Mgso4 7h2o

For observing AF-derived fluorescence, we prepared PDA supplemented with 3 g·L⁻¹ α-CD (FUJIFILM Wako Pure Chemical, Osaka, Japan) and 0.3 g·L⁻¹ activated carbon (Table 1). We also prepared PD broth (Merck) with activated carbon and without α-CD as the liquid culture medium. Aliquots (495 μL) of the liquid medium were dispensed into 1-mL pipette tips that were sealed with laboratory film (PM-996, Bemis, Neenah, WI, USA) and used for incubation [27 (link)]. The total volume was adjusted to 500 μL by adding the spore solution. To prepare trace element solutions, (CH₃CHOCOO)₂Ca·5H₂O, MgSO₄·7H₂O, FeCl₃·6H₂O, and [CH₃CH(OH)COO]₃Al (FUJIFILM Wako Pure Chemical) were dissolved in distilled water. The trace element solutions were added to the liquid culture media. The final concentrations of Ca, Mg, and Fe were set to 5 mg·L⁻¹ and that of Al was set to 3 mg·L⁻¹. The fungal spore solution (5 μL) was dispensed onto a culture plate or into the liquid culture medium. After liquid cultivation, the laboratory film was removed from each pipette tip, and the tips were centrifuged at 3000× g for 15 min. The AF concentrations were measured in the obtained culture solutions. Mycelia with spores in the incubation tip were dried in a dehydrator (SLI-450N, TOKYO RIKAKIKAI, Tokyo, Japan) at 65 °C overnight, and the fungal mass was calculated using the tip masses before and after incubation.

Pseudomonas aeruginosa strains used in this study are listed in Table 1. Bacteria were grown in an Air-Jacketed Incubator IC802 (Yamato Scientific Co., Ltd., Tokyo, Japan) at 37 °C under aerobic conditions, as previously described [12 (link)]. Unless otherwise indicated, bacteria were cultured using lysogeny broth, Lennox (LB)-agar prepared fresh from 1.0% Bacto^TM Tryptone (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), 0.5% Bacto^TM yeast extract (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), and 0.5% NaCl (FUJIFILM Wako Pure Chemical Corp., Osaka, Japan). Pseudomonas agar F was prepared from 2.0% Bacto^TM proteose peptone no. 3 (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), 1.0% Bacto^TM Casitone (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), 1.0% glycerol (Nacalai Tesque Inc., Kyoto, Japan), 0.15% K₂HPO₄ (FUJIFILM Wako Pure Chemical Corp., Osaka, Japan), 0.073% MgSO₄⋅7H₂O (FUJIFILM Wako Pure Chemical Corp.), and 1.5% agar (FUJIFILM Wako Pure Chemical Corp.), and used as a solid medium in STAR SDish9015 ver.2 petri dishes (Rikaken Co., Ltd., Nagoya, Japan) for assays of pyoverdine production. Bacterial growth was quantified by measuring the optical density at 600 nm (OD₆₀₀) using a WPA CO8000 Cell Density Meter (Biochrom Ltd., Cambridge, UK).

The H. beimenensis NTU-111 strain used in this study was originally isolated from brine samples from the Beimen saltern in southern Taiwan^{22 (link)}. H. beimenensis was grown in basal medium containing 5 g/L yeast extract (Bacto, BD), 5 g/L Casamino acids (Bacto, BD) and 5 g/L MgSO₄·7H₂O (Wako), pH 7.5. To determine the optimal growth conditions for H. beimenensis, the bacteria were grown in basal medium with various concentrations of NaCl, including 0%, 5%, 10%, 15%, 20%, and 25% NaCl (w/v), and were incubated at 37 °C with shaking at 220 rpm. The concentration of H. beimenensis (OD600) was monitored by a spectrophotometer (Libra S4, Biochrom) every 6 h until 72 h with three replicates.