Enzyme activities in a limed agricultural soil

 This study assessed the effect of eight lime application rates, with four field replications, on the activities of 14 enzymes involved in C, N, P, and S cycling in soils. The enzymes were assayed at their optimal pH values. The soil used was a Kenyon loam located at the Northeast Research Center in Nashua, Iowa. Lime was applied in 1984 at rates ranging from 0 to 17,920 kg effective calcium carbonate equivalent (ha^–1), and surface samples (0–15 cm) were taken after 7 years. Results showed that organic C and N were not significantly affected by lime application, whereas the soil pH was increased from 4.9 to 6.9. The activities of the following enzymes were assayed: α- and β-glucosidases, α- and β-galactosidases, amidase, arylamidase, urease, l-glutaminase, l-asparaginase, l-aspartase, acid and alkaline phosphatases, phosphodiesterase, and arylsulfatase. With the exception of acid phosphatase, which was significantly (P<0.001) but negatively correlated with soil pH (r=–0.69), the activities of all the other enzymes were significantly (P<0.001)and positively correlated with soil pH, with r values ranging from 0.53 for the activity of α-galactosidase to 0.89 for alkaline phosphatase and phosphodiesterase. The Δ activity/Δ pH values ranged from 4.4 to 38.5 for the activities of the glycosidases, from 1.0 to 107 for amidohydrolases and arylamidase, 97 for alkaline phosphatase, 39.4 for phosphodiesterase, and 11.2 for arylsulfatase. This value for acid phosphatase was –35.0. The results support the view that soil pH is an important indicator of soil health and quality. Received: 3 May 1999     

Upland rice seedling wilt and microbial biomass and enzyme activities of compost-treated soils

Rice seedling wilt frequently occurs in upland nurseries under well-aerated conditions and causes considerable economic loss. Whether the wilt is pathogenic or edaphic is not known. We hypothesize the use of composts to alleviate seedling wilt. The severity level of upland rice seedling wilt was significantly (p < 0.05) positively correlated with soil pH (r = 0.499; n = 19), but negatively correlated with soil organic matter (r = −0.745), microbial biomass C (r = −0.669), activities of dehydrogenase (r = −0.589), arylsulfatase (r = −0.272), fluorescein diacetate hydrolysis (r = −0.466), and β-glucosidase (r = −0.280). Correlations between severity level and soil inorganic N and exchangeable potassium K were not significant. Contents of Fe, Zn, Cu, and Mn in healthy seedlings were not significantly (p < 0.05) different from those in infected seedlings. These data suggest that seedling wilts are not associated with nutrient constraints. Compost amendment at the rate of 3% or above in pot experiments significantly improved seedling growth and reduced the wilt symptoms. Field trials further showed that aboveground weight of seedlings in compost-amended treatment ranged from 11.5 to 14.9 mg per plant, significantly higher than the range from 6.38 to 12.1 mg per plant in the control treatment; in addition to rice growth compost significantly increased microbial biomass and enzyme activities of soils. Soil fumigation significantly increased rice growth and alleviation symptoms in 11 out of 19 soils, suggesting the involvement of pathogens. It is concluded that upland seedling wilt is a pathogen-associated disease. Probably high soil pH and low soil biochemical activities may favor pathogen activities.     

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.     

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     

Variability in Azolla sporulation in response to phosphorus application

 Phosphorus application decreased the sporulation frequency and number of sporocarps per plant in all the three Azolla species and 21 A. pinnata strains evaluated in this study. The number of megasporocarps tended to be more depressed than the number of microsporocarps. Nevertheless, the sporulation of A. caroliniana was less sensitive to P than that of A. pinnata and A. microphylla. Its sporulation frequency in the mineral medium did not decrease at 2.5 μg P ml^–1 and remained unaffected between 5 and 20 μg P ml^–1. The sporulation frequency and sporocarp number in this species in the soil culture also were not significantly affected by an increase in the dose of P from 10.7 to 21.4 or 21.4 to 32.1 mg pot^–1. Large variations in the degree of inhibition of sporulation due to the application of P (21.4 mg pot^–1) also occurred among the A. pinnata strains tested. Received: 20 October 1999     

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.     

