Characterization of Root Exudates at Different Growth Stages of Ten Rice (Oryza sativa L.) Cultivars

Abstract: Plant root exudates play important roles in the rhizosphere. We tested three media (nutrient solution, deionized water and CaSO₄ solution) for three periods of time (2, 4 and 6 h) for collecting root exudates of soil‐grown rice plants. Nutrient culture solution created complications in the analyses of exudates for total organic C (TOC) by the wet digestion method and of organic acids by HPLC due to the interference by its components. Deionized water excluded such interference in analytical analyses but affected the turgor of root cells; roots of four widely different rice cultivars excreted 20 to 60 % more TOC in deionized water than in 0.01 M CaSO₄. Furthermore, the proportion of carbohydrates in TOC was also enhanced. Calcium sulfate solution maintained the osmotic environment for root cells and did not interfere in analytical procedures. Collection for 2 h avoided under‐estimation of TOC and its components exuded by rice roots, which occurred during prolonged exposure. By placing plants in 0.01 M CaSO₄ for 2 h, root exudates of soil‐grown traditional, tall rice cultivars (Dular, B40 and Intan), high‐yielding dwarf cultivars (IR72, IR52, IR64 and PSBRc 20), new plant type cultivars (IR65598 and IR65600) and a hybrid (Magat) were collected at seedling, panicle initiation, flowering and maturity and characterized for TOC and organic acids. The exudation rates were, in general, lowest at seedling stage, increased until flowering but decreased at maturity. Among organic acids, malic acid showed the highest concentration followed by tartaric, succinic, citric and lactic acids. With advancing plant growth, exudation of organic acids substituted exudation of sugars. Root and shoot biomass were positively correlated with carbon exudation suggesting that it is driven by plant biomass. As root exudates provide substrates for methanogenesis in rice fields, large variations in root exudation by cultivars and at different growth stages could greatly influence CH₄ emissions. Therefore, the use of high‐yielding cultivars with lowest root excretions, for example IR65598 and IR65600, would mediate low exudate‐induced CH₄ production. The screening of exciting rice cultivars and breeding of new cultivars with low exudation rates could offer an important option for mitigation of CH₄ emission from rice agriculture to the atmosphere.     

CH4 emission with differences in atmospheric CO2 enrichment and rice cultivars in a Japanese paddy soil

A pot experiment was conducted to investigate CH₄ emissions from a sandy paddy soil as influenced by rice cultivars and atmospheric CO₂ elevation. The experiment with two CO₂ levels, 370 μL L⁻¹ (ambient) and 570 μL L⁻¹ (elevated), was performed in a climatron, located at the National Institute for Agro‐Environmental Sciences, Tsukuba, Japan. Four rice cultivars were tested in this experiment, including IR65598, IR72, Dular and Koshihikari. Tiller number, root length and grain yield were clearly larger under elevated CO₂ than under ambient CO₂. IR72 and Dular showed significantly higher tiller number, root length and grain yield than Koshihikari and IR65598. Average daily CH₄ fluxes under elevated CO₂ were significantly larger by 10.9–23.8% than those under ambient CO₂, and varied with the cultivars in the sequence Dular ≧ IR72>IR65598 ≧ Koshihikari. Dissolved organic C (DOC) content in the soil was obviously higher under elevated CO₂ than under ambient CO₂ and differed among the cultivars, in the sequence IR72>Dular>Koshihikari>IR65598. The differences in average daily CH₄ fluxes between CO₂ levels and among the cultivars were related to different root exudation as DOC content, root length and tiller number. This study indicated that Koshihikari should be a potential cultivar for mitigating CH₄ emission and simultaneously keeping stable grain yield, because this cultivar emitted lowest CH₄ emission and produced medium grain yield.     

