Cleavage of the diketonitrile derivative of the herbicide isoxaflutole by extracellular fungal oxidases

Isoxaflutole is a herbicide activated in soils and plants to its diketonitrile derivative, the active herbicide principle. The diketonitrile derivative undergoes cleavage to the inactive benzoic acid analogue. In this paper, it is established that an oxidative mechanism implicating two successive reactions in the presence of dimethyldioxirane can chemically initiate the cleavage of the diketonitrile. It is also shown that two white rot strains, Phanerochaete chrysosporium and Trametes versicolor, are able to convert the diketonitrile to the acid when cultured in liquid media. This main metabolite amounts to 24.6 and 15.1% of initial herbicide content after 12-15 days of culture. Another polar metabolite represents <3.7% of the parent compound amount during the same period. Oxidative enzymes produced by the fungi show a time course similar to that of diketonitrile degradation. Purified laccase (EC 1. 10.3.2), in the presence of 2 mM 2, 2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) acting as a redox mediator at pH 3 supports the reaction with rates of 0.3-0.4 nmol h(-)(1) unit(-)(1).     

Effect of soil properties on the degradation of isoxaflutole and the sorption-desorption of isoxaflutole and its diketonitrile degradate

The transformation of isoxaflutole (ISOX) to its herbicidally active diketonitrile degradate (DKN) was significantly enhanced in the presence of soil and occurred more rapidly in systems containing soil with a greater soil pH. Sorption-desorption of ISOX and DKN in five soils collected within a field revealed both ISOX and DKN were more readily sorbed to soils with greater organic matter, clay content, and lower soil pH. Sorption of ISOX residues occurred within 2 h, and extracts contained similar concentrations of ISOX and DKN at 24 h, suggesting the 24-h sorption coefficients for ISOX-treated systems were actually for mixed ISOX residues. Freundlich sorption coefficients were 3 and 4 times greater for ISOX than for DKN. On the basis of the Freundlich organic carbon sorption constants, ISOX and DKN can be categorized in the very high and high mobility classes, suggesting their potential to leach in the soils needs to be evaluated.     

Reaction pathways of the diketonitrile degradate of isoxaflutole with hypochlorite in water

Isoxaflutole (IXF; Balance) belongs to a new class of isoxazole herbicides. Isoxaflutole has a very short half-life in soil and rapidly degrades to a stable and phytotoxic degradate, diketonitrile (DKN). DKN was previously discovered to rapidly react with hypochlorite (OCl-) in tap water, yielding the benzoic acid (BA) degradate as a major product, but the complete reaction pathway and mechanism have not been elucidated. Thus, the objectives of this work were to (1) determine the stoichiometry of the reaction between DKN and OCl-; (2) identify products in addition to BA; and (3) propose a complete pathway and reaction mechanism for oxidation of DKN by OCl-. Stoichiometry of the reaction showed a molar ratio of OCl-/DKN of 2. In addition, two previously uncharacterized chlorinated intermediates were identified under conditions in which OCl- was the limiting reactant. The proposed chemical structure of a chlorinated benzoyl intermediate was inferred from a series of HPLC/MS and HPLC/MS/MS experiments and the use of mass spectral simulation software. A chlorinated ketone intermediate was also identified using ion trap GC/MS. Two additional end products were also identified: cyclopropanecarboxylic acid (CPCA) and dichloroacetonitrile (DCAN). On the basis of the reaction stoichiometry, the structure of the chlorinated intermediates, and the identification of the products, two reaction pathways are proposed. Both pathways involve a two-step nucleophilic attack and oxidation of the diketone structure of DKN, leading to formation of BA, DCAN, and CPCA.     

Fate of isoxaflutole in soil under controlled conditions

Isoxaflutole (IFT, 5-cyclopropyl-1,2-oxazol-4yl-alpha,alpha,alpha-trifluoro-2-mesyl-p-tolyl ketone) is a new pre-emergence proherbicide used in maize and sugarcane. Its two main derivatives are a diketonitrile derivative, 2-cyano-3-cyclopropyl-1-(2-methanesulfonyl-4-trifluoromethylphenyl)propane-1,3-dione, called DKN, and a benzoic acid derivative, 2-methanesulfonyl-4-trifluoromethylbenzoic acid, called BA. Few data are available of the factors influencing the degradation of IFT in soil, and the purpose of the present work was to determine the relative importance of, and factors affecting, the degradation of IFT in soil. Experiments were conducted on five soils with distinct physicochemical characteristics, at different temperatures and moisture contents in biotic and abiotic conditions. The isomerization of IFT to DKN is rapid, increasing with higher moisture contents and higher temperatures. It depends strongly on pH and is governed by chemical processes. The degradation of DKN to BA appeared to be essentially due to the biological activity of the soil.     

