土壤CH4氧化与土壤甲烷营养菌及主要研究方法

甲烷营养菌(methanotrophs)是一类以CH4为唯一碳源和能源的细菌,广泛分布在水稻土、森林土、苔原土、泥炭地、海洋与湖泊底泥、堆肥、垃圾填埋场及地下水等环境中,并作为大气甲烷(CH4)唯一的生物汇(库),在全球温室效应研究中备受关注。目前,关于土壤甲烷营养菌的研究主要包括菌株的多样性、生态分布以及环境因素对微生物氧化CH4过程的影响。本文从甲烷营养菌的分类入手,概述稻田土壤CH4的氧化与释放、旱地土壤CH4的氧化以及影响土壤CH4氧化的因素等方面的研究进展,同时介绍了土壤甲烷营养菌研究领域的几种主要的分子研究技术,以期为甲烷营养菌相关的研究提供参考。     

长期定位试验对水稻田土壤甲烷氧化活性和甲烷排放通量的影响

通过对长期定位施肥的黄松稻田土的甲烷氧化活性和甲烷排放通量的研究表明,长期定位施肥对稻田土壤的好氧甲烷氧化活性和甲烷排放通量有显著性的影响(|t|t0.05,P0.05),而对稻田土壤的厌氧甲烷氧化活性有影响但未达显著性水平(|t|t0.05,P0.05)。施加有机肥能显著增加稻田土壤的甲烷氧化活性和甲烷排放通量;当有机肥和无机肥混合施用时,其促进作用明显大于单施有机肥或无机肥。施加尿素后,稻田土壤的甲烷氧化活性及甲烷排放量都有所下降,但钾肥和磷肥能缓解由尿素引起的抑制作用。施肥后耕作的稻田甲烷氧化活性和甲烷排放通量高于施肥后不耕作的稻田;耕作而不施肥的稻田甲烷氧化活性和甲烷排放通量要低于不施肥也不耕作的稻田。     

基于核酸DNA/RNA同位素示踪技术的水稻土甲烷氧化微生物研究

稳定性同位素示踪复杂土壤中微生物DNA/RNA的技术难点是13C-DNA/RNA的鉴定。本研究针对我国六种典型水稻土,利用稳定性同位素13CH4示踪活性的甲烷氧化菌,超高速密度梯度离心获得不同浮力密度DNA/RNA后,以甲烷氧化菌独有的pmo A功能基因和16S r RNA特异基因作为分子标靶,通过半定量凝胶电泳技术评价了特异基因作为分子标靶判定13C-DNA/RNA的可行性,进一步利用克隆文库技术研究水稻土中的活性甲烷氧化菌群落结构。结果表明:甲烷氧化菌功能基因pmo A作为分子标靶,能够准确鉴别13C-DNA,而甲烷氧化菌特异的16S r RNA基因则能较好地区分12C和13C标记的RNA,但13C-RNA中的非目标微生物污染高于13C-DNA示踪技术。进一步以13C-DNA和13C-RNA为模板,分别构建了pmo A和16S r RNA基因的克隆文库,系统发育分析表明I型菌主导了土壤甲烷氧化过程,其中江西鹰潭和黑龙江五常土壤中活性甲烷氧化菌全部属于Ia型,而四川资阳、浙江嘉兴、江苏常熟和江都土壤中Ia型和Ib型甲烷氧化菌均有发现,并且后者比例较低。这些结果表明分子标靶基因能够有效判定复杂土壤中的甲烷氧化菌13C-DNA/RNA,在DNA和RNA水平的结果基本一致,我国典型水稻土中活性甲烷氧化菌可能存在一定的地理分异规律。     