Microbial mechanism for rice variety control on methane emission from rice field soil

Rice variety is one of the key factors regulating methane (CH₄) production and emission from the paddy fields. However, the relationships between rice varieties and populations of microorganisms involved in CH₄ dynamics are poorly understood. Here we investigated CH₄ dynamics and the composition and abundance of CH₄‐producing archaea and CH₄‐oxidizing bacteria in a Chinese rice field soil planted with three types of rice. Hybrid rice produced 50–60% more of shoot biomass than Indica and Japonica cultivars. However, the emission rate of CH₄ was similar to Japonica and lower than Indica. Furthermore, the dissolved CH₄ concentration in the rhizosphere of hybrid rice was markedly lower than Indica and Japonica cultivars. The rhizosphere soil of hybrid rice showed a similar CH₄ production potential but a higher CH₄ oxidation potential compared with the conventional varieties. Terminal restriction fragment length polymorphism analysis of the archaeal 16S rRNA genes showed that the hydrogenotrophic methanogens dominated in the rhizosphere whereas acetoclastic methanogens mainly inhabited the bulk soil. The abundance of total archaea as determined by quantitative (real‐time) PCR increased in the later stage of rice growth. However, rice variety did not significantly influence the structure and abundance of methanogenic archaea. The analysis of pmoA gene fragments (encoding the α‐subunit of particulate methane monooxygenase) revealed that rice variety also did not influence the structure of methanotrophic proteobacteria, though variable effects of soil layer and sampling time were observed. However, the total copy number of pmoA genes in the rhizosphere of hybrid rice was approximately one order of magnitude greater than the two conventional cultivars. The results suggest that hybrid rice stimulates the growth of methanotrophs in the rice rhizosphere, and hence enhances CH₄ oxidation which attenuates CH₄ emissions from the paddy soil. Hybrid rice is becoming more and more popular in Asian countries. The present study demonstrated that planting of hybrid rice will not enhance CH₄ emissions albeit a higher grain production than the conventional varieties.     

Colonization of rice roots with methanogenic archaea controls photosynthesis‐derived methane emission

The methane emitted from rice fields originates to a large part (up to 60%) from plant photosynthesis and is formed on the rice roots by methanogenic archaea. To investigate to which extent root colonization controls methane (CH₄) emission, we pulse‐labeled rice microcosms with ¹³CO₂ to determine the rates of ¹³CH₄ emission exclusively derived from photosynthates. We also measured emission of total CH₄ (¹²⁺¹³CH₄), which was largely produced in the soil. The total abundances of archaea and methanogens on the roots and in the soil were analysed by quantitative polymerase chain reaction of the archaeal 16S rRNA gene and the mcrA gene coding for a subunit of the methyl coenzyme M reductase respectively. The composition of archaeal and methanogenic communities was determined with terminal restriction fragment length polymorphism (T‐RFLP). During the vegetative growth stages, emission rates of ¹³CH₄ linearly increased with the abundance of methanogenic archaea on the roots and then decreased during the last plant growth stage. Rates of ¹³CH₄ emission and the abundance of methanogenic archaea were lower when the rice was grown in quartz‐vermiculite with only 10% rice soil. Rates of total CH₄ emission were not systematically related to the abundance of methanogenic archaea in soil plus roots. The composition of the archaeal communities was similar under all conditions; however, the analysis of mcrA genes indicated that the methanogens differed between the soil and root. Our results support the hypothesis that rates of photosynthesis‐driven CH₄ emission are limited by the abundance of methanogens on the roots.     

Effects of free-air CO2 enrichment (FACE) on CH4 emission from a rice paddy field

Methane (CH₄) is a particularly potent greenhouse gas with a radiative forcing 23 times that of CO₂ on a per mass basis. Flooded rice paddies are a major source of CH₄ emissions to the Earth's atmosphere. A free‐air CO₂ enrichment (FACE) experiment was conducted to evaluate changes in crop productivity and the crop ecosystem under enriched CO₂ conditions during three rice growth seasons from 1998 to 2000 in a rice paddy at Shizukuishi, Iwate, Japan. To understand the influence of elevated atmospheric CO₂ concentrations on CH₄ emission, we measured methane flux from FACE rice fields and rice fields with ambient levels of CO₂ during the 1999 and 2000 growing seasons. Methane production and oxidation potentials of soil samples collected when the rice was at the tillering and flowering stages in 2000 were measured in the laboratory by the anaerobic incubation and alternative propylene substrates methods, respectively. The average tiller number and root dry biomass were clearly larger in the plots with elevated CO₂ during all rice growth stages. No difference in methane oxidation potential between FACE and ambient treatments was found, but the methane production potential of soils during the flowering stage was significantly greater under FACE than under ambient conditions. When free‐air CO₂ was enriched to 550 ppmv, the CH₄ emissions from the rice paddy field increased significantly, by 38% in 1999 and 51% in 2000. The increased CH₄ emissions were attributed to accelerated CH₄ production potential as a result of more root exudates and root autolysis products and to increased plant‐mediated CH₄ emissions because of the larger rice tiller numbers under FACE conditions.     

