Involvement of Bradyrhizobium japonicum denitrification in symbiotic nitrogen fixation by soybean plants subjected to flooding

Denitrification by Bradyrhizobium japonicum bacteroids contributes to nitric oxide (NO) production within soybean nodules in response to flooding conditions. However, the physiological relevance of NO production by denitrification in B. japonicum-Glycine max symbiosis is still unclear. In this work, soybean plants were inoculated with B. japonicum strains lacking the nirK or norC genes which encode the copper-containing nitrite reductase and the c-type nitric oxide reductase enzymes, respectively. 14 days flooding increased nodule number of plants inoculated with the WT and norC strains, but not of plants inoculated with the nirK mutant. However, nodule dry weight was not affected by 14 days flooding regardless of the strain used for inoculation. Supporting this observation, individual nodule growth was significantly higher in plants inoculated with nirK than those inoculated with WT or norC after 14 days flooding. Nodule functioning was strongly inhibited by flooding since leghemoglobin content of the nodules induced by any of the strains was significantly decreased after 7 or 14 days flooding compared to control plants. However, this effect was more relevant in nodules of plants inoculated with the WT or norC mutant than in those inoculated with the nirK mutant. Nitrogen fixation was also estimated by analyzing nitrogen content derived from biological nitrogen fixation in shoots, using the ¹⁵N isotope dilution technique. By using this approach, we observed that the negative effect of 14 days flooding on nitrogen fixation was more pronounced in plants inoculated with the norC mutant. However, nitrogen fixation of plants inoculated with nirK showed the highest tolerance to 14 days flooding. These findings allowed us to demonstrate the previously proposed hypothesis which suggests that NO formed by copper-containing nitrite reductase in soybean nodules, in response to flooding, has a negative effect on nitrogenase activity. We propose that inoculation of soybeans with a B. japonicum nirK mutant, which does not produce NO from nitrate, increases the tolerance of symbiotic nitrogen fixation to flooding.     

Response of soybean cultivars toward inoculation with three arbuscular mycorrhizal fungi and Bradyrhizobium japonicum in the alluvial soil

The aim of this study was to assess the comparative efficacy of three arbuscular mycorrhizal fungi (AMF) combined with cultivar specific Bradyrhizobium japonicum (CSBJ) in soybean under greenhouse conditions. Soybean seeds of four cultivars namely JS 335, JS 71-05, NRC 2 and NRC 7 were inoculated with three AM fungi (Glomus intraradices, Acaulospora tuberculata and Gigaspora gigantea) and CSBJ isolates, individually or in combination, and were grown in pots using autoclaved alluvial soil of a non-legume cultivated field of Ajmer (Rajasthan). Assessment of the data on nodulation, plant growth and seed yield revealed that amongst the single inoculations of three AMF, G. intraradices produced the largest increases in the parameters studied followed by A. tuberculata and G. gigantea indicating that plant acted selectively on AMF symbiosis. The dual inoculation with AMF + CSBJ further improved these parameters demonstrating synergism between the two microsymbionts. Among all the dual treatments, G. intraradices + B. japonicum brought about the largest increases in the studied characteristics particularly in seed weight per plant that increased up to 115.19%, which suggested that a strong selective synergistic relationship existed between AMF and B. japonicum. The cv. JS 335 exhibited maximum positive response towards inoculation. The variations in efficacy of different treatments with different soybean cultivars indicate the specificity of the inoculation response. These results provide a basis for selection of an appropriate combination of specific AMF and Bradyrhizobium which could further be utilized for verifying the symbiotic effectiveness and competitive ability of microsymbionts under field conditions of Ajmer region.     

