Intrinsic antibiotic resistance and competition in fast- and slow-growing soybean rhizobia on a hybrid of Asian and US cultivars

Summary Six fast-growing soybean rhizobia (Rhizobium fredii) and thirteen slow-growing soybean rhizobia (Bradyrhizobium japonicum) were examined for resistance to 10 antibiotics. Axenic studies were carried out to determine the competitiveness of dual-strain inocula consisting of fast- and slow-growing rhizobia isolated from subtropical-tropical soils for nodule occupancy on a hybrid of Asian and US soybean cultivars. Nodule occupancy was determined by intrinsic resistance to erythromycin and neomycin. The results showed wide variability in resistance to 10 antibiotics for fast- and slow-growing rhizobia. The intrinsic antibiotic resistance of fast- and slow-growing rhizobia was extremely high against nalidixic acid (400 g ml^–1) and penicillin (200 g ml^–1). The competitive ability of inoculant strains for nodule occupancy varied for different combination sets and with the plant growing media. Our results show that fast-growing rhizobia nodulate a hybrid of Asian and US soybean cultivars. Fast-growing soybean rhizobia did not completely exclude nodulation by the slow-growing strains, which formed 0–79% nodules, depending on the strain used in the inoculum.     

Competition among strains of Bradyrhizobium and vesicular-arbuscular mycorrhizae for groundnut (Arachis hypogaea L.) root infection and their effect on plant growth and yield

Summary Strains of Bradyrhizobium influenced root colonization by a species of vesicular-arbuscular mycorrhizae (VAM), and species of VAM influenced root nodulation by strains of Bradyrhizobium in pot experiments. In a field experiment, the effects of VAM on competition amongst inoculated bradyrhizobia were less evident, but inoculation with Bradyrhizobium strains increased root colonization by VAM. Certain VAM/Bradyrhizobium inoculum strain combinations produced higher nodule numbers. Plants grown without Bradyrhizobium and VAM, but supplied with ammonium nitrate (300 g ml^–1) and potassium phosphate (16 g ml^–1), produced higher dry-matter yields than those inoculated with both symbionts in the pot experiment. Inoculation with either symbiont in the field did not result in higher pod and haulm yields at harvest.ICRISAT Journal Article No. 886     

Effect of heavy metals on growth and survival of Macrophomina phaseolina (Tassi) Goid.

Summary The effects of Cd, Co, Ni and Zn on growth and survival of Macrophomina phaseolina were studied in vitro. Cd, Ni and Co at 500 g ml^–1 inhibited growth by 78.8%, 73.6% and 11.8%, respectively, after 4 days at 25 ± 1°C. The mycelial dry weight yield was enhanced by 2.1% at 100 g ml^–1 Zn. The population of the pathogen declined in soil amended separately with Cd, Co, Ni and Zn. Cd (4000 mg kg^–1) proved to be the most toxic, by completely inhibiting the survival of the pathogen in soil after 20 days.     

Influence of oxygen on denitrification and aerobic respiration in soil

Summary The influence of the partial pressure of oxygen on denitrification and aerobic respiration was investigated at defined P02 values in a mull rendzina soil. The highest denitrification and respiration rates obtained in remoistened, glucose- and nitrate-amended soil were 43 1 N₂0 h^–1g^–1 soil and 130 1 O₂ h^–1g^–1 soil, respectively. At -55 kPa matric water potential, corresponding to 40% water saturation, N₂0 was produced only below P0₂ 40 hPa. The K _m, for O₂ was 3.0 x 10^–6 M. Formation of N₂O and consumption of O₂ occurred simultaneously with half maximum rates at P0₂ 6.7–13.3 hPa. Nitrite accumulated in soil below 40 hPa and increased with decreasing pO₂. The upper threshold for N₂0 formation in amended soil was P0₂ 33–40 hPa (39-47 M O₂).     

