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A Simple in situ Assay to Assess Plant-Associative Bacterial Nitrogenase Activity

Assessment of plant-associative bacterial nitrogen (N) fixation is crucial for selection and development of elite diazotrophic inoculants that could be used to supply cereal crops with nitrogen in a sustainable manner. Although diazotrophic bacteria possess diverse oxygen tolerance mechanisms, most...

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Autores principales: Haskett, Timothy L., Knights, Hayley E., Jorrin, Beatriz, Mendes, Marta D., Poole, Philip S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8261070/
https://www.ncbi.nlm.nih.gov/pubmed/34248916
http://dx.doi.org/10.3389/fmicb.2021.690439
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author Haskett, Timothy L.
Knights, Hayley E.
Jorrin, Beatriz
Mendes, Marta D.
Poole, Philip S.
author_facet Haskett, Timothy L.
Knights, Hayley E.
Jorrin, Beatriz
Mendes, Marta D.
Poole, Philip S.
author_sort Haskett, Timothy L.
collection PubMed
description Assessment of plant-associative bacterial nitrogen (N) fixation is crucial for selection and development of elite diazotrophic inoculants that could be used to supply cereal crops with nitrogen in a sustainable manner. Although diazotrophic bacteria possess diverse oxygen tolerance mechanisms, most require a sub 21% oxygen environment to achieve optimal stability and function of the N-fixing catalyst nitrogenase. Consequently, assessment of N fixation is routinely carried out on “free-living” bacteria grown in the absence of a host plant and such experiments may not accurately divulge activity in the rhizosphere where the availability and forms of nutrients such as carbon and N, which are key regulators of N fixation, may vary widely. Here, we present a modified in situ acetylene reduction assay (ARA), utilizing the model cereal barley as a host to comparatively assess nitrogenase activity in diazotrophic bacteria. The assay is rapid, highly reproducible, applicable to a broad range of diazotrophs, and can be performed with simple equipment commonly found in most laboratories that investigate plant-microbe interactions. Thus, the assay could serve as a first point of order for high-throughput identification of elite plant-associative diazotrophs.
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spelling pubmed-82610702021-07-08 A Simple in situ Assay to Assess Plant-Associative Bacterial Nitrogenase Activity Haskett, Timothy L. Knights, Hayley E. Jorrin, Beatriz Mendes, Marta D. Poole, Philip S. Front Microbiol Microbiology Assessment of plant-associative bacterial nitrogen (N) fixation is crucial for selection and development of elite diazotrophic inoculants that could be used to supply cereal crops with nitrogen in a sustainable manner. Although diazotrophic bacteria possess diverse oxygen tolerance mechanisms, most require a sub 21% oxygen environment to achieve optimal stability and function of the N-fixing catalyst nitrogenase. Consequently, assessment of N fixation is routinely carried out on “free-living” bacteria grown in the absence of a host plant and such experiments may not accurately divulge activity in the rhizosphere where the availability and forms of nutrients such as carbon and N, which are key regulators of N fixation, may vary widely. Here, we present a modified in situ acetylene reduction assay (ARA), utilizing the model cereal barley as a host to comparatively assess nitrogenase activity in diazotrophic bacteria. The assay is rapid, highly reproducible, applicable to a broad range of diazotrophs, and can be performed with simple equipment commonly found in most laboratories that investigate plant-microbe interactions. Thus, the assay could serve as a first point of order for high-throughput identification of elite plant-associative diazotrophs. Frontiers Media S.A. 2021-06-23 /pmc/articles/PMC8261070/ /pubmed/34248916 http://dx.doi.org/10.3389/fmicb.2021.690439 Text en Copyright © 2021 Haskett, Knights, Jorrin, Mendes and Poole. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Haskett, Timothy L.
Knights, Hayley E.
Jorrin, Beatriz
Mendes, Marta D.
Poole, Philip S.
A Simple in situ Assay to Assess Plant-Associative Bacterial Nitrogenase Activity
title A Simple in situ Assay to Assess Plant-Associative Bacterial Nitrogenase Activity
title_full A Simple in situ Assay to Assess Plant-Associative Bacterial Nitrogenase Activity
title_fullStr A Simple in situ Assay to Assess Plant-Associative Bacterial Nitrogenase Activity
title_full_unstemmed A Simple in situ Assay to Assess Plant-Associative Bacterial Nitrogenase Activity
title_short A Simple in situ Assay to Assess Plant-Associative Bacterial Nitrogenase Activity
title_sort simple in situ assay to assess plant-associative bacterial nitrogenase activity
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8261070/
https://www.ncbi.nlm.nih.gov/pubmed/34248916
http://dx.doi.org/10.3389/fmicb.2021.690439
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