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The microbial community in anoxic rice field soil produces CH4 over a wide temperature range up to 55°C. However, at temperatures higher than about 40°C, the methanogenic path changes from CH4 production by hydrogenotrophic plus acetoclastic methanogenesis to exclusively hydrogenotrophic methanogenesis and simultaneously, the methanogenic community consisting of Methanosarcinaceae, Methanoseataceae, Methanomicrobiales, Methanobacteriales and Rice Cluster I (RC-1) changes to almost complete dominance of RC-1. We studied changes in structure and function of the methanogenic community with temperature to see whether microbial members of the community were lost or their function impaired by exposure to high temperature. We characterized the function of the community by the path of CH4 production measuring δ13C in CH4 and CO2 and calculating the apparent fractionation factor (αapp) and the structure of the community by analysis of the terminal restriction fragment length polymorphism (T-RFLP) of the microbial 16S rRNA genes. Shift of the temperature from 45°C to 35°C resulted in a corresponding shift of function and structure, especially when some 35°C soil was added to the 45°C soil. The bacterial community (T-RFLP patterns), which was much more diverse than the archaeal community, changed in a similar manner upon temperature shift. Incubation of a mixture of 35°C and 50°C pre-incubated methanogenic rice field soil at different temperatures resulted in functionally and structurally well-defined communities. Although function changed from a mixture of acetoclastic and hydrogenotrophic methanogenesis to exclusively hydrogenotrophic methanogenesis over a rather narrow temperature range of 42-46°C, each of these temperatures also resulted in only one characteristic function and structure. Our study showed that temperature conditions defined structure and function of the methanogenic microbial community.
Most of the methane (CH4) emission from rice fields is derived from plant photosynthates,
which are converted to CH4. Rice cluster I (RC-1) archaea colonizing the
rhizosphere were found to be the methanogens responsible for this process. Hence, RC-1
methanogens seem to play a crucial role in emission of the greenhouse gas CH4. We
determined the community composition and activity of methanogens colonizing the
roots of eight different rice cultivars after growth on both Italian rice soil and river bank
soil, which contained different communities of methanogenic archaea. The community
composition was analyzed by terminal restriction fragment length polymorphism and
cloning/sequencing of the archaeal 16S rRNA gene and the mcrA gene coding for a
subunit of the methyl coenzyme M reductase. When grown on rice field soil, the
methanogenic community of the different rice cultivars was always dominated by RC-1
methanogens. In contrast, roots were colonized by Methanomicrobiales when grown on
river bank soil, in which RC-1 methanogens were initially not detectable. Roots
colonized with Methanomicrobiales compared with RC-1 exhibited lower CH4 production
and CH4 emission rates. The results show that the type of methanogens colonizing
rice roots has a potentially important impact on the global CH4 cycle.
The microbial community structure was investigated together with the path of methane production in Italian rice field soil incubated at moderate (35 °C) and high (45 °C) temperature using terminal restriction fragment length polymorphism and stable isotope fractionation. The structure of both the archaeal and bacterial communities differed at 35 °C compared with 45 °C, and acetoclastic and hydrogenotrophic methanogenesis dominated, respectively. Changing the incubation of the 45 °C soil to different temperatures (25, 30, 35, 40, 45, 50 °C) resulted in a dynamic change of both microbial community structure and stable isotope fractionation. In all treatments, acetate first accumulated and then decreased. Propionate was also transiently produced and consumed. It is noteworthy that acetate was also consumed at thermophilic conditions, although archaeal community composition and stable isotope fractionation indicated that acetoclastic methanogenesis did not operate. Instead, acetate must have been consumed by syntrophic acetate oxidizers. The transient accumulation and subsequent consumption of acetate at thermophilic conditions was specifically paralleled by terminal restriction fragments characteristic for clostridial cluster I, whereas those of clostridial clusters I and III, Acidaminococcaceae and Heliobacteraceae, paralleled the thermophilic turnover of both acetate and propionate.