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Title: Simultaneous Discovery of Positive and Negative Interactions Among Rhizosphere Bacteria Using Microwell Recovery Arrays

Abstract

Understanding microbe-microbe interactions is critical to predict microbiome function and to construct communities for desired outcomes. Investigation of these interactions poses a significant challenge due to the lack of suitable experimental tools available. Here we present the microwell recovery array (MRA), a new technology platform that screens interactions across a microbiome to uncover higher-order strain combinations that inhibit or promote the function of a focal species. One experimental trial generates 104 microbial communities that contain the focal species and a distinct random sample of uncharacterized cells from plant rhizosphere. Cells are sequentially recovered from individual wells that display highest or lowest levels of focal species growth using a high-resolution photopolymer extraction system. Interacting species are then identified and putative interactions are validated. Using this approach, we screen the poplar rhizosphere for strains affecting the growth of Pantoea sp. YR343, a plant growth promoting bacteria isolated from Populus deltoides rhizosphere. In one screen, we montiored 3,600 microwells within the array to uncover multiple antagonistic Stenotrophomonas strains and a set of Enterobacter strains that promoted YR343 growth. The later demonstrates the unique ability of the platform to discover multi-membered consortia that generate emergent outcomes, thereby expanding the range of phenotypes that canmore » be characterized from microbiomes. This knowledge will aid in the development of consortia for Populus production, while the platform offers a new approach for screening and discovery of microbial interactions, applicable to any microbiome.« less

Authors:
; ; ; ; ; ; ;
Publication Date:
Research Org.:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
OSTI Identifier:
1755778
Alternate Identifier(s):
OSTI ID: 1756240
Grant/Contract Number:  
SC0018579; AC05-00OR22725
Resource Type:
Published Article
Journal Name:
Frontiers in Microbiology
Additional Journal Information:
Journal Name: Frontiers in Microbiology Journal Volume: 11; Journal ID: ISSN 1664-302X
Publisher:
Frontiers Research Foundation
Country of Publication:
Switzerland
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; microdevice; microbiome; microbial communities; microbial interactions; consortia; plant growth promoting rhizobacteria; high throughput screening; microwell

Citation Formats

Barua, Niloy, Herken, Ashlee M., Stern, Kyle R., Reese, Sean, Powers, Roger L., Morrell-Falvey, Jennifer L., Platt, Thomas G., and Hansen, Ryan R. Simultaneous Discovery of Positive and Negative Interactions Among Rhizosphere Bacteria Using Microwell Recovery Arrays. Switzerland: N. p., 2021. Web. doi:10.3389/fmicb.2020.601788.
Barua, Niloy, Herken, Ashlee M., Stern, Kyle R., Reese, Sean, Powers, Roger L., Morrell-Falvey, Jennifer L., Platt, Thomas G., & Hansen, Ryan R. Simultaneous Discovery of Positive and Negative Interactions Among Rhizosphere Bacteria Using Microwell Recovery Arrays. Switzerland. https://doi.org/10.3389/fmicb.2020.601788
Barua, Niloy, Herken, Ashlee M., Stern, Kyle R., Reese, Sean, Powers, Roger L., Morrell-Falvey, Jennifer L., Platt, Thomas G., and Hansen, Ryan R. Tue . "Simultaneous Discovery of Positive and Negative Interactions Among Rhizosphere Bacteria Using Microwell Recovery Arrays". Switzerland. https://doi.org/10.3389/fmicb.2020.601788.
@article{osti_1755778,
title = {Simultaneous Discovery of Positive and Negative Interactions Among Rhizosphere Bacteria Using Microwell Recovery Arrays},
author = {Barua, Niloy and Herken, Ashlee M. and Stern, Kyle R. and Reese, Sean and Powers, Roger L. and Morrell-Falvey, Jennifer L. and Platt, Thomas G. and Hansen, Ryan R.},
abstractNote = {Understanding microbe-microbe interactions is critical to predict microbiome function and to construct communities for desired outcomes. Investigation of these interactions poses a significant challenge due to the lack of suitable experimental tools available. Here we present the microwell recovery array (MRA), a new technology platform that screens interactions across a microbiome to uncover higher-order strain combinations that inhibit or promote the function of a focal species. One experimental trial generates 104 microbial communities that contain the focal species and a distinct random sample of uncharacterized cells from plant rhizosphere. Cells are sequentially recovered from individual wells that display highest or lowest levels of focal species growth using a high-resolution photopolymer extraction system. Interacting species are then identified and putative interactions are validated. Using this approach, we screen the poplar rhizosphere for strains affecting the growth of Pantoea sp. YR343, a plant growth promoting bacteria isolated from Populus deltoides rhizosphere. In one screen, we montiored 3,600 microwells within the array to uncover multiple antagonistic Stenotrophomonas strains and a set of Enterobacter strains that promoted YR343 growth. The later demonstrates the unique ability of the platform to discover multi-membered consortia that generate emergent outcomes, thereby expanding the range of phenotypes that can be characterized from microbiomes. This knowledge will aid in the development of consortia for Populus production, while the platform offers a new approach for screening and discovery of microbial interactions, applicable to any microbiome.},
doi = {10.3389/fmicb.2020.601788},
journal = {Frontiers in Microbiology},
number = ,
volume = 11,
place = {Switzerland},
year = {Tue Jan 05 00:00:00 EST 2021},
month = {Tue Jan 05 00:00:00 EST 2021}
}

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