Tpp6 well tissue culture plates

Strains were grown in MB and diluted 10,000-fold and grown stagnant in 5 mL IOCG in TPP6-well tissue culture plates (Merck) at 25°C. Plates were incubated in a humidity chamber to prevent desiccation. Samples were taken after 17 (exponential phase) and 72 h (stationary phase) for metabolome analysis, and after 72 h for transcriptomics and proteomics. Experiments were carried out with five biological replicates for metabolomics and proteomics, and in three biological replicates for transcriptomics. CFU determination and microscopy was performed for each sample. Transcriptomics, proteomics, and metabolomics workflows can be found in Supplemental Text S1.

Phaeobacter strains were grown in MB and diluted 10,000-fold and grown in IOCG in TPP6-well tissue culture plates (Merck) at 25°C in biological triplicates. Plates were incubated in a humidity chamber to prevent desiccation. For chemical complementation, samples were supplemented with 50% sterile-filtered WT or mutant supernatant of 3-day-old cultures grown in IOCG stagnant at 25°C. For genetic complementations, strains were grown in IOCG with 200 μg/mL kanamycin and incubated in the humidity chamber, stagnant at 25°C for 7 days. Cell morphology was assessed through brightfield microscopy using a Nikon ECLIPSE Ti2 inverted microscope at ×60 magnification. Cell length measurements were carried out using ImageJ (83 (link)). For each replicate, three pictures were taken and five random cells within each picture were measured for statistical comparisons.

Phaeobacter strains were grown in MB and diluted to OD₆₀₀ = 0.00001 in IOCG and incubated at 25°C stagnant in TPP6-well tissue culture plates (Merck) with 5 mL media per well. The experiment was done in biological triplicates and growth followed by colony counts that were log transformed and plotted using ggplot2 (82 ) in Rstudio.