Methane-oxidizing activities and methanotrophic populations associated with wetland rice plants

Acetylene up to 500 μl l^–1 did not affect methane formation in anoxic soil up to 12 h, but further incubation for 1 week showed strong inhibition of methanogenesis. To ascertain the extent of the oxidation of methane produced from rice-planted pots, the effect of acetylene on methane emission was studied. Two rice varieties (Toyohatamochi and Yamahikari) were grown in a greenhouse in submerged soil in pots. At about maximum tillering, heading, and grain-forming stages, methane fluxes were measured. Flux measurement was performed for 3 h from 6 pm, then acetylene at 100 μl l^–1 was added to some of the pots. At 6 a.m. the following day, methane fluxes were again measured for 3 h. Only at maximum tillering stage of the variety Toyohatamochi was a significant increase (1.4 times) in methane flux caused by acetylene observed, whereas in the other treatments no significant increase in methane fluxes by acetylene could be defected. To ascertain the activity of methane monooxygenase (MMO), propylene oxide (PPO) formation from propylene was measured with excised roots and a basal portion of stems of the rice plants grown on the submerged soil. A level of 0.1–0.2 μmol PPO h^–1 plant^–1 was recorded. The roots showed the highest PPO formation per gram dry matter, followed by basal stems. Methane oxidation was roughly proportional to PPO formation. Soluble MMO-positive methanotroph populations were measured by plate counts. The number of colony-forming units per gram dry matter was 10⁶–10⁵ in roots, and 10⁴–10³ in basal stems. These results indicate the possibility of methane oxidation in association with wetland rice plants. Received: 26 October 1995     

Methane emission from two Indian soils planted with different rice cultivars

In a greenhouse study, methane emissions were measured from two diverse Indian rice-growing soils planted to five rice cultivars under similar water regimes, fertilizer applications and environmental conditions. Significant variations were observed in methane emitted from soils growing different cultivars. Total methane emission varied between 8.04 and 20.92gm^–2 from IARI soil (Inceptisol) and between 1.47 and 10.91gm^–2 from Raipur soil (Vertisol) planted to rice. In all the cultivars, emissions from IARI soil were higher than from Raipur soil. The first methane flux peak was noticed during the reproductive phase and the second peak coincided with the grain-ripening stage of the rice cultivars. Received: July 7, 1996     

土壤中钙结合和铁铝结合腐殖质的组成及结构特征

Calcium-bound and iron- and aluminium-bound humus extracted from different soils collected from north to south of China were characterized by chemical and spectroscopic methods. Meaningful differences in the composition and structure between them were revealed by ¹³C NMR, visible spectroscopy and elemental analysis. Results showed that the contents of carbon, hydrogen and nitrogen were higher in iron- and aluminium-bound humus than in calcium-bound humus while oxygen content in calcium-bound humus was shown to be higher. The calcium-bound humus had higher C/N and O/C ratios than iron- and aluminium-bound humus. The calcium-bound humic acid (HA1) showed higher E₄/E₆ ratios than iron- and aluminum-bound humic acid (HA2) while iron- and aluminum-bound fulvic acid (FA2) showed higher E₄/E₆ ratios than calcium-bound fulvic acid (FA1). An inverse relationship between E₄/E₆ ratios and aromaticity as determined by ¹³C NMR spectra was observed for HA and FA from black soil. The ¹³C NMR spectroscopy revealed that HA2 was more aromatic than HA1. On the other hand, FA1 exhibited a higher aromaticity than FA2.     

Ammonium oxidation coupled to dissimilatory reduction of iron under anaerobic conditions in wetland soils

In exploring the dynamics of iron and nitrogen cycling in sediments from riparian forests we have observed a redox reaction that has not been previously described. During incubations of soil slurries under strictly anaerobic conditions, we repeatedly measured an unexpected production of both nitrite () and ferrous iron [Fe(II)]. Using this indirect evidence we hypothesize that, under anaerobic conditions, there is a biological process that uses ferric iron [Fe(III)] as an electron acceptor while oxidizing ammonium () to for energy production. This oxidation under iron reducing anaerobic conditions is thermodynamically feasible and is potentially a critical component of the N cycle in saturated sediments.     