Trace gas emissions from soil of the central highlands of Mexico as affected by natural vegetation: a laboratory study

In the central highlands of Mexico, mesquite (Prosopis laevigata) and huisache (Acacia schaffneri), N₂-fixing trees or shrubs, dominate the vegetation and are currently used in a reforestation program to prevent erosion. We investigated how natural vegetation or cultivation of soil affected oxidation of CH₄, and production of N₂O. Soil was sampled under the canopy of mesquite (MES treatment) and huisache trees (HUI treatment), outside their canopy (OUT treatment) and from fields cultivated with maize (ARA treatment) at three different sites while production of CO₂, and dynamics of CH₄, N₂O and inorganic N (NH₄⁺, and NO₃^–) were monitored in an aerobic incubation. The production of CO₂ was 2.3 times higher and significantly greater in the OUT treatment, 3.0 times higher in the MES treatment and 4.0 times higher in the HUI treatment compared to the ARA treatment. There was no significant difference in oxidation of CH₄ between the treatments, which ranged from 0.019 g CH₄–C kg^–1 day^–1 for the HUI treatment to 0.033 CH₄–C kg^–1 day^–1 for the MES treatment. The production of N₂O was 30 g N₂O–N kg^–1 day^–1 in the MES treatment and >8 times higher compared to the other treatments. The average concentration of NO₃^– was 2 times higher and significantly greater in the MES treatment than in the HUI treatment, 3 times greater than in the OUT treatment and 10 times greater than in the ARA treatment. It was found that cultivation of soil decreased soil organic matter content, C and N mineralization, but not oxidation of CH₄ or production of N₂O.     

Agricultural factors affecting methane oxidation in arable soil

CH₄ oxidation activity in a sandy soil (Ardoyen) and agricultural practices affecting this oxidation were studied under laboratory conditions. CH₄ oxidation in the soil proved to be a biological process. The instantaneous rate of CH₄ consumption was in the order of 800 mol CH₄ kg^–1 day^–1 (13 mg CH₄ kg^–1 day^–1) provided the soil was treated with ca. 4.0 mmol CH₄ kg^–1 soil. Upon repeated supplies of a higher dose of CH₄, the oxidation was accelerated to a rate of at least 198 mg CH₄ kg^–1 day^–1. Addition of the plant-growth promoting rhizopseudomonad strains Pseudomonas aeruginosa 7NSK2 and Pseudomonas fluorescens ANP15 significantly decreased the CH₄ oxidation by 20 to 30% during a 5-day incubation. However, with further incubation this suppression was no longer detectable. Growing maize plants prevented the suppression of CH₄ oxidation. The numbers of methanotrophic bacteria and fungi increased significantly after the addition of CH₄, but there were no significant shifts in the population of total bacteria and fluorescent pseudomonads. Drying and rewetting of soil for at least 1 day significantly reduced the activity of the indigenous methanotrophs. Upon rewetting, their activity was regained after a lag phase of about 3 days. The herbicide dichlorophenoxy acetic acid (2,4-D) had a strong negative effect on CH₄ oxidation. The application of 5 ppm increased the time for CH₄ removal; at concentrations above 25 ppm 2,4-D CH₄–oxidizing activity was completely hampered. After 3 days of delay, only the treatments with below 25 ppm 2,4-D showed recovery of CH₄–oxidizing activity. This finding suggests that it can be important to include a CH₄–removal bioassay in ecotoxicology studies of the side effects of pesticides. Changes in the native soil pH also affected the CH₄–oxidizing capacity. Permanent inhibition occurred when the soil pH was altered by 2 pH units, and partial inhibition by 1 pH unit change. A rather narrow pH range (5.9–7.7) appeared to allow CH₄ oxidation. Soils pre-incubated with NH ₄ ⁺ had a lower CH₄–removal capacity. Moreover, the nitrification inhibitor 2-chloro-6-trichloromethyl pyridine (nitrapyrin) strongly inhibited CH₄ oxidation. Probably methanotrophs rather than nitrifying microorganisms are mainly responsible for CH₄ removal in the soil studied. It appears that the causal methanotrophs are remarkably sensitive to soil environmental disturbances.     

Molybdenum reserves of seed,and growth and N2 fixation by Phaseolus vulgaris L.