Pre-inoculation with arbuscular mycorrhizal fungi suppresses root knot nematode (<Emphasis Type="Italic">Meloidogyne incognita</Emphasis>) on cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis>)

A pot experiment was conducted to evaluate the influence of pre-inoculation of cucumber plants with each of the three arbuscular mycorrhizal (AM) fungi Glomus intraradices, Glomus mosseae, and Glomus versiforme on reproduction of the root knot nematode Meloidogyne incognita. All three AM fungi tested significantly reduced the root galling index, which is the percentage of total roots forming galls. Numbers of galls per root system were significantly reduced only in the G. intraradices + M. incognita treatment. The number of eggs per root system was significantly decreased by AM fungus inoculation, no significant difference among the three AM fungal isolates. AM inoculation substantially decreased the number of females, the number of eggs g⁻¹ root and of the number of eggs per egg mass. The number of egg masses g⁻¹ root was greatly reduced by inoculation with G. mosseae or G. versiforme. By considering plant growth, nutrient uptake, and the suppression of M. incognita together, G. mosseae and G. versiforme were more effective than G. intraradices.     

Phosphate solubilization potentials of soil Acinetobacter strains

Many phosphate solubilizing microorganisms (PSM) require external pyrroloquinoline quinone (PQQ) for strong phosphorus (P) solubilization in vitro. The objective of this study was to isolate efficient and PQQ-independent PSM. A total of 21 PSM were isolated from the rhizosphere soil of wheat and maize grown in the pots. Acinetobacter strains were the only PQQ-independent and most effective solubilizers of tricalcium phosphate containing agar. The mean P dissolved in liquid cultures of Acinetobacter strains in a 5-day incubation ranged from 167 to 888 μg/ml P. The pH dropped to below 4.7 from 7.8 in six isolates, which produced gluconic acid in concentrations ranging between 27.5 and 37.5 mM. There was a linear regression between soluble P and gluconic acid concentrations in the bacterial cultures (P < 0.05; R ² = 0.59). Inoculation with Acinetobacter sp. WR922 significantly (P < 0.05) increased wheat (Triticum aestivum L.) P content by 27% at 15 days after emergence (DAE) and dry matter by 15% at 30 DAE compared to the control. The plant P content in inoculated plants at 30 DAE was linearly correlated with soluble P of the bacterial cultures (P < 0.05; R ² = 0.69). Gluconic acid production directly affected phosphate solubilization in vitro, which in turn influenced plant P content of inoculated plants in PQQ-independent P-solubilizing Acinetobacter strains.     

Quantitative assessment of hydrolase production and persistence in soil

The aim of this work was to calculate indices of hydrolase production (Pr) and persistence (Pe) through simple arithmetical calculations. Changes in acid and alkaline phosphomonoesterase, phosphodiesterase, urease, protease, and β-glucosidase activities were monitored under controlled conditions in seven soils with a wide range of properties, in which microbial growth was stimulated by adding glucose and nitrogen. Glucose mineralization was monitored by CO₂–C evolution, and microbial growth was quantified by determining the soil adenosine triphosphate (ATP) content. Hydrolase Pr and Pe indices were numerically quantified by the following relationships: Pr = H / t _H and Pe = (r / H)Δt, respectively, where H indicates the peak value of each measured hydrolase activity, t _H is the time of the peak value, r indicates the residual activity value, and Δt is the time interval t _r − t _H, where t _r is the time of the residual activity value. Addition of glucose and N-stimulated soil respiration increased ATP content and stimulated the production of the measured hydrolase activities in all soils; the measured variable reached a maximum value and then decreased, returning to the value of the control soil. Apart from β-glucosidase activity, whose activity was not stimulated by glucose and N addition, the other measured hydrolase activities showed a trend that allowed us to calculate the Pr and Pe indices using the above-mentioned equations. Acid phosphomonoesterase and protease Pr values were significantly higher in soils under forest or set aside management; the alkaline phosphomonoesterase and phosphodiesterase Pr values were generally higher in the neutral and alkaline soils, and the urease Pr values showed no obvious relationships with soil pH or management. Concerning the persistence of enzyme activities, Pe values of the acid phosphomonoesterase activity were significantly higher in the acidic soils, and those of urease activity were higher in acidic soils and the Bordeaux neutral soil. No relationships were observed between Pe values of alkaline phosphomonoesterase, phosphodiesterase, or protease activities and soil pH or management. The different responses of hydrolases were discussed in relation to soil properties, microbial growth, and regulation at the enzyme molecular level.     

