Release, movement and recovery of 3,4-dimethylpyrazole phosphate (DMPP), ammonium, and nitrate from stabilized nitrogen fertilizer granules in a silty clay soil under laboratory conditions

 The composition of soil microbiota in four heated (350  °C, 1 h) soils (one Ortic Podsol over sandstone and three Humic Cambisol over granite, schist or limestone) inoculated (1.5 μg chlorophyll a g^–1 soil or 3.0 μg chlorophyll a g^–1 soil) with cyanobacteria (Oscillatoria PCC9014, Nostoc PCC9025, Nostoc PCC9104, Scytonema CCC9801, and a mixture of the four) was studied by cultural methods. The aims of the work were to investigate the potential value of cyanobacteria as biofertilizers for accelerating soil recolonization after fire as well as promoting microbiotic crust formation and to determine the microbial composition of such a crust. The inoculated cyanobacteria proliferated by 5 logarithmic units in the heated soils which were colonized very quickly and, after 2 months of incubation, the cyanobacterial filaments and associated fungal hyphae made up a matrix in which surface soil particles were gathered into crusts of up to 1.0 cm in thickness. These crusts were composed, on average, of 2.5×10¹⁰ cyanobacteria, 2.8×10⁶ algae, 6.1×10¹⁰ heterotrophic bacteria (of which 1.2×10⁸ were acidophilic, 1.3×10⁶ were Bacillus spp. and 1.5×10⁸ were actinomycetes) and 77.8 m fungal mycelium (1.4×10⁶ were fungal propagules) g^–1 crust. Counts of most microbial groups were positively correlated to cyanobacterial numbers. The efficacy of treatment depended on both the class of inoculum and the type of soil. The best inoculum was the mixture of the four strains and, whatever the inoculum used, the soil over lime showed the most developed crust followed by the soils over schist, granite and sandstone; however, the latter was comparatively the most favoured by the amendment. In the medium term there were no significant differences between the two inocula rates used. Biofertilization increased counts of cyanobacteria by 8 logarithmic units while heterotrophic bacteria, actinomycetes, algae and fungal propagules rose by >4 logarithmic units, acidophilic bacteria and Bacillus spp. by around 3 logarithmic units and fungal mycelia showed an 80-fold increase. The results showed that inoculation of burned soils with particle-binding diazotrophic cyanobacteria may be a means of both improving crust formation and restoring microbial populations. Received: 8 March 2000     

Spatial changes of soil fungal and bacterial biomass from a sub-alpine coniferous forest to grassland in a humid, sub-tropical region

 Fungal and bacterial biomass were determined across a gradient from a forest to grassland in a sub-alpine region in central Taiwan. The respiration-inhibition and ergosterol methods for the evaluation of the microbial biomass were compared. Soil fungal and bacterial biomass both significantly decreased (P<0.05) with the shift of vegetation from forest to grassland. Fungal and bacterial respiration rates (evolved CO₂) were, respectively, 89.1 μl CO₂ g^–1 soil h^–1 and 55.1 μl CO₂ g^–1 soil h^–1 in the forest and 36.7 μl CO₂ g^–1 soil h^–1 and 35.7 μl CO₂ g^–1 soil h^–1 in the grassland surface soils (0–10 cm). The fungal ergosterol content in the surface soil decreased from the forest zone (108 μg g^–1) to the grassland zone (15.9 μg g^–1). A good correlation (R ²=0.90) was exhibited between the soil fungal ergosterol content and soil fungal CO₂ production (respiration) for all sampling sites. For the forest and grassland soil profiles, microbial biomass (respiration and ergosterol) declined dramatically with depth, ten- to 100-fold from the surface organic horizon to the deepest mineral horizon. With respect to fungal to bacterial ratios for the surface soil (0–10 cm), the forest zone had a significantly (P<0.05) higher ratio (1.65) than the grassland zone (1.05). However, there was no fungal to bacterial ratio trend from the surface horizon to the deeper mineral horizons of the soil profiles. Received: 30 March 2000     

