Microbial metabolism of dinitramine

The microbial metabolism of N³,N³-diethyl 2,4-dinitro-6-trifluoromethyl-m-phenylenediamine (dinitramine), N-sec-butyl-4-tert-butyl-2,6-dinitroaniline (A-820), N-n-propyl-N-cyclopropylmethyl-4-trifluoromethyl-2,6-dinitroaniline (CGA-10832), and α,α,α-trifluoro-2,6-dinitro-N,N-dipropyl-p-toluidine (trifluralin) by Aspergillus fumigatus Fres., Fusarium oxysporum Schlecht and Paecilomyces sp. was investigated. The dinitrodiamine and dinitroanilines were most readily metabolized by Paecilomyces sp. The metabolism of dinitramine was examined in detail, and four metabolites were isolated: N³-ethyl 2,4-dinitro-6-trifluoromethyl-m-phenylenediamine; 2,4-dinitro-6-tri-fluoromethyl-m-phenylenediamine; 6-amino-1-ethyl-2-methyl-7-nitro-5-trifluoromethylbenzimidazole and 6-amino-2-methyl-7-nitro-5-trifluoromethylbenzimidazole. The presence of a dinitramine-degrading enzyme system in A. fumigatus was demonstrated. The enzyme dealkylates dinitramine and requires NADPH and Fe²⁺ as cofactors.     

Microbial metabolism of fenoxaprop-ethyl

Fenoxaprop-ethyl was found to be metabolized by a mixed microbial population isolated from a leachate from a landfill. Fenoxaprop acid and 6-chloro-2(3H)-benzoxazolone were detected as metabolic products in the culture broths. It was shown that the microbial population could utilize fenoxapropethyl as a source of carbon and nitrogen, but the presence of an additional source of carbon increased the metabolism rate.     

Metabolism of the pesticide metabolites 4-nitrophenol and 3,4-dichloroaniline in carrot (Daucus carota) cell suspension cultures

The metabolism of [ 14 C]-4-nitrophenol and [ 14 C]-3,4-dichloroaniline (the xenobiotics are degradation products of parathion and propanil, respectively) was studied in cell suspension cultures of carrot (Daucus carota L.). 4-Nitrophenol was transformed almost quantitatively to water-soluble conjugates with minor amounts of non-extractable residues. The conjugates identified were 1-(O-β-D-glucopyranosyl)-4-nitrobenzene and 1-(6′-O-malonyl-O-β-D-glucopyranosyl)-4-nitrobenzene. In addition, two unidentified metabolites were observed, possibly a disaccharide and another malonylated glycoside of 4-nitrophenol. Time-course studies demonstrated that 4-nitrophenol was rapidly taken up and conjugated; all metabolites remained associated with the cells rather than nutrient medium. 3,4-Dichloroaniline was transformed quantitatively to water-soluble conjugates and bound residues (3.6%). The water-soluble metabolites were identified as 6′-O-malonyl-N-(β-D-glucopyranosyl)-3,4-dichloroaniline, N-(β-D-glucopyranosyl)-3,4-dichloroaniline and N-malonyl-3,4-dichloroaniline. A time-course study showed that the glucosides were formed initially, then decreased, possibly due to hydrolysis. This decrease was paralleled by an increase of the main metabolite, N-malonyl-3,4-dichloroaniline, which was predominantly recovered from the medium.     

Propanil metabolism in rice: A comparison of propanil amidase activities in rice plants and callus cultures

Propanil amidase is an enzyme found in rice, which hydrolyzes the herbicide 3,4-dichloropropionanilide, propanil. The activity of the enzyme as measured in rice plants was found to be two to four times greater in plants with four leaves than in plants with fewer than four leaves. The higher amidase activity of the four-leaf plants was localized in the unexpanded leaves.Rice root callus suspension in cultures also demonstrated propanil amidase activity. In vivo experiments indicated that the herbicide was metabolized by the tissue culture. Propanil amidase activity as determined in vitro, however, was detected only after the culture had developed to late stationary phase.     