Ecological pre-release risk assessment of two genetically engineered, bioluminescent Rhizobium meliloti strains in soil column model systems

In order to identify potential ecological risks associated with the environmental release of two Rhizobium meliloti strains, genetically engineered with the firefly-derived luciferase gene (luc), a pre-release greenhouse investigation was conducted. The upper 4 cm of soil columns (30 cm diameter; 65 cm depth), which were filled according to the horizons of an agricultural field (loamy sand), were inoculated with seeds of Medicago sativa (alfalfa) and R. meliloti cells at approximately 5×10⁶ cells·g^–1 soil. Four treatments were tested: inoculation with a non-engineered wild type strain (2011), strain L33 (luc ⁺), strain L1(luc ⁺, recA^–) and non-inoculated controls. The fate of the engineered strains was followed by two methods: (1) selective cultivation and subsequent detection of bioluminescent colonies and (2) PCR detection of the luc gene in DNA, directly extracted from soil. Strain R. meliloti L33 declined to 9.0×10⁴ cfu·g^–1 soil within 24 weeks and to 2.8×10³ cfu·g^–1 soil within 85 weeks in the upper 25 cm of the soil columns. Decline rates for R. meliloti L1 were not significantly different. Vertical distribution analysis of the recombinant cells after 37 weeks revealed that in three of four columns tested, the majority of cells (>98%) remained above 10 cm soil depth and no recombinant cells occurred below 20 cm depth. However, in one column all horizons below 20 cm were colonized with 2.2×10⁴ to 6.8×10⁴ cfu g^–1 soil. Ecological monitoring parameters included organic substance, total nitrogen, ammonium and nitrate, microbial biomass, culturable bacteria on four different growth media and the immediate utilization of 95 carbon sources (BiologGN) by soil-extracted microbial consortia. None of the parameters was specifically affected by the genetically engineered cells. Received: 6 December 1996     

Effectiveness of foreign bradyrhizobia strains in enhancing nodulation, dry matter and seed yield of soybean (Glycine max L.) cultivars in Nigeria

 Field experiments were conducted to investigate the performance of three soybean cultivars with five foreign bradyrhizobia strains in different regions. The experiments at the two sites were designed with soybean (Glycine max L.) cultivars as the main factor and bradyrhizobia strains (USDA 136, TAL 122, USDA 6, TAL 377 and TAL 102) as the sub-factor. The experiments were arranged in randomised complete block design with four replications. Results show that nodule number, nodule dry weight and shoot dry weight, total N and seed yield were significantly increased when soybean cultivars were inoculated with foreign bradyrhizobia in two locations in the south east of Nigeria. At 63 days after planting the percentage increase in nodule number and dry weight after inoculation of soybean cultivars with bradyrhizobia strains ranged from 71 to 486% and from 0 to 200%, respectively. The percentage increase in shoot dry matter, %N and total N after bradyrhizobia inoculation ranged between 2–130%, 18–62% and 35–191%, respectively at Awka, and at the Igbariam site the percentage increase in shoot dry weight, %N and total N ranged between 3–76%, 0–43% and 19–125%, respectively. Seed yields after bradyrhizobia inoculation of soybean cultivar TGX 1485–1D at Igbariam ranged between 1.20 and 2.18 t ha^–1 against the uninoculated plants, which had seed yields of 1.05 t ha^–1. The poorest yield response after inoculation with bradyrhizobia strains was observed in soybean cultivar M-351, with a seed yield ranging from 0.60 to 0.98 t ha^–1. The fact that foreign bradyrhizobia strains were more effective than the indigenous strains for all the parameters studied suggests that there is a need to use bradyrhizobia inoculants for increased soybean production in Nigeria. The variations in the strain performance with the different soybean cultivars at the two sites, emphasises the need for careful Bradyrhizobium spp. strain selection. The fact that inoculation response was cultivar- and site-specific suggests that strategies for improving inoculation response in soybean cultivars should also consider the soil environment where the soybean is to be produced. Received: 25 May 1999     

Diversity of arbuscular mycorrhizal fungi associated with the rhizosphere of tea growing in ‘natural’ and ‘cultivated’ ecosites