恶臭假单胞菌P861(Gus)在油菜根部定殖的生态研究

本研究采用Gus 基因标记技术和常规方法跟踪考察了恶臭假单胞菌P861(Gus) 在缩影系统油菜根圈的定殖情况,以及缩影系统内土壤类型、土壤含水量对根部定殖的影响。土壤含水量分别为60% FC和75% FC时,P861(Gus) 在砂姜黑土中的定殖水平高于50% FC的,不但能散布至种子下8cm 以内的根段部位,且定殖水平分别为7.5×10²和2.8×10³cfu·g^-1。在灰潮土缩影中,P861(Gus) 在油菜根圈的定殖动态表现为在油菜播种后3 ～6 天,定殖密度可达最高水平(5.5×10⁶cfu·g^-1) ,然后急速下降,最后保持在一个相对稳定的较低水平(7.6×10²cfu·g^-1) 。P861(Gus) 在不同根段部位的定殖密度并无从上到下逐渐递减的规律。     

Control of cotton <Emphasis Type="Italic">Verticillium</Emphasis> wilt and fungal diversity of rhizosphere soils by bio-organic fertilizer

Cotton Verticillium wilt is a destructive soil-borne disease affecting cotton production. In this study, application of bio-organic fertilizer (BIO) at the beginning of nursery growth and/or at the beginning of transplanting was evaluated for its ability to control Verticillium dahliae Kleb. The most efficient control of cotton Verticillium wilt was achieved when the nursery application of BIO was combined with a second application in transplanted soil, resulting in a wilt disease incidence of only 4.4%, compared with 90.0% in the control. Denaturing gradient gel electrophoresis patterns showed that the consecutive applications of BIO at nursery and transplanting stage resulted in the presence of a unique group of fungi not found in any other treatments. Humicola sp., Metarhizium anisopliae, and Chaetomium sp., which were considered to be beneficial fungi, were found in the BIO treatment, whereas some harmful fungi, such as Alternaria alternate, Coniochaeta velutina, and Chaetothyriales sp. were detected in the control. After the consecutive applications of BIO at nursery and transplanting stage, the V. dahliae population in the rhizosphere soil in the budding period, flowering and boll-forming stage, boll-opening stage, and at harvest time were 8.5 × 10², 3.1 × 10², 4.6 × 10², and 1.7 × 10² colony-forming units per gram of soil (cfu g⁻¹), respectively, which were significantly lower than in the control (6.1 × 10³, 3.4 × 10³, 5.2 × 10³, and 7.0 × 10³ cfu g⁻¹, respectively). These results indicate that the suggested application mode of BIO could effectively control cotton Verticillium wilt by significantly changing the fungal community structure and reducing the V. dahliae population in the rhizosphere soil.     

覆盖模式及小麦根系对土壤微生物区系的影响

采用平皿分离培养法研究了5种栽培模式和小麦根系对土壤细菌、真菌及放线菌数量的影响。连续2年的定位测定结果表明:覆膜有利于土壤微生物数量增加。5种栽培模式中,小麦根区、根外土壤细菌数量均以覆膜模式下最高,分别为116.8×106cfu·g-1和86.7×106cfu·g-1;土壤真菌和放线菌数量均以垄沟覆膜(垄上覆膜、垄沟播种)模式下最高,分别为3.0×103cfu·g-1、1.4×103cfu·g-1和18.9×105cfu·g-1、19.7×105cfu·g-1。不同模式下小麦根系对土壤细菌和真菌数量影响较大,表现为根区高于根外;而根系对放线菌影响较小,只有补灌和覆膜2种模式为根区高于根外。多重比较结果显示,覆膜与其他模式之间细菌数量差异极显著,根区土壤细菌和真菌数量与根外存在显著差异。覆盖和根系能大幅度增加根区细菌、真菌和放线菌的数量,强化小麦根区根外细菌和真菌的数量差异。     