Effect of ducks on CH4 emission from paddy soils and its mechanism research in the rice-duck ecosystem

The effect of ducks on CH₄ emission from paddy soils and its mechanism were probed in order to decide the optimum number of ducks in the rice-duck ecosystem. Methane emission fluxes from paddy soils were measured by the static box technique. The correlations between methane emission and soil physical and chemical characteristics were also analyzed. The results showed that significant differences (p < 0.01) existed in the dissolved oxygen content of water body in the treatment fields, and the more the ducks, the higher the dissolved oxygen content. Secondly, the soil redox matter content and methanogenic bacteria population of the rice-duck ecosystem reduced more sharply than those of the no-duck rice farming, resulting in a lower methane production. Thirdly, the amount of methane emission differed between the treatments—the more the ducks, the less the methane emission. Other related analyses showed that the negative correlation was significant (p < 0.001) between the methane emission flux and dissolved oxygen content of water body. However, CH4 emission flux had significantly positive correlation (p < 0.01) with the soil redox matter content and rice field methanogenic population.     

Methane sources in arctic thermokarst lake sediments on the North Slope of Alaska

The permafrost on the North Slope of Alaska is densely populated by shallow lakes that result from thermokarst erosion. These lakes release methane (CH₄) derived from a combination of ancient thermogenic pools and contemporary biogenic production. Despite the potential importance of CH₄ as a greenhouse gas, the contribution of biogenic CH₄ production in arctic thermokarst lakes in Alaska is not currently well understood. To further advance our knowledge of CH₄ dynamics in these lakes, we focused our study on (i) the potential for microbial CH₄ production in lake sediments, (ii) the role of sediment geochemistry in controlling biogenic CH₄ production, and (iii) the temperature dependence of this process. Sediment cores were collected from one site in Siqlukaq Lake and two sites in Sukok Lake in late October to early November. Analyses of pore water geochemistry, sedimentary organic matter and lipid biomarkers, stable carbon isotopes, results from CH₄ production experiments, and copy number of a methanogenic pathway‐specific gene (mcrA) indicated the existence of different sources of CH₄ in each of the lakes chosen for the study. Analysis of this integrated data set revealed that there is biological CH₄ production in Siqlukaq at moderate levels, while the very low levels of CH₄ detected in Sukok had a mixed origin, with little to no biological CH₄ production. Furthermore, methanogenic archaea exhibited temperature‐dependent use of in situ substrates for methanogenesis, and the amount of CH₄ produced was directly related to the amount of labile organic matter in the sediments. This study constitutes an important first step in better understanding the actual contribution of biogenic CH₄ from thermokarst lakes on the coastal plain of Alaska to the current CH₄ budgets.     

Methane Biogeochemistry and Methanogen Communities in Two Northern Peatland Ecosystems,New York State