Mobility and degradation of the herbicide imazosulfuron in lysimeters under field conditions

The mobility and degradation of imazosulfuron, labeled with carbon-14 at the imidazole ([imi-(14)C]imazo) or pyrimidine ring ([pyr-(14)C]imazo), in lysimeters with 1 m(2) surface and 110 cm depth were investigated for three years. One lysimeter was treated with [imi-(14)C]imazo in two successive years at the rate of 50 g of active ingredient (ai)/ha each. The other two lysimeters were treated once with [pyr-(14)C]imazo and a mixture (1:1, w/w) of the two labeled imazosulfurons, respectively (50 g of ai/ha). In the first and second years of monitoring, the yearly mean concentration of (14)C in the leachate water was <0.10 microg/L in each lysimeter. Although in the third year the concentration of (14)C in the leachate water was 0.17 microg/L for the lysimeter treated twice with [imi-(14)C]imazo, the concentration of imazosulfuron and its degradation products in the leachate water ranged from 0.01 to 0.06 microg/L. At the study termination, the main portion of (14)C recovered was found in the upper 30 cm soil layer in each lysimeter, and no (14)C was detected below a depth of 50 cm. These findings indicated that imazosulfuron and its degradation products in soils translocated into groundwater only slightly.     

Factors affecting aggregate instability of Greek agricultural soils

Abstract

The aggregate stability of several surface soils from Thessaly plain, Central Greece, was studied using an instability index which classified the soils according to the stability of their aggregates to water. This index was correlated to the basic soil properties influencing aggregate stability. It was found that soil aggregate stability was positively correlated with clay content, total specific surface area, cation exchange capacity, base saturation, and free silicon oxides content. Silt, very fine and fine sand, and the ratio (sand+silt)/clay influenced negatively the aggregate stability. Organic matter content and sesquioxides have no effect on aggregate stability. It was also observed that the fraction of carbonates with silt dimension influenced negatively the aggregate stability. From the results of this study it was concluded that the factors affecting water aggregate stability were specific surface area and cation exchange capacity of the soils.     

Recolonization of methyl bromide sterilized soils under four different field conditions

Summary The course of recovery in biological activity was assessed in the top 5 cm of undisturbed soil cores (29.7 cm diameter, 30 cm deep) that had been fumigated in the laboratory with methyl bromide. The cores were returned to their original pasture and forest sites, two with a moderate and two with a high rainfall, and untreated soils at all sites served as baselines. Sampling took place over 166 days (midsummer to midwinter). Microbial biomass (as measured by fumigation-extraction and substrate-induced respiration procedures) and dehydrogenase activity both recovered rapidly, but remained consistently lower in the fumigated than in untreated samples at both forest sites and at the moister of the two pasture sites. Bacterial numbers also recovered rapidly. Fungal hyphal lengths were, on average over 166 days, 25% lower in the fumigated soils. Levels of mineral N were initially highest in the fumigated soils, but declined with time. Fumigation generally had no detectable effects on the subsequent rates of net N mineralization and little effect on nitrification rates. Fumigation almost totally eliminated protozoa, with one to three species being recovered on day 0; the numbers recovered most rapidly in the moist forest soil and slowly in the dry pasture soil. The recoionization rate of protozoan species was similar in all soils, with species numbers on day 110 being 33 and 34 in the fumigated and untreated soils, respectively. Nematodes were eliminated by fumigation; recolonization was first detected on day 26 but by day 166, nematode numbers were still lower in fumigated than in untreated soils, the abundance being 10 and 62 g^-1 soil and diversity 10 and 31 species, respectively. Overall, the results suggest that protozoan and nematode populations and diversities could provide a useful medium-term ecological index of the recovery in comprehensive soil biological activity following major soil pollution or disturbance.     

Estimation of the total gaseous nitrogen losses from clay soils under laboratory and field conditions

Acetylene blockage was evaluated as a method for measuring losses of N₂O + N₂ from two Denchworth series clay soils. The denitrification potential in anaerobic, dark incubations at 20°C with nitrate (equivalent to 100 kg N ha^?1 0–20 cm depth), maximum water holding capacity, and acetylene (1%), was equivalent to 32 ± 11 and 39 ± 6 kg N ha^?1 per day for the two 0–20 cm soils and was positively correlated with carbon content (r= 0.98). After 4 days N₂O was reduced to N₂ in the presence of C₂H₂. In April 1980 following irrigation (24 mm) and applications of ammonium nitrate (70 kg N ha^?1) and acetylene, the mean nitrous oxide flux from soil under permanent grass was 0.05 ± 0.01 kg N₂O-N ha^?1 per day for 8 days. In June 1980, the losses of nitrogen from cultivated soils under winter wheat after irrigation (36 mm) and acetylene treatment were 0.006 ± 0.002 and 0.04–0.07 ± 0.01 kg N ha^?1 per day respectively before and after fertilizer application (70 kg N ha^?1). The nitrous oxide flux in the presence of acetylene decreased briefly, indicating that nitrification was rate determining in drying soil.     