Ability of free-living cells ofBradyrhizobium japonicum to denitrify in soils

Summary Experiments to assess the ability of free-living cells of six strains of soybean rhizobia (Bradyrhizobium japonicum USDA 76, 94, 110, 122, 123, and 135) to denitrify nitrate in five soils showed that although some strains ofB. japonicum have the capacity to rapidly denitrify nitrate in soils under anaerobic conditions, it is unlikely that the numbers of soybean rhizobia commonly found under field conditions are sufficient to significantly influence either the extent or the products of denitrification in soil. It is our general conclusion that the advantages, if any, that the ability to denitrify conveys to rhizobia or to the rhizobia-legume symbiosis are not offset by increased losses of plant-available N when denitrifying strains of rhizobia are present as free-living cells in soil.     

Size, symbiotic effectiveness and genetic diversity of field pea rhizobia (Rhizobium leguminosarum bv. viciae) populations in South Australian soils

Field pea (Pisum sativum L.) is widely grown in South Australia (SA), often without inoculation with commercial rhizobia. To establish if symbiotic factors are limiting the growth of field pea we examined the size, symbiotic effectiveness and diversity of populations of field pea rhizobia (Rhizobium leguminosarum bv. viciae) that have become naturalised in South Australian soils and nodulate many pea crops. Most probable number plant infection tests on 33 soils showed that R. l. bv. viciae populations ranged from undetectable (six soils) to 32×10³ rhizobia g⁻¹ of dry soil. Twenty-four of the 33 soils contained more than 100 rhizobia g⁻¹ soil. Three of the six soils in which no R. l. bv. viciae were detected had not grown a host legume (field pea, faba bean, vetch or lentil). For soils that had grown a host legume, there was no correlation between the size of R. l. bv. viciae populations and either the time since a host legume had been grown or any measured soil factor (pH, inorganic N and organic C). In glasshouse experiments, inoculation of the field pea cultivar Parafield with the commercial Rhizobium strain SU303 resulted in a highly effective symbiosis. The SU303 treatment produced as much shoot dry weight as the mineral N treatment and more than 2.9 times the shoot dry weight of the uninoculated treatment. Twenty-two of the 33 naturalised populations of rhizobia (applied to pea plants as soil suspensions) produced prompt and abundant nodulation. These symbioses were generally effective at N₂ fixation, with shoot dry weight ranging from 98% (soil 21) down to 61% (soil 30) of the SU303 treatment, the least effective population of rhizobia still producing nearly double the growth of the uninoculated treatment. Low shoot dry weights resulting from most of the remaining soil treatments were associated with delayed or erratic nodulation caused by low numbers of rhizobia. Random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) fingerprinting of 70 rhizobial isolates recovered from five of the 33 soils (14 isolates from each soil) showed that naturalised populations were composed of multiple (5-9) strain types. There was little evidence of strain dominance, with a single strain type occupying more than 30% of trap host nodules in only two of the five populations. Cluster analysis of RAPD PCR banding patterns showed that strain types in naturalised populations were not closely related to the current commercial inoculant strain for field pea (SU303, ≥75% dissimilarity), six previous field pea inoculant strains (≥55% dissimilarity) or a former commercial inoculant strain for faba bean (WSM1274, ≥66% dissimilarity). Two of the most closely related strain types (≤15% dissimilarity) were found at widely separate locations in SA and may have potential as commercial inoculant strains. Given the size and diversity of the naturalised pea rhizobia populations in SA soils and their relative effectiveness, it is unlikely that inoculation with a commercial strain of rhizobia will improve N₂ fixation in field pea crops, unless the number of rhizobia in the soil is very low or absent (e.g. where a legume host has not been previously grown and for three soils from western Eyre Peninsula). The general effectiveness of the pea rhizobia populations also indicates that reduced N₂ fixation is unlikely to be the major cause of the declining field pea yields observed in recent times.     

