Methane production potential and methanogenic archaeal community structure in tropical irrigated Indian paddy soils

Soil characteristics regulate various belowground microbial processes including methanogenesis and, consequently, affect the structure and function of methanogenic archaeal communities due to change in soil type which in turn influences the CH₄ production potential of soils. Thus, five different soil orders (Alfisol, Entisol, Inceptisol, Podzol and Vertisol) were studied to assess their CH₄ production potential and also the methanogenic archaeal community structure in dryland irrigated Indian paddy soils. Soil incubation experiments revealed CH₄ production to range from 178.4 to 431.2 μg CH₄ g^-1 dws in all soil orders as: Vertisol<Inceptisol<Entisol<Podzol<Alfisol. The numbers of methanogens as quantified using real-time quantitative polymerase chain reaction (qPCR) targeting mcrA genes varied between 0.06 and 72.97 (×10⁶ copies g^-1 dws) and were the highest in Vertisol soil and the least in Alfisol soil. PCR-denaturing gradient gel electrophoresis (DGGE)-based approach targeting 16S rRNA genes revealed diverse methanogenic archaeal communities across all soils. A total of 43 DGGE bands sequenced showed the closely related groups to Methanomicrobiaceae, Methanobacteriaceae, Methanocellales, Methanosarcinaceae, Methanosaetaceae and Crenarchaeota. The composition of methanogenic groups differed among all soils and only the Methanocellales group was common and dominant in all types of soils. The highest diversity of methanogens was found in Inceptisol and Vertisol soils. Methane production potential varied significantly in different soil orders with a positive relationship (p?<?0.05) with methanogens population size, permanganate oxidizable C (POXC) and CO₂ production. The present study suggested that CH₄ production potential of different soils depends on physicochemical properties, methanogenic archaeal community composition and the population size.     

Dynamics of the methanogenic archaeal community in anoxic rice soil upon addition of straw

Addition of rice straw, which is a common practice in rice agriculture, generally results in enhanced production and emission of the greenhouse gas methane (CH₄). However, it is unclear whether straw addition affects only the activity or also the composition of the methanogenic microbial community. It is also unclear to what extent methanogenic archaea would be able to proliferate in the soil. Anoxic slurries of Italian rice‐field soil produced CH₄ after a lag, during which ferric iron and sulfate were reduced. Addition of rice straw slightly decreased this lag and greatly enhanced the subsequent production of CH₄. At the same time, addition of rice straw enhanced the intermediate production of H₂ and acetate that served as the methanogenic substrates. Compared with the unamended control, the addition of rice straw resulted in an increased concentration of phospholipid fatty acids in the soil. Quantitative ‘real‐time’ PCR targeting the 16S rRNA gene also showed increased copy numbers of both Bacteria and Archaea in the straw‐amended soil at the end of the experiment. The composition of the archaeal community was followed over time by terminal restriction length polymorphism (T‐RFLP) analysis of the archaeal 16S rRNA genes extracted from straw‐amended soil and the control. Rice Cluster‐I (RC‐I) methanogens and Methanosarcinaceae were the most abundant methanogenic populations, followed by Methanobacteriales, Methanomicrobiales and Methanosaetaceae. Addition of rice straw resulted in a relative increase of Methanosarcinaceae and Methanobacteriales and a relative decrease of RC‐I methanogens and Methanomicrobiales. Our results revealed a dynamic methanogenic community in anoxic rice‐field soil and showed that addition of organic matter selectively enhanced the growth of particular methanogenic populations, which were apparently better adapted to the presence of straw than the others. The extent of archaeal growth was consistent with that expected theoretically from the ambient Gibbs free energies of hydrogenotrophic and acetoclastic methanogenesis.     