水稻田土壤甲烷氧化活性及其环境影响因子的研究

报道环境因子土壤含水量、温度和pH对发育于河流沉积物母质的水稻田土黄松泥田土氧化外源甲烷活性影响的研究结果。表明土壤中存在有氧和无氧两个甲烷氧化系统。有氧甲烷氧化系统 (AMOS)最适条件下的氧化甲烷最大活性比无氧甲烷氧化系统(AAMOS)最适条件下的氧化甲烷最大活性高 1至 2倍。在土壤通气良好的条件下 ,AMOS占主导地位 ,在无氧或极微氧的土壤中 ,AAMOS起主要作用。影响活性的主要因子是土壤的分子氧含量、甲烷含量、水含量、温度和pH值。分子氧对AAMOS氧化甲烷的活性具有一定抑制作用 ,土壤中甲烷量和含水量对AAMOS氧化甲烷活性的影响比对AMOS氧化甲烷活性的影响更为强烈。土壤氧化甲烷的活性对温度较为敏感 ,其最适氧化甲烷的温度范围在 2 5～ 35℃之间。当土壤在 5 0℃培养的时间超过 6h后 ,土壤氧化外源甲烷的活性全部丧失 ,且不能在2 8℃下得到恢复。pH是另一个影响土壤氧化甲烷的重要环境因子。其最适pH范围在 6～ 7之间 ,pH低于 3或pH高于 9时 ,几乎完全丧失氧化外源甲烷的活性。     

放牧对黄土高原典型草原冬季牧场甲烷氧化菌类群及活性的影响

土壤甲烷氧化菌是大气甲烷（CH4）氧化的唯一生物汇。放牧能通过影响甲烷氧化菌的丰度和多样性来调控草地土壤的甲烷氧化活性。采集黄土高原典型草地冬季牧场中不同强度连续放牧的土壤样本,通过室内培养和高通量测序的方法测定不同放牧强度下土壤甲烷氧化活性以及土壤甲烷氧化菌的组成和丰度变化规律。结果表明,该草地是CH4汇,中放牧强度（MG）和高放牧强度（HG）增加了甲烷氧化速率。同时,与未放牧（CK）相比, MG和HG的甲烷氧化菌的平均丰度也显著增加。高通量测序结果显示,放牧对甲烷氧化菌的多样性有显著影响,不同放牧强度下均以USCγ为优势类群,同时存在少量的甲基暖菌属（Methylocaldum）和甲基孢囊菌属（Methylocystis）。皮尔森（Pearson）相关性分析表明,甲烷氧化速率（MOR）与土壤的水分含量和硝态氮的含量存在显著正相关（P < 0.05）,与USCγ的绝对数量存在极显著的正相关关系（P < 0.01）,这说明USCγ在该草地甲烷吸收过程中起主要作用。本研究证明了放牧可提升黄土高原典型草地的甲烷汇功能。     

FACE对水稻土产甲烷菌和甲烷氧化菌种群及其活性的影响

利用江都市小记镇的稻-麦轮作FACE平台,采用最大可能(MPN)法,在2005年水稻生长季研究了不同施肥(常规N量和低N量)、不同秸秆还田(秸秆全还田和秸秆不还田)处理土壤中的产甲烷菌和甲烷氧化菌数量在大气CO2浓度升高(FACE)条件下随时间的变化情况,并且借助气相色谱测定了土壤的产甲烷潜力和甲烷氧化潜力。结果表明:在秸秆全还田情况下,FACE对于产甲烷菌在分蘖期具有促进作用,而在抽穗期与收获期具有抑制作用,这种作用在低N条件下达到显著性(P<0.05)水平。而秸秆不还田情况下,FACE对产甲烷菌无明显促进作用;在低量N的施用情况下,FACE对于土壤甲烷氧化菌的活性具有刺激作用,在水稻抽穗期土壤甲烷氧化菌数量明显地高于对照,达到显著性水平(P<0.05);而常规施N量秸秆全还田的情况下,在水稻的分蘖期、拔节期和收获期FACE土壤中的甲烷氧化菌数量却受到一定程度的抑制。土壤的产甲烷潜力测定结果表明,FACE能促进土壤的CH4释放,尤其是在常规N量施用条件下。当底物(加入外源CH4)充足时,FACE条件下能使土壤具有较高的氧化CH4的能力,其CH4氧化潜力明显大于对照土壤,并且这种作用在常规N肥施用条件下尤为明显,达极显著性水平(P<0.01)。     

不同离子对水稻田土壤甲烷氧化活性影响的研究

研究了矿质元素不同的阴离子和阳离子与黄松泥田水稻土壤氧化甲烷活性之间的相关性。结果表明 ,不同的阴离子和阳离子对甲烷氧化活性的影响有显著性差异 ,不同浓度的同一种阴离子或阳离子对甲烷氧化活性的影响也有显著性差异。Na+较K+对土壤甲烷氧化活性具有更强的抑制作用。NH4+和NO2-可与甲烷氧化竞争土壤中的分子氧而有明显的抑制作用。Cr3+对微生物具毒性而影响土壤的甲烷氧化。阴离子PO43-和CO32-无明显影响。这种影响的差异性不仅与阴离子和阳离子本身的理化特征有关 ,而且与土壤对阳离子的吸附力及阴离子与土壤的相互作用强度有关。土壤理化特性同样影响阴离子和阳离子对甲烷氧化活性影响的强弱     

