Fig 1.
As illustrated here with images of Bsal, chytrid fungi have a biphasic life cycle characterized by a stationary growth phase called a sporangium (top) and a motile dispersal phase called a zoospore (bottom). Images taken at 100X using differential interference contrast (DIC) microscopy.
Table 1.
Antibiotic concentrations used to select for gene expression in select eukaryotes.
This table lists the key features of the antibiotics used in this study: the drug class, the target, known resistance genes, the current listed price per gram from Millipore Sigma, and the concentrations used in select eukaryotes. Species include representatives from plants (Arabidopsis thaliana and Chlamydomonas reinhardtii), protozoa (Trypanosoma brucei), amoebae (Dictyostelium discoideum), fungi (Aspergillus spp., Schizosaccharomyces pombe, Saccharomyces cerevisiae), and animals (human) in addition to the two species tested in this study. The lowest concentrations of each antibiotic which inhibited growth in liquid and solid media for Bd and Bsal are listed from our findings in this study. These concentrations were used to calculate the cost per liter of growth media for both Bd and Bsal.
Fig 2.
Inhibition of Bd growth in liquid media.
Percent of Bd growth in liquid media supplemented with (A) Hygromycin, (B), Zeocin, (C) Blasticidin, (D) Puromycin, and (E) Neomycin as compared to an antibiotic free control for three temporally isolated replicates (circle, square, and triangle, shades of blue). Orange symbols indicate concentrations at which no growth occurred after three days in all three replicates.
Fig 3.
Inhibition of Bsal growth in liquid media.
Percent of Bsal growth in liquid media supplemented with (A) Hygromycin, (B), Zeocin, (C) Blasticidin, (D) Puromycin, and (E) Neomycin as compared to an antibiotic free control for three temporally isolated replicates (circle, square, and triangle, shades of blue). Orange symbols indicate concentrations at which no growth occurred after four days in all three replicates.
Fig 4.
Inhibition of Bd growth on solid media.
(A) Examples of Bd growth after three days on antibiotic selection plates. The ‘+’ demonstrates the relative zoospore activity of each plate compared to an antibiotic-free control plate. The box highlights zoospores, which appear as small dots while the bracket highlights sporangia. The zoospores in the ‘0’ image are immotile (see S1 Video). Scale bar 50 μm. (B) Bd growth on antibiotic selection plates. Concentrations highlighted in bold and orange are the lowest concentrations that prevent growth for at least 14 days post zoospore plating.
Fig 5.
Inhibition of Bsal growth on solid media.
(A) Examples of Bsal growth after four days on antibiotic selection plates. The ‘+’ demonstrates the relative zoospore activity of each plate compared to a no antibiotic control plate. The box highlights zoospores, which appear as small dots while the bracket highlights sporangia. The zoospores in the ‘0’ image are immotile (see S1 Video). Scale bar 50 μm. (B) Bsal growth on antibiotic selection plates. Concentrations highlighted in bold and orange are the lowest concentrations that prevent growth for at least 14 days post zoospore plating.