Methane in pristine and impaired mangrove soils and its possible effect on establishment of mangrove seedlings

Pristine and impaired mangrove soils (from road construction, aquaculture, and sewage) in Baja California Sur, Mexico were investigated for methane dynamics, related soil properties, and their impact on initial establishment of black mangrove propagules. All soils (Salic Fluvisols and Histosols) had neutral to alkaline pH, were saline, and had variable organic carbon content, and redox potentials. Most pristine mangrove soils showed low methane concentration, low methane production rates, and no methane emission. Impaired mangrove soil (from aquaculture) and mangrove soil affected by sewage water showed high methane concentration, high methane production rates, and high methane emission, thus acting as a methane source. Elevated methane concentrations, similar to levels detected in the impaired mangrove soil, reduce the growth of seedlings under closed chamber conditions. Addition of sulfate to the soil reversed the trend. These results indicate that impaired mangrove soils in dry climatic regions produce and emit methane and that elevated methane concentration in the vicinity of propagules may affect establishment of mangrove seedlings in impaired mangrove soils. This paper is in memory of the late mangrove researcher Dr. Gina Holguin of Mexico.     

Hydrological parameterization of piping in loess‐rich soils in the Bergisches Land,Nordrhein‐Westfalen,Germany

Pedohydrological properties were investigated on a piped slope in the Bergisches Land in the Rhenish Slate Mountains (Nordrhein‐Westfalen, Germany). The study confirmed that genesis and development of pipes decisively depend on hydrological conditions in the soils. Vertical water permeability of saturated samples was very high. This promoted fast seepage. Many macropores produced by earthworms also caused high transport capacity for soil water. Even more efficient were the burrows of moles and mice, enabling immediate infiltration and direct vertical and lateral water movement. On the contrary, the horizontal saturated permeability was low indicating no correlation with piping. Porosity of piped soils was not different to that of other soils of the region without pipes.     

Characteristics of microbial biomass and soil organic matter in newly reclaimed wetland rice soils

Microbial biomass C, N, total organic C, N and mineralizable N were measured in newly reclaimed wetland sandy loam rice soil with a very low nutrient status. Microbial biomass C increased 5.4–10.4 times due to application of barnyard manure, but decreased drastically to 24–27% during rice cultivation. Organic C and N contents also decreased during cultivation, but to a lesser extent to 59–76%. At the tillering stage of the rice plant, microbial biomass N was highly correlated with mineralizable N (r=0.986).     

Interpreting pH and nitrate concentration in solutions drained from acid soils. Possible artefacts of lysimeters

Abstract. Solutions collected from lysimeters of acid soils can show pH values close to or even above neutral. Laboratory experiments on an acid soil from Burundi were planned to test if denitrification or CO₂ degassing might explain such a paradox. In the first experiment, soil profiles were reconstituted in columns and leached with 55 μ m Ca(NO₃)₂ solutions at 30 °C and 4 °C. Two drainage regimes were applied: intermittent suction or no suction at the bottom of the columns. In the second experiment, pH values were measured in solutions drained from different horizons at 30 °C, before and after equilibration with ambient air. Sterilized soil was also tested in the same way. Results from experiment 1 showed that despite the accumulation of water in the bottom of soil profiles when no suction was applied, aeration still existed so that reduction reactions, namely denitrification, are not expected to affect greatly the percolate composition. Indeed nitrate concentration was similar in both drainage regimes and was close to the input value. The pH values in percolates were close to 7 at 30 °C and they dropped to about 5.5 when the columns were at 4 °C. In experiment 2, equilibration of percolates with ambient air resulted in pH increase which was greater for the top horizon (C-rich) but negligible when the soil was first sterilized. These convergent results illustrate the very important effect of CO₂ degassing on pH of drained solutions when microbial activity is stimulated at high temperatures, in C-rich soil. This is of prime importance when interpreting results from lysimeter experiments. By chance, this study also showed that large quantities of nitrate can be produced in soil at low temperatures.     