Rates of methane (CH₄) production vary considerably among northern peat-forming wetlands, and it is not clear whether variability is caused by environmental factors affecting CH₄ production or differences in methanogen communities. We investigated CH₄ production and emission dynamics concomitantly with 16S rRNA gene sequence-based community analysis of Archaea in two contrasting peat-forming northern wetlands, an ombrotrophic bog and a minerotrophic conifer swamp. Individual measurements of CH₄ emissions to the atmosphere followed a lognormal distribution pattern in both sites, and mean rates were 30× greater in the bog site. Rates of CH₄ production measured in vitro were initially 3× greater in the bog than in the conifer swamp; although, after 30 days of incubation, production rates were similar suggesting that in situ environmental conditions limited production in the conifer swamp. Amplified ribosomal DNA restriction analysis (ARDRA) and rarefaction techniques indicated that both sites had similar levels of archaeal richness, with 27 unique taxa in the bog and 23 taxa in the conifer swamp. However, the bog had more pronounced dominance of a few taxa, whereas the conifer swamp had more even distribution among taxa. A 16S rRNA gene sequence-based phylogenetic analysis indicated high levels of diversity with similarity to known methanogenic families Methanosarcinaceae, Methanosaetaceae, Methanobacteriaceae, and likely Methanomicrobiaceae as well as two additional lineages previously characterized as groups of yet uncultivated Euryarchaeota commonly occurring in flooded rice soils. Therefore, sites with low and high rates of CH₄ production supported very diverse methanogenic communities.     

Functional-Structural Analysis of Nitrogen-Cycle Bacteria in a Hypersaline Mat from the Omani Desert

Anaerobic microbial activity in northern peat soils most often results in more carbon dioxide (CO ₂ ) production than methane (CH₄) production. This study examined why methanogenic conditions (i.e., equal molar amounts of CH₄ production and CO₂ production) prevail so infrequently. We used peat soils from two ombrotrophic bogs and from two rheotrophic fens. The former two represented a relatively dry bog hummock and a wet bog hollow, and the latter two represented a forested fen and a sedge-dominated fen. We quantified gas production rates in soil samples incubated in vitro with and without added metabolic substrates (glucose, ethanol, H₂/CO₂). None of the peat soils exhibited methanogenic conditions when incubated in vitro for a short time (< 5 days) and without added substrates. Incubating some samples > 50 days without added substrates led to methanogenic conditions in only one of four experiments. The anaerobic CO₂:CH₄ production ratio ranged from 5:1 to 40:1 in peat soil without additions and was larger in samples from the dry bog hummock and forested fen than the wet bog hollow and sedge fen. Adding ethanol or glucose separately to peat soils led to methanogenic conditions within 5 days after the addition by stimulating rates of CH₄ production, suggesting CH₄ production from both hydrogenotrophic and acetoclastic methanogenesis. Our results suggest that methanogenic conditions in peat soils rely on a constant supply of easily decomposable metabolic substrates. Sample handling and incubation procedures might obscure methanogenic conditions in peat soil incubated in vitro.     

Higher yields and lower methane emissions with new rice cultivars

Breeding high‐yielding rice cultivars through increasing biomass is a key strategy to meet rising global food demands. Yet, increasing rice growth can stimulate methane (CH₄) emissions, exacerbating global climate change, as rice cultivation is a major source of this powerful greenhouse gas. Here, we show in a series of experiments that high‐yielding rice cultivars actually reduce CH₄ emissions from typical paddy soils. Averaged across 33 rice cultivars, a biomass increase of 10% resulted in a 10.3% decrease in CH₄ emissions in a soil with a high carbon (C) content. Compared to a low‐yielding cultivar, a high‐yielding cultivar significantly increased root porosity and the abundance of methane‐consuming microorganisms, suggesting that the larger and more porous root systems of high‐yielding cultivars facilitated CH₄ oxidation by promoting O₂ transport to soils. Our results were further supported by a meta‐analysis, showing that high‐yielding rice cultivars strongly decrease CH₄ emissions from paddy soils with high organic C contents. Based on our results, increasing rice biomass by 10% could reduce annual CH₄ emissions from Chinese rice agriculture by 7.1%. Our findings suggest that modern rice breeding strategies for high‐yielding cultivars can substantially mitigate paddy CH₄ emission in China and other rice growing regions.     

