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      Regulation of indole‐3‐acetic acid biosynthesis and consequences of auxin production deficiency in Serratia plymuthica

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          Abstract

          Indole‐3‐acetic acid (IAA) is emerging as a key intra‐ and inter‐kingdom signal molecule that modulates a wide range of processes of importance during plant–microorganism interaction. However, the mechanisms by which IAA carries out its functions in bacteria as well as the regulatory processes by which bacteria modulate auxin production are largely unknown. Here, we found that IAA synthesis deficiency results in important global transcriptional changes in the broad‐range antibiotic‐producing rhizobacterium Serratia plymuthica A153. Most pronounced transcriptional changes were observed in various gene clusters for aromatic acid metabolism, including auxin catabolism. To delve into the corresponding molecular mechanisms, different regulatory proteins were biochemically characterized. Among them, a TyrR orthologue was essential for IAA production through the activation of the ipdc gene encoding a key enzyme for IAA biosynthesis. We showed that TyrR specifically recognizes different aromatic amino acids which, in turn, alters the interactions of TyrR with the ipdc promoter. Screening of mutants defective in various transcriptional and post‐transcriptional regulators allowed the identification of additional regulators of IAA production, including PigP and quorum sensing‐related genes. Advancing our knowledge on the mechanisms that control the IAA biosynthesis in beneficial phytobacteria is of biotechnological interest for improving agricultural productivity and sustainable agricultural development.

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          A new mathematical model for relative quantification in real-time RT-PCR.

          M. Pfaffl (2001)
          Use of the real-time polymerase chain reaction (PCR) to amplify cDNA products reverse transcribed from mRNA is on the way to becoming a routine tool in molecular biology to study low abundance gene expression. Real-time PCR is easy to perform, provides the necessary accuracy and produces reliable as well as rapid quantification results. But accurate quantification of nucleic acids requires a reproducible methodology and an adequate mathematical model for data analysis. This study enters into the particular topics of the relative quantification in real-time RT-PCR of a target gene transcript in comparison to a reference gene transcript. Therefore, a new mathematical model is presented. The relative expression ratio is calculated only from the real-time PCR efficiencies and the crossing point deviation of an unknown sample versus a control. This model needs no calibration curve. Control levels were included in the model to standardise each reaction run with respect to RNA integrity, sample loading and inter-PCR variations. High accuracy and reproducibility (<2.5% variation) were reached in LightCycler PCR using the established mathematical model.
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            The Phyre2 web portal for protein modeling, prediction and analysis.

            Phyre2 is a suite of tools available on the web to predict and analyze protein structure, function and mutations. The focus of Phyre2 is to provide biologists with a simple and intuitive interface to state-of-the-art protein bioinformatics tools. Phyre2 replaces Phyre, the original version of the server for which we previously published a paper in Nature Protocols. In this updated protocol, we describe Phyre2, which uses advanced remote homology detection methods to build 3D models, predict ligand binding sites and analyze the effect of amino acid variants (e.g., nonsynonymous SNPs (nsSNPs)) for a user's protein sequence. Users are guided through results by a simple interface at a level of detail they determine. This protocol will guide users from submitting a protein sequence to interpreting the secondary and tertiary structure of their models, their domain composition and model quality. A range of additional available tools is described to find a protein structure in a genome, to submit large number of sequences at once and to automatically run weekly searches for proteins that are difficult to model. The server is available at http://www.sbg.bio.ic.ac.uk/phyre2. A typical structure prediction will be returned between 30 min and 2 h after submission.
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              Experiments in Molecular Genetics

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                Author and article information

                Contributors
                miguel.matilla@eez.csic.es
                Journal
                Microb Biotechnol
                Microb Biotechnol
                10.1111/(ISSN)1751-7915
                MBT2
                Microbial Biotechnology
                John Wiley and Sons Inc. (Hoboken )
                1751-7915
                22 June 2023
                August 2023
                : 16
                : 8 , Special Issue Part 1: End Hunger: Enhancing Crop Yields with Microbes ( doiID: 10.1111/mbt2.v16.8 )
                : 1671-1689
                Affiliations
                [ 1 ] Department of Biotechnology and Environmental Protection, Estación Experimental del Zaidín Consejo Superior de Investigaciones Científicas Granada Spain
                Author notes
                [*] [* ] Correspondence

                Miguel A. Matilla, Department of Biotechnology and Environmental Protection, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Prof. Albareda 1, Granada 18008, Spain.

                Email: miguel.matilla@ 123456eez.csic.es

                Author information
                https://orcid.org/0000-0002-0126-1478
                https://orcid.org/0000-0002-9040-3166
                https://orcid.org/0000-0002-8468-9604
                Article
                MBT214296 MICROBIO-2023-142-RA.R1
                10.1111/1751-7915.14296
                10364317
                37345981
                ea85f6a1-6322-4901-854f-97a3b92f0b2e
                © 2023 The Authors. Microbial Biotechnology published by Applied Microbiology International and John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 02 June 2023
                : 10 April 2023
                : 04 June 2023
                Page count
                Figures: 9, Tables: 2, Pages: 19, Words: 12893
                Funding
                Funded by: CSIC grant
                Award ID: 2023AEP002
                Funded by: JAE Intro grant of the CSIC
                Award ID: JAEINT21_EX_0605
                Funded by: Junta de Andalucía , doi 10.13039/501100011011;
                Award ID: P18‐FR‐1621
                Funded by: Secretaría General de Universidades, Investigación y Tecnología de la Junta de Andalucía
                Award ID: AND21_EEZ_M2_044
                Funded by: Spanish Ministry for Science and Innovation
                Award ID: PID2019‐103972GA‐I00
                Award ID: PID2020‐112612GB‐I00
                Categories
                Research Article
                Research Articles
                Custom metadata
                2.0
                August 2023
                Converter:WILEY_ML3GV2_TO_JATSPMC version:6.3.2 mode:remove_FC converted:24.07.2023

                Biotechnology
                Biotechnology

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