Soil-released carbon dioxide from microbial biomass carbon in the cultivated soils of karst areas of southwest China

 Soil microbial biomass and the emission of CO₂ from the soil surface were measured in yellow soils (Ultisols) of the karst areas of southwest China. The soils are relatively weathered, leached and impoverished, and have a low input of plant residues. The measurements were made for a 1-year period and show a reciprocal relationship between microbial biomass and surface CO₂ efflux. The highest (42.6±2.8 mg CO₂-C m^–2 h^–1) and lowest (15.6±0.6 mg CO₂-C m^–2 h^–1) CO₂ effluxes are found in the summer and winter, respectively. The cumulative CO₂ efflux is 0.24 kg CO₂-C m^–2 year^–1. There is also a marked seasonal variation in the amount of soil microbial biomass carbon, but with the highest (644±71 μg C g^–1 soil) and lowest (270±24 μg C g^–1 soil) values occurring in the winter and summer, respectively. The cumulative loss of soil microbial biomass carbon in the top 10 cm of the soil was 608 μg C g^–1 year^–1 soil over 17 sampling times. The mean residence time of microbial biomass is estimated at 105 days, suggesting that the carbon in soil microbial biomass may act as a source of the CO₂ released from soils. Received: 13 July 1999     

Critical sulphur concentration and sulphur requirement of microbial biomass in a glucose and cellulose-amended regosol

 The critical S concentration and S requirement of the soil microbial biomass of a granitic regosol was examined. S was applied at the rate of 0, 5, 10, 20, 30 and 50 μg S as MgSO₄·7H₂O, together with either 3000 μg glucose-C or 3333 μg cellulose-C, 400 μg N, and 200 μg P g ^–1 soil and 200 μg K g^–1 soil. Microbial biomass, inorganic SO₄ ^2–-S, and CO₂ emission were monitored over 30 days during incubation at 25  °C. Both glucose and cellulose decomposition rates responded positively to the S made available for microbial cell synthesis. The amounts of microbial biomass C and S increased with the level of applied S up to 10 μg S g^–1 soil and 30 μg S g^–1 soil in the glucose- and cellulose-amended soil, respectively, and then declined. Incorporated S was found to be concentrated within the microbial biomass or partially transformed into soil organic matter. The concentration of S in the microbial biomass was higher in the cellulose- (4.8–14.2 mg g^–1) than in the glucose-amended soil (3.7–10.9 mg g^–1). The microbial biomass C:S ratio was higher in the glucose- (46–142 : 1) than in the cellulose-amended soil (36–115 : 1). The critical S concentration in the microbial biomass (defined as that required to achieve 80% of the maximum synthesis of microbial biomass C) was estimated to be 5.1 mg g^–1 in the glucose- and 10.9 mg g^–1 in the cellulose-amended soil. The minimum requirement of SO₄ ^2–-S for microbial biomass formation was estimated to be 11 μg S g^–1 soil and 21 μg S g^–1 soil for glucose- and cellulose-amended soil, respectively. The highest levels of activity of the microbial biomass were observed at the SO₄ ^2–-S concentrations of 14 μg S g^–1 soil and 17 μg S g^–1 soil, for the glucose and cellulose amendments, respectively, and were approximately 31–54% higher during glucose than cellulose decomposition. Received: 20 October 1999     

Natural compounds enhancing growth and survival of rhizobial inoculants in vermicompost-based formulations

The present study demonstrates the usefulness of natural microbial growth-promoting compounds for improving the stability and life of vermicompost-based (both granular and its aqueous extract) bioformulations. Granular vermicompost maintained the number of cells of Rhizobium meliloti Rmd 201 up to 5.9 × 10⁸ after 180 days at 28°C compared with 2.1 × 10⁸ in charcoal (powdered), while aqueous extract of the vermicompost supported the 5.6 × 10⁷ rhizobia numbers even after 270 days. The addition of 25 μL/mL cow urine and 0.01 mM calliterpinone, a natural plant growth promoter, increased the rhizobia number significantly in granular vermicompost and its aqueous extract, respectively.     