Importance of hydrogenotrophic, aceticlastic and methylotrophic methanogenesis for methane production in terrestrial, aquatic and other anoxic environments: A mini review

Microbial methanogenesis is a major source of the greenhouse gas methane(CH4).It is the final step in the anaerobic degradation of organic matter when inorganic electron acceptors such as nitrate,ferric iron,or sulfate have been depleted.Knowledge of this degradation pathway is important for the creation of mechanistic models,prediction of future CH4 emission scenarios,and development of mitigation strategies.In most anoxic environments,CH4 is produced from either acetate(aceticlastic methanogenesis)or hydrogen(H2)plus carbon dioxide(CO2)(hydrogenotrophic methanogenesis).Hydrogen can be replaced by other CO2-type methanogenesis,using formate,carbon monoxide(CO),or alcohols as substrates.The ratio of these two pathways is tightly constrained by the stoichiometry of conversion processes.If the degradation of organic matter is complete(e.g.,degradation of straw in rice paddies),then fermentation eventually results in production of acetate and H2 at a ratio of>67%aceticlastic and<33%hydrogenotrophic methanogensis.However,acetate production can be favored when heterotrophic or chemolithotrophic acetogenesis is enhanced,and H2 production can be favored when syntrophic acetate oxidation is enhanced.This typically occurs at low and elevated temperatures,respectively.Thus,temperature can strongly influence the methanogenic pathway,which may range from 100%aceticlastic methanogenesis at low temperatures to 100%hydrogenotrophic methanogenesis at high temperatures.However,if the degradation of organic matter is not complete(e.g.,degradation of soil organic matter),the stoichiometry of fermentation is not tightly constrained,resulting,for example,in the preferential production of H2,followed by hydrogenotrophic methanogenesis.Preferential production of CH4 by either aceticlastic or hydrogenotrophic methanogenesis can also happen if one of the methanogenic substrates is not consumed by methanogens but is,instead,accumulated,volatilized,or utilized otherwise.Methylotrophic methanogens,which can use methanol as a substrate,are widespread,but it is unlikely that methanol is produced in similar quantities as acetate,CO2,and H2.Methylotrophic methanogenesis is important in saline environments,where compatible solutes are degraded to methyl compounds(trimethyl amine and dimethyl sulfide)and then serve as non-competitive substrates,while acetate and hydrogen are degraded by non-methanogenic processes,e.g.,sulfate reduction.     

Effect of rice straw application on stable carbon isotopes,methanogenic pathway,and fraction of CH4 oxidized in a continuously flooded rice field in winter season

Straw application is a common practice in rice agriculture, but its effect on stable carbon isotopes (δ¹³C) in each process of CH₄ emission, relative contribution of acetate to CH₄ production (F_ac), and fraction of the CH₄ that is oxidized (F_ox) in the winter season is not well known. We investigated CH₄ production and oxidation potentials in paddy soil, CH₄ concentrations in soil pore water and floodwater, and CH₄ emission as well as their δ¹³C from a continuously flooded paddy field with rice straw application (RSA) during the 2007 and 2008 winter seasons and quantified the F_ac and F_ox using the isotopic data. RSA significantly increased CH₄ emission (p < 0.05) but made δ¹³CH_{4 (emission)} lower by 5–8‰. RSA obviously increased CH₄ concentrations in soil pore water and floodwater, but it played a slight role in porewater δ¹³CH₄ while caused floodwater δ¹³CH₄ 4‰ lower. A relatively low contribution of acetate-dependent methanogenesis (about 50%) was observed, and RSA slightly increased F_ac (about 0–10%) at a relatively low temperature whereas greatly decreased it (about 10–20%) at a high temperature. Floodwater CH₄ was much more ¹³C-enriched than porewater CH₄ (p < 0.05), suggesting that about 60–70% of the CH₄ was oxidized at the soil–water interface. F_ox generally decreased in the winter season and it was reduced by RSA (about 10%). Soil temperature was positively correlated with CH₄ flux and CH₄ flux was negatively related to δ¹³CH_{4 (emission)}, suggesting that temperature might be an important factor influencing F_ox, thus CH₄ emission and δ¹³CH_{4 (emission)}. The findings suggest that RSA significantly increased CH₄ production while little affected CH₄ oxidation, thus decreasing F_ox. Moreover, the effect of RSA on methanogenic pathway might be controlled by soil temperature, and RSA generally reduced F_ac and F_ox was the main reason for the emitted CH₄ depleted in ¹³C.     

