RT Journal Article
SR Electronic
T1 FlowPot axenic plant growth system for microbiota research
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 254953
DO 10.1101/254953
A1 James M. Kremer
A1 Bradley C. Paasch
A1 David Rhodes
A1 Caitlin Thireault
A1 John E. Froehlich
A1 Paul Schulze-Lefert
A1 James M. Tiedje
A1 Sheng Yang He
YR 2018
UL http://biorxiv.org/content/early/2018/01/30/254953.abstract
AB The presence of resident microbiota on and inside plants is hypothesized to influence many phenotypic attributes of the host. Likewise, host factors and microbe-microbe interactions are believed to influence microbial community assembly. Rigorous testing of these hypotheses necessitates the ability to grow plants in the absence or presence of resident or defined microbiota. To enable such experiments, we developed the scalable and inexpensive FlowPot growth platform. FlowPots have a sterile peat substrate amenable to colonization by microbiota, and the platform supports growth of the model plant Arabidopsis thaliana in the absence or presence of soil-derived microbial communities. Mechanically, the FlowPot system is unique in that it allows for total-saturation of the sterile substrate by “flushing” with water and/or nutrient solution via an irrigation port. The irrigation port also facilitates passive drainage of the substrate, preventing root anoxia. Materials to construct an individual FlowPot total ∼$2. A simple experiment with 12 FlowPots requires ∼4.5 h of labor following peat and seed sterilization. Plants are grown on FlowPots within a standard tissue culture microbox after inoculation, thus the Flowpot system is modular and does not require a sterile growth chamber. Here, we provide a detailed assembly and microbiota inoculation protocol for the FlowPot system. Collectively, this standardized suite of tools and colonization protocols empowers the plant microbiome research community to conduct harmonized experiments to elucidate the rules microbial community assembly, the impact of microbiota on host phenotypes, and mechanisms by which host factors influence the structure and function of plant microbiota.