Response of field-grown bean (Phaseolus vulgaris L.) to Rhizobium inoculation and nitrogen fertilization in two Cerrados soils

 Most soils sown with field beans (Phaseolus vulgaris L.) contain indigenous rhizobia which might interfere with the establishment of inoculated strains. As a consequence, the benefits of bean inoculation are usually questioned, and the use of N fertilizer is gradually becoming a common practice. The present study had the objective of evaluating the effectiveness of inoculation and N fertilization in field soil with (site 1) and without (site 2) a previous bean-cropping history. At site 1, which had a rhizobial population of 7×10² cells g^–1 soil, inoculation had no effect on nodulation or yield, whereas at site 2 (<10 cells g^–1 soil) inoculation increased nodulation, nodule occupancy by the inoculated strain and grain yield. N fertilizer decreased nodulation at both sites, but increased grain yield at site 1 but not at site 2, indicating that the response to inoculation and N fertilization depends on the cropping history. When bean was cultivated for the first time, indigenous populations of rhizobia were low and high yields were accomplished solely with seed inoculation, with no further response to N fertilizer. In contrast, previous cultivation of bean increases soil rhizobia, preventing nodule formation by inoculated strains, and N fertilizer may be necessary for maximum yields. A significant interaction effect between N fertilizer and inoculation was detected for serogroup distribution only at site 2, with N fertilizer decreasing nodule occupancy by the inoculated strain and increasing the occurrence of indigenous strains. Consequently, although no benefits were obtained by the combination of inoculation and N fertilizer, this practice may be feasible with the selection of appropriate N-tolerant strains from the indigenous rhizobial population. Received: 26 May 1999     

Determination of competitive abilities of Bradyrhizobium japonicum strains in soils from soybean production regions in South Africa

Bradyrhizobium japonicum strain CB 1809 was recently chosen to replace strain WB 1 in commercial soybean [Glycine max (L.) Merr.] inoculants in South Africa, the selection criterion being N₂-fixing effectiveness. Nodulation competitiveness is an additional characteristic required of inoculants and was determined for CB 1809 and WB 1 as well as two other strains, USDA 110 and a Brazilian strain 965, using the gusA marker gene to identify strains. Initial experiments with plants grown in sterile sand showed that the competitive index of strain WB 1 was less than that of the other strains. Further comparisons used plants grown in five soils containing established populations of B. japonicum. When strains were applied in peat inoculum to seed at a rate of 1,000 cells per seed in a soil containing 300 rhizobia g^–1, significant differences in nodule occupancy were detected and strains ranked in the order 965>CB 1809>USDA 110>WB 1. The remaining four soils each contained about 10⁶ rhizobia g^–1 and 5×10⁶ cells were applied per seed. Nodule occupancy by inoculant strains ranged from 22% to 81% between soils. In this experiment, WB 1 was consistently the poorest performer and its competitiveness was significantly less than CB 1809. The competition results supported the recent decision to replace WB 1 with CB 1809 in commercial inoculants. Although WB 1 had been used in inoculants over a period of 19 years, this strain was detected in only one soil, where it comprised 8% of isolates. In contrast, a substantial proportion (32–78%) of isolates from the soils corresponded serologically to a former inoculant strain WB 66, which had been discontinued in 1966. This illustrates the difficulty of replacing a resident population with an introduced strain. The effect of naturalized populations on the establishment of CB 1809 in South African soils will need monitoring Received: 23 November 1999     

