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      Interactions between plant‐beneficial microorganisms in a consortium: Streptomyces microflavus and Trichoderma harzianum

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          Abstract

          The construction of microbial consortia is challenging due to many variables to be controlled, including the cross‐compatibility of the selected strains and their additive or synergistic effects on plants. In this work, we investigated the interactions in vitro, in planta, and at the molecular level of two elite biological control agents (BCAs), that is Streptomyces microflavus strain AtB‐42 and Trichoderma harzianum strain M10, to understand their attitude to cooperate in a consortium. In vitro, we observed a strong cross‐antagonism between AtB‐42 and M10 in agar plates due to diffusible metabolites and volatile organic compounds. In liquid co‐cultures, M10 hindered the growth of AtB‐42 very likely because of secondary metabolites and strong competition for the nutrients. The interaction in the co‐culture induced extensive transcriptional reprogramming in both strains, especially in the pathways related to ribosomes, protein synthesis, and oxidoreductase activity, suggesting that each strain recognized the counterpart and activated its defence responses. The metabolome of both strains was also significantly affected. In contrast, in the soil, M10 growth was partially contrasted by AtB‐42. The roots of tomato seedlings inoculated with the consortium appeared smaller than the control and single‐strain‐inoculated plants, indicating that plants diverted some energy from the development to defence activation, as evidenced by the leaf transcriptome. The consortium induced a stronger transcriptional change compared to the single inoculants, as demonstrated by a higher number of differentially expressed genes. Although the cross‐antagonism observed in vitro, the two strains exerted a synergistic effect on tomato seedlings by inducing resistance responses stronger than the single inoculants. Our observations pose a question on the usefulness of the sole in vitro assays for selecting BCAs to construct a consortium. In vivo experiments should be preferred, and transcriptomics may greatly help to elucidate the activity of the BCAs beyond the phenotypic effects on the plant.

          Abstract

          The construction of microbial consortia is challenging due to many variables to be controlled, including the cross‐compatibility of the selected strains and their additive or synergistic effects on plants. In this work, we investigated the interactions in vitro, in planta, and at the molecular level of two elite biological control agents (BCAs), i.e., Streptomyces microflavus strain AtB‐42 and Trichoderma harzianum strain M10, to understand their attitude to cooperate in a consortium. Microorganisms, including BCAs, compete against every other microorganism to preserve the species and their niche. Although we observed a cross‐antagonism in vitro, these two strains exerted synergistic effects in vivo on tomatoes. Whenever possible, hence, it would be more advantageous to study the microorganisms in vivo. Finally, the analysis of the plant transcriptome represented a valuable tool for interpreting the phenotypic effects of the BCAs on the crop.

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          Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2

          In comparative high-throughput sequencing assays, a fundamental task is the analysis of count data, such as read counts per gene in RNA-seq, for evidence of systematic changes across experimental conditions. Small replicate numbers, discreteness, large dynamic range and the presence of outliers require a suitable statistical approach. We present DESeq2, a method for differential analysis of count data, using shrinkage estimation for dispersions and fold changes to improve stability and interpretability of estimates. This enables a more quantitative analysis focused on the strength rather than the mere presence of differential expression. The DESeq2 package is available at http://www.bioconductor.org/packages/release/bioc/html/DESeq2.html. Electronic supplementary material The online version of this article (doi:10.1186/s13059-014-0550-8) contains supplementary material, which is available to authorized users.
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            Fast gapped-read alignment with Bowtie 2.

            As the rate of sequencing increases, greater throughput is demanded from read aligners. The full-text minute index is often used to make alignment very fast and memory-efficient, but the approach is ill-suited to finding longer, gapped alignments. Bowtie 2 combines the strengths of the full-text minute index with the flexibility and speed of hardware-accelerated dynamic programming algorithms to achieve a combination of high speed, sensitivity and accuracy.
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              Cytoscape: a software environment for integrated models of biomolecular interaction networks.

              Cytoscape is an open source software project for integrating biomolecular interaction networks with high-throughput expression data and other molecular states into a unified conceptual framework. Although applicable to any system of molecular components and interactions, Cytoscape is most powerful when used in conjunction with large databases of protein-protein, protein-DNA, and genetic interactions that are increasingly available for humans and model organisms. Cytoscape's software Core provides basic functionality to layout and query the network; to visually integrate the network with expression profiles, phenotypes, and other molecular states; and to link the network to databases of functional annotations. The Core is extensible through a straightforward plug-in architecture, allowing rapid development of additional computational analyses and features. Several case studies of Cytoscape plug-ins are surveyed, including a search for interaction pathways correlating with changes in gene expression, a study of protein complexes involved in cellular recovery to DNA damage, inference of a combined physical/functional interaction network for Halobacterium, and an interface to detailed stochastic/kinetic gene regulatory models.
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                Author and article information

                Contributors
                giovanninicola.bubici@cnr.it
                Journal
                Microb Biotechnol
                Microb Biotechnol
                10.1111/(ISSN)1751-7915
                MBT2
                Microbial Biotechnology
                John Wiley and Sons Inc. (Hoboken )
                1751-7915
                18 July 2023
                December 2023
                : 16
                : 12 , Part 2: End Hunger: Enhancing Crop Yields with Microbes ( doiID: 10.1111/mbt2.v16.12 )
                : 2292-2312
                Affiliations
                [ 1 ] Istituto per la Protezione Sostenibile delle Piante Consiglio Nazionale delle Ricerche Bari Italy
                [ 2 ] Istituto per la Protezione Sostenibile delle Piante Consiglio Nazionale delle Ricerche Portici Italy
                [ 3 ] Dipartimento di Agraria Università degli Studi di Napoli Federico II Portici Italy
                [ 4 ] Dipartimento di Medicina Veterinaria e Produzioni Animali Università degli Studi di Napoli Federico II Naples Italy
                Author notes
                [*] [* ] Correspondence

                Giovanni Bubici, Istituto per la Protezione Sostenibile delle Piante, Consiglio Nazionale delle Ricerche, via 7 Amendola 165/A, Bari 70126, Italy.

                Email: giovanninicola.bubici@ 123456cnr.it

                Author information
                https://orcid.org/0000-0002-6913-5232
                https://orcid.org/0000-0002-8946-1609
                https://orcid.org/0000-0002-5090-8127
                https://orcid.org/0000-0001-9143-2711
                Article
                MBT214311 MICROBIO-2023-143-RA.R1
                10.1111/1751-7915.14311
                10686133
                37464583
                8d9208de-5421-463e-998a-7a0264e34f1c
                © 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
                : 19 June 2023
                : 11 April 2023
                : 25 June 2023
                Page count
                Figures: 8, Tables: 0, Pages: 21, Words: 14273
                Funding
                Funded by: PROdotti, servizi e TEcnologie innovative per il ConTrollo bIOlogico e la difesa ecososteNibile in agricoltura (PROTECTION)
                Award ID: F/050421/01‐03/X32
                Funded by: European Union Next‐Generation EU (PIANO NAZIONALE DI RIPRESA E RESILIENZA (PNRR) – MISSIONE 4 COMPONENTE 2, INVESTIMENTO 1.4 – D.D. 1032 17/06/2022, CN00000022)
                Award ID: AgritechNationalResearchCenter
                Categories
                Research Article
                Special Issue: Part 2: End Hunger: Enhancing Crop Yields with Microbes
                Research Articles
                Custom metadata
                2.0
                December 2023
                Converter:WILEY_ML3GV2_TO_JATSPMC version:6.3.5 mode:remove_FC converted:29.11.2023

                Biotechnology
                Biotechnology

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