Summary Plants grown from seed with high (1.5–7.3 g Mo seed^-1) and low (0.07–1.4 g Mo seed^-1) Mo contents were grown in the presence and absence of Mo in growth media (perlite) or in a flowing-solution culture, in a controlled environment. Neither the high (1.5 g Mo seed^-1) nor the low (0.1 g Mo seed^-1) Mo content in seed from a small-seeded genotype (BAT 1297) was able to prevent Mo deficiency (reduced shoot, root and nodule dry weight, N₂ fixation and seed production) in growth media without an external supply of Mo, whereas both the high (7.3 g Mo seed^-1) and the low (0.07 g Mo seed^-1) contents in seed were able to prevent Mo deficiency in a large-seeded genotype (Canadian Wonder). Responses to Mo treatment by the Two genotypes were inconsistent between the growth media and solution culture experiments. Seed with a large Mo content (3.5 g Mo seed^-1) from the Canadian Wonder genotype was unable to prevent Mo deficiency (reduced shoot and nodule dry weight and N₂-fixation) in a solution culture without an external source of Mo, whereas both the large (1.7 g Mo seed^-1) and the small (0.13 g Mo seed^-1) contents in seed prevented a deficiency in BAT 1297. Growing plants from seed with a small Mo content, without additional Mo, reduced the seed Mo content by 83–85% and seed production by up to 38% in both genotypes. Changes in seed size and increases in shoot, root and nodule dry weight occurred, but varied with the genotype and growth conditions. These effects were also observed in some cases where plants were grown with additional Mo, demonstrating that the amount of Mo in the seed sown can influence plant nutrition irrespective of the external Mo supply. Nodule dry weight, total N content of shoots and seed production were improved by using seed with a small Mo content (1.64–3.57 g Mo seed^-1) on acid tropical soils in Northern Zambia. Plants of both the large- and small-seeded genotypes grown from seed with a small Mo content (<1.41 g Mo seed^-1) had a smaller nodule weight, accumulated less N and produced less seed. The viability of seed with a small Mo content was lower (germination up to 50% less) than that of seed with a large Mo content.     

Phosphate solubilization by <Emphasis Type="Italic">Penicillium</Emphasis> spp. closely associated with wheat roots

In this study we found that Penicillium spp. exhibiting P-solubilizing activity are common both on and in the roots of wheat plants grown in southern Australian agricultural soils. From 2,500 segments of washed and surface-disinfested root pieces, 608 and 223 fungi were isolated on a selective medium, respectively. All isolates were screened for P solubilization on solid medium containing hydroxyapatite (HA); 47 isolates (5.7%) solubilized HA and were identified as isolates of Penicillium or its teleomorphs. These isolates were evaluated for solubilization of Idaho rock phosphate (RP) in liquid culture. Penicillium bilaiae strain RS7B-SD1 was the most effective, mobilizing 101.7 mg P l^–1 after 7 days. Other effective isolates included Penicillium simplicissimum (58.8 mg P l^–1), five strains of Penicillium griseofulvum (56.1–47.6 mg P l^–1), Talaromyces flavus (48.6 mg P l^–1) and two unidentified Penicillium spp. (50.7 and 50 mg P l^–1). A newly isolated strain of Penicillium radicum (KC1-SD1) mobilized 43.3 mg P l^–1. RP solubilization, biomass production and solution pH for P. bilaiae RS7B-SD1, P. radicum FRR4718 or Penicillium sp. 1 KC6-W2 was determined over time. P. bilaiae RS7B-SD1 solubilized the greatest amount of RP (112.7 mg P l^–1) and had the highest RP-solubilizing activity per unit of biomass produced (up to 603.2 g P l^–1 mg biomass^–1 at 7 days growth). This study has identified new isolates of Penicillium fungi with high mineral phosphate solubilizing activity. These fungi are being investigated for the ability to increase crop production on strong P-retaining soils in Australia.     