不同温度下尿素对水稻土甲烷产生及相关古菌群落的影响

水稻田是大气甲烷的重要排放源。尿素氮肥施用是提升水稻产量和品质的重要措施,但其对稻田土壤中产甲烷古菌的影响规律仍不清楚。通过模拟水稻生长季节可能的田间温度变化,本研究设置水稻土施加尿素(N,400 mg/kg干土)与未施加尿素两个处理,在15℃、25℃、37℃以及50℃下进行为期100天的厌氧培养,定期测定了培养过程中甲烷累积量以及土壤理化因子如p H、NH_4~+-N以及有机碳的变化,并运用基于16S r RNA基因的T-RFLP(末端限制性片段多态性分析)技术分析了产甲烷过程中古菌群落结构随时间的变化情况。结果表明:在中低温范围内(15~37℃),尿素对水稻土产甲烷有抑制作用,但在50℃高温下尿素对水稻土产甲烷量没有显著影响。尿素可能通过改变产甲烷古菌群落结构来影响甲烷的产生,在15~37℃范围内,尿素降低了水稻土产甲烷古菌群落的稳定性,增大了其在不同时间的差异性;而在50℃高温时,尿素对水稻土产甲烷古菌稳定性和差异性的影响不明显。不同温度下,尿素均降低了甲烷八叠球菌(Methanosarcinaceae)的丰度,且随着温度的变化,尿素对水稻土产甲烷机制的改变可能没有影响。     

风干土保存时间和湿土培育时间对黄淮海平原潮土酶活性的影响

以经历18年不同施肥管理的土壤为研究对象,阐明它们经过4个不同时间保存或处理后的土壤脲酶、转化酶、脱氢酶、及FDA酶活性的动态变化。处理包括:风干保存30天或鲜土状态、风干保存210天、风干土湿润至田间持水量(25℃)条件下分别培育15天和51天;同时评估这些酶活性的变化程度与土壤本身有机碳含量之间的关系。结果表明,风干土保存时间和风干土湿润后短期培育均对脲酶活性影响很小,但风干土湿润培养51天后其活性则显著降低;随风干土保存时间延长,转化酶活性显著降低;与鲜土相比,风干土湿润培养15天后,脱氢酶活性显著提高,但继续湿润培养至51天后,其活性又降至与鲜土相当,因此风干土湿润培育一定时间后测定的脱氢酶活性可用来代表其田间自然湿度时的状态;FDA酶活性的变异程度最大,与其从鲜土状态至风干状态的活性急剧下降有关。土壤本身有机碳含量与脲酶和脱氢酶的活性变化程度成显著负相关关系,说明土壤有机碳含量是决定它们随环境条件改变而变化的主要因素之一。另外,土壤NH4+-N、NO3--N和可溶性有机碳含量对上述4种处理的响应程度也存在差异。其中风干状态土壤经湿润培育处理后,NH4+-N含量呈先降后升趋势,正好与脲酶活性变化趋势相反;而NO3--N含量整体上呈上升趋势,可溶性有机碳含量则正好相反。     

生物质炭提高稻田甲烷氧化活性

为了揭示生物质炭输入对稻田根际土壤产甲烷和甲烷氧化活性的影响,该文通过1a 的田间试验,研究了2种原料制备的生物质炭（竹炭和水稻秸秆炭）对水稻根际土壤产甲烷和甲烷氧化活性的影响。结果表明,2种生物质炭因理化性质的不同,对水稻根际土壤产甲烷活性和甲烷氧化活性的影响存在较大差别。秸秆炭的输入可以显著提高水稻苗期根际土壤产甲烷活性,而竹炭在水稻的整个生长期对根际土壤产甲烷活性均没有显著性影响。竹炭和秸秆炭不稳定易降解组分含量的差异,使其对稻田土壤产甲烷微生物产生不同程度的影响,进而导致稻田根际土壤产甲烷活性响应差别。除抽穗期竹炭处理和成熟期秸秆炭处理,尿素施加并未显著改变生物质炭对根际土壤产甲烷活性的影响趋势。在水稻整个生长期,秸秆炭和竹炭对稻田土壤甲烷氧化活性都有促进作用,但只有秸秆炭在苗期和成熟期表现出显著性的差异。尿素对苗期和抽穗期根际土壤甲烷氧化活性有促进作用。与竹炭相比,秸秆炭输入在改善土壤通气条件、提高土壤pH值和电导率EC、以及K、P元素含量等方面更为有效,同时可能是秸秆炭对水稻根际土壤甲烷氧化活性产生显著性促进作用的潜在机理。     