Effect of rice plants on methane production and rhizospheric metabolism in paddy soil

In order to elucidate the effects of rice plants on CH₄ production, we conducted experiments with soil slurries and planted rice microcosms. Methane production in anoxic paddy soil slurries was stimulated by the addition of rice straw, of unsterile or autoclaved rice roots, and of the culture fluid in which rice plants had axenically been cultivated. The addition of these compounds also increased the concentrations of acetate and H₂, precursors of CH₄ production, in the soil. Planted compared to unplanted paddy soil microcosms exhibited lower porewater CH₄ concentrations but higher CH₄ emission rates. They also exhibited higher sulfate concentrations but similar nitrate concentrations. Concentrations of acetate, lactate and H₂ were not much different between planted and unplanted microcosms. Pulse labeling of rice plants with¹⁴CO₂ resulted during the next 5 days in transient accumulation of radioactive lactate, propionate and acetate, and after the second day of incubation in the emission of¹⁴CH₄. Most of the radioactivity (40–70%) was incorporated into the above-ground biomass of rice plants. However, during a total incubation of 16 days about 3–6% of the applied radioactivity was emitted as¹⁴CH₄, demonstrating that plant-derived carbon was metabolized and significantly contributed to CH₄ production. The sequence of the appearance of radioactive products and their specific radioactivities indicate that CH₄ was produced from root exudates by a microbial community consisting of fermenting and methanogenic bacteria.     

Effects of N-fertilisation on CH4 oxidation and production, and consequences for CH4 emissions from microcosms and rice fields

The world's growing human population causes an increasing demand for food, of which rice is one of the most important sources. In rice production nitrogen is often a limiting factor. As a consequence increasing amounts of fertiliser will have to be applied to maximise yields. There is an ongoing discussion on the possible effects of fertilisation on CH₄ emissions. We therefore investigated the effects of N‐fertiliser (urea) on CH₄ emission, production and oxidation in rice microcosms and field experiments. In the microcosms, a substantial but short‐lived reduction of CH₄ emission was observed after N‐addition to 43‐d‐old rice plants. Methane oxidation increased by 45%, demonstrated with inhibitor measurements and model calculations based on stable carbon isotope data (δ¹³CH₄). A second fertilisation applied to 92‐d‐old plants had no effect on CH₄ emission rates. The positive effect of additional N on methanotrophic bacteria was also found in vitro for potential CH₄ oxidation rates in soil and root samples from the microcosm and field experiments, indicated by elevated initial oxidation rates and reduced lag‐phases. Fertilisation did not affect methane production in the microcosms. In the field, the effects were diverse: methane production was inhibited in the topsoil, but stimulated instead in the bulk soil. Stimulation occurred probably in the anaerobic food chain at the level of hydrolytic or fermenting bacteria, because acetate, a methanogenic precursor, increased simultaneously. Combining field, microcosm and laboratory experiments we conclude that any agricultural treatment improving the N‐supply to the rice plants will also be favourable for the CH₄ oxidising bacteria. However, N‐fertilisation had only a transient influence and was counter‐balanced in the field by an elevated CH₄ production. A negative effect of the fertilisation was a transient increase of N₂O emissions from the microcosms. However, integrating over the season the global warming potential (GWP) of N₂O emitted after fertilisation was still negligible compared to the GWP of emitted CH₄.     

Transgenic Bt rice has adverse impacts on CH4 flux and rhizospheric methanogenic archaeal and methanotrophic bacterial communities

Background and Aims

The effect of transgenic insect-resistant crops on soil microorganisms has become an issue of public concern. The goal of this study was to firstly realize the variation of in situ methane (CH₄) emission flux and methanogenic and methanotrophic communities due to planting transgenic Bt rice (Bt) cultivar.

Methods

CH₄ emitted from paddy soil was collected by static closed chamber technique. Denaturing gradient gel electrophoresis and real-time PCR methods were employed to analyze methanogenic archaeal and methanotrophic bacterial community structure and abundance.

Results

Results showed that planting Bt rice cultivar effectively reduced in situ CH₄ emission flux and methanogenic archaeal and methanotrophic bacterial community abundance and diversity. Data analysis showed that in situ CH₄ emission flux increased significantly with the increase of methanogenic archaeal abundance (R ² ?=?0.839, p?<?0.001) and diversity index H′ (R ² ?=?0.729, p?<?0.05), whereas was not obviously related to methanotrophic bacterial community.

