectively. Finally, we discuss how the CRISPR/Cas9 system can contribute to bridging the gap between crop and inoculant improvement, and its possible role in producing a new generation of climate-smart inoculants.
publishDate 2024
dc.date.none.fl_str_mv 2024-08-22T10:19:47Z
2024-08-22T10:19:47Z
2024-04
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/20.500.12123/19060
https://link.springer.com/article/10.1007/s11240-024-02764-y
https://doi.org/10.1007/s11240-024-02764-y
url http://hdl.handle.net/20.500.12123/19060
https://link.springer.com/article/10.1007/s11240-024-02764-y
https://doi.org/10.1007/s11240-024-02764-y
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/restrictedAccess
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 restrictedAccess
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 Springer
publisher.none.fl_str_mv Springer
dc.source.none.fl_str_mv Plant Cell, Tissue and Organ Culture (PCTOC) 157 : 35 (Published online: 22 April 2024)
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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