Influence of nitrogen on atrazine and 2, 4 dichlorophenoxyacetic acid mineralization in blackwater and redwater forested wetland soils

 Microcosms were used to determine the influence of N additions on active bacterial and fungal biomass, atrazine and dichlorophenoxyacetic acid (2,4-D) mineralization at 5, 10 and 15 weeks in soils from blackwater and redwater wetland forest ecosystems in the northern Florida Panhandle. Active bacterial and fungal biomass was determined by staining techniques combined with direct microscopy. Atrazine and 2,4-D mineralization were measured radiometrically. Treatments were: soil type, (blackwater or redwater forested wetland soils) and N additions (soils amended with the equivalent of 0, 200 or 400 kg N ha^–1 as NH₄NO₃). Redwater soils contained higher concentrations of C, total N, P, K, Ca, Mn, Fe, B and Zn than blackwater soils. After N addition and 15 weeks of incubation, active bacterial biomass in redwater soils was lower when N was added. Active bacterial biomass in blackwater soils was lower when 400 kg N ha^–1, but not when 200 kg N ha^–1, was added. Active fungal biomass in blackwater soils was higher when 400 kg N ha^–1, but not when 200 kg N ha^–1, was added. Active fungal biomass in redwater soils was lower when 200 kg N ha^–1, but not when 400 kg N ha^–1, was added. After 15 weeks of incubation 2,4-D degradation was higher in redwater wetland soils than in blackwater soils. After 10 and 15 weeks of incubation the addition of 200 or 400 kg N ha^–1 decreased both atrazine and 2,4-D degradation in redwater soils. The addition of 400 kg N ha^–1 decreased 2,4-D degradation but not atrazine degradation in blackwater soils after 10 and 15 weeks of incubation. High concentrations of N in surface runoff and groundwater resulting from agricultural operations may have resulted in the accumulation of N in many wetland soils. Large amounts of N accumulating in wetlands may decrease mineralization of toxic agricultural pesticides. Received: 26 June 1998     

Evaluation of alternative substrates for determining methane-oxidizing activities and methanotrophic populations in soils

The magnitude of methane emission is a net result of methane production and the oxidation rate. The possibility of measuring oxidized products of alternative substrates of methane monooxygenase was examined to determine methane-oxidizing ability of soils, and to count methanotrophic populations in soils. Wetland rice soils were incubated under methane containing air to enirch the methanotrophs. Methane loss and oxygen uptake were inhibited by acetylene, dimethylether, and nitrapyrin (N-Serve). Acetylene was used routinely, because it inhibited methane oxidation even at a low concentration of 0.03 to 0.06 l ml^-1 in the incubation headspace. Propylene at 10 kPa was used as an alternative substrate of methane monooxygenase, and the formation of propylene oxide was measured. When soils were incubated under methane, their methane-oxidizing activity increased. Propylene oxide formation increased simultaneously. Acetylene also blocked propylene oxidation. The results of several experiments and propylene oxide formation (r=0.87 after long-transformation). These results indicate that propylene oxide formation can be used as a semiquantitative measure of the methane-oxidizing activity of soils. The colonies of soluble methane monooxygenase-forming methanotrophs were counted on Cu-deficient methanotroph agar medium by the formation of naphthol from haphthalene. The counts increased from 10⁴ (0 days) to 10⁷ (21 days) g^-1 soil during oxic incubation under methane.     