Survival and persistence of genetically modified Sinorhizobium meliloti in soil

Studies were conducted to evaluate the survival and persistence of Sinorhizobium meliloti 104A14 and two acid phosphatase-negative mutants in Kirkland (fine, mixed, thermic Udertic Paleustolls) silt loam soils with various fertility levels, and to assess the impact of inoculation on nodule occupancy and soil microbial community structure in the inoculated alfalfa (Medicago sativa L.) rhizosphere. Recovery of the inoculated strains was 100% (in the order of 10⁸ cells g⁻¹ soil) immediately following inoculation to soils, but decreased from 10⁸ cells g⁻¹ soil to undetectable levels in a nutrient-poor soil within 32 days. In a nutrient-rich soil, approximately 2–3% (4.7–7.43×10⁶ cells g⁻¹ soil) of the mutants and 23% (5.84×10⁷ cells g⁻¹ soil) of the wild-type inocula persisted for more than 64 days. Survivability and persistence of the wild-type S. meliloti were significantly greater than that of the genetically modified acid phosphatase negative mutants in all the soils tested. The persistence and nodule occupancy of the introduced S. meliloti in sterile and non-sterile soils were also tested for two repeated alfalfa growth periods in the same plant growth units, with a 1 month interval in between and no additional inoculation for the second period. Nodule occupancy of the introduced S. meliloti in non-sterile soils ranged from 30 to 60% for the first period and 85 to 100% for the second period. Our results suggest that survival and persistence of S. meliloti was enhanced by alfalfa cultivation and increased soil fertility, but impaired by mutation of acid phosphatase genes regardless of phosphorus nutritional levels. Moreover, inoculation with genetically modified S. meliloti strain 104A14 promoted indigenous bacterial growth in soil (increased bacterial population from 1.4×10⁶ to 4.3×10⁶ cells g⁻¹ soil), but not the growth of fungi and yeast. However, inoculation of the wild-type S. meliloti or genetically modified mutants did not result in significant changes in microbial community structure as indicated by EP indices and ratios of r/K strategists.     

Nitrogen mineralization in soil layers, soil particles and macro-organic matter under grassland

 A study was conducted to determine mineralization rates in the field and in different soil layers under three grassland managements (viz. a reseeded sward, a permanent sward with a conventional N management, and a long-term grass sward with 0 N (0-N) input). Potential mineralization rates of soil particles (sand, silt and clay) and macro-organic matter fractions of different sizes (i.e. 0.2–0.5, 0.5–2.0 and >2 mm) were also determined in the laboratory. In the reseeded plots, net mineralization was unchanged down to 40 cm depth. In the undisturbed conventional-N swards, mineralization rates were substantially higher in the top layer (0–10 cm) than in the deeper layers. In plots which had received no fertilizer N, mineralization was consistently low in all the layers. There was more macro-organic matter (MOM) in the 0-N plots (equivalent to 23 g kg^–1 soil for 0–40 cm) than in the two fertilized plots (i.e. conventional-N and reseeded) which contained similar amounts (ca. 15 g kg^–1 soil). C and N contents of separated soil particles did not differ amongst the treatments, but there were large differences with depth. Potential mineralization in the bulk soil was greatest in the 0–10 cm layers and gradually decreased with depth in all the treatments. Separated sand particles had negligible rates of potential mineralization and the clay component had the highest rates in the subsurface layers (10–40 cm). MOMs had high potential rate of mineralization in the surface layer and decreased with soil depth, but there was no clear pattern in the differences between different size fractions. Received: 17 November 1997     

