5. Conclusion
Our study showed that rice field soils have the potential for chemolithotrophic acetogenesis, but only when H2/CO2 are added as external substrates. Only then very low δ13C of acetate was observed for all incubations irrespectively of the incubation temperature. However, the very low δ13C of acetate was transient and reverted to values closer to the δ13C of organic matter as soon as the added H2/CO2 was consumed. Without addition of H2/CO2, chemolithotrophic acetogenesis was insignificant and could not be detected using isotopic values of acetate. Importance of acetate production may be higher at low versus medium temperatures, but the relatively high δ13C of acetate indicated production by fermentation (or maybe by heterotrophic acetogenesis) rather than by chemolithotrophic acetogenesis. It was possible to determine production of acetate together with the isotopic signatures without simultaneous consumption when aceticlastic methanogenesis was inhibited with BES. However, this approach was not possible at high temperatures when acetate was presumably consumed by syntrophic acetate oxidizers which were not inhibited by BES. Addition of KCN as putative inhibitor of acetogens was not helpful. The quantification of fhs, as characteristic gene for acetogens, also turned out to be inconclusive. Hence, discrimination of acetogenically formed acetate using the described techniques is still a problem for future research. Currently, it is only safe to determine the potential for chemolithotrophic acetogenesis through stimulation by H2/CO2 and evaluating the δ13C of acetate.