Using the spermosphere model technique to describe the dominant nitrogen-fixing microflora associated with wetland rice in two Egyptian soils

Summary This study is an attempt to describe the dominant N₂-fixing microflora associated with the roots of wetland rice. Rice cultivar Giza 171 was grown in a phytotron on two alluvial Egyptian soils for 8 days, a stage when the nitrogenase activity of undisturbed plants reached a level of 245 × 10^–6 mol C₂H₄ h^–1 g^–1 dry weight of leaf. The roots and rhizosphere soils were then used for counting and isolating dominant diazotrophs. Counts and initial enrichment steps were carried out on a selective medium made of an axenic rice plantlet, the spermosphere model, incubated under 1 % acetylene. The counts were very high, exceeding 10⁸ bacteria g^–1 dry weight of rhizosphere soil. Enterobacteriaceae were dominant; most isolates were Enterobacter cloacae belonging to different biotypes in the two soils. Enterobacter agglomerans, Citrobacter freundii and Klebsiella planticola were also present as members of the dominant microflora. Azospirillum brasilense and Azospirillum lipoferum were present as well, but less abundant.     

Impact of land use changes on the hydrological properties of volcanic ash soils in South Ecuador

Abstract. The effect of land use on the water retention capacity of Umbric Andosols in south Ecuador was studied. The objective was to acquire a better insight into the hydrological processes of the ecosystem and the role of the soil, in order to assess the impact of changing soil properties due to land use change on the hydrology of the high Andes region. Field data on the water retention capacity at wilting point of Umbric Andosols were collected for both cultivated field conditions and original bush vegetation. The pH in water and in NaF, texture, organic matter content and dry bulk density were measured to show which physicochemical soil characteristics are responsible for the water retention of the Umbric Andosols and for the irreversible loss in water retention due to air drying. Organic matter content appears to be very important and certainly more important than allophane clay content. Water retention of the organic litter layer was calculated to be 16 mm, this would be lost when vegetation was cleared and the land cultivated.     

Localization of denitrification activity in macropores of a riparian wetland

Soil structure heterogeneity in the form of macropores and preferential flow channels can complicate efforts to quantify the physical and biological characteristics of wetland systems. We collected soil cores from two riparian wetlands to determine whether soil associated with macropores had elevated denitrification potentials compared to bulk soil from the same core. Cores were inspected for obvious macropores, which were distinguished as visible holes in the core, sometimes with decaying root matter, or as highly unconsolidated layers that appeared to have a substantially lower bulk density than the surrounding soil. Denitrification potentials were significantly higher in pores (P<0.05) for six of the 16 cores that were obtained from the Cheraw State Park site. In cores obtained from a second site, denitrification potentials were significantly higher in pores for six of 20 cores and the trend of higher denitrification in pores was present in the majority of cores that had measurable activity. In cores with significant differences, denitrification was often 1-2 orders of magnitude greater in soil surrounding the macropore than in the bulk soil. Denitrification potentials of the bulk soils were similar in magnitude to the potentials measured in composited cores from previous studies. It is possible that the difference between macropore and bulk denitrification rates developed due to preferential flow of nitrate-rich water through the macropores. Previous work showed that water entering these riparian systems in groundwater and storm runoff had elevated levels of NO₃⁻.     

Plant species traits regulate methane production in freshwater wetland soils

Ecosystem and biogeochemical responses to anthropogenic stressors are the result of complex interactions between plants and microbes. A mechanistic understanding of how plant traits influence microbial processes is needed in order to predict the ecosystem-level effects of natural or anthropogenic change. This is particularly true in wetland ecosystems, where plants alter the availability of both electron donors (e.g., organic carbon) and electron acceptors (e.g., oxygen and ferric iron), thereby regulating the total amount of anaerobic respiration and the production of methane, a highly potent greenhouse gas. In this study, we examined how plant traits associated with plant inputs of carbon (photosynthesis and biomass) and oxygen (root porosity and ferric iron on roots) to mineral soils relate to microbial competition for organic carbon and, ultimately, methane production. Plant productivity was positively correlated with microbial respiration and negatively correlated to methane production. Root porosity was relatively constant across plant species, but belowground biomass, total biomass, and the concentration of oxidized (ferric) iron on roots varied significantly between species. As a result the size of the total root oxidized iron pool varied considerably across plant species, scaling with plant productivity. Large pools of oxidized iron were related to high CO₂:CH₄ ratios during microbial respiration, indicating that as plant productivity and biomass increased, microbes used non-methanogenic respiration pathways, most likely including the reduction of iron oxides. Taken together these results suggest that increased oxygen input from plants with greater biomass can offset any potential stimulation of methanogenic microbes from additional carbon inputs. Because the species composition of plant communities influences both electron donor and acceptor availability in wetland soils, changes in plant species as a consequence of anthropogenic disturbance have the potential to trigger profound effects on microbial processes, including changes in anaerobic decomposition rates and the proportion of mineralized carbon emitted as the greenhouse gas methane.     