Bioaccumulation of selenium by floating aquatic plants

The degree to which floating aquatic plants concentrate Se in tissues was determined for four species grown in solutions containing various levels of Se. Results of this greenhouse study showed that all four plant species, Azolla caroliniana, Eichhornia crassipes, Salvinia rotundi folia, and Lemna minor absorbed Se quickly upon exposure to Se in water as concentrated as 2.5 g Se mL^–1, and attained maximum tissue concentrations within 1 to 2 weeks. Azolla absorbed Se to the highest tissue concentration (about 1000 g Se g^–1 dry matter) from the 2.5 g Se mL^–1 solution, followed by Salvinia (700 g Se g^–1), Lemna (500 g Se g^–1),and Eichhornia (300 g Se g^–1). Plant growth appeared unaffected by solution Se concentrations lower than about 1.25 g mL^–1. These results indicate potential for rapid Se movement from water into aquatic food chains, and for use of aquatic plants for Se removal in wastewater treatment systems.     

Microbial biomass and respiratory activity in soil aggregates of different sizes from three beechwood sites on a basalt hill

We studied the effects of aggregates of different sizes on the soil microbial biomass. The distribution of aggregate size classes (<2, 2–4, 4–10, >10 mm) in the upper mineral soil horizon (A_h layer) was very different in three sites (upper, intermediate, lower) in a beechwood (Fagus sylvatica) on a basalt hill (Germany). Aggregates of different sizes (<2, 2–4, 4–10 mm) contained different amounts of C and N but the C:N ratios were similar. C and N contents were generally higher in smaller aggregates. The maximum initial respiratory response by microorganisms in intact aggregates and in aggregates passed through a 1-mm sieve declined with the aggregate size, but the difference was more pronounced in intact aggregates. Disruption of aggregates generally increased this response, particularly in 4- to 10-mm aggregates in the lower site. Basal respiration differed strongly among sites, but was similar in each of the aggregate size classes. Aggregate size did not significantly affect the specific respiration (g O₂ g^–1 microbial C h^–1) nor the microbial: organic C ratio, but these parameters differed among sites. Microbial growth was increased strongly by passing the soil through a 1-mm sieve in each of the aggregate materials. The growth of microorganisms in disrupted aggregates was similar, and the effect of aggregate disruption depended on the growth of microorganisms in intact aggregates.     

External phosphorus requirement of mycorrhizal and non-mycorrhizal barley and soybean plants

An experiment was conducted under greenhouse conditions to evaluate the effects of vesicular arbuscular mycorrhizal (VAM) fungi on the external P requirements of barley and soybeans. The plants were grown in pots containing a P-deficient soil. A range of 10 P levels was obtained by adding 0, 20, 30, 40, 50, 60, 70, 110, 160, or 310 mg P kg^-1 as NaH₂PO₄ 2H₂O. Half of the pots were inoculated with the VAM fungus Glomus intraradices. The P concentration in the soil solution was determined using an adsorption isotherm and plotted against the relative yield. Barley did not respond to mycorrhizal inoculation and we concluded that P nutrition was not the limiting factor on the growth of this lowmycotrophic plant. In contrast, mycorrhizal inoculation stimulated the growth of soybeans. The external P requirements were 0.110 g ml^-1 for mycorrhizal and 0.148 g ml^-1 for non-mycorrhizal soybeans to obtain 80% of the maximum yield. In terms of P fertilization this corresponds to a saving of 222 kg P₂O₅ ha^-1. The mycorrhizal dependency of the soybean was highly correlated with the P concentration in the soil solution and it is proposed that both values should be displayed together.     

Quantification and potential availability of non-symbiotically fixed 15N in soil

Summary Non-symbiotic N₂ fixation was studied under laboratory conditions in two soils from Pakistan (Hafizabad silt loam and Khurrarianwala silt loam) and one from Illinois, USA (Drummer silty clay loam) incubated in a ¹⁵N-enriched atmosphere. N₂ fixation was greatest with the Drummer soil (18–122 g g^–1 soil, depending upon the soil treatment) and lowest with the Khurrarianwala soil (4–81 g g^–1 soil). Fixation was increased by the addition of glucose, a close correlation being observed between the amount of glucose added and the amount of N₂ fixed in the three soils (r = 0.96). Efficiency of N₂ fixation varied with soil type and treatment and was greatest in the presence of added inorganic P. Application of Mo apparently had a negative effect on the amount and efficiency of N₂ fixation in all the soils. The percentage of non-symbiotically fixed ¹⁵N in potentially mineralizable form (NH ₄ ⁺ -N released in soil after a 15-day incubation period under anaerobic conditions) was low (2%–18%, depending upon the soil treatment), although most of the fixed N (up to 90%) was recovered as forms hydrolysable with 6N HCl. Recovery in hydrolysable forms was much greater for the fixed N than for the native soil N, indicating that the former was more available for uptake by plants.     