Mycorrhizal alleviation of acid soil stress in the sweet potato (Ipomoea batatas)

It is not known why sweet potato (Ipomoea batatas) cultivated in tropical regions tolerates acid soil. Here, we report the involvement of mycorrhizal symbiosis in this tolerance. Plants were grown in root-boxes filled with either acidic soil (pH 4.2) or the same soil amended with lime (pH 5.2) for 30 d in a growth chamber. In the inoculated treatments, the percentage of root length colonized by Gigaspora margarita was not affected by soil pH (23±9% at pH 4.2 vs. 30±12% at pH 5.2). The root and shoot dry weights of the non-mycorrhizal plants at pH 4.2 were 27 and 35%, respectively, of those at pH 5.2. The root and shoot dry weights of the mycorrhizal plants at pH 4.2 were 70 and 51% of those at pH 5.2. Growth promotion in mycorrhizal plants was significant only at pH 4.2 (2-fold increase in whole plant dry weight), but not at pH 5.2. As a result, no significant difference was detected in whole plant dry weight between the mycorrhizal plants at pH 4.2 and non-mycorrhizal plants at pH 5.2. The mycorrhizal plants at pH 4.2 showed reduced toxic symptoms of Mn (brown specks on mature leaves) and Al (poor root growth) compared to non-mycorrhizal ones, but tissue concentrations of P, K and Ca did not increase in mycorrhizal plants. We assume that the mycorrhizal colonization can reduce toxic effects of those elements while the exact mechanisms should be further investigated.     

The abundance and efficacy of Rhizobium leguminosarum bv. viciae in cultivated soils of the eastern Canadian prairie

For optimum production, the use of commercial rhizobial inoculant on pea (Pisum sativum L.) at seeding is necessary in the absence of compatible rhizobial strains or when rhizobial soil populations are low or symbiotically ineffective. Multiple site experiments were conducted to characterize the abundance and effectiveness of resident populations of Rhizobium leguminosarum bv. viciae (Rlv) in eastern Canadian prairie soils. A survey of 20 sites across a broad geographical range of southern Manitoba was carried out in 1998 and was followed by more intensive study of five of the sites in 1999 and 2000. Appreciable nodulation of uninoculated pea was observed at all sites which had previously grown inoculated pea. However, uninoculated pea grown at two sites, which had not previously grown pea, had negligible nodulation. Likewise, wild Lathyrus sp. and Vicia sp. plants collected from uncultivated areas adjacent to agricultural sites were poorly nodulated. In the more intensively studied sites, there was a tendency towards higher nodulation in pea plants receiving commercial inoculant containing Rlv strain PBC108 across all site-years (e.g., 4.7% in nodulation and 22% in nodule mass), but the effect was significant at only 2 of 10 site-years. Despite a relatively high range of soil pH (6-8), regression analysis indicated that decreasing soil pH resulted in lower nodulation rates. Likewise, electrical conductivity (EC) was correlated to nodulation levels, however the effect of EC was likely more indicative of the influence of soil texture and organic matter than salinity. As with nodulation, commercial inoculation tended to increase above-ground dry matter (DM) and fixed-N (estimated by the difference method) at the early pod-filling stage, but again the effects were significant at only 2 of 10 site-years. Specifically, above-ground DM and fixed-N levels were up to 29 and 51% greater, respectively, in inoculated compared to non-inoculated treatments at these sites. Addition of N-fertilizer at a rate of 100 kg N ha⁻¹ decreased nodulation at almost all site-years (by as much as 70% at one site), but rarely resulted in increases in above-ground DM compared to inoculated plots. The study indicates for the first time that populations of infective, and generally effective strains of Rlv occur broadly in agricultural soils across the eastern Canadian prairie, but that there is a tendency for increased symbiotic efficiency with the use of commercial inoculant.     

