Synthetic biology toolbox for nitrogen-fixing soil microbes.
Autor(a) principal: | |
---|---|
Data de Publicação: | 2023 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
Texto Completo: | http://www.alice.cnptia.embrapa.br/alice/handle/doc/1159849 https://doi.org/10.1021/acssynbio.3c00414 |
Resumo: | The soil environment adjacent to plant roots, termed the rhizosphere, is home to a wide variety of microorganisms that can significantly affect the physiology of nearby plants. Microbes in the rhizosphere can provide nutrients, secrete signaling compounds, and inhibit pathogens. These processes could be manipulated with synthetic biology to enhance the agricultural performance of crops grown for food, energy, or environmental remediation, if methods can be implemented in these nonmodel microbes. A common first step for domesticating nonmodel organisms is the development of a set of genetic engineering tools, termed a synthetic biology toolbox. A toolbox comprises transformation protocols, replicating vectors, genome engineering (e.g., CRISPR/Cas9), constitutive and inducible promoter systems, and other gene expression control elements. This work validated synthetic biology toolboxes in three nitrogen-fixing soil bacteria: Azotobacter vinelandii, Stutzerimonas stutzeri (Pseudomonas stutzeri), and a new isolate of Klebsiella variicola. All three organisms were amenable to transformation and reporter protein expression, with several functional inducible systems available for each organism. S. stutzeri and K. variicola showed more reliable plasmid-based expression, resulting in successful Cas9 recombineering to create scarless deletions and insertions. Using these tools, we generated mutants with inducible nitrogenase activity and introduced heterologous genes to produce resorcinol products with relevant biological activity in the rhizosphere. |
id |
EMBR_c083ed36c6a277b14e910fc89ca0f6f8 |
---|---|
oai_identifier_str |
oai:www.alice.cnptia.embrapa.br:doc/1159849 |
network_acronym_str |
EMBR |
network_name_str |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
repository_id_str |
2154 |
spelling |
Synthetic biology toolbox for nitrogen-fixing soil microbes.CRISPRCas9Caixa de ferramentasBiologia sintéticaEdição de genomaSoloNitrogenaseSynthetic biologyGenomeThe soil environment adjacent to plant roots, termed the rhizosphere, is home to a wide variety of microorganisms that can significantly affect the physiology of nearby plants. Microbes in the rhizosphere can provide nutrients, secrete signaling compounds, and inhibit pathogens. These processes could be manipulated with synthetic biology to enhance the agricultural performance of crops grown for food, energy, or environmental remediation, if methods can be implemented in these nonmodel microbes. A common first step for domesticating nonmodel organisms is the development of a set of genetic engineering tools, termed a synthetic biology toolbox. A toolbox comprises transformation protocols, replicating vectors, genome engineering (e.g., CRISPR/Cas9), constitutive and inducible promoter systems, and other gene expression control elements. This work validated synthetic biology toolboxes in three nitrogen-fixing soil bacteria: Azotobacter vinelandii, Stutzerimonas stutzeri (Pseudomonas stutzeri), and a new isolate of Klebsiella variicola. All three organisms were amenable to transformation and reporter protein expression, with several functional inducible systems available for each organism. S. stutzeri and K. variicola showed more reliable plasmid-based expression, resulting in successful Cas9 recombineering to create scarless deletions and insertions. Using these tools, we generated mutants with inducible nitrogenase activity and introduced heterologous genes to produce resorcinol products with relevant biological activity in the rhizosphere.MAYA VENKATARAMANAUDREY YÑIGEZ-GUTIERREZVALENTINA INFANTEAPRIL MACINTYREPAULO IVAN FERNANDES JUNIOR, CPATSAJEAN-MICHEL ANÉBRIAN PFLEGER.VENKATARAMAN, M.YÑIGEZ-GUTIERREZ, A.INFANTE, V.MACINTYRE, A.FERNANDES JUNIOR, P. I.ANÉ, J.-M.PFLEGER, B.2023-12-18T13:32:30Z2023-12-18T13:32:30Z2023-12-182023info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleACS Synthetic Biology, v. 12, n. 12, p. 3623-3634, 2023.http://www.alice.cnptia.embrapa.br/alice/handle/doc/1159849https://doi.org/10.1021/acssynbio.3c00414enginfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice)instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)instacron:EMBRAPA2023-12-18T13:32:31Zoai:www.alice.cnptia.embrapa.br:doc/1159849Repositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestopendoar:21542023-12-18T13:32:31falseRepositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestcg-riaa@embrapa.bropendoar:21542023-12-18T13:32:31Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)false |