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     

Relationships between ammonia oxidizers and N2O and CH4 fluxes in agricultural fields with different soil types

Nitrous oxide (N₂O) is a greenhouse gas that contributes to the destruction of stratospheric ozone, and agricultural soil is an important source of N₂O. Aerobic soils are sinks for atmospheric methane (CH₄), a greenhouse gas. Ammonia monooxygenase (AMO) can oxidize CH₄, but CH₄ is mostly oxidized by methane monooxygenase (MMO), and CH₄ oxidation by AMO is generally negligible in the soil. We monitored the N₂O and CH₄ fluxes after urea application in fields containing different soils using an automated sampling system to determine the effects of environmental and microbial factors on the N₂O and CH₄ fluxes. The soil types were Low-humic Andosol (Gleyic Haplic Andosol), yellow soil (Gleyic Haplic Alisol) and gray lowland soil (Entric Fluvisol). Cumulative N₂O emissions from the yellow soil were higher than those from other soil types, although the difference was not significant. The CH₄ uptake level by Andosol was one order of magnitude higher than that by other soils. There were significant relationships between the ammonia oxidation potential, AOB and AOA amoA copy numbers, and the CH₄ uptake. In contrast, the gene copy numbers of methane-oxidizing bacteria (MOB) pmoA were below the detection limit. Our results suggested that the AMOs of AOB and AOA may have more important roles than those previously considered during CH₄ oxidation in agricultural soils treated with N fertilizers.     

Microbial ethylene production and inhibition of methanotrophic activity in a deciduous forest soil

Production of C₂H₄, but not of CH₄, was observed in anoxically incubated soil samples (cambisol on loamy sand) from a deciduous forest. Ethylene production was prevented by autoclaving, indicating its microbial origin. Ethylene production gradually decreased from 4 to 12 cm soil depth and was not affected by moisture or addition of methionine, a possible precursor of C₂H₄. Oxidation of atmospheric CH₄ in soil samples was inhibited by C₂H₄. Ethylene concentrations of 3, 6 and 10 μl l⁻¹ decreased CH₄ uptake by 21, 63 and 98%, respectively. Methionine and methanethiol, a possible product of methionine degradation, also inhibited CH₄ oxidation. Under oxic conditions, C₂H₄ was consumed in the soil samples. Ethylene oxidation kinetics exhibited two apparent K_m values of 40 μl l⁻¹ and 12,600 μl l⁻¹ suggesting the presence of two different types of C₂H₄-oxidizing microorganisms. Methanotrophic bacteria were most probably not responsible for C₂H₄ oxidation, since the maximum of C₂H₄ oxidation activity was localized in soil layers (2-8 cm depth) above those (8-10 cm depth) of CH₄ oxidation activity. Our observations suggest that C₂H₄ production in the upper soil layers inhibits CH₄ oxidation, thus being one reason for the localization of methanotrophic activity in deeper soil layers.     

Water-soluble Al inhibits methane oxidation at atmospheric concentration levels in Japanese forest soil

The effect of aluminium on methane oxidation was examined from incubation experiments involving the addition of several concentrations of Al solution (0.1, 0.2, 0.5, 1, 3 and 5 mM) to two soil samples that possessed different CH₄ oxidation potential. Atmospheric CH₄ oxidation activity was inhibited by the addition of as little as 0.1 mM Al solution (approximately 0.5 μg of Al per gram dry weight soil) to a forest soil that contained low water-soluble Al and possessed a high CH₄ oxidation potential. Our results indicate that Al inhibition of CH₄ oxidation activity is concentration-dependant after a certain time and the inhibition increases gradually over time until at least 96 h have elapsed. We also found that relatively small amounts of Al additions, such as 10-20 μg per gram dry weight of soil, halved the CH₄ oxidation rate compared to the control, regardless of the original CH₄ oxidation potential of the soil. Since the Al concentrations used in our experiment are often observed in forest soils, we can assume that Al acts as an important inhibitor of CH₄ oxidation in forest soils under natural conditions. The sharp falls and a continuous decrease in CH₄ oxidation rate in other forest samples with the addition of deionized water implies that the water-soluble Al contained in soils contributes to the inhibition of CH₄ oxidation rate. This result suggests that precipitation causes a relatively prolonged inhibition of CH₄ oxidation in soils containing a high concentration of water-soluble Al.     