Nitrate removal from drained and reflooded fen soils affected by soil N transformation processes and plant uptake

Knowledge about nitrate transformation processes and how they are affected by different plants is essential in order to reduce the loss of valuable N fertiliser as well as to prevent environmental pollution due to nitrate leaching or N₂O emission after fertilisation or the reflooding of degraded fens with nitrate-containing municipal sewage. Therefore four microcosm ¹⁵N tracer experiments were performed to evaluate the effect of common wetland plants (Phalaris arundinacea, Phragmites australis) combined with different soil moisture conditions (from dry to reflooded) on nitrate turnover processes. At the end of experiment, the total formation of gaseous N compounds was calculated using the ¹⁵N balance method. In two experiments (wet and reflooded soil conditions) the N₂O and N₂ emissions were also directly determined.Our results show that in degraded fen soils, which process mainly takes place—denitrification or transformation into organic N compounds—is determined by the soil moisture conditions. Under dry soil moisture conditions (water filled pore space: 31%) up to 80% of the ¹⁵N nitrate added was transformed into organic N compounds. This transformation process is not affected by plant growth. Under reflooded conditions (water filled pore space: 100%), the total gaseous N losses were highest (77-95% of the ¹⁵N-nitrate added) and the transformation into organic N compounds was very low (1.8% of ¹⁵N nitrate added). Under almost all soil conditions plant growth reduced the N losses by 20-25% of the ¹⁵N nitrate added due to plant uptake. The N₂ emissions exceeded the N₂O emissions by a factor of 10-20 in planted soil, and as much as 30 in unplanted soil. In the treatments planted with Phragmites australis, N₂O emission was about two times higher than in the corresponding unplanted treatment. 15% of the N₂O and N₂ formed was transported via the Phragmites shoots from the soil into the atmosphere. By contrast, Phalaris arundinacea did not affect N₂O emissions and no emission via the shoots was observed.     

Phylogeny and nitrogen fixation potential of Bradyrhizobium species isolated from the legume cover crop Pueraria phaseoloides (Roxb.) Benth. in Eastern Cameroon

Pueraria is an herbaceous, perennial legume crop originating in Asia. Pueraria phaseoloides (Roxb.) Benth. (tropical kudzu) is frequently introduced into production systems and is used as green manure, a cover crop and a forage plant, making it important economically. We used P. phaseoloides as a trap crop to study and characterize soil rhizobia in Eastern Cameroon. Bacteria were isolated from fresh nodules collected from field-grown P. phaseoloides roots. The 16S-23S rRNA internal transcribed spacer (ITS) sequences from 30 bacterial isolates were amplified by polymerase chain reaction (PCR) and the reaction products were sequenced. Phylogenetic analysis revealed that all isolates were ascribed to the genus Bradyrhizobium and were grouped into three clusters of Bradyrhizobium sp. strains, one cluster of B. yuanmingense strains, and one cluster of B. elkanii strains. Acetylene reduction assay (ARA) results indicated that the B. yuanmingense strains had significantly higher nitrogen fixation potential and that they could be used as inoculants to enhance nitrogen fixation in Pueraria grown in Eastern Cameroon.     

Nitrogen fixation by biological soil crusts and heterotrophic bacteria in an intact Mojave Desert ecosystem with elevated CO2 and added soil carbon

Fixation of N by biological soil crusts and free-living heterotrophic soil microbes provides a significant proportion of ecosystem N in arid lands. To gain a better understanding of how elevated CO₂ may affect N₂-fixation in aridland ecosystems, we measured C₂H₂ reduction as a proxy for nitrogenase activity in biological soil crusts for 2 yr, and in soils either with or without dextrose-C additions for 1 yr, in an intact Mojave Desert ecosystem exposed to elevated CO₂. We also measured crust and soil δ¹⁵N and total N to assess changes in N sources, and δ¹³C of crusts to determine a functional shift in crust species, with elevated CO₂. The mean rate of C₂H₂ reduction by biological soil crusts was 76.9±5.6 μmol C₂H₄ m⁻² h⁻¹. There was no significant CO₂ effect, but crusts from plant interspaces showed high variability in nitrogenase activity with elevated CO₂. Additions of dextrose-C had a positive effect on rates of C₂H₂ reduction in soil. There was no elevated CO₂ effect on soil nitrogenase activity. Plant cover affected soil response to C addition, with the largest response in plant interspaces. The mean rate of C₂H₂ reduction in soils either with or without C additions were 8.5±3.6 μmol C₂H₄ m⁻² h⁻¹ and 4.8±2.1 μmol m⁻² h⁻¹, respectively. Crust and soil δ¹⁵N and δ¹³C values were not affected by CO₂ treatment, but did show an effect of cover type. Crust and soil samples in plant interspaces had the lowest values for both measurements. Analysis of soil and crust [N] and δ¹⁵N data with the Rayleigh distillation model suggests that any plant community changes with elevated CO₂ and concomitant changes in litter composition likely will overwhelm any physiological changes in N₂-fixation.     