Effect of potassium phosphate fertilization on production and emission of methane and its C-stable isotope composition in rice microcosms

Rice fields are an important source for atmospheric CH₄, but the effects of fertilization are not well known. We studied the turnover of CH₄ in rice soil microcosms without and with addition of potassium phosphate. Height and tiller number of rice plants were higher in the fertilized than in the unfertilized microcosms. Emission rates of CH₄ were also higher, but porewater concentrations of CH₄ were lower. The δ¹³C values of the emitted CH₄ and of the CH₄ in the porewater were both 2-6% higher in the fertilized microcosms than in the control. Potassium phosphate did not affect rate and isotopic signature of CH₄ production in anoxic soil slurries. On the other hand, roots retrieved from fertilized microcosms at the end of incubation exhibited slightly higher CH₄ production rates and slightly higher CH₄-δ¹³C values compared to roots from unfertilized plants. Addition of potassium phosphate to excised rice roots generally inhibited CH₄ production and resulted in increasingly lower δ¹³C values of the produced CH₄. Fractionation of ¹³C during plant ventilation (i.e. δ¹³C in pore water CH₄ versus CH₄ emitted) was larger in the fertilized microcosms than in the control. Besides plant ventilation, this difference may also have been caused by CH₄ oxidation in the rhizosphere. However, calculation from the isotopic data showed that less than 27% of the produced CH₄ was oxidized. Collectively, our results indicate that potassium phosphate fertilization stimulated CH₄ emission by enhancing root methanogenesis, plant ventilation and/or CH₄ oxidation, resulting in residence times of CH₄ in the porewater in the order of hours.     

Influences of indigenous phototrophs on methane emissions from a straw-amended paddy soil

A pot experiment was conducted to investigate the influences of indigenous phototrophs on methane (CH₄) emissions from a paddy soil where rice straw was incorporated or was surface-applied. During the cultivation, half of the pots were covered with aluminum foil, except for the minimum space for rice plants, to prevent ambient light reaching the floodwater or the soil surface. Growth of oxygen-producing phototrophs was hardly observed in the unilluminated plots, whereas intensive growth of algae, duckweed and hydrophytes was found in the illuminated ones. Plant growth was not affected by the different treatments. Seasonal changes in CH₄ emission determined by a closed chamber method indicated that illumination had no or only minor effects on CH₄ emissions when rice straw was incorporated or was not applied, but significantly reduced CH₄ emissions when rice straw was surface-applied. Methanogenesis occurring in the soil-floodwater interface was further investigated in two lab-scale model experiments measuring methanogenic activity. As a result, more activated methanogenesis was found in the surface-applied rice straw and the soil around the straw compared with the soil incubated without rice straw. The magnitude of the methanogenic activity in the rice straw incubated under illuminated conditions was significantly lower than that incubated in the dark. Consequently, this study demonstrates that methanogenesis in paddy soil occurs even in the soil-floodwater interface if plant residues like rice straw exist, and such methanogenesis is likely to be suppressed by growth of indigenous phototrophs under illumination.     

Effects of rice‐straw and phosphorus application on production and emission of methane from tropical rice soil

Rice‐straw amendment increased methane production by 3‐fold over that of unamended control. Application of P as single superphosphate at 100 μg (g soil)^–1 inhibited methane (CH₄) production distinctly in flooded alluvial rice soil, in the absence more than in the presence of rice straw. CH₄ emission from rice plants (cv. IR72) from alluvial soil treated with single superphosphate as basal application, in the presence and absence of rice straw, and held under non‐flooded and flooded conditions showed distinct variations. CH₄ emission from non‐flooded soil amended with rice straw was high and almost similar to that of flooded soil without rice‐straw amendment. The cumulative CH₄ efflux was highest (1041 mg pot^–1) in rice‐straw‐amended flooded soil. Appreciable methanogenic reactions in rice‐straw‐amended soils were evident under both flooded and non‐flooded conditions. Rice‐straw application substantially altered the balance between total aerobic and anaerobic microorganisms even in non‐flooded soil. The mitigating effects of single‐superphosphate application or low‐moisture regime on CH₄ production and emission were almost nullified due to enhanced activities of methanogenic archaea in the presence of rice straw.     