The colonization and diversity of arbuscular mycorrhizal fungi (AMF) associated with the rhizosphere of tea [Camellia sinensis (L.) O. Kuntze] growing under ‘natural’ as well as ‘cultivated’ conditions in the Kumaun region of Uttaranchal Himalaya (India), during the periods of active growth and dormancy were investigated. Root and rhizosphere soil samples, collected from both the ecosites (natural and cultivated), were monitored for root colonization. While the percent root colonization was quite high (77.66 ± 4.40 and 86.40 ± 3.02%, in the natural and cultivated tea, respectively) during the period of active growth in both the ecosites, relatively higher colonization (97.33 ± 0.78 and 98.13 ± 0.80%, in the natural and cultivated tea, respectively) was recorded during the period of dormancy. The rhizosphere of cultivated tea bushes was found to be dominated by Glomus morhpotypes (88.89% of the total isolates) along with three morphotypes of Acaulospora; occurrence of 35 morphotypes belonging to four genera viz. Acaulospora (11.43%), Gigaspora (11.43%), Glomus (68.57%) and Scutellospora (8.57%) was recorded in the rhizosphere of tea plants from the natural ecosite. A total of 51 AMF morphotypes were detected. Shannon–Weaver index of diversity was higher (1.80 ± 0.13 and 2.05 ± 0.10 during periods of active growth and dormancy, respectively) at the species level for the natural ecosite over its counterparts from the cultivated ecosite. Values for the diversity indices of natural and cultivated ecosites did not show much variation in the period of dormancy. These data suggest that collectively, various cultural practices negatively affect AMF diversity at the genus level in tea plantations of the colder regions.     

Metabolic and physiological characteristics of salt-tolerant strains of Bradyrhizobium spp.

 Biochemical and physiological characteristics of salt-tolerant (88 mM, 264 mM and 440 mM NaCl) strains of Bradyrhizobium were evaluated according to their capacity for using different carbon sources, growth rate, resistance to antibiotics, plasmid profile and exopolysaccharide production. Salt-tolerant strains significantly enhance their capacity to oxidize C sources (about 75–85 compounds) by increasing growth rate and exopolysaccharide production involved in adhesion, resulting in a greater adapting capacity to colonize unfavorable saline environments. However, salt stress could work as a curing agent and thus the gene stability would become critical for the biological nitrogen-fixation information present in plasmids, as is the case in Rhizobium. Received: 5 January 1998     

Nodulation of rooibos (<Emphasis Type="Italic">Aspalathus linearis</Emphasis> Burm. f.), an indigenous South African legume,by members of both the α-Proteobacteria and β-Proteobacteria

Rooibos (Aspalathus linearis) has been reported to be nodulated by rhizobia belonging to members of the genus Bradyrhizobium but based solely on slow growth rate on growth media in vitro. Because there is very little information about the rhizobia that nodulate and fix nitrogen in rooibos, the characterization of rhizobial strains and their ability to nodulate A. linearis was investigated in this study. Soils intially collected from the rhizosphere of different Aspalathus populations were used in a baiting experiment to trap rhizobia by rooibos roots. The rhizobia trapped in the nodules were re-isolated and used in Koch’s postulate experiment using the Leonard jar assembly in the glasshouse. The strains that formed on the average between five and 12 indeterminate pink nodules per plant resulted in statistically significant (P = 0.05) increase in shoot and root dry weights. Phylogenetic analysis of the 16S ribosomal RNA sequence of the isolates from the root nodules revealed for the first time that A. linearis is nodulated by different groups of rhizobia belonging to members of both the α-Proteobacteria and the β-Proteobacteria. It was also found that only 2% of the total rhizobia isolated from the root nodules of rooibos were represented by the genus Bradyrhizobium. The finding that rooibos is nodulated by different groups of α-rhizobia and β-rhizobia provides valuable information both in the study of the microbial ecology of rooibos and in the selection of highly efficient nitrogen fixing strains for the commercial cultivation of this indigenous legume.     