The effect of growing soybean (Glycine max. L.) on N2O emission from soil

A pot experiment was conducted to investigate the effect of growing soybean on N₂O emission from soil. When soybean was growing in pots, the cumulative N₂O emission during the growing season was 2.26 mg N pot⁻¹, which was 5.9 times greater than that from the identical but unplanted pots (CK). However, the difference in N₂O fluxes between the two treatments was not significant until the grain-filling stage. Of the total N₂O emission, 94% took place during the period from grain-filling to ripening. Premature harvesting of the aerial parts of the plants at various growth stages substantially stimulated N₂O emission from the soil. These results implied that the process of symbiotic N fixation per se does not stimulate N₂O production or emission, but rather senescence and decomposition of the roots and nodules in the late growth stage. Therefore, additional N₂O would be emitted from the soil after harvesting of soybean with roots, litter, and residues left in situ.     

Elevated CO2 concentration and nitrogen fertilisation effects on N2O and CH4 fluxes and biomass production of Phleum pratense on farmed peat soil

The effects of elevated CO₂ supply on N₂O and CH₄ fluxes and biomass production of Phleum pratense were studied in a greenhouse experiment. Three sets of 12 farmed peat soil mesocosms (10 cm dia, 47 cm long) sown with P. pratense and equally distributed in four thermo-controlled greenhouses were fertilised with a commercial fertiliser in order to add 2, 6 or 10 g N m⁻². In two of the greenhouses, CO₂ concentration was kept at atmospheric concentration (360 μmol mol⁻¹) and in the other two at doubled concentration (720 μmol mol⁻¹). Soil temperature was kept at 15 °C and air temperature at 20 °C. Natural lighting was supported by artificial light and deionized water was used to regulate soil moisture. Forage was harvested and the plants fertilised three times during the basic experiment, followed by an extra fertilisations and harvests. At the end of the experiment CH₄ production and CH₄ oxidation potentials were determined; roots were collected and the biomass was determined. From the three first harvests the amount of total N in the aboveground biomass was determined. N₂O and CH₄ exchange was monitored using a closed chamber technique and a gas chromatograph. The highest N₂O fluxes (on average, 255 μg N₂O m⁻² h⁻¹ during period IV) occurred just after fertilisation at high water contents, and especially at the beginning of the growing season (on average, 490 μg N₂O m⁻² h⁻¹ during period I) when the competition of vegetation for N was low. CH₄ fluxes were negligible throughout the experiment, and for all treatments the production and oxidation potentials of CH₄ were inconsequential. Especially at the highest rates of fertilisation, the elevated supply of CO₂ increased above- and below-ground biomass production, but both at the highest and lowest rates of fertilisation, decreased the total amount of N in the aboveground dry biomass. N₂O fluxes tended to be higher under doubled CO₂ concentrations, indicating that increasing atmospheric CO₂ concentration may affect N and C dynamics in farmed peat soil.     