Properties of an Aspergillus nidulans propanil hydrolase

Aspergillus nidulans is able to hydrolyze the herbicide propanil (3′,4′-dichloropropionanilide) with liberation of 3′,4′-dichloroaniline. When the fungus is grown with or without propanil, the hydrolytic activity is identical, but can be increased by starving the mycelium either for carbon, nitrogen, or both carbon and nitrogen. The enzyme which is responsible of this activity is of the aryl acylamidase type (EC 3.5.1 aryl acylamine amidohydrolase). It is also active on propionanilide and acetanilide, two structural analogs of propanil. A value of K_m = 0.13 mM has been obtained for propanil. Temperature optimum is 40°C, when assayed with propanil as substrate. Although the pH optimum is 8, there is a relatively high enzyme activity over a wide range of pH values between 7.8 and 10.2. Carbaryl has been found to effectively inhibit the enzyme activity on propanil (K_i = 0.03 mM). The results indicated that the properties of this aryl acylamidase from A. nidulans are very similar to those of enzymes isolated from a variety of organisms such as rice, mammals and the fungus Fusarium solani.     

Peroxidase activity and propanil degradation in soil*

The herbicide N-(3,4-dichloropxhenyl)-propionamide (propanil) and a metabolite of propanil, 3,4-dichloroaniline (DCA), were mixed with Nixon loam soil which was subjected to moisture and air-drying treatments. Degradation of propanil was altered by subjecting the treated soil samples to storage conditions of moisture, drying and chloroform. The peroxidase activity in fresh soil was very low when soil samples were collected during the cold season. The amount of 3,3′,4,4′- tetrachloroazobenzene (TCAB) produced from DCA increased with a simultaneous increase in the peroxidase activity in preincubated soil where carbon and nitrogen sources were added.     

3,5-二氯苯胺在水中的光解及水解特性

3,5-二氯苯胺(3,5-DCA)是二甲酰亚胺类杀菌剂(DCFs)在环境和植物中的主要降解代谢产物,比其母体化合物具有更强的毒性和持久性。本研究通过室内模拟试验,利用气相色谱-质谱联用仪(GC-MS)和高效液相色谱(HPLC),研究了3,5-DCA的光解和水解特性。结果表明：初始质量浓度为5 mg/L的3,5-DCA在氙灯和紫外灯下光解的半衰期分别为49.5和11.6 min;在中性、酸性和碱性条件下光解的半衰期分别为9.9、168和10.7 min;在甲醇、乙腈、正己烷中光解的半衰期分别为4.10、2.69和0.58 h。进一步研究发现,3,5-DCA在正己烷中的光解产物为单脱氯产物。3,5-DCA在中性、酸性和碱性条件下的水解半衰期分别为40.8、77.0和86.6 d。不同浓度的表面活性剂十二烷基磺酸钠(SDS)和十六烷基三甲基溴化铵(CTAB)溶液均可抑制3,5-DCA的水解,其中CTAB的抑制效果强于SDS。研究结果有助于更全面地了解二甲酰亚胺类杀菌剂的环境归趋,可为其合理使用及环境安全性评价提供数据支持。     

