Effects of chemical and heat treatments on ethylene production in soil

Factors influencing C₂H₄ production in a silt loam were investigated in an effort to determine the source of this gas in soil. Air-dried samples of soil in glass vials were moistened to about ?10 kPa, sealed with rubber septa, and incubated at 30 or 35°C with an original atmosphere of air or O₂-free N₂. C₂H₄ concentrations in the vials were determined by gas chromatography.Addition of the antibacterial agents chloramphenicol or novobiocin to the soil inhibited C₂H₄ production, whereas the antifungal agent cycloheximide had no effect. Sodium azide and sodium cyanide also reduced C₂H₄ production. Treatment of the soil with moist heat (i.e. passing a steam-air mixture through it) at 80°C for 30 min failed to reduce the ability of the soil to produce C₂H₄ during subsequent incubation at 30°C, but autoclaving it twice at 121°C prevented C₂H₄ production. As with nonheated soil, C₂H₄ production from soil treated at 80°C was prevented by novobiocin but not by cycloheximide. Only about 10% of the bacteria isolated from nontreated soil were spore-formers. In contrast, 95–98% and possibly more of the bacteria isolated from heat-treated soil were spore-formers, including those in soil which was heat-treated and then incubated moist at 30°C for an additional 3 days before dilution plating. Addition of methionine had no effect on the production of C₂H₄ in anaerobic soil, whereas ethionine, chlorogenic acid, and ethylenediaminetetraacetic acid (EDTA) all enhanced C₂H₄ production. Ethionine, but not chlorogenic acid or EDTA, also resulted in considerable C₂H₄ accumulation in autoclaved soil; the C₂H₄ detected in ethionine-amended soil was apparently nonmicrobial in origin. Soil samples incubated at constant temperatures of 30, 50, or 70°C all produced C₂H₄.The results collectively indicate that C₂H₄ in soil is most likely produced by facultative or strictly anaerobic bacteria which are probably spore-formers and may also be thermophilic. Several isolates of spore-forming bacteria were inoculated into autoclaved soil, but none produced appreciable amounts of C₂H₄ under the test conditions.