Temperature sensitivity of respiration differs among forest floor layers in a Pinus resinosa plantation

Decomposer microorganisms contribute to carbon loss from the forest floor as they metabolize organic substances and respire CO₂. In temperate and boreal forest ecosystems, the temperature of the forest floor can fluctuate significantly on a day-to-night or day-to-day basis. In order to estimate total respiratory CO₂ loss over even relatively short durations, therefore, we need to know the temperature sensitivity (Q₁₀) of microbial respiration. Temperature sensitivity has been calculated for microbes in different soil horizons, soil fractions, and at different depths, but we would suggest that for some forests, other ecologically relative soil portions should be considered to accurately predict the contribution of soil to respiration under warming. The floor of many forests is heterogeneous, consisting of an organic horizon comprising a few more-or-less distinct layers varying in decomposition status. We therefore determined at various measurement temperatures the respiration rates of litter, F-layer, and H-layer collected from a Pinus resinosa plantation, and calculated Q₁₀ values for each layer. Q₁₀ depended on measurement temperature, and was significantly greater in H-layer than in litter or F-layer between 5 and 17 °C. Our results indicate, therefore, that as the temperature of the forest floor rises, the increase in respiration by the H-layer will be disproportionate to the increase by other layers. However, change in respiration by the H-layer associated with change in temperature may contribute minimally or significantly to changes of total forest floor respiration in response to changes in temperature depending on the depth and thickness of the layer in different forest ecosystems.     

Nitrous oxide production and consumption in serially diluted soil suspensions as related to in situ N2O emission in submerged soils

A simple method for characterizing soil microbial community composition relevant to N₂O production and consumption was proposed. Ten-fold series soil dilution was prepared. Nitrate or N₂O was provided as the sole electron acceptor. Nitrous oxide concentration in the headspace gas across the serially diluted soil suspensions was measured against controls. Results showed that the patterns of N₂O production and consumption across the soil suspensions provided useful information on the microbial community composition relevant to N₂O production and consumption in these soils. An independent method, to that proposed here, was also employed to characterize denitrifier community compositions of the same soils. Data indicated that information on the soil microbial community composition characterized by both methods were compatible or mutually supporting and apparently related to in situ N₂O emissions. Soil samples from manure (applied with animal manure plus chemical fertilizer) plots had higher denitrification rates than the samples from normal fertilizer (applied with chemical fertilizer only) plots. It was concluded that functional characteristics of soil microbial communities relevant to N₂O production and consumption could be characterized at ecological levels and may potentially affect N₂O emissions.     

Net nitrification rate and presence of Nitrosospira cluster 2 in acid coniferous forest soils appear to be tree species specific

The impact of four coniferous tree species and their corresponding soil factors on N transformation rates and presence of ammonia-oxidising bacteria (AOB) was studied in an acid pine forest soil (Appelscha, The Netherlands). Pine soil had a relatively low net nitrification rate, while spruce, fir and larch soils showed high net nitrification rates. 16S rRNA and amoA sequences were only found in soils with high nitrification rates and belonged solely to Nitrosospira cluster 2. We conclude that tree species, possibly through their effects on soil C/N ratios, determines the presence of Nitrosospira cluster 2. Whenever AOB are present, however, the AOB community composition appears to be similar.     

Physiological responses to acid stress of an acid-soil tolerant and an acid-soil sensitive strain of Rhizobium leguminosarum biovar trifolii

Physiological responses to acid stress in two strains of Rhizobium leguminosarum bv trifolii of differing acid-soil tolerance were compared. Acidity affected the size and morphology of the acid-tolerant strain, WSM409, but not of the acid-sensitive strain, TA1. Acid grown cells of WSM409 and TA1 had less cell-associated Ca and Mg and more P than cells grown at pH 7.0. Potassium content was lower in acid grown cells; WSM409 was less affected by pH than that in TA1. WSM409 was more tolerant of pH shock at pH 3.5 when grown at pH 4.8 than when grown at pH 7.0. TA1 was more sensitive to pH shock when grown at pH 4.8 than when grown at pH 7.0. WSM409 shows a characteristic adaptive acid tolerance response, whereas TA1 shows an acid sensitive response.     