Arylsulfatase activity of microbial biomass in soils as affected by cropping systems

 The impacts of crop rotations and N fertilization on different pools of arylsulfatase activity (total, intracellular, and extracellular) were studied in soils of two long-term field experiments in Iowa to assess the contibution of the microbial biomass to the activity of this enzyme. Surface-soil samples were taken in 1996 and 1997 in corn, soybeans, oats, or meadow (alfalfa) plots that received 0 or 180 kg N ha^–1 before corn, and an annual application of 20 kg P ha^–1 and 56 kg K ha^–1. The arylsulfatase activity in the soils was assayed at optimal pH (acetate buffer, pH 5.8) before and after chloroform fumigation; microbial biomass C (C_mic) and N (N_mic) were determined by chloroform-fumigation methods. All pools of arylsulfatase activity in soils were significantly affected by crop rotation and plant cover at sampling time, but not by N fertilization. Generally, the highest total, intracellular, and extracellular arylsulfatase activities were obtained in soils under cereal-meadow rotations, taken under oats or meadow, and the lowest under continuous cropping systems.Total, intracellular, and extracellular arylsulfatase activities were significantly correlated with C_mic (r>0.41, P<0.01) and N_mic (r>0.38, P<0.01) in soils. The averages of specific activity values, i.e., of arylsulfatase activity of the microbial biomass, expressed per milligram C_mic, ranged from 315 to 407 μg p-nitrophenol h^–1. The total arylsulfatase activity was significantly correlated with the intracellular activity, with r values >0.79 (P<0.001). In general, about 45% of the total arylsulfatase activity was extracellular, and 55% was associated with the microbial biomass in soils, indicating the importance of the microflora as an enzyme source in soils. Received: 23 April 1998     

Comparative Study of Lead Accumulation in Different Organs of the Freshwater Crab <Emphasis Type="Italic">Zilchiopsis oronensis</Emphasis>

Lead (Pb) is known as an important aquatic contaminant with different toxic effects on various organisms. Until now, only few quantitative investigations have been published comparing Pb content in different organs of adult freshwater crabs. Their capacity to bioaccumulate other heavy metals is already known, and they can potentially transfer Pb to the terrestrial systems, as they are frequent trophic items of reptiles and birds, even humans. The objectives of this study were to assess Pb accumulation in the gills, carapace, digestive gland, and quela muscle of the freshwater crab Zilchiopsis oronensis, and to correlate bioaccumulation with morphometric data and sex. The crabs were manually caught in unpolluted ponds of the middle Paraná River alluvial valley (Santa Fe, Argentina). After the acclimation period, they were individually and randomly exposed per quadruplicate to three Pb experimental doses: 20, 40, and 80 mg Pb/L, in plastic cages during 15 days. After dissecting the crabs, the tissues were analyzed for lead in a Perkin Elmer Analyst 800 atomic absorption spectrometer. We found significant differences (p < 0.05) between the control and each one of the treatments but not between treatments (p > 0.05) and highly significant differences (p < 0.0001) between Pb concentration in organs. The Tukey posttest showed significant differences (p < 0.05) between gills–carapace, gills–digestive gland, and gills–quela muscle. The weight of the crabs only showed a negative correlation with Pb in the quela muscle (r = −0.53; p = 0.03). Pb in the carapace (but not in the other tissues) was positively correlated with the width (p = 0.571) and length (p = 0.616). Males accumulated more Pb than females, though not significantly. The present paper is aimed to contribute to our knowledge on Pb accumulation in freshwater crabs and select the better indicator organisms for biomonitoring.     

Decolorization and Metabolism of Anthraquionone-Type Dye by Laccase of White-Rot Fungi Polyporus sp. S133

Utilization of microbes including white-rot fungi and bacterial strains for decolorization of synthetic dyes is one promising strategy of an environmentally friendly and cost-competitive alternative to physico-chemical decomposition processes for treating industrial effluents. In this study, the biodegradation ability of the white-rot fungi Polyporus sp. S133 that produce high laccase was investigated in order to decolorize anthraquinone-type dye. Parameter including pH, temperature, and non-ionic surfactant were used to comparatively study the decolorizing effects on Remazol Brilliant Blue R (RBBR). The purified laccase totally decolorized 200 mg L⁻¹ initial concentration of RBBR dye when only 1.5 U L⁻¹ of laccase was used in the reaction mixture. The optimal decolorization rates were achieved at pH 5 and at a temperature of 50°C. N-hydroxybenzotriazole, a small molecular weight redox mediator, was found to accelerate the decolorization. Tween 20 inhibited the decolorization while Tween 80 and Brij 35 showed no inhibition effect. Two compounds were identified as the intermediates (m/z 304.3 and m/z 342.2). These results suggest that laccase from Polyporus sp. S133 is a powerful tool for the decolorization of anthraquinone dyes. A pathway for the metabolism of the RBBR by laccase of Polyporus sp S133 was proposed. These proposed pathways could contribute to a better comprehension of the mechanisms used by oxidative enzymes to transform organic compounds.     