Microbial communities of Lumbricus terrestris L. middens: structure, activity, and changes through time in relation to earthworm presence

Background, aim, and scope  Earthworms make a major contribution to decomposition in ecosystems where they are present, mainly acting in the drilosphere, that is, galleries, burrows, casts, and middens. Earthworm middens are hot-spots of microbial activity and nutrient dynamics and represent a suitable model for studying earthworm-mediated influences on soil microbial communities by alteration of the patch structure of the microbial environment. We studied the structure and activity of the microbial communities in the soil system formed by middens of Lumbricus terrestris and the soil below and surrounding them and the role of earthworms in maintaining these structures through time. Material and methods  We set up an experiment in which middens were either left (control) or removed from their original place (translocated) and left in a nearby area free of earthworm activity for 2 months. After 1 and 2 months we sampled middens, soil below them, and surrounding soil. We analyzed the phospholipid fatty acid (PLFA) profiles and measured respiratory fluxes of CO₂ and CH₄. Results  Microbial communities of middens clearly differed from those of soil below and surrounding soil samples, showing higher bacterial and fungal PLFAs (p < 0.0001 and p < 0.01, respectively); furthermore, changes in microbial communities were stronger in control middens than in translocated middens. Moreover, gram positive and negative bacterial PLFAs were greater in translocated than control middens (p < 0.0001 and p < 0.001, respectively), as well as total organic carbon (p < 0.001). Microbial activity was higher in middens than in soil below and surrounding soil samples both for CO₂ (p < 0.0001) and CH₄ (p < 0.0001). Discussion  Soil bioturbation by the earthworm L. terrestris was strong in their middens, but there was not any effect on soil below and surrounding soil. Microbial communities of middens maintain their biomass and activity when earthworms were not present, whereas they decreased their biomass and increased their activity when earthworms were present. Conclusions  Earthworms strongly enhanced microbial activity measured as CO₂ production in middens, which indicates that there are hot spots for soil microbial dynamics and increasing habitat heterogeneity for soil microorganisms. Moreover, our data strongly support the fact that the impact of this earthworm species in this soil is restricted to their middens and increasing soil heterogeneity. Recommendations and perspectives  Our data indicate that it is not clear if earthworms enhance or depress microbial communities of middens since the microbial activity increased, but did not modify their biomass and this was not dependent on soil organic C content. These results indicate no competence for C pools between this anecic earthworm and microorganisms, which has been found for other earthworm species, mainly endogeics. Conversely, they suggest some type of facilitation due to the release of additional nutrient pools in middens when earthworms are present, through the digestion of middens' material or the addition of casts produced from other food sources.     

Earthworm bioturbation stabilizes carbon in non-flooded paddy soil at the risk of increasing methane emissions under wet soil conditions

Studies on earthworms in rice-based ecosystems tend to focus on some pest species, while the potential of these important soil engineers for beneficially affecting carbon storage and cycling is widely ignored. We carried out a microcosm experiment to quantify the impact of the tropical earthworm Pheretima sp. on the C turnover in paddy soils under different conditions of water saturation and N fertilization. The soil was sampled at the lowland farm of the International Rice Research Institute (Philippines). In the absence of earthworms, soil respiration showed a distinct hump-shaped maximum at intermediate levels of water saturation (4-fold higher than in hand-dry soil) and increased 1.5-fold with increasing amounts of N fertilization. Amounts of CH₄ emitted, in contrast, were small at low to moderate soil humidity and became very high under conditions of water saturation (80-fold higher than hand-dry soil). No response to nitrogen addition was observed. Earthworms suppressed both the respiration maximum at intermediate saturation levels (by a factor of 1.4) and the stimulating impact of N fertilization (1.7-fold at maximum fertilizer level). On the other hand, earthworms strongly increased CH₄ release under conditions of high water saturation (3-fold). No consistent response of the soil microflora (bacterial abundance, soil enzymes) to earthworm activity could be established. Our findings suggest that the stabilization of soil organic C via earthworm bioturbation is confined to the range of soil humidity that allows high activity of Pheretima sp. Under conditions of intensive agriculture, the stabilizing effect of the worms may even be augmented by the fact that they offset the positive effect of N fertilization on microbial respiration. Earthworms may thus play a vital role in reducing the CO₂ flush from paddy soils after the conversion to non-flooded crops such as aerobic rice or maize. Acceleration of methane emission in very humid soils nevertheless points to a certain risk that is associated with increasing earthworm abundance in production systems that are still exposed to temporary flooding during the wet season.     