The wood-decaying fungus Hygrophoropsis aurantiaca increases P availability in acid forest humus soil, while N addition hampers this effect

We evaluated the influence of the brown rot fungus Hygrophoropsis aurantiaca on P solubility in the humus layer of a podzolic forest soil. This fungus is known to exude large amounts of oxalic acid that may stimulate weathering of minerals and increase dissolution of humus, which in turn may increase P availability in the soil surrounding the fungus. Humus was inoculated using small wooden pieces colonised by the fungus. The presence of the fungus resulted in elevated concentration of PO₄⁻ in the humus solution. In a second experiment birch seedlings grown in the same humus were able to utilise the PO₄⁻ mobilised by the fungus to increase their internal P content. The factor determining this increased P uptake and the increased available P might be oxalate produced by fungus. The acid may directly dissolve P or change organic forms of P making it more susceptible to reaction with phosphatases. This fungal effect on P solubility diminished when N was added to the soil in the form of a slow release N fertilizer (methyl urea), or when a soil with a higher soil N concentration was used. We found a strong correlation between NH₄⁺ concentration and total organic carbon in the soil solution at high NH₄⁺ concentrations, suggesting the dissolution of humus as a result of the high NH₄⁺ content in the solution. This study demonstrates that the wood-decaying fungus H. aurantiaca influences nutrient turnover in forest soil, and thereby nutrient uptake by forest trees. An intensified harvest of forest products such as whole tree harvesting may decrease the active biomass of the wood decomposers and may thereby change the availability of P and the leaching of N.     

Effect of intermittent drainage in reduction of methane emission from paddy soils in Hokkaido,northern Japan

ABSTRACT

Emission of methane (CH₄), a major greenhouse gas, from submerged paddy soils is generally reduced by introducing intermittent drainage in summer, which is a common water management in Japan. However, such a practice is not widely conducted in Hokkaido, a northern region in Japan, to prevent a possible reduction in rice grain yield caused by cold weather. Therefore, the effects of intermittent drainage on CH₄ emission and rice grain yield have not been investigated comprehensively in Hokkaido. In this study, we conducted a three-year field experiment in Hokkaido and measured CH₄ and nitrous oxide (N₂O) fluxes and rice grain yield to elucidate whether the reduction in CH₄ emission can be achieved in Hokkaido as well as other regions in Japan. Four experimental treatments, namely, two soil types [soils of light clay (LiC) and heavy clay (HC) textures] and two water management [continuous flood irrigation (CF), and intermittent drainage (ID)], were used, and CH₄ and N₂O fluxes were measured throughout the rice cultivation periods from 2016 to 2018. Cumulative CH₄ emissions in 2016 were markedly low, suggesting an initially low population of methanogens in the soils presumably due to no soil submergence or crop cultivation in the preceding years, which indicates a possible reduction in CH₄ emission by introducing paddy-upland crop rotation. Cumulative CH₄ emissions in the ID-LiC and ID-HC plots were 21–91% lower than those in the CF-LiC and CF-HC plots, respectively, whereas the cumulative N₂O emissions did not significantly differ between the different water managements. The amount of CH₄ emission reduction by the intermittent drainage was largest in 2018, with a comparatively long period of the first drainage for 12 days in summer. Rice grain yields did not significantly differ between the different water managements for the entire 3 years, although the percentage of well-formed rice grains was reduced by the intermittent drainage in 2018. These results suggest that CH₄ emission from paddy fields can be reduced with no decrease in rice grain yield by the intermittent drainage in Hokkaido. In particular, the first drainage for a long period in summer is expected to reduce CH₄ emission markedly.     

