Synthetic biology toolbox for nitrogen-fixing soil microbes.

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dc.contributorMAYA VENKATARAMAN
dc.contributorAUDREY YÑIGEZ-GUTIERREZ
dc.contributorVALENTINA INFANTE
dc.contributorAPRIL MACINTYRE
dc.contributorPAULO IVAN FERNANDES JUNIOR, CPATSA
dc.contributorJEAN-MICHEL ANÉ
dc.contributorBRIAN PFLEGER.
dc.creatorVENKATARAMAN, M.
dc.creatorYÑIGEZ-GUTIERREZ, A.
dc.creatorINFANTE, V.
dc.creatorMACINTYRE, A.
dc.creatorFERNANDES JUNIOR, P. I.
dc.creatorANÉ, J.-M.
dc.creatorPFLEGER, B.
dc.date2023-12-18T13:32:30Z
dc.date2023-12-18T13:32:30Z
dc.date2023-12-18
dc.date2023
dc.date.accessioned2026-07-07T05:58:25Z
dc.descriptionThe 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.
dc.identifierACS Synthetic Biology, v. 12, n. 12, p. 3623-3634, 2023.
dc.identifierhttp://www.alice.cnptia.embrapa.br/alice/handle/doc/1159849
dc.identifierhttps://doi.org/10.1021/acssynbio.3c00414
dc.identifier.urihttp://hdl.handle.net/123456789/503662
dc.languageeng
dc.rightsopenAccess
dc.subjectCRISPR
dc.subjectCas9
dc.subjectCaixa de ferramentas
dc.subjectBiologia sintética
dc.subjectEdição de genoma
dc.subjectSolo
dc.subjectNitrogenase
dc.subjectSynthetic biology
dc.subjectGenome
dc.titleSynthetic biology toolbox for nitrogen-fixing soil microbes.
dc.typeArtigo de periódico

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