Organic carbon and stable C isotope in conservation agriculture and conventional systems

Conservation agriculture might have the potential to increase soil organic C content compared to conventional tillage based systems. The present study quantified soil organic carbon (SOC) and soil C derived from C₃ (wheat) and C₄ (maize) plant species using δ¹³C stable isotope. Soil with 16 y of continuous application of zero tillage (ZT) or conventional tillage (CT), monoculture (M) or rotation (R) of wheat and maize, and with (+r) and without retention (−r) in the field of crop residues were studied in the central highlands of Mexico. The highest SOC content was found in the 0-5 cm layer under ZTM and ZTR with residues retention. The soil cultivated with maize showed a higher SOC content in the 0-10 cm layer with residue retention than without residue. In the 10-20 cm layer, the highest SOC content was found in the CT treatment with residue retention. The SOC stock expressed as equivalent soil mass was greatest in the 0-20 cm layer of the ZTM (wheat and maize) and ZTR cultivated treatments with residue retention. After 16 y, the highest content of soil δ¹³C was found in ZTM + r and CTM + r treated soil cultivated with maize; −16.56‰ and −18.08‰ in the 0-5 cm layer, −18.41‰ and −18.02‰ in the 5-10 cm layer and −18.59‰ and −18.72‰ in the 10-20 cm layer respectively. All treatments had a higher percentages of C-C₃ (derived from wheat residues or the earlier forest) than C-C₄ (derived from maize residues). The highest percentages of C-C₄, was found in ZTM + r and CTM + r treated soil cultivated with maize, i.e. 33.0% and 13.0% in 0-5 cm layer, 9.1% and 14.3% in the 5-10 cm layer and 5.0% and 6.8% in 10-20 cm layer, respectively. The gross SOC turnover was lower in soil with residue retention than without residues. It was found that the ZT system with residue retention and rotation with wheat is a practice with a potential to retain organic carbon in soil.     

Anaerobic Eury- and Crenarchaeota inhabit ectomycorrhizas of boreal forest Scots pine

Members of the euryarchaeotal genera Methanolobus and Halobacterium as well as group 1.1c Crenarchaeota were enriched from ectomycorrhizal samples and cultured under anaerobic conditions. 16S rRNA gene sequences of Methanolobus were obtained in a H₂ + CO₂ atmosphere and autofluorescent putatively methanogenic microbial cells were detected by epifluorescence microscopy of the anaerobic methane-producing enrichment cultures. Halobacterium and group 1.1c Crenarchaeota grew anaerobically when either H₂ or CH₄ was added to the atmosphere. Group 1.1c Crenarchaeota were also enriched under aerobic conditions on mineral media, but only when methane or methanol was added as carbon sources. The 16S rRNA gene sequences of 1.1c Crenarchaeota grown under both anaerobic and aerobic conditions were highly similar. Our study demonstrates the growth of group 1.1c Crenarchaeota and Halobacteria derived from non-extreme soil environment in non-saline enrichments under anaerobic conditions. The results suggest that 1.1c Crenarchaeota may play a role in the cycling of C-1 substrates in the boreal forest soil ecosystem.     

Temporal variation in methanogenic community structure and methane production potential of tropical rice ecosystem

Temporal variations in diversity of methanogenic community and CH₄ production potential were analyzed in an Indian tropical rice ecosystem. Laboratory incubations showed that methane production varied from 20.86 to 134.11 μg CH₄ g⁻¹ d.w.s. during the two consecutive years, 2009 and 2010. CH₄ production potential was high at the flowering stage of the rice crop followed by ripening, tillering, post-harvest and pre-plantation stage. Phylogenetic analysis of 16S rRNA genes of methanogenic community indicated that flowering and ripening stages comprised of Methanomicrobiaceae, Methanosarcinaceae, Methanosaetaceae and RC I methanogenic groups, while only the members of Methanomicrobiaceae and RC I were present in the remaining stages. Further, the dominance of RC I was observed in all stages. This study demonstrates that flowering and ripening stages of rice crop offer relatively favorable ecological niche for methanogenic community. The overall analyses suggest that the temporal change in diversity of methanogens regulates CH₄ production potential in rice field soils.     