Effect of soil storage on propanil degradation

Sassafras sandy loam soil was subjected to various storage conditions after collection and 3′,4′-dichloropropionanilide (propanil) was used for testing the biodegradation potential of the stored soil. Cleavage of propanil was affected drastically after 5 days’ moist storage at 0, 10, 25 or –15° C and by 5 days’ air-drying. In 1 week approximately 90% of the propanil was degraded when fresh soil samples were used. However, 60–80% of residual propanil was found in soil which had been stored under various conditions from 5 to 30 days. Samples air-dried for 10 days were mildly affected as compared with the samples moist-stored at various temperatures, indicating that restricted gas exchange had a detrimental effect on biodegradation in moist-stored samples. These findings show that any biodegradation study should be conducted with fresh soil samples. Effet de la conservation du sol sur la degradation du propanil Un sol argilo sableux a été soumis apres prélèvement à diverses conditions de conservation et du 3,4-dichloropro-pionanilide (propanil) a été utilisé pour estimer le potentiel de biodégradation du sol ainsi conservé. La dégradation du propanil aété profondément affectée aprés 5 jours de conservation à l'humidité, à 0, 10, 25 ou –15° C et par 5 jours de séchage à l'air. En une semaine, 90 % environ du propanil aété dégrade dans le cas oil des échantillons frais de sol ontété utilises. Toutefois, 60 a 80 % de propanil résiduel aété retrouvé dans un sol conservé pendant 5 à 30 jours dans des conditions variées. Les eéchantillons séchés k l'air pendant 10 jours ontété moyennement affectés en compar-aison avec les échantillons conserves ä l'humiditéä des températures variées; ceci indique que la restriction des échanges gazeux a un effet défavorable sur la biodégradation dans les échantillons conserves à rhumidité. Ces résultats montrent que toute étude de biodégradation devrait être conduite avec des échantillons de sol frais. Wirkung der Lagerung von Boden auf den Abbau von Propanil Boden (Sassafras-sandiger-Lehm) wurde nach der Probennahme unter verschiedenen Bedingungen gelagert und, seine biologische Abbaufahigkeit mit 3′,4′-Dichlorpro-pionanilid (Propanil) untersucht. Der Abbau von Propanil wurde nach fünftägiger Lagerung des Bodens in feuchtem Zustand bei 0, 10, 25 oder – 25°C und fünftägiger Lufttrocknung sehr stark beeinflusst. Wenn frische Bodenproben verwendet wurden, waren nach einer Woche etwa 90% des Herbizids abgebaut. Dagegen waren noch 60–80% des Propanils vorhanden, wenn der Boden unter verschiedenen Bedingungen 5–30 Tage gelagert worden war. Proben die 10 Tage lang luftgetrocknet wurden, waren, verglichen mit den bei verschiedenen Temperaturen feucht gelagerten Bodenproben, nur wenig beeinflusst. Das weist darauf hin, da ein verringerter Gasaustausch sich schädlich auf die im feuchten Zustand gelagerten Proben auswirkte. Diese Ergebnisse zeigen, dass jede Untersuchung des biologischen Abbaus von Herbiziden im Boden mit frischen Bodenproben durchgefuhrt werden sollte.     

Transformation of the herbicides propanil and chlorpropham by micro-algae

Two species of green algae and four of blue-green algae hydrolysed the acylanilide herbicide propanil to the aniline derivative, 3,4-dichloroaniline. Of the cultures tested, only the blue-green alga Anacystis nidulans was shown to be capable of converting the phenylcarbamate herbicides propham and chlorpropham to the corresponding anilines. The green alga Ulothrix fimbriata was apparently unable to hydrolyse propanil or chlorpropham.     

Resistance mechanism of Leptochloa chinensis Nees to propanil

The resistance mechanism of Leptochloa chinensis Nees to propanil was investigated, based on propanil metabolism, aryl acylamidase activity, and chlorophyll fluorescence at the 8 week growth stage of L. chinensis. The concentration of propanil in the leaf and culm extracts of the resistant (R) and susceptible (S) biotypes, as measured by gas chromatography (GC), was found to increase after propanil treatment. The concentration of propanil in the leaf and culm extracts of the S biotype at 72 h was 1.55 and 0.49 µg mL^?1, respectively. However, a lower concentration of propanil was observed in the R biotype, as compared to that in the S biotype. The residue of 3,4‐dichloroaniline, as measured by GC, was detected only in the leaf extracts of the R biotype. In contrast, no residue of 3,4‐dichloroaniline was observed in the S biotype. The level of aryl acylamidase in the leaf tissue extracts of the R biotype was ～140% higher than that in the S biotype. The fluorescence studies showed that propanil inhibited the quantum efficiency of the photosystem II in both the R and S biotypes after 2 h of incubation time. However, when the leaf disks were transferred and incubated in deionized water for 48 h, the quantum efficiency increased in the R biotype but decreased in the S biotype. These results suggest that propanil metabolism, enhanced by aryl acylamidase activity, is the most likely factor contributing towards the mechanism of propanil resistance in L. chinensis plants at the 8 week growth stage.     