Comparison of nitrogen mineralized under anaerobic and aerobic conditions for some agricultural and forest soils of washington

Abstract

Seven agricultural soils and eight forest soils from Washington state were tested for mineralizable nitrogen using both anaerobic and aerobic incubation procedures. Each procedure had been used previously to. develop nitrogen indices for agricultural and forested ecosystems. Forest soils mineralized less nitrogen under anaerobic than aerobic conditions, while the opposite was true for agricultural soils. There were statistically significant correlations between the two methods for each of the time periods tested. Experimental variations were consistently lower than previously reported.     

Permethrin and its two metabolite residues in seven agricultural crops

Metabolite residues of permethrin are not reported in the literature for most agricultural crops. This paper reports residues of permethrin and its 2 metabolites (dichlorovinyl acid and metaphenoxybenzyl alcohol) in 7 different agricultural crops (Chinese cabbage, spinach, asparagus, raspberries, green peas, turnip roots, and turnip greens). Permethrin residues declined approximately 85% within 7 days after treatment in all crops. In most cases, the acid metabolite residues peaked at 3 days, and declined after that. Translocation of residues into turnip roots was very slight; the average was less than 0.05 ppm for permethrin and alcohol metabolite residues and none was detected for the acid metabolite residue. Permethrin residues in the turnip greens averaged approximately 2 ppm for the 0.112 kg ai/ha treatment, and 6 ppm for the 0.224 kg ai/ha treatment.     

Microbial immobilisation and turnover of C labelled substrates in two arable soils under field and laboratory conditions

Microbial biomass C immobilisation and turnover were studied under field and laboratory conditions in soils of high yield (HY) and low yield (LY) areas within an agricultural field. We compared the size and activity of soil microbial biomass (SMB) in the soils of the different yield areas under field and laboratory conditions. Soils were amended with ¹³C labelled mustard (Sinapis alba) residues (both experiments) and labelled glucose (laboratory only) at 500 μg C g⁻¹ dry soil. SMB-C, dissolved organic carbon (DOC) and total C content were monitored in the field and the laboratory. CO₂-efflux was also measured in laboratory treatments. Isotope ratios were determined for SMB in both experiments, but other variables only in the laboratory treatments. A positive priming effect was measured in three of four laboratory treatments. Priming was induced after a significant increase of soil derived C in the microbial biomass. Thereafter, the total C loss through priming was always smaller than or equal to the decline in microbial biomass C. In field and laboratory experiments SMB in the HY soil immobilised less of the added substrate C than LY soil SMB. Calculated turnover times in the laboratory glucose amendment were 0.24 (HY) and 0.31 y (LY), in the laboratory mustard treatment 0.58 (HY) and 0.44 y (LY) and in the field mustard amendments 1.09 (HY) and 1.25 y (LY). In both the field mustard and laboratory glucose treatments turnover in the HY soil tended to exceed that in the LY soil. These turnover times as well as the reaction of SMB-C to drying-rewetting and substrate addition, indicated that the HY soil possessed a more active microbial community with a more rapid C turnover than the LY soil. As C turnover is considered to be closely linked to nutrient cycles, faster turnover in the HY soil may involve a better nutrient supply for crops resulting in higher agricultural yield.     

Microbial immobilisation and turnover of N labelled substrates in two arable soils under field and laboratory conditions

Microbial biomass N dynamics were studied under field and laboratory conditions in soils of high yield (HY) and low yield (LY) areas in an agricultural field. The objective of the study was to determine the size and activity of soil microbial biomass in the soils of the different yield areas and to compare these data obtained under field and laboratory conditions. Soils were amended with ¹⁵N labelled mustard (Sinapis alba) residues (both experiments) and labelled nitrate (laboratory only) at 30 μg N g⁻¹ dry soil. Soil microbial biomass (SMB) N, mineral N (N_min) and total N content was monitored both in the field and in the laboratory. N₂O efflux was additionally measured in laboratory treatments. Isotope ratios were determined for SMB in both experiments, for all other parameters only in the laboratory treatments. In the laboratory less amounts of added substrate N were immobilised by the SMB in HY soils compared to LY soils, whereas in the field immobilisation of added N by SMB was higher in HY soils initially and slightly lower after 40 days of incubation. Calculated turnover times in the laboratory nitrate, laboratory mustard and field mustard amendments were 0.18, 0.27 and 0.74 years (HY) and 0.22, 0.61 and 1.01 years (LY), respectively. The turnover times of added substrate N always showed the trend to be faster in HY soils compared to LY soils. A faster turnover of nutrients in the HY soils may involve a better nutrient supply of the plants, which coincides with the higher agricultural yield observed in these areas.     