C signature of CO2 evolved from incubated maize residues and maize-derived sheep faeces

Analyses of the spatial and temporal variations in the natural abundance of ¹³C are frequently employed to study transformations of plant residues and soil organic matter turnover on sites where long continued vegetation with the C₃-type photosynthesis pathway has been replaced with a C₄-type vegetation (or vice versa). One controversial issue associated with such analyses is the significance of isotopic fractionation during the microbial turnovers of C in complex substrates. To evaluate this issue, C₃-soil and quartz sand were amended with maize residues and with faeces from sheep feed exclusively on maize silage. The samples were incubated at 15 °C for 117 days (maize residues) or 224 days (sheep faeces). CO₂ evolved during incubation was trapped in NaOH and analysed for C isotopic contents. At the end of incubation, 63 and 50% of the maize C was evolved as CO₂ in the soil and sand, respectively, while 32% of the faeces C incubated with soil and with sand was recovered as CO₂. Maize and faeces showed a similar decomposition pattern but maize decomposed twice as fast as faeces. The δ¹³C of faeces was 0.3‰ lower than that of the maize residue (δ¹³C −13.4‰), while the δ¹³C of the C₃-soil used for incubation was −31.6‰. The δ¹³C value of the CO₂ recovered from unamended C₃-soil was similar or slightly lower (up to −1.5‰) than that of the C₃-soil itself except for an initial flush of ¹³C enriched CO₂. The δ¹³C values of the CO₂ from sand-based incubations typically ranged −15‰ to −17‰, i.e. around −3‰ lower than the δ¹³C measured for maize and faeces. Our study clearly demonstrates that the decomposition of complex substrates is associated with isotopic fractionation, causing evolved CO₂ to be depleted in ¹³C relative to substrates. Consequently the microbial products retained in the soil must be enriched in ¹³C.     

Effect of soil water status and mulching on N<Subscript>2</Subscript>O and CH<Subscript>4</Subscript> emission from lowland rice field in China

Cultivation of rice in unsaturated soils covered with mulch is receiving more attention in China because of increasingly serious water shortage; however, greenhouse gas emission from this cultivation system is still poorly understood. A field experiment was conducted in 2001 to compare nitrous oxide (N₂O) and methane (CH₄) emission from rice cultivated in unsaturated soil covered with plastic or straw mulch and the traditional waterlogged production system. Trace gas fluxes from the soil were measured weekly throughout the entire growth period using a closed chamber method. Nitrous oxide emissions from unsaturated rice fields were large and varied considerably during the rice season. They were significantly affected by N fertilizer application rate. In contrast, N₂O emission from the waterlogged system was very low with a maximum of 0.28 mg N₂O m^–2 h^–1. However, CH₄ emission from the waterlogged system was significantly higher than from the unsaturated system, with a maximum emission rate of 5.01 mg CH₄ m^–2 h^–1. Our results suggested that unsaturated rice cultivation with straw mulch reduce greenhouse gas emissions.     

Growth of hydrogenotrophic and acetoclastic methanogens on substrate from rice plant callus cells in anaerobic soil: an estimation to the role of slough-off root cap cells to their growth

Flooded paddy fields are the major anthropogenic sources of methane (CH₄) emission, and organic materials of rice plant origin were estimated to be important as its source. This study used rice (Oryza sativa L. cv, Yukihikari) callus cells as a model material for slough-off root cap cells, and carbon-13 (¹³C)-labelled callus cells were subjected to decomposition in aerobic and anaerobic soil microcosms for 56 days. DNA was extracted from a soil incubated with carbon-12 (¹²C)- and ¹³C-callus cells and subjected to buoyant density gradient centrifugation to identify methanogenic archaeal species that assimilated carbon from the callus cells. ¹³C-labelled 16S rRNA gene (16S rDNA) fragments from methanogenic archaea were not polymerase chain reaction (PCR)-amplified in heavy fractions under aerobic soil conditions, while they were successfully done from day 3 onwards under anaerobic soil conditions. Eighty-four denaturing gradient gel electrophoresis (DGGE) bands in heavy fractions were sequenced, revealing that they were members of Methanosarcina spp. (20 clones), Methanosaeta spp. (18 clones), Methanocella spp. (25 clones), Methanomicrobiales (10 clones), Methanobacterium spp. (7 clones) and Cluster ZC-I (2 clones). They included hydrogenotrophic and acetoclastic methanogens and were phylogenetically different from those residing in rice roots and, presumably, from those assimilating root exudate and mucilage-derived carbon. This study indicates that carbon of slough-off root cap cells propagates specific methanogenic species in rice rhizosphere under anaerobic soil conditions and thus augments the diversity of the total rhizospheric methanogenic community.     