Effect of cropping systems on nitrogen mineralization in soils

 Understanding the effect of cropping systems on N mineralization in soils is crucial for a better assessment of N fertilizer requirements of crops in order to minimize nitrate contamination of surface and groundwater resources. The effects of crop rotations and N fertilization on N mineralization were studied in soils from two long-term field experiments at the Northeast Research Center and the Clarion-Webster Research Center in Iowa that were initiated in 1979 and 1954, respectively. Surface soil samples were taken in 1996 from plots of corn (Zea mays L.), soybean (Glycine max (L.) Merr.), oats (Avena sativa L.), or meadow (alfalfa) (Medicago sativa L.) that had received 0 or 180 kg N ha^–1 before corn and an annual application of 20 kg P and 56 kg K ha^–1. N mineralization was studied in leaching columns under aerobic conditions at 30  °C for 24 weeks. The results showed that N mineralization was affected by cover crop at the time of sampling. Continuous soybean decreased, whereas inclusion of meadow increased, the amount of cumulative N mineralized. The mineralizable N pool (N _o) varied considerably among the soil samples studied, ranging from 137 mg N kg^–1 soil under continuous soybean to >500 mg N kg^–1 soil under meadow-based rotations, sampled in meadow. The results suggest that the N _o and/or organic N in soils under meadow-based cropping systems contained a higher proportion of active N fractions. Received: 10 February 1999     

Effect of coffee residues on growth and reproduction of Hyperiodrilus africanus (Oligochaeta, Eudrilidae) in Ivory Coast

Hyperiodrilus africanus (Beddard) is a 12-cm to 16-cm-long earthworm, which is widely distributed in West and Central Africa. It lives in the upper 10–20 cm of the soil, and feeds on a mixture of soil and above-ground litter. Cocoons obtained in the laboratory hatched on average 17 days after deposition and produced two juveniles on average. Paired individuals fed soil amended with 2% coffee residues grew significantly (P<0.05) faster than those in the control soil. Daily individual weight increments were respectively 6.1 mg worm^–1 day^–1 and 1.0 mg worm^–1 day^–1 in supplemented and control soil. The generation time was short, and cocoon production reached 9.6 month^–1 (i.e. 115 cocoons adult^–1 year^–1). When H. africanus collected from the field were raised in the laboratory, they grew slowly, laid fewer cocoons and mortality was high. Demographic parameters indicated an improvement when H. africanus were raised in batches rather than individually. Mating enhanced cocoon production although parthenogenesis was possible. Received: 4 April 1997     

Emission of N2O from rye grass (Lolium perenne L.)

 The possibility of an additional N₂O emission pathway via plants was investigated in a soil-rye-grass (Lolium perenne L.) system. The N₂O emission rate of the system varied between 0.8 and 13.3 mg N₂O-N m^–2 day^–1. Comparing the N₂O emission rate of the system before and immediately after cutting the rye grass allowed us to calculate the contribution of the rye grass to the N₂O emission from the soil-plant system. It was found that, depending on the type of fertilization and the growing period of the plants, the N₂O released from the rye grass varied between 0 and 2.8 mg N₂O-N m^–2 day^–1. N ₂ O emission mediated by the rye grass increased towards the end of the growing period. An exponential correlation [R²=0.93, y=(8×10^–6 x ² )–(2×10^–5 x)+0.21] was observed between the N₂O emission (y) from the rye grass and its NO₃ ^––N content (x). However, it was not clear whether N₂O was produced by the plants themselves or whether the rye grass served as a conduit for N₂O produced in the soil. Received: 18 March 1998     

Changes in CO<Subscript>2</Subscript>, <Superscript>13</Superscript>C abundance,inorganic nitrogen, β-glucosidase,and oxidative enzyme activities of soil during the decomposition of switchgrass root carbon as affected by inorganic nitrogen additions

This study was conducted to investigate the effect of inorganic nitrogen (N) and root carbon (C) addition on decomposition of organic matter (OM). Soil was incubated for 200 days with nine treatments (three levels of N (no addition (N0) = 0, low N (NL) = 0.021, high N (NH) = 0.083 mg N g⁻¹ soil) × three levels of C (no addition (C0) = 0, low C (CL) = 5, high C (CH) = 10 mg root g⁻¹ soil)). The carbon dioxide (CO₂) efflux rates, inorganic N concentration, pH, and potential activities of β-glucosidase and oxidative enzyme were measured during incubation. At the beginning and the end of incubation, the native soil organic carbon (SOC) and root-derived SOC were quantified by using a natural labeling technique based on the differences in δ ¹³C between C3 and C4 plants. Overall, the interaction between C and N was not significant. The decomposition of OM in the NH treatment decreased. This could be attributed to the formation of recalcitrant OM by N because the potentially mineralizable C pool was significantly lower in the NH treatment (3.1 mg C g⁻¹) than in the N0 treatment (3.6 mg C  g⁻¹). In root C addition treatments, the CO₂ efflux rate was generally in order of CH > CL > C0 over the incubation period. Despite no differences in the total SOC concentration among C treatments, the native SOC in the CH treatment (18.29 mg C g⁻¹) was significantly lower than that in the C0 treatment (19.16 mg C g⁻¹).     