易利用态有机物质对水稻土甲烷排放的激发作用

为探讨外源有机物质对淹水稻田土壤CH4排放的激发作用,对比不同外源有机物质对土壤CH4排放的贡献差别,本研究选取3种标记的易利用态有机物(葡萄糖、乙酸和草酸)分别加入水稻土,进行了为期1个月的培养。结果表明:培养30 d后不同处理CH4的累计排放量差异显著(P0.05),其中,乙酸葡萄糖草酸对照;双因素方差分析结果显示,外源有机物质的添加加速了土壤易利用态有机质的矿化(即产生正激发效应);不同处理条件下激发作用产生的CH4分别占各处理CH4总累计排放量的73.3%(葡萄糖处理)、71.5%(乙酸处理)和40.9%(草酸处理),且CH4排放量与CH4激发效应之间极显著正相关关系说明土壤CH4排放主要要来自于土壤原有机质的分解,外源有机物质可能主要对土壤微生物活性及代谢途径有影响。     

Effect of silicate fertilizer on reducing methane emission during rice cultivation

Slag-type silicate fertilizer, which contains high amount of active iron oxide, a potential source of electron acceptor, was applied at the rate of 0, 2, 6, 10, and 20 Mg ha⁻¹ to reduce methane (CH₄) emission from rice planted in potted soils. Methane emission rates measured by closed chamber method decreased significantly with increasing levels of silicate fertilizer application during rice cultivation. Soil redox potential (Eh) decreased rapidly after flooding, but floodwater pH and soil pH increased significantly with increasing levels of silicate fertilizer application. Iron concentrations in potted soils and in percolated water significantly increased with the increasing levels of silicate fertilizer application, which acted as oxidizing agents and electron acceptors, and thereby suppressed CH₄ emissions. Silicate fertilization significantly decreased CH₄ production activity, while it increased carbon dioxide (CO₂) production activity. Rice plant growth, yield parameters, and grain yield were positively influenced by silicate application levels. The maximum increase in grain yield (17% yield increase over the control) was found with 10 Mg ha⁻¹ silicate application along with 28% reduction in total CH₄ flux during rice cultivation. It is, therefore, concluded that slag-type silicate fertilizer could be a suitable soil amendment for reducing CH₄ emissions as well as sustaining rice productivity and restoring the soil nutrient balance in rice paddy soil.     

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.     

The role of rice plants in regulating mechanisms of methane missions

 Rice plants play a pivotal role in different levels of the methane (CH₄) budget of rice fields. CH₄ production in rice fields largely depends on plant-borne material that can be either decaying tissue or root exudates. The quantity and quality of root exudates is affected by mechanical impedance, presence of toxic elements, nutrient deficiencies, water status of growing medium, and nitrogenase activity in the rhizosphere. CH₄ oxidation in rice fields is localized in the rhizosphere where the concentration gradients of CH₄ and oxygen overlap. CH₄ oxidation capacity is a function of the downward transport of oxygen through the aerenchyma, which, in turn, also acts as a conduit for CH₄ from the soil to the atmosphere. The decisive step in the passage of CH₄ through rice plant is the transition from root to stem. However, rice plants show an enormous variety of morphological and physiological properties, including differences in root exudation and gas transfer capacity. Comparative studies on different cultivars are deemed crucial for accomplishing a better understanding of the mechanisms of CH₄ consumption in the rhizosphere and CH₄ transport through the rice plant as well as the interaction of these processes. The results of such studies are considered tools for devising mitigation options. Received: 7 April 1999