Short-term measurements of uptake of nitrogen fixed in the rhizospheres of sorghum (Sorghum bicolor) and millet (Pennisetum americanum)

Summary In a series of short-term experiments root systems of young sorghum and millet plants inoculated with N₂-fixing bacteria were exposed to ¹⁵N₂-enriched atmospheres for 72 h. The plants were grown in a normal atmosphere for up to 22 days after the end of the exposure to allow them to take up the fixed N₂. Environmental conditions and genotypes of sorghum and millet were selected to maximise N₂-fixation in the rhizosphere. Detectable amounts of fixed N (> 16 g/plant) were rapidly incorporated into sorghum plants grown in a sand/farmyard manure medium, but measurable fixation was found on only one occasion in plants grown in soil. N₂ fixation was detectable in some experiments with soil-grown millet plants but the amounts were small (2–4 g/plant) and represented less than 1 % of plant N accumulated over the same period. In many cases there was no detectable ¹⁵N₂ incorporation despite measurable increases in ethylene concentration found during an acetylene reduction assay.Published as ICRISAT Journal Article No. JA 740     

Reduction of bacterial growth by a vesicular-arbuscular mycorrhizal fungus in the rhizosphere of cucumber (Cucumis sativus L.)

Summary Cucumber was grown in a partially sterilized sand-soil mixture with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus fasciculatum or left uninoculated. Fresh soil extract was places in polyvinyl chloride tubes without propagules of mycorrhizal fungi. Root tips and root segments with adhering soil, bulk soil, and soil from unplanted tubes were sampled after 4 weeks. Samples were labelled with [³H]-thymidine and bacteria in different size classes were measured after staining by acridine orange. The presence of VAM decreased the rate of bacterial DNA synthesis, decreased the bacterial biomass, and changed the spatial pattern of bacterial growth compared to non-mycorrhizal cucumbers. The [³H]-thymidine incorporation was significantly higher on root tips in the top of tubes, and on root segments and bulk soil in the center of tubes on non-mycorrhizal plants compared to mycorrhizal plants. At the bottom of the tubes, the [³H]-thymidine incorporation was significantly higher on root tips of mycorrhizal plants. Correspondingly, the bacterial biovolumes of rods with dimension 0.28–0.40×1.1–1.6 m, from the bulk soil in the center of tubes and from root segments in the center and top of tubes, and of cocci with a diameter of 0.55–0.78 m in the bulk soil in the center of tubes, were significantly reduced by VAM fungi. The extremely high bacterial biomass (1–7 mg C g^-1 dry weight soil) was significant reduced by mycorrhizal colonization on root segments and in bulk soil. The incorporation of [³H]-thymidine was around one order of magnitude lower compared to other rhizosphere measurements, probably because pseudomonads that did not incorporate [³H]-thymidine dominated the bacterial population. The VAM probably decreased the amount of plant root-derived organic matter available for bacterial growth, and increased bacterial spatial variability by competition. Thus VAM plants seem to be better adapted to compete with the saprophytic soil microflora for common nutrients, e.g., N and P, compared to non-mycorrhizal plants.     

Contribution of denitrification and autotrophic and heterotrophic nitrification to nitrous oxide production in andosols