Effect of the herbicide butachlor on methane emission and ebullition flux from a direct-seeded flooded rice field

 Application of a commercial formulation of the herbicide butachlor (N-butoxymethyl-2-chloro-2′,6′-diethyl acetanilide) at 1 kg a.i. ha^–1 to an alluvial soil planted with direct-seeded flooded rice (cv. Annada), significantly inhibited both crop-mediated emission and ebullition fluxes of methane (CH₄). Over a cropping period of 110 days, the crop-mediated cumulative emission flux of CH₄ was lowered by ∼20% in butachlor-treated field plots compared with that of an untreated control. Concurrently, ebollition flux of CH₄ was also retarded in butachlor-treated field plots by about 81% compared with that of control plots. Significant relationships existed between CH₄ emission and redox potential (E _h) and Fe²⁺ content of the flooded soil. Application of butachlor retarded a drop in soil redox potential as well as accumulation of Fe²⁺ in treated field plots. Methanogenic bacterial population, counted at the maturity stage of the crop, was also low in butachlor-treated plots, indicating both direct and indirect inhibitory effects of butachlor on methanogenic bacterial populations and their activity. Results indicate that butachlor, even at field-application level, can effectively abate CH₄ emission and ebollition from flooded soils planted to rice whilst maintaining grain yield. Received: 15 March 2000     

Dynamics of methanogenesis and methanotrophy in tropical paddy soils as influenced by elevated CO2 and temperature interaction

Response of methanogenesis and methanotrophy to elevated carbon dioxide (CO₂) could be affected by changes in soil moisture content and temperature. In soil microcosms contained in glass bottles and incubated under laboratory conditions, we assessed the impact of elevated CO₂ and temperature interactions on methanogenesis and methanotrophy in alluvial and laterite paddy soils of tropical origin. Soil samples were incubated at ambient (370 μmol mol⁻¹) and elevated (600 μmol mol⁻¹) CO₂ concentrations at 25, 35 and 45 °C under non-flooded and flooded conditions for 60 d. Under flooded condition, elevated CO₂ significantly increased methane (CH₄) production while under non-flooded condition, only marginal increase in CH₄ production was observed in both the soils studied and the increase was significantly enhanced by further rise in temperature. Increased methanogenesis as a result of elevated CO₂ and temperature interaction was mostly attributed to decreased soil redox potential, increased readily mineralizable carbon, and also noticeable stimulation of methanogenic bacterial population. In contrast to CH₄ production, CH₄ oxidation was consistently low under elevated CO₂ concentration and the decrease was significant with rise in temperature. The low affinity and high affinity CH₄ oxidation were faster under non-flooded condition as compared to flooded condition. Admittedly, decreased low and high affinity CH₄ oxidation as a result of elevated CO₂ and temperature interaction was related to unfavorable lower redox status of soil and the inhibition of CH₄-oxidizing bacterial population.     

Natural N abundances of inorganic nitrogen in soil treated with fertilizer and compost under changing soil moisture regimes

