Effect of temperature on the rate limiting step in the methanogenic degradation pathway in rice field soil

Temperature is an important factor controlling CH₄ production in rice field soils. However, it is unknown which step in the methanogenic degradation of organic matter is the limiting one that is controlled by temperature. Soil slurries prepared from Italian rice field soil were anaerobically incubated in the dark at six different temperatures between 10 and 37 °C until quasi-steady state was reached. Then, the potential and actual rates of polysaccharide hydrolysis and of CH₄ production from different immediate (acetate, H₂) and distal (glucose, propionate) methanogenic substrates were determined. Potential activities of exo-glucanase and glucosidase were always higher than the actual rates of polysaccharide hydrolysis indicating that the availability of the polysaccharide substrate was limiting at all temperatures. The actual rates of CH₄ production were always lower than those predicted from glucose release during polysaccharide hydrolysis indicating that a substantial amount of the released glucose was assimilated into microbial biomass. Addition of the different methanogenic substrates stimulated CH₄ production at all temperatures >10 °C, but only at >20 °C to values higher than rates of polysaccharide hydrolysis. Under steady state conditions, however, hydrolysis of organic polymers was the rate-limiting step at all temperatures >10 °C.