Influence of N and non-N salts on atmospheric methane oxidation by upland boreal forest and tundra soils

 The short-term (24 h) and medium-term (30 day) influence of N salts (NH₄Cl, NaNO₃ and NaNO₂) and a non-N salt (NaCl) on first-order rate constants, k (h^–1) and thresholds (C_Th) for atmospheric CH₄ oxidation by homogenized composites of upland boreal forest and tundra soils was assessed at salt additions ranging to 20 μmol g^–1 dry weight (dw) soil. Additions of NH₄Cl, NaNO₃ and NaCl to 0.5 μmol g^–1 dw soil did not significantly decrease k relative to watered controls in the short term. Higher concentrations significantly reduced k, with the degree of inhibition increasing with increasing dose. Similar doses of NH₄Cl and NaCl gave comparable decreases in k relative to controls and both soils showed low native concentrations of NH₄ ⁺-N (≤1 μmol g^–1dw soil), suggesting that the reduction in k was due primarily to a salt influence rather than competitive inhibition of CH₄ oxidation by exogenous NH₄ ⁺-N or NH₄ ⁺-N released through cation exchange. The decrease in k was consistently less for NaNO₃ than for NH₄Cl and NaCl at similar doses, pointing to a strong inhibitory effect of the Cl^– counter-anion. Thresholds for CH₄ oxidation were less sensitive to salt addition than k for these three salts, as significant increases in C_Th relative to controls were only observed at concentrations ≥1.0 μmol g^–1 dw soil. Both soils were more sensitive to NaNO₂ than to other salts in the short term, showing a significant decrease in k at an addition of 0.25 μmol NaNO₂ g^–1 dw soil that was clearly attributable to NO₂ ^–. Soils showed no recovery from NaCl, NH₄ ⁺-N or NaNO₃ addition with respect to atmospheric CH₄ oxidation after 30 days. However, soils amended with NaNO₂ to 1.0 μmol NaNO₂ g^–1 dw showed values of k that were not significantly different from controls. Recovery of CH₄-oxidizing ability was due to complete oxidation of NO₂ ^–-N to NO₃ ^–-N. Analysis of soil concentrations of N salts necessary to inhibit atmospheric CH₄ oxidation and regional rates of N deposition suggest that N deposition will not decrease the future sink strength of upland high-latitude soils in the atmospheric CH₄ budget. Received: 30 April 1999     

Response of CH<Subscript>4</Subscript> oxidation and methanotrophic diversity to NH<Subscript>4</Subscript><Superscript>+</Superscript> and CH<Subscript>4</Subscript> mixing ratios

