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All parental lines were maintained at low density on standard cornmeal-molasses medium in bottle cultures to limit confounding environmental variation.
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Put in bottle.
The initial AR80 concentration in the culture medium in each bottle was 1.0 × 106 cells/mL.
Birnessite (final concentration = 0.3 mg/mL) was suspended with the culture medium in serum bottles sealed with blue butyl rubber stoppers (for anaerobic) and in flasks (for aerobic) with total volume of 80 mL.
Prior to placing the bottles in the anaerobic jar, caps were loosened to help exchange of gases between the jar and the medium in the bottles.
For culture a volume of 0.25 ml of each decontaminated processed sputum specimen was inoculated onto slants of Lowenstein-Jensen (LJ) medium in culture bottles (Becton Dickenson Microbiology Systems, Cockeysville, MD, USA).
The effects of H2O2 and catalase (Sigma-Aldrich) on the intrinsic H2O2 concentrations and flowering time were determined using plants grown on MS medium in glass bottles (5 cm high, 2 and 1.5 cm wide diametrically at top and bottom).
Anaerobiosis was achieved by nitrogen sparging and sealing with butyl-rubber stoppers of the serum bottles (50 mL medium in 100 mL bottles).
Cells of BY4742 strain and its PHO8-expressing derivatives were grown in 100 ml of YPD medium in Erlenmeyer flasks (bottle size – 300 ml) overnight and then used to inoculate a 20 ml of YNB medium with 10% glucose in 50 ml Erlenmeyer flasks.
Each generation was reared on a standard medium in 250 ml bottles, for a total of 10 bottles.
All experimental flies were reared from the eggs laid by females on malt medium in half-pint bottles.
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