To quantify the contribution of denitrification and autotrophic and heterotrophic nitrification to N₂O production in Andosols with a relatively high organic matter content, we first examined the effect of C₂H₂ concentrations on N₂O production and on changes in mineral N contents. The optimum C₂H₂ concentration for inhibiting autotrophic nitrification was 10 Pa. Secondly, and Andosol taken from an arable field was incubated for 32 days at 30°C at 60, 80, and 100% water-holding capacity with or without the addition of NH ₄ ⁺ or NO _inf3 ^sup- (200 mg N kg^-1), and subsamples collected every 4–8 days were further incubated for 24 h with or without C₂H₂ (10 Pa). At 60 and 80% water-holding capacity with NH ₄ ⁺ added, 87–92% of N₂O produced (200–250 g N₂O–N kg^-1) was derived from autotrophic nitrification. In contrast, at 100% water-holding capacity with or without added NO _inf3 ^sup- , enormous amounts of N₂O (29–90 mg N₂O–N kg^-1) were produced rapidly, mostly by denitrification (96–98% of total production). Thirdly, to examine N₂O production by heterotrophic nitrification, the Andosol was amended with peptone or NH ₄ ⁺ (both 1000 mg N kg^-1)+citric acid (20 g C kg^-1) and with or without dicyandiamide (200 mg N kg^-1). Treatment with citric acid alone or with citric acid+dicyandiamide suppressed N₂O production. In contrast, peptone increased N₂O production (5.66 mg N₂O–N kg^-1) mainly by denitrification (80% of total production). However, dicyandiamide reduced N₂O production to 1.1 mg N₂O–N kg^-1. These results indicate that autotrophic nitrification was the main process for N₂O production except at 100% water-holding capacity where denitrification became dominant and that heterotrophic nitrification had a lesser importance in the soils examine.Dedicated to Professor J. C. G. Ottow on the occasion of his 60th birthday     

Uptake and toxicity of copper and zinc for the African earthworm,Eudrilus eugeniae (Oligochaeta)

Growth rate change in earthworms is considered to be a suitable endpoint when determining sublethal effects. In this study we evaluated growth and maturation in the vermicomposting earthworm speciesEudrilus eugeniae as marker of sublethal toxicity of copper and zinc. We also compared routes of uptake. Apart from exposing worms experimentally for 73 days to contaminated food, a series of contact filter paper tests was also performed to determine LD₅₀ for copper and zinc. Both copper and zinc at sublethal concentrations affected growth and maturation in worms exposed to contaminated food. These worms had a copper content of 34.5 g g^–1 after 73 days and a zinc content of 184.9 g g^–1, showing a differential uptake. Copper was more toxic than zinc. Also in the contact test worms did take up more zinc than copper and the LD₅₀ (48 h) for copper was 0.011 mg cm^–2 and for zinc 0.066 mg cm^–2, which translated to body burdens of 6 g g^–1 for copper and 131 g g^–1 for zinc. Indications were that a regulatory mechanism existed for both metals. Both metals were taken up through the body wall at a relatively fast rate. This study indicated that the skin was the major route of metal uptake. This study also showed a poor relation between the two types of tests for purposes of evaluating lethality of zinc and copper.     

Mineral-fixed ammonium in clay- and silt-size fractions of soils incubated with 15N-ammonium sulphate for five years

Summary Four soils with 6, 12, 23, and 47% of clay were incubated for 5 years with ¹⁵N-labeled (NH_{4 2}SO₄ and hemicellulose. The incubations took place at 20°C and 55% water-holding capacity. Samples of whole soils, and clay- (<2 m) and silt-(2–20 m) size fractions (isolated by ultrasonic dispersion and gravity sedimentation) were analysed for labeled and native mineral-fixed ammonium. Mineral-fixed ammonium in non-incubated soil samples accounted for 3.4%–8.3% of the total N and showed a close positive correlation with the soil clay content (r ² = 0.997). After 5 years of incubation, the content of mineral-fixed ammonium in the clay fraction was 255–430 g N g^–1, corresponding to 71%–82% of the mineral-fixed ammonium in whole soils. Values for silt were 72–166 g N g^–1 (14%–33% of whole soil content). In the soils with 6% and 12% clay, less than 1 % of the labeled clay N was present as mineral-fixed ammonium. In the soil with 23% clay, 3% of the labeled N in the clay was mineral-fixed ammonium. Labeled mineral-fixed ammonium was not detected in the silt fractions. For whole soils, and clay and silt fractions, the proportion of native N present as mineral-fixed ammonium varied between 3% and 6%. In contrast, the proportion of labeled N found as mineral-fixed ammonium in the soil with 4701o clay was 23%, 38% and 31% for clay, silt, and whole-soil samples, respectively. Corresponding values for native mineral-fixed ammonium were 12%, 16%, and 10%. Consequently, studies based on soil particle-size fractions and addressing the N turnover in clay-rich soils should consider the pool of mineral-fixed ammonium, especially when comparing results from different size fractions with those from fractions isolated from soils of a widely different textural composition.     