Methane oxidising activity and community structure of 11, specifically targeted, methanotrophic species have been examined in an arable soil. Soils were sampled from three different field plots, receiving no fertilisation (C), compost (G) and mineral fertiliser (M), respectively. Incubation experiments were carried out with and without pre-incubation at elevated CH₄ mixing ratios (100 ml CH₄ l⁻¹) and with and without ammonium (100 mg N kg⁻¹) pre-incubation. Four months after fertilisation, plots C, G and M did not show significant differences in physicochemical properties and CH₄ oxidising activity. The total number of methanotrophs (determined as the sum the 11 specifically targeted methanotrophs) in the fresh soils was 17.0×10⁶, 13.7×10⁶ and 15.5×10⁶ cells g⁻¹ for treatment C, G and M, respectively. This corresponded to 0.11 to 0.32% of the total bacterial number. The CH₄ oxidising activity increased 10⁵-fold (20–26 mg CH₄ g⁻¹ h⁻¹), the total number of methanotrophs doubled (28–76×10⁶ cells g⁻¹) and the methanotrophic diversity markedly increased in treatments with a pre-incubation at elevated CH₄ concentrations. In all soils and treatments, type II methanotrophs (62–91%) outnumbered type I methanotrophs (9–38%). Methylocystis and Methylosinus species were always most abundant. After pre-incubation with ammonium, CH₄ oxidation was completely inhibited; however, no change in the methanotrophic community structure could be detected.     

Isolation and selection of indigenous Azospirillum spp. from a subtropical island, and effect of inoculation on growth of lowland rice under several levels of N application

 Thirty-five Azospirillum strains (13 strains from plant roots and 22 strains from soils) were isolated from Ishigaki island, Japan, which has a subtropical climate. These strains were different from each other according to polymerase-chain-reaction band patterns obtained by using a random primer (OPT-08). Two Azospirillum strains (AZ43 and AZ92-2) were also examined for use in further experiments. Inoculation of lowland rice with these strains enhanced early growth of rice to various degrees. Inoculation of strains VIII.P1-2, AZ92-2, V.S2-2, and V.P5 in sterilized soil yielded higher shoot dry weights than the application of 90 μg N g^–1 soil without inoculation. Only inoculation with strains AZ92-2 and VIII.P1-2 caused higher N uptake than the application of 90 μg N g^–1 soil. Three strains were selected for the next experiment based on the results of their effect on the early growth of rice. An investigation was conducted to determine the ability of two indigenous Azospirillum strains (V.S2-2 and VIII.P1-2) and one stock strain (AZ92-2) to promote growth and nutrient-uptake of lowland rice in unsterilized soil under several levels of N application (0, 80, 160, and 240 mg N pot^–1). Inoculation with these strains without N application increased shoot dry weight by 12–15% compared to the uninoculated treatment. Inoculation with Azospirillum V.S2-2 together with the application of 160 mg N pot^–1 resulted in a shoot dry weight as high as that obtained in the treatment with 240 mg N pot^–1 without inoculation. Thus, in this former case, the amount of N applied could be reduced by 80 mg pot^–1 due to the effect of the microbial inoculum without a significant change in the high, targeted, yield.     

Effect of plant residue return on the development of surface soil pH gradients

 In the field, surface soil pH gradients were observed under senescing plants over late spring and summer. A soil incubation experiment was conducted (119 days, 20  °C) to provide direct evidence of the influence of plant residue incorporation on soil pH. This was investigated in terms of plant residue type (wheat and subterranean clover) and dry matter addition rate (0, 6.25, 12.5 and 25.0 g kg^–1), as well as the soil layer of incorporation (0–2.5 and 7.5–10 cm) and moisture regime (continuously moist and moist-dry cycles). During incubation, moist unamended soils slowly acidified. In contrast, the addition of plant residue resulted in a rapid (day 0–7) increase of soil pH due to the association, and particularly oxidation, of added organic anions. This was followed by a gradual (day 7–119) pH decline attributed to the mineralization and subsequent nitrification of added organic N. The addition of 12.5–25.0 g kg^–1 of cereal crop residues, and 6.25–25.0 g kg^–1 of legume-based pasture residues, resulted in a net alkalization of the surface 2.5 cm of soil. It was therefore concluded that surface soil pH gradients observed in the field were largely attributable to an increase of pH at the surface 2.5 cm in response to plant residue return. The magnitude of such gradients will be particularly large with the return of large quantities of plant residues of high ash alkalinity in soils of relatively low initial pH and biological activity, and when the surface of the soil is exposed to moist-dry cycles. Received: 11 October 1999