Conclusions

Our results suggested that the lower in situ CH₄ emission flux from Bt soil may result from lower methanogenic archaeal community abundance and diversity, lower methanogenic activity and higher methanotrophic activity. Moreover, our results inferred that specific functional microorganisms may be a more sensitive indicator than the total archaeal, bacterial or fungal population to assess the effects of transgenic insect-resistant plants on soil microorganisms.     

UV-B辐射增强对抗除草剂转基因水稻 CH4排放的影响

在大田条件下,研究了UV-B(ultraviolet-B)辐射增强对抗除草剂转基因水稻及亲本常规水稻甲烷(CH4)排放的影响。UV-B辐射设2水平,即对照(CK,自然光),增强(Elevated,14.4 kJ·m-·2d-1),相当于南京地区大气臭氧耗损25%的辐射剂量。结果表明,UV-B辐射增强并没有改变稻田CH4排放通量的季节性变化规律。与对照相比,UV-B辐射增强显著提高CH4排放通量和累积排放量。水稻分蘖期CH4累积排放量最高,占全生育累积排放量的51.55%—61.01%;其次是拔节至孕穗期,占20.00%—26.64%。抗除草剂转基因水稻的CH4排放通量和累积排放量显著低于亲本常规水稻。研究说明,UV-B辐射增强下种植抗除草剂转基因水稻对于减缓稻田甲烷排放有积极意义。     

Agronomic aspects of wetland rice cultivation and associated methane emissions

Wetland rice cultivation is one of the major sources of atmospheric methane (CH₄). Global rice production may increase by 65% between 1990 and 2025, causing an increase of methane emissions from a 92 Tg CH₄ y^–1 now to 131 Tg in 2025.Methane production depends strongly on the ratio oxidizing: reducing capacity of the soil. It can be influenced by e.g. addition of sulphate, which inhibits methanogenesis. The type and application mode of mineral fertilizers may also affect methane emissions. Addition of organic matter in the form of compost or straw causes an increase of methane emissions, but methane production is lower for materials with a low C/N ratio.High percolation rates in wetland rice soils and occasional drying up of the soil during the cultivation period depresses methane release. Water management practices aimed at reducing emissions are only feasible during specific periods in the rice growing season in flat lowland irrigated areas with high security of water availability and good control of the water supply. Intermittent drying of soils may not be possible on terraced rice lands.Assuming a 10 to 30% reduction in emission rates per unit harvested area, the global emission may amount to 93 Tg CH₄ y^– in 2025. A reduction of global emissions seems very difficult. To develop techniques for reducing CH₄ emissions from wetland rice fields, research is required concerning interactions between soil chemical and physical properties, and soil, water and crop management and methanogenesis. Such techniques should not adversely affect rice yields.     

Processes involved in formation and emission of methane in rice paddies

The seasonal change of the rates of production and emission of methane were determined under in-situ conditions in an Italian rice paddy in 1985 and 1986. The contribution to total emission of CH₄ of plant-mediated transport, ebullition, and diffusion through the flooding water was quantified by cutting the plants and by trapping emerging gas bubbles with funnels. Both production and emission of CH₄ increased during the season and reached a maximum in August. However, the numbers of methanogenic bacteria did not change. As the rice plants grew and the contribution of plant-mediated CH₄ emission increased, the percentage of the produced CH₄ which was reoxidized and thus, was not emitted, also increased. At its maximum, about 300 ml CH₄ were produced per m² per hour. However, only about 6% were emitted and this was by about 96% via plant-mediated transport. Radiotracer experiments showed that CH, was produced from H₂/CO₂. (30–50%) and from acetate. The pool concentration of acetate was in the range of 6–10 mM. The turnover time of acetate was 12–16 h. Part of the acetate pool appeared to be not available for production of CH₄ or CO₂     

Functional and structural response of the methanogenic microbial community in rice field soil to temperature change

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 (α_app) 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.     

