Ecosystem Fabrication (EcoFAB) Protocols for The Construction of Laboratory Ecosystems Designed to Study Plant-microbe Interactions

Gao, Jian; Sasse, Joelle; Lewald, Kyle M.; Zhalnina, Kateryna; Cornmesser, Lloyd T.; Duncombe, Todd A.; Yoshikuni, Yasuo; Vogel, John P.; Firestone, Mary K.; Northen, Trent R.

doi:10.3791/57170

Title: Ecosystem Fabrication (EcoFAB) Protocols for The Construction of Laboratory Ecosystems Designed to Study Plant-microbe Interactions

Journal Article · Tue Apr 10 00:00:00 EDT 2018 · Journal of Visualized Experiments

DOI:https://doi.org/10.3791/57170· OSTI ID:1506304

Gao, Jian ^[1]; Sasse, Joelle ^[1]; Lewald, Kyle M. ^[1]; Zhalnina, Kateryna ^[1]; Cornmesser, Lloyd T. ^[1]; Duncombe, Todd A. ^[2]; Yoshikuni, Yasuo ^[3]; Vogel, John P. ^[3]; Firestone, Mary K. ^[4]; Northen, Trent R. ^[1]

Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Environmental Genomics and Systems Biology Division; USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States)
Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States)
USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States)
Univ. of California, Berkeley, CA (United States). Dept. of Environmental Science Policy and Management

Beneficial plant-microbe interactions offer a sustainable biological solution with the potential to boost low-input food and bioenergy production. A better mechanistic understanding of these complex plant-microbe interactions will be crucial to improving plant production as well as performing basic ecological studies investigating plant-soil-microbe interactions. Here, a detailed description for ecosystem fabrication is presented, using widely available 3D printing technologies, to create controlled laboratory habitats (EcoFABs) for mechanistic studies of plant-microbe interactions within specific environmental conditions. Two sizes of EcoFABs are described that are suited for the investigation of microbial interactions with various plant species, including Arabidopsis thaliana, Brachypodium distachyon, and Panicum virgatum. These flow-through devices allow for controlled manipulation and sampling of root microbiomes, root chemistry as well as imaging of root morphology and microbial localization. This protocol includes the details for maintaining sterile conditions inside EcoFABs and mounting independent LED light systems onto EcoFABs. Detailed methods for addition of different forms of media, including soils, sand, and liquid growth media coupled to the characterization of these systems using imaging and metabolomics are described. Together, these systems enable dynamic and detailed investigation of plant and plant-microbial consortia including the manipulation of microbiome composition (including mutants), the monitoring of plant growth, root morphology, exudate composition, and microbial localization under controlled environmental conditions. We anticipate that these detailed protocols will serve as an important starting point for other researchers, ideally helping create standardized experimental systems for investigating plant-microbe interactions.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); LBNL Laboratory Directed Research and Development (LDRD) Program

Grant/Contract Number:: AC02-05CH11231; SC0014079

OSTI ID:: 1506304

Journal Information:: Journal of Visualized Experiments, Vol. 2018, Issue 134; ISSN 1940-087X

Publisher:: MyJoVE Corp.Copyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 28 works

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Need for Laboratory Ecosystems To Unravel the Structures and Functions of Soil Microbial Communities Mediated by Chemistry Zhalnina, Kateryna; Zengler, Karsten; Newman, Dianne mBio, Vol. 9, Issue 4 https://doi.org/10.1128/mbio.01175-18	journal	July 2018

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Title: Ecosystem Fabrication (EcoFAB) Protocols for The Construction of Laboratory Ecosystems Designed to Study Plant-microbe Interactions

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