Physiological and symbiotic characteristics of Rhizobium fredii isolated from subtropical-tropical soils

Summary Physiological and symbiotic characteristics were identified in Rhizobium fredii isolated from subtropical-tropical soils. The generation times of R. fredii Taiwan isolated-SB 357 and -SB 682 were 1.7 and 2.5 h, respectively. These strains were associated with acid production in yeast-extract mannitol medium. They were able to use hexoses, pentose, sucrose, trehalose and raffinose. Strain SB 357 can resist a high concentration of kanamycin (100 g ml^–1 and penicillin (400 g ml^–1). It can tolerate up to 2.34% NaCl and 1031.3 mosmol kg^–1 (23.4 bars). The growth rate of R. fredii SB 357 under the concentration of approximately 450 mosmol kg^–1 (10.2 bars) was not affected by salinity, but responded to osmotic pressure. Both strains (SB 357 and SB 682) isolated from subtropical-tropical soils were able to form an effective N₂-fixing symbiosis with the US soybean cv Clark lanceolate leaflet.     

The effects of rice (Oryza sativa L. ssp. japonica) husk biochar on nitrogen dynamics during the decomposition of hairy vetch in two soils under high-soil moisture condition

ABSTRACT

Legumes, including hairy vetch (Vicia villosa Roth), are widely used as green manures. They fix nitrogen (N) and provide the N to other crops when they decompose, and thus are considered alternatives for chemical N fertilizers. However, N-rich plant residues, including hairy vetch, are also sources of soil nitrous oxide (N₂O) emissions, a greenhouse gas. On one hand, rice (Oryza sativa L. ssp. japonica) husk biochar is widely used as a soil conditioner in Japan and has been reported as a tool to mitigate soil N₂O emissions. We conducted a soil core incubation experiment (1.5 months) to compare the N₂O emissions during the decomposition of surface-applied hairy vetch (0.8 kg dried hairy vetch m^?2 soil) under semi-saturated soil moisture conditions (~100% water-filled pore space (WFPS)), using two soil types, namely Andosol and Fluvisol. Throughout the incubation period, the use of biochar suppressed soil NH₄⁺-N concentrations in Andosol, whereas the effect of biochar on NH₄⁺-N was not clear in Fluvisol. Biochar increased the nitrate (NO₃^?-N) levels both in Andosol and Fluvisol, suggesting a negative influence on denitrification and/or a positive influence on nitrification. Biochar application did not influence the cumulative N₂O emissions. Our study suggests that rice husk biochar is not a good option to mitigate N₂O emissions during the decomposition of surface-applied hairy vetch, although this study was performed under laboratory conditions without plants. However, the trends of the inorganic-N concentration changes followed by the addition of hairy vetch and biochar were markedly different between the two soil types. Thus, factors behind the differences need to be further studied.     

Rehabilitation of saline soil with biogas digestate,humic acid,calcium humate and their amalgamations

ABSTRACT

Amelioration of saline soil is a requisite in order to increase crop productivity. A soil incubation study was performed for 60 days using digestate, humic acid, calcium humate and their combinations to investigate the influence on physical, chemical, microbial and enzyme activities of saline soil. Overall, digestate combined with calcium humate followed by humic acid treatments have shown their potency in decreasing the soil pH, electrical conductivity (EC), and sodium ion (Na⁺) concentration, and increase in potassium ion (K ⁺), calcium ion (Ca ²⁺), magnesium ion (Mg ²⁺), mean weight diameter (MWD), soil enzyme activities, microbial biomass carbon (MBC), MBC: microbial biomass nitrogen (MBN) and soil respiration than control. The digestate, humic acid individually and their amalgamation evidenced greater MBN among all the treatments. The digestate alone efficiently improved the soil properties than humic acid and calcium humate individual groups except for the MWD where it is pronounced more in the latter groups. The greater metabolic quotient (qCO₂) was observed in control than organic matter amended treatments indicating the stress conditions. The increase in water-extractable organic matter (WEOM) with minimal aromaticity (specific ultraviolet absorbance at 254 nm-Suva ₂₅₄) in integrated amendments comprising groups, laid the ground reason to improve the properties of saline soil. Therefore, this study concludes that the fusion of fresh and humified substrates could facilitate reclamation.