The carbon and hydrogen stable isotope composition of bacteriogenic methane: A laboratory study using a landfill inoculum

Anaerobic bacterial degradation of landfill waste produces a globally significant source of the greenhouse gas methane. Stable isotopic measurements of methane [δ^I3C(CH₄) and δD(CH₄)] can often fingerprint different sources of methane (natural vs. anthro‐pogenic) and help identify the bacterial processes involved in methane production. Landfill microbial communities are complex and diverse, and hence so too is the biogeochem‐istry of methane formation. To investigate the influence of (l) the methane formation pathway (acetoclastic methanogenesis and CO₂ reduction), and (2) SD of water on the stable isotopic composition of landfill methane, two model butyrate‐degrading landfill systems were established. The systems were inoculated with domestic refuse from a landfill and incubated in the laboratory for 92 days. Both systems were identical except δD of water initially added to system 2 was 118% heavier than system 1. Between days 39 and 72 the systems were resupplemented with butyrate. Production of CH₄ and CO₂ and changes in volatile fatty acid concentration confirmed that active methanogenic populations had been established. CH₄ became ¹³C enriched in both incubations with time. Interpreting changes in acetate, butyrate, and propionate concentration during incubation is complicated, but these observations and other information suggest that the dominant methanogenic substrate changed front CO₂/H₂ to acetate as the experiment progressed. This is also consistent with the observed ¹³C enrichment of CH₄, as ¹³C discrimination during methane production from acetate is less than from CO₂. In contrast, δD(CH₄) remained relatively constant, suggesting that this measurement may not provide a reliable basis for distinguishing between CH₄ from CO₂ reduction and acetoclastic methanogenesis, as has previously been suggested.     

Pattern and Amount of Aerenchyma Relate to Variable Methane Transport Capacity of Different Rice Cultivars

Abstract: Aerenchyma, developed in both root and aboveground parts of rice plants, is predominantly responsible for plant‐mediated transfer of methane (CH₄) from the soil to the atmosphere. To clarify the pathways of CH₄ transport through the rice plant and find differences that may determine the large¡variation in the patterns of methane transport capacity (MTC) of rice cultivars, we examined the appearance, the distribution pattern, and the density of aerenchyma in different parts of rice¡plants of three widely varying rice cultivars during panicle initiation, flowering, and maturity stages. The data on the amount and density of small (> 1 ¡Á 10³? 5 ¡Á 10³Ìm²), medium (> 5 ¡Á 10³? 20 ¡Á 10³Ìm²) and large aerenchyma lacunae (> 20 ¡Á 10³Ìm²) were collected using a computer assisted image‐analyzing system. The brightfield optical microscopy of roots of all tested rice plants demonstrated the continuity of aerenchyma channels in the roots that function as conduits for bi‐directional transport of gases. The aerenchyma channels of primary roots showed direct connection with those of culms. Intercalary meristems were found at the transition zone of rootaCculm aerenchyma connections. Well‐developed aerenchyma lacunae present in the internodal region of the culm base were uniformly distributed in the peripheral cortical zone. The nodal region had relatively fewer and smaller aerenchyma lacunae that showed a non‐uniform distribution pattern. As a result, few aerenchyma channels continued from the internodal region through to the nodal region. The aerenchyma in the cortex zone of the culm expanded along with the growing secondary tiller, developing continuity between the culm and the secondary tiller. The micrographs of longitudinal sections of different specimens of culmaCleaf sheath intersection showed the continuity of aerenchyma channels from the culm to the leaf. The amount of medium and large aerenchyma lacunae in the leaf sheath was respectively 2 and 33 times greater as compared to those of the tiller. The proportion of the large lacunae in the total amount of aerenchyma in leaf sheath was 75 % as compared to only 8 % in the tiller, revealing higher number and larger size of aerenchyma in the former. There were significant differences in amount and density of aerenchyma between individual cultivars at a given growth stage, as well as in the development patterns. While the amount and density of medium and small aerenchyma lacunae in the internodal region of the culm base did not show any relationship with MTC of rice cultivars, large aerenchyma lacunae exhibited highly significant correlations with MTC of different cultivars, suggesting that the wide variation in MTC of rice plants during different growth stages are related to these structural features.