Photodegradation of rotenone in soils under environmental conditions

An environmental fate study was performed to analyze the effects of soil components on the photochemical behavior of rotenone. Photodegradation experiments were carried out on three types of soil collected in southern Italy, Valenzano (VAL), Turi (TUR), and Conversano (CON), from April to June 2006. Soil thin-layer plates (1 mm thick) were spiked with 1.5 mg/kg of rotenone and exposed under natural conditions of sunlight and temperature. The plates were removed from the sunlight at predetermined intervals of continuous irradiation. Other soil samples, control and sterilized, were kept in the dark to evaluate possible effects of chemical and microbiological degradation during the irradiation experiment. The time for 50% loss of the initially applied rotenone varied from 5 to 7 h, following the order TUR < CON < VAL. In environmental studies, changes in temperature and/or moisture affected the degradation rate and caused deviations from first-order kinetics. The photolysis reaction fit the two compartment or the multiple compartment model pathways better. A fast initial decrease during the first 5 h of rotenone irradiation was followed by a much slower decline, which clearly indicates the rather complex chemical process of rotenone photodegradation on soil surfaces. Also, the degradation was shown to be directly related to the soil concentration of clay and organic matter. Rotenolone (12abeta-hydroxyrotenone) was detected by HPLC/DAD/MS analysis as the only photodegradation byproduct of rotenone in soil thin layers. Results provide additional insights on the rates and the mechanisms of rotenone degradation, aiming to describe more clearly the degradation performance of chemical residues in the environment.     

Microbial community structure and activity in agricultural soils under different management

For the development of management strategies in sustainable agriculture it is necessary to describe and predict the role of soil microbes in different management systems. The classical approach uses the microbial biomass as the key parameter for the entire system, but for ecological purposes the variability of biotic parameters in time and space has to be better described. Moreover, the biomass active in the total soil profile or its most active zones should be used as a basis for the assessment of soil activity. The sum of adenylates was found to be more closely related to the microbial biomass than was ATP, which however appeared to be a better indicator for the microbial activity. Fatty acids from phospholipids were highly correlated with the soil microbial biomass. The pattern of fatty acids from soils under different long-term management indicated a high potential to typify the microbial community in soils and special organism populations. To overcome the problem, that only a small portion of the soil inhabiting microbes can be cultivated, first steps to use serological and genetical methods to directly identify or localize specific populations in the rhizosphere are shown.     

Improved HPLC-MS/MS method for determination of isoxaflutole (balance) and its metabolites in soils and forage plants

A robust multi-residue procedure is needed for the analysis of the pro-herbicide isoxaflutole and its degradates in soil and plant materials at environmentally relevant (<1 microg kg-1) levels. An analytical method using turbo-spray and heat-nebulizer high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed for the analysis of isoxaflutole (IXF) and its two metabolites, diketonitrile (DKN) and the benzoic acid metabolite (BA), at sub-microgram per kilogram levels in soil and plant samples. The average recoveries of the three compounds in spiked soil and plant samples ranged from 84 to 110% and 94 to 105%, respectively. The limits of quantification were validated at 0.06 microg kg-1 for soil and 0.3 microg kg-1 for plant samples. The limits of detection (LOD) for soil analysis were 0.01, 0.002, and 0.01 microg kg-1 for IXF, DKN, and BA, respectively. Corresponding LOD for the plant analysis method were 0.05, 0.01, and 0.05 microg kg-1. The developed method was validated using forage grass and soil samples collected from a field lysimeter study in which IXF was applied to each of four forage treatments. Forage plants and soils were sampled for analyses 25 days after IXF application to the soil. In soils, IXF was not detected in any treatment, and DKN was the predominant metabolite found. In forage plants, the concentrations of DKN and BA were 10-100-fold higher than that in soil samples, but IXF was not detected in any forage plants. The much higher proportion of BA to DKN in plant tissues (23-53%), as compared to soils (0-5%), suggested that these forages were capable of detoxifying DKN. The developed methods provided LODs at sub-microgram per kilogram levels to determine the fate of IXF and its metabolites in soils and forage plants, and they also represent considerable improvements in extraction recovery rates and detection sensitivity as compared to previous analytical methods for these compounds.     