Response of soil constituents to freeze-thaw cycles in wetland soil solution

Soil freeze-thaw cycles in the winter-cold zone can substantially affect soil carbon, nitrogen and phosphorus cycling, and deserve special consideration in wetlands of cold climates. Semi-disturbed soil columns from three natural wetlands (Carex marsh, Carex marshy meadow and Calamagrostis wet grassland) and a soybean field that has been reclaimed from a wetland were exposed to seven freeze-thaw cycles. The freeze-thaw treatments were performed by incubating the soil columns at −10 °C for 1 d and at 5 °C for 7 d. The control columns were incubated at 5 °C for 8 d. After each freeze-thaw cycle, the soil solution was extracted by a solution extractor installed in each soil layer of the soil column, and was analyzed for dissolved organic carbon (DOC), NH₄⁺-N, NO₃⁻-N and total dissolved phosphorus (TDP). The results showed that freeze-thaw cycles could increase DOC, NH₄⁺-N and NO₃⁻-N concentrations in soil solutions, and decrease TDP concentrations. Moreover, the changes of DOC, NH₄⁺-N, NO₃⁻-N and TDP concentrations in soil solutions caused by freeze-thaw cycles were different in various sampling sites and soil layers. The increments of DOC concentrations caused by freeze-thaw cycles were greater in the wetland soil columns than in the soybean field soil columns. The increments of NH₄⁺-N concentrations caused by freeze-thaw cycles decreased with the increase of soil depth. The depth variation in the increments of NO₃⁻-N concentrations caused by freeze-thaw cycles in the wetland soil columns was different from that in the soybean field soil columns. The decrements of TDP concentrations caused by freeze-thaw cycles were greater in columns of Carex marsh and Carex marshy meadow than in columns of Calamagrostis wet grassland and the soybean field. The study results provide information on the timing of nutrient release related to freezing and thawing in natural versus agronomic soils, and have implications for the timing of nutrient application in farm fields in relation to water quality protection.     

The effects of changes in soil moisture on nitrogen cycling in acid wetland types of the New Jersey Pinelands (USA)

Wetlands are subject to changes in soil moisture as a result of both short-term seasonal climate variations and long-term changes in regional water resource management, both of which can modify the dynamics of ground and surface water inputs. In the New Jersey Pinelands, forested wetlands that differ in both plant communities and soil structure occur along a topographic and hydrological gradient associated with an unconfined aquifer. Proposed groundwater withdrawals may affect water content of soils along this gradient. We hypothesized that prolonged changes in soil moisture would alter net nitrogen mineralization and nitrification rates in proportion to the amount of moisture change, and that these changes would be similar for the different soils along the drainage catena. Soils from two catenary sequences of wetlands, including pine-dominated (driest landscape position), hardwood-dominated, and Atlantic white-cedar-dominated (wettest landscape position) communities were used in long-term laboratory incubations (36 weeks). Production of NH₄⁺-N, NO₃⁻-N, and dissolved organic N were measured under two sets of conditions: constant moisture levels of 100%, 60% and 30% water-holding capacity (WHC), and fluctuating moisture levels (alternating 2 week periods at 100% and 30% WHC). In soils from most of the wetlands, we observed increases in net mineralization and nitrification when constant low-moisture conditions were established, but not under fluctuating conditions. Contrary to expectations, responses to the drying treatments varied between wetland types and between replicate wetlands of each type. Under constant-moisture conditions, nitrification increased more in cedar swamps than in either type of pine wetland. Under all conditions, soils from all the wetlands within one of the catenas produced more inorganic and organic soluble N than did the wetlands from the other catena, suggesting that area-wide effects are as important as wetland type in regulating production of soluble N. Within both catenas, pine-hardwood wetlands generated more soluble N under all moisture conditions than did either pine-dominated or cedar wetlands. Our results suggest that changes in soil moisture due to management of water resources will affect N cycling in wetland soils, but that the magnitude of the effects, and the potential for large releases of nitrate, will depend on the specific soil properties of affected wetlands.     