Abundance of soil mites under four agroforestry tree species with contrasting litter quality

 Populations of soil-dwelling mites were monitored in monoculture plots of four agroforestry tree species, Gliricidia sepium, Leucaena leucocephala, Dactyladenia barteri and Treculia africana, and compared to those in grass and secondary forest plots in the dry season (December 1993 to January 1994) and in the wet season (April to June 1994) in southwest Nigeria. Mite populations were very low in all plots during the dry season (500–3000 m^–2), compared to those during the wet season (10 000–30 000 m^–2). The highest mite population was observed in Gliricidia plots (3 044 m^–2) for the dry season and Leucaena plots (30 240 m^–2) for the wet season. Mite genera that were dominant in all the experimental plots were Annectacarus, Haplozetes, Machadobelba, Scheloribates and members of the Galumnidae, Dermanyssidae and Parasitidae. The community structure of mites was similar in the soil for Treculia and Gliricidia plots and for Leucaena and Dactyladenia plots. There were more taxonomic groups of mites under Leucaena than in the other agroforestry plots. Based on the density, diversity and complexity of the mite communities, Leucaena was considered to be better than other agroforestry species in encouraging the growth of mite populations. Received: 28 April 1998     

Repair of a compacted Oxisol by the earthworm Pontoscolex corethrurus (Glossoscolecidae, Oligochaeta)

The potential of Pontoscolex corethrurus to repair the physical degradation of a compacted Oxisol was studied. The Oxisol from Kingaroy, Queensland, Australia was uniformly packed to four treatments of different bulk densities (0.95, 1.15, 1.25, 1.35 Mg m^–3) in pots of 0.24 m diameter and 0.22 m deep. Each pot was inoculated with 12 earthworms (equivalent to 300 m^–2) and maintained close to field capacity water content for 3 months, after which selected soil physical (dry bulk density, penetration resistance, water infiltration), worm activity (cast production, worm weight) measurements and image analysis were carried out. Results showed that worm numbers were maintained at the initial levels in all the treatments except in the 1.35 Mg m^–3 treatment, where there was a 33% decrease. The weight of surface casts per surviving worm was the greatest in the 1.35 Mg m^–3 treatment compared to the lower density treatments. Final soil density was lessened in all treatments to a depth of 0.2 m. Surface cast production was positively correlated with the percentage reduction in bulk density. The greatest percentage reduction in bulk density was in the 1.35 Mg m^–3 treatment and was equivalent to a doubling of soil aeration (to 18.4%). Penetration resistance to the 0.2-m layer was also reduced and values were less than 2 MPa. Slumping of the surface soil was evident in both the lowest and the highest bulk densities resulting in low rates of water infiltration. Image analysis supported the soil physical properties showing an abundance of both fine pores and worm channels in the three lowest bulk densities, and lesser activity (concentrated in the immediate soil surface) in the 1.35 Mg m^–3 treatment. Received: 6 November 1996     

Promotion of mycorrhiza formation and growth of willows by the bacterial strain <Emphasis Type="Italic">Sphingomonas</Emphasis> sp. 23L on fly ash

Re-vegetation of fly ash, the principal by-product of coal fired power stations, is hampered by its unfavourable chemical and physical properties for plant growth. In the present study, we evaluated the use of inoculation with a mycorrhiza-associated bacterial strain (Sphingomonas sp. 23L) to promote mycorrhiza formation and plant growth of three willow clones (Salix spp.) on fly ash from an over-burdened dump in a pot experiment. The high pH_H2O (8.7) and low nitrogen content (N_t = 0.1 g kg⁻¹) in combination with hydrophobicity of the particle surfaces caused low plant growth. Inoculation of the willows with Sphingomonas sp. 23L improved the nitrogen uptake by plants, increased plant growth and stimulated formation of ectomycorrhizae with an autochthonous Geopora sp. strain on all three willow clones. The ectomycorrhiza formed by the Geopora sp. was morphologically and anatomically described. The inoculation significantly increased the shoot growth of two Salix viminalis clones and the root growth of a S. viminalis x caprea hybrid clone. We conclude that inoculation with mycorrhiza promoting bacterial strains might be a suitable approach to support mycorrhiza formation with autochtonous site-adapted ectomycorrhizal fungi in fly ash and thereby to improve re-vegetation of fly ash landfills with willows.