Effect of chemical composition on the release of nitrogen from agricultural plant materials decomposing in soil under field conditions

Summary In two field experiments, plant materials labelled with ¹⁵N were buried separately within mesh bags in soil, which was subsequently sown with barley. In the first experiment, different parts of white clover (Trifolium repens), red clover (T. pratense), subterranean clover (T. subterraneum), field bean (Vicia faba), and timothy (Phleum pratense) were used, and in the second, parts of subterranean clover of different maturity. The plant materials were analysed for their initial concentrations of total N, ¹⁵N, C, ethanol-soluble compounds, starch, hemicellulose, cellulose, lignin, and ash. After the barley had been harvested, the bags were collected and analysed for their total N and ¹⁵N. In the first experiment the release of N was highest from white clover stems + petioles (86%) and lowest from field bean roots (20%). In stepwise regression analysis, the release of N was explained best by the initial concentrations of lignin, cellulose, hemicellulose, and N (listed according to decreasing partial correlations). Although the C/N ratio of the plant materials varied widely (11–46), statistically the release of N was not significantly correlated with this variable. The results of the second experiment using subterranean clover of different maturity confirmed those of the first experiment.     

Hydrolysis of ammonium polyphosphate in soils under aerobic and anaerobic conditions

Summary The hydrolysis of fertilizer-grade solid and liquid ammonium polyphosphate (APP) was studied on an aluvial (Entisol, Typic Ustifluvent), a sodic (Entisol Aquic Ustifluvent), and a laterite (Oxisol, Aquox) soil under aerobic and anaerobic conditions. The hydrolysis rate was the fastest on laterite, intermediate on sodic, and slowest in the alluvial soil. Ammonium polyphosphate hydrolyzed more rapidly under anaerobic than under aerobic conditions. The hydrolysis of liquid ammonium polyphospahte was faster than that of solid ammonium polyphosphate in all soils. The half-life values for the polyforms of P in liquid ammonium polyphosphate ranged from 1.6 to 2.0 days under anaerobic and from 5.2 to 8.7 days under aerobic conditions. The corresponding values for solid ammonium polyphosphate were 3.9 to 9.2 days under anaerobic and 12.5 to 27.0 days under aerobic conditions.     

Comparison of different methodologies for field measurement of net nitrogen mineralization in pasture soils under different soil conditions

 Net mineralization was measured in free-draining and poorly drained pasture soils using three different field incubation methodologies. Two involved the use of enclosed incubation vessels (jar or box) containing C₂H₂ as a nitrification inhibitor. The third method confined soil cores in situ in an open tube in the ground, with an anion-exchange resin at the base to retain leached NO₃ ^– (resin-core technique, RCT). Measurements were made on three occasions on three free-draining pastures of different ages and contrasting organic matter contents. In general, rates of net mineralization increased with pasture age and organic matter content (range: 0.5–1.5 kg N ha^–1 day^–1) and similar rates were obtained between the three techniques for a particular pasture. Coefficients of variation (CVs) were generally high (range: 10.4–98.5%), but the enclosed incubation methods were rather less variable than the RCT and were considered overall to be the more reliable. The RCT did not include C₂H₂ and, therefore, newly formed NO₃ ^– may have been lost through denitrification. In a poorly drained pasture soil, there were discrepancies between the two enclosed methods, especially when the soil water content approached field capacity. The interpretation of the incubation measurements in relation to the flux of N through the soil inorganic N pool is discussed and the drawbacks of the various methodologies are evaluated. Received: 18 November 1999     

Effect of inoculation with Azotobacter chroococcum on nitrogenase activity of Zea mays roots grown in agricultural soils under aseptic and non-sterile conditions

Summary Inoculated and non-inoculated seedlings of Zea mays were grown in agricultural soils under aseptic and non-sterile conditions. Acetylene reduction activity and microbial counts were determined after 7 and 30 days of growth. Irrespective of the soil type Azotobacter spp. were commonly isolated under maize cultivation. Inoculation of agricultural soils with a suspension of A. chroococcum led to an increase in Azotobacter numbers, although this effect diminished with time. Nitrogenase activity was detected on maize roots and increased in response to the inoculation with A. chroococcum, showing that this associative growth could be of primary importance for the plant. The results of assays for acetylene reduction activity indicated that the nitrogenase activity was associated only with the root systems.