Influence of phosphorus or phosphorus-potassium fertilization on biomass and dinitrogen fixation of the stem-nodulating green-manure legume Sesbania rostrata in different marginally productive wetland rice soils

The performance of Sesbania rostrata varies widely from site to site. This makes it difficult to predict the N yield and biomass of this plant in marginally productive soils, and to arouse the interest of farmers in green manure technology. Three consecutive pot experiments were conducted in a greenhouse at the International Rice Research Institute (IRRI) to evaluate growth, nodulation, N₂ fixation (C₂H₂ reduction assay and ¹⁵N dilution method), and N yield of 6-week-old S. rostrata on 13 physicochemically different wetland rice soils of the Philippines and on three artificial substrates. The performance of S. rostrata on the unfertilized controls was compared with two fertilizer treatments containing either P (100 mg P kg^-1 dry soil) or P+K (100 mg P kg^-1 and 200 mg K kg^-1 dry soil). In the control soils and substrates, the N yield of S. rostrata varied between 20 and 470 mg N per pot, with the N rate from N₂ fixation ranging between 0 and 95%. In three of the nutritionally poor soils even Mn toxicity symptoms apparently occurred with S. rostrata. P application alleviated these symptoms and increased the overall N yield considerably, mainly through increased biological N₂ fixation. An additional increase in N yield was obtained by the PK treatment. Multiple regression analysis between soil characteristics and the N yield of S. rostrata showed that the original level of P (Olsen-extracted) and Mn in the soil accounted for 73% of the variance in biomass production by S. rostrata among the unfertilized soils and substrates.     

Humic acids as electron acceptors in wetland decomposition

Decomposition of organic matter in inundated wetland soils requires a number of interdependent microbial processes that ultimately generate CO₂ and CH₄. Largely as the result of anaerobic decomposition, wetland soils store globally significant amounts of organic carbon and are currently net sources of CH₄ to the atmosphere. Given the importance of wetlands in the global carbon cycle, it is important to understand controls on anaerobic decomposition in order to predict feedbacks between wetland soils and global climate change. One perplexing pattern observed in many wetland soils is the high proportion of CO₂ resulting from anaerobic decomposition that cannot be explained by any measured pathway of microbial respiration. Recent studies have hypothesized that humic substances, and in particular solid-phase humic substances in wetland soils, can support anaerobic microbial respiration by acting as organic electron acceptors. Humic substances may thus account for much of the currently unexplained CO₂ measured during decomposition in wetland soils. Here we demonstrate that humic acids extracted from a variety of wetland soils act as either electron donors or electron acceptors and alter the ratio of CO₂:CH₄ produced during anaerobic laboratory incubations. Our results suggest that soil-derived humic substances may play an important, and currently unexplored, role in anaerobic decomposition in wetland soils.     

Enzyme activity and carbon dioxide evolution from upland and wetland rice soils under three agricultural practices in hilly regions

Summary Dehydrogenase activity (a measure of microbial biomass), urease activity and CO₂ evolution were measured in soils planted to rice (Oryza sativa L.) under three different agricultural practices prevalent in hill regions. The effects of hill slope, terrace and valley agriculture were investigated for two cropping seasons. The valleys and terraces were kept flooded during each cropping season while the hill slopes were cultivated with dryland practices. The type of agricultural practice and the date of observation had a significant influence on enzyme activity and CO₂ evolution. A positive correlation was observed between dehydrogenase and urease activity and soil moisture content. Dehydrogenase and urease activity and soil respiration were positively correlated among themselves. The activity of both enzymes and CO₂ evolution were highest in valley soils followed by terrace and hill-slope sites.