This study was conducted to examine whether the applications of N-inputs (compost and fertilizer) having different N isotopic compositions (δ¹⁵N) produce isotopically different inorganic-N and to investigate the effect of soil moisture regimes on the temporal variations in the δ¹⁵N of inorganic-N in soils. To do so, the temporal variations in the concentrations and the δ¹⁵N of NH₄⁺ and NO₃⁻ in soils treated with two levels (0 and 150 mg N kg⁻¹) of ammonium sulfate (δ¹⁵N=−2.3‰) and compost (+13.9‰) during a 10-week incubation were compared by changing soil moisture regime after 6 weeks either from saturated to unsaturated conditions or vice versa. Another incubation study using ¹⁵N-labeled ammonium sulfate (3.05 ¹⁵N atom%) was conducted to estimate the rates of nitrification and denitrification with a numerical model FLUAZ. The δ¹⁵N values of NH₄⁺ and NO₃⁻ were greatly affected by the availability of substrate for each of the nitrification and denitrification processes and the soil moisture status that affects the relative predominance between the two processes. Under saturated conditions for 6 weeks, the δ¹⁵N of NH₄⁺ in soils treated with fertilizer progressively increased from +2.9‰ at 0.5 week to +18.9‰ at 6 weeks due to nitrification. During the same period, NO₃⁻ concentrations were consistently low and the corresponding δ¹⁵N increased from +16.3 to +39.2‰ through denitrification. Under subsequent water-unsaturated conditions, the NO₃⁻ concentrations increased through nitrification, which resulted in the decrease in the δ¹⁵N of NO₃⁻. In soils, which were unsaturated for the first 6-weeks incubation, the δ¹⁵N of NH₄⁺ increased sharply at 0.5 week due to fast nitrification. On the other hand, the δ¹⁵N of NO₃⁻ showed the lowest value at 0.5 week due to incomplete nitrification, but after a subsequence increase, they remained stable while nitrification and denitrification were negligible between 1 and 6 weeks. Changing to saturated conditions after the initial 6-weeks incubation, however, increased the δ¹⁵N of NO₃⁻ progressively with a concurrent decrease in NO₃⁻ concentration through denitrification. The differences in δ¹⁵N of NO⁻₃ between compost and fertilizer treatments were consistent throughout the incubation period. The δ¹⁵N of NO₃⁻ increased with the addition of compost (range: +13.0 to +35.4‰), but decreased with the addition of fertilizer (−10.8 to +11.4‰), thus resulting in intermediate values in soils receiving both fertilizer and compost (−3.5 to +20.3‰). Therefore, such differences in δ¹⁵N of NO₃⁻ observed in this study suggest a possibility that the δ¹⁵N of upland-grown plants receiving compost would be higher than those treated with fertilizer because NO₃⁻ is the most abundant N for plant uptake in upland soils.     

Effects of inorganic fertilizers and micronutrients on methane production from wetland rice (Oryza sativa L.)

We compared the effects of adding different forms of nitrogenous fertilizers on the production of CH₄ in soil and on CH₄ emission from rice plants, Urea and diammonium phosphate gave the highest rates of CH₄ production from the soil and emission through rice plants, followed by (NH₄)₂SO₄. NaNO₃ was the least effective. The effects of micronutrients like Mo, Ni, or B were more prominent than those of Fe, Mn, Zn, V, or Co. It is concluded that CH₄ emission from rice paddies is influenced by both macro- and micronutrients, through effects on both microbial methanogenesis in soil and elimination through rice plants as a consequence of the effects on plant growth.     

Compound-specific δC and δH analyses of plant and soil organic matter: A preliminary assessment of the effects of vegetation change on ecosystem hydrology

Here we present δ¹³C and δ²H data of long-chained, even-numbered (C₂₇-C₃₁) n-alkanes from C3 (trees) and C4 (grasses) plants and from the corresponding soils from a grassland-woodland vegetation sequence in central Queensland, Australia. Our data show that δ¹³C values of the C4 grassland species were heavier relative to those of C3 tree species from the woodland (Acacia leaves) and woody grassland (Atalaya leaves). However, n-alkanes from the C4 grasses had lighter δ²H values relative to the Acacia leaves, but showed no significant difference in δ²H values when compared with C3 Atalaya leaves. These results differ from those of previous studies, showing that C4 grasses had heavier δ²H values relative to C3 grasses and trees. Those observations have been explained by C4 plants accessing the more evaporation-influenced and isotopically heavier surface water and tree roots sourcing deeper, isotopically lighter soil water (“Two-layered soil-water system”). By comparison, our data suggest that ecosystem changes (vegetation “thickening”) can significantly alter the soil hydrological characteristics. This is shown by the heavier δ²H values in the woodland soil compared with lighter δ²H values in the grassland soil, implying that the recent vegetation change (increased tree biomass) in the woodland had altered soil hydrological conditions. Estimated δ²H values of the source-water for vegetation in the grassland and woodland showed that both trees and grasses in open settings accessed water with lighter δ²H values (avg. −46‰) compared with water accessed by trees in the woodland vegetation (avg. −7‰). These data suggest that in semi-arid environments the “two-layer” soil water concept might not apply. Furthermore, our data indicate that compound-specific δ²H and δ¹³C analyses of n-alkanes from soil organic matter can be used to successfully differentiate between water sources of different vegetation types (grasses versus trees) in natural ecosystems.