dc.title.none.fl_str_mv |
Synthetic biology toolbox for nitrogen-fixing soil microbes. |
title |
Synthetic biology toolbox for nitrogen-fixing soil microbes. |
spellingShingle |
Synthetic biology toolbox for nitrogen-fixing soil microbes. VENKATARAMAN, M. CRISPR Cas9 Caixa de ferramentas Biologia sintética Edição de genoma Solo Nitrogenase Synthetic biology Genome |
title_short |
Synthetic biology toolbox for nitrogen-fixing soil microbes. |
title_full |
Synthetic biology toolbox for nitrogen-fixing soil microbes. |
title_fullStr |
Synthetic biology toolbox for nitrogen-fixing soil microbes. |
title_full_unstemmed |
Synthetic biology toolbox for nitrogen-fixing soil microbes. |
title_sort |
Synthetic biology toolbox for nitrogen-fixing soil microbes. |
author |
VENKATARAMAN, M. |
author_facet |
VENKATARAMAN, M. YÑIGEZ-GUTIERREZ, A. INFANTE, V. MACINTYRE, A. FERNANDES JUNIOR, P. I. ANÉ, J.-M. PFLEGER, B. |
author_role |
author |
author2 |
YÑIGEZ-GUTIERREZ, A. INFANTE, V. MACINTYRE, A. FERNANDES JUNIOR, P. I. ANÉ, J.-M. PFLEGER, B. |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
MAYA VENKATARAMAN AUDREY YÑIGEZ-GUTIERREZ VALENTINA INFANTE APRIL MACINTYRE PAULO IVAN FERNANDES JUNIOR, CPATSA JEAN-MICHEL ANÉ BRIAN PFLEGER. |
dc.contributor.author.fl_str_mv |
VENKATARAMAN, M. YÑIGEZ-GUTIERREZ, A. INFANTE, V. MACINTYRE, A. FERNANDES JUNIOR, P. I. ANÉ, J.-M. PFLEGER, B. |
dc.subject.por.fl_str_mv |
CRISPR Cas9 Caixa de ferramentas Biologia sintética Edição de genoma Solo Nitrogenase Synthetic biology Genome |
topic |
CRISPR Cas9 Caixa de ferramentas Biologia sintética Edição de genoma Solo Nitrogenase Synthetic biology Genome |
description |
The soil environment adjacent to plant roots, termed the rhizosphere, is home to a wide variety of microorganisms that can significantly affect the physiology of nearby plants. Microbes in the rhizosphere can provide nutrients, secrete signaling compounds, and inhibit pathogens. These processes could be manipulated with synthetic biology to enhance the agricultural performance of crops grown for food, energy, or environmental remediation, if methods can be implemented in these nonmodel microbes. A common first step for domesticating nonmodel organisms is the development of a set of genetic engineering tools, termed a synthetic biology toolbox. A toolbox comprises transformation protocols, replicating vectors, genome engineering (e.g., CRISPR/Cas9), constitutive and inducible promoter systems, and other gene expression control elements. This work validated synthetic biology toolboxes in three nitrogen-fixing soil bacteria: Azotobacter vinelandii, Stutzerimonas stutzeri (Pseudomonas stutzeri), and a new isolate of Klebsiella variicola. All three organisms were amenable to transformation and reporter protein expression, with several functional inducible systems available for each organism. S. stutzeri and K. variicola showed more reliable plasmid-based expression, resulting in successful Cas9 recombineering to create scarless deletions and insertions. Using these tools, we generated mutants with inducible nitrogenase activity and introduced heterologous genes to produce resorcinol products with relevant biological activity in the rhizosphere. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-12-18T13:32:30Z 2023-12-18T13:32:30Z 2023-12-18 2023 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
ACS Synthetic Biology, v. 12, n. 12, p. 3623-3634, 2023. http://www.alice.cnptia.embrapa.br/alice/handle/doc/1159849 https://doi.org/10.1021/acssynbio.3c00414 |
identifier_str_mv |
ACS Synthetic Biology, v. 12, n. 12, p. 3623-3634, 2023. |
url |
http://www.alice.cnptia.embrapa.br/alice/handle/doc/1159849 https://doi.org/10.1021/acssynbio.3c00414 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa) instacron:EMBRAPA |
instname_str |
Empresa Brasileira de Pesquisa Agropecuária (Embrapa) |
instacron_str |
EMBRAPA |
institution |
EMBRAPA |
reponame_str |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
collection |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) |
repository.name.fl_str_mv |
Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa) |
repository.mail.fl_str_mv |
cg-riaa@embrapa.br |
_version_ |
1794503553962737664 |