Age-related mechanisms of propanil tolerance in Jungle-rice,Echinochloa colona

Uptake and metabolism of propanil were measured in both susceptible (S) and resistant (R) biotypes of Jungle-rice, Echinochloa colona (L.) link at different growth stages. Results showed that there was no significant difference in uptake between S and R biotypes of E. colona at any given growth stage, but that uptake was significantly reduced at older plant growth stages in all biotypes studied. Metabolism of propanil was more rapid in R biotypes than in S biotypes at all growth stages studied. Specific and total aryl acylamidase activity, responsible for the first stage of propanil metabolism, was higher in R biotypes than in S at all growth stages, but declined to about 50% of the maximum at older growth stages, confirming the importance of this enzyme in conferring resistance to this herbicide. The area of necrosis that developed around a single drop of propanil deposited on the adaxial leaf surface was used to assess the degree of propanil resistance; it was found that resistance increased at older E. colona growth stages in contrast to the rate of propanil metabolism and amidase activity. Treatment of leaves with the amidase inhibitors, carbaryl or piperophos, simultaneously with propanil, caused a decrease in resistance at growth stages where amidase activity was greatest. This treatment was less effective at older growth stages. These results show that, in E. colona, propanil metabolism is important for conferring resistance in younger plants (four-six-leaf stage). It is suggested that restricted uptake confers resistance in older plants.     

Effects of propanil on growth and cell constituents of Nostoc calcicola

The rice field herbicide, propanil, was toxic to the nitrogen-fixing cyanobacterium Nostoc calcicola. A decrease in growth was observed with the increasing concentrations of propanil, 30 μg/ml being lethal. Since toxicity of the herbicide could be reversed by exogeneous supplementation of assimilable organic carbon glucose, it is suggested that carbon fixation was sensitive to the herbicide. The herbicide inhibited heterocyst differentiation and nitrogen fixation. There was a rapid decrease in total protein, nucleic acids (DNA, RNA), and carbohydrate content accompanied by a loss of photosynthetic pigments. The phycocyanin: chlorophyll a ratio showed positive correlation with increased dosages of the herbicide, suggesting the inhibition of chlorophyll a.     

Association between elevated activity of aryl acylamidase and propanil resistance in Jungle-rice,Echinochloa colona

Aryl acylamidase (aryl-acylamine amidohydrolase, EC 3.5.1.13) activity has been measured in crude extracts from leaves of propanil-susceptible (S) and propanil-resistant (R) biotypes of the grass weed. Echinochloa colona (L.) Link from Columbia. Both specific and total amidase activity increased with plant age up to 15 days (four-leaf stage), then decreased beyond 20 days to about 50% of the maximum at 36 days in both R and S E. colona biotypes. Specific activity with propanil in the R biotype was about 80% of that obtained for rice (Oryza sativa L.), compared to 25% in the susceptible biotype. The specific activity of the propanil amidase was three-fold higher in the R biotype than in the S. Partially purified amidase extracts from rice and both S and R biotypes of E. colona were compared biochemically. Both rice and E. colona amidases had a pH optimum of 7.5 and native relative molecular masses, estimated by gel filtration, of 179 000 and 181 000, respectively. Out of six substrates tested, three produced appreciable activity (propanil, 4-chloroacetanilide and acetanilide) in both rice and E. colona. Michaelis constants showed that the rice amidase had a higher affinity for propanil (0.36 mM) than had the E. colona enzyme (1.1 mM). Carbamates and organo-phosphorus pesticides were shown to inhibit amidase activity in partially purified rice and E. colona extracts. Additional preliminary data have implicated peroxidase in the next step of propanil metabolism in vitro. These data demonstrate that increased aryl acylamidase activity contributes to resistance to the herbicide propanil in E. colona weeds. Also, a biochemical comparison of purified aryl acylamidases from S and R biotypes of E. colona is presented for the first time.     