Nitrogen mineralization and reorganization in casts of the geophagous tropical earthworm Pontoscolex corethrurus (Glossoscolecidae)

Summary Mineral N concentrations ranged from 133.1 to 167.8 g g^-1 dry soil in fresh casts of the endogeic earthworm Pontoscolex corethrurus fed on an Amazonian Ultisol; this was approximately five times the concentration in non-ingested soil. Most of this N was in the form of NH _inf4 ^sup+ . N also accumulated in microbial biomass, which increased from a control value of 10.5–11.3 to 67.5–74.1 g g^-1 in fresh casts. During a 16-day incubation, part of the NH _inf4 ^sup+ -N was nitrified and/or transferred to the microbial biomass. Total labile N (i.e., mineral+biomas N) decreased sharply at first (ca. 50% in the first 12 h), and then more slowly. The exact fate of this N (microbial metabolites, denitrification, or volatilization) is not known. After 16 days, the overall N content of the casts was still 28% higher than that of the control soil. Incubation of the soil before ingestion by the earthworms significantly increased the production of NH _inf4 ^sup+ in casts. We calculate that in a humid tropical pasture, 50–100 kg mineral N may be produced annually in earthworm casts. Part of this N may be conserved in the compact structure of the cast where the cast is not in close contact with plant roots.     

Development of cyanobacterial blooms in Valencian rice fields

Seasonal variation of biomass, photosynthetic pigments and C and N contents of the cyanobacterial blooms developed during the crop cycles of 1998, 1999 and 2000 at two locations in Valencian rice fields (Spain) were studied to find their potential contribution to soil fertility. Blooms of Gloeotrichia sp., Gloeocapsa sp., Microchaete sp., and Nostoc sp. were small and dispersed and appeared only for a few weeks in the experimental fields of the Tancat de Malta location during the crop season of 1998. The biomass of these blooms ranged from 4.4 to 12.8 mg dry wt cm^–2. A bloom of Anabaena sp. was found in the same location during most of the crop cycles of 1998 and 1999, covering up to 80% of the experimental fields. The biomass of this bloom ranged from 2.7 to 11.4 mg dry wt cm^–2. In the moment of its maximal extension (July 1998) we estimate a total biomass of 2,100 kg dry wt for this bloom, equivalent to 420 kg dry wt ha^–1. In the Sueca location a bloom of Microchaete sp. was observed in August 1999 and during most of the crop cycle of 2000, when it covered a wide area (60–70%) of the experimental field. The biomass of this bloom ranged from 0.9 to 5.6 mg dry wt cm^–2. Carbon contents of the blooms ranged from 150 to 310 g mg^–1 dry wt and N contents ranged from 11 to 30.2 g mg^–1 dry wt. Taking into account their N content and their extension we estimate that the incorporation of cyanobacterial blooms could add 4–12 kg N ha^–1 to soil. From an agronomical point of view this means that, in spite of their impressive aspect, cyanobacterial blooms can only fulfill a small proportion of the N requirements of rice plants.     

Nitrogen-fixing stem nodules on Aeschynomene afraspera

Summary Aeschynomene afraspera is a wild annual legume growing in periodically waterlogged soils in western Africa. This legume is characterized by a profuse stem nodulation. Nodules are formed on the stem at the emergence of lateral root primordia, called nodulation sites. These sites are irregularly distributed on vertical rows all along the stem and branches. Stem nodules are hemispherically shaped. Their outside is dark green and they contain a red-pigmented central zone. Stem nodules exhibit a high nitrogen-fixing potential. Acetylene reduction assays result in stem nodule activity of 309 mol C₂H₄ g^–1 dry nodule h^–1. Field-grown stem nodulated Aeschynomene accumulated more N (51 g N m^–2 in 10 weeks) than the root nodulated one. Because of this nitrogenfixing potential and its ability to grow in waterlogged conditions, A. afraspera could probably be introduced into tropical rice cropping systems.