Kinetics and products of the hydrolysis of 3,4-dihydroprecocene I 3,4-epoxide in aqueous organic solvents

The hydrolysis of 3, 4-dihydroprecocene I 3, 4-epoxide (3, 4-dihydro-7-methoxy-2, 2-dimethyl-3, 4-epoxy-2H-benzo[b]pyran), the putative ultimate cytotoxin of the insect growth regulator precocene I (7-methoxy-2, 2-dimethyl-2H-benzo[b]pyran), has been studied and found to exhibit first-order kinetics [k = 0.17 s^?1 in 10 mm-phosphate buffer pH 7.0, containing 1, 4-dioxane (1 + 1 by volume), ionic strength 0.1]. Plots of log k versus pH, and k versus buffer concentration, suggest that the reaction is subject to both specific and general acid catalysis. High-performance liquid chromatography showed the reaction products to be predominantly the corresponding stereoisomeric diols (3, 4-dihydro-7-methoxy-2, 2-dimethyl-2H-benzo[b]pyran-3, 4-diol), the trans : cis ratio of which varied from 1.8: 1 to 2.2: 1 but was constant over the pH range 6-8, at a given buffer concentration. The results indicate that acid-catalysed hydration of 3, 4-dihydroprecocene I 3, 4-epoxide is an S_N1 reaction, involving a trigonally hybridised carbocation at C4, even at physiological pH. Similar studies on 3, 4-dihydroiso-precocene I 3, 4-epoxide (3, 4-dihydro-6-methoxy-2, 2-dimethyl-3, 4-epoxy-2H-benzo-[b]pyran), a biologically inactive isomer of 3, 4-dihydroprecocene I 3, 4-epoxide suggest that an S_NI mechanism also contributes to its hydrolysis, but the rate constant is 4000 times lower than that for 3, 4-dihydroprecocene I 3, 4-epoxide. Knowledge of the reactivity and mechanism of reaction of such compounds forms an important part of the basis for rational prediction of biological activity in precocene analogues, and hence their possible use as pest control agents.     

微生物除草剂与生物安全

随着人类环境意识的提高和农业可持续发展的需要,高效、环保微生物除草剂的研究越来越显示其重要的社会意义和经济价值。本文概述了微生物除草剂的种类、剂型及特点,介绍了利用微生物及其天然产物防治杂草的潜在优势和目前国内外的研究进展,尤其是对微生物除草剂的生物安全性进行了评价。     

Toxicological and metabolic parameters of the teleost fish (Leporinus obtusidens) in response to commercial herbicides containing clomazone and propanil

Pesticides, such as herbicides can affect the metabolic and toxicological parameters on fish. For this reason, an experiment was carried out with the objective of to evaluate the effects of commercial formulations of clomazone and propanil herbicides on acetylcholinesterase (AChE), thiobarbituric acid-reactive substances (TBARS), catalase (CAT) and metabolic parameters in teleost fish (Leporinus obtusidens). Fish were exposed during 90 days to field measured concentration of the herbicides clomazone and propanil (376 and 1644 μg/L, respectively) on rice paddy water. Specific AChE activity in the brain and muscle decreased and TBARS levels decreased in brain, muscle and liver tissues. Liver catalase decreased after exposure to both herbicides. Metabolic parameters in the liver and white muscle showed different changes after exposure to both herbicides. In summary, the results showed that clomazone and propanil affects toxicological and metabolic parameters of piavas. These results suggest that environmentally relevant herbicides concentrations are toxic to Leporinus obtusidens.     

微生物农药常见术语及释义

本文参照FAO、WHO及国内有关微生物农药的准则和标准,并结合微生物分类、鉴定等方面的研究进展,归纳、总结了微生物农药常见的术语及释义,为科学认识微生物农药和推进产业规范化发展提供参考。     

Soil persistence studies with bromoxynil, propanil and [14C]dicamba in herbicidal mixtures

The persistence of bromoxynil (3,5-dibromo-4-hydroxybenzonitrile), [¹⁴C]dicamba (3,6-dichloro-2-methoxybenzoic-7-¹⁴C acid) and propanil [N-(3,4-dichlorophenyl)propionamide] at rates equivalent to 1 kg ha^?1, were studied under laboratory conditions in a clay loam, a heavy clay and a sandy loam at 85% of field capacity and at 20±1°C, both singly and in the presence of herbicides normally applied with these chemicals as tank-mix or split-mix components. The degradation of bromoxynil was rapid with over 90% breakdown occurring within a week in the heavy clay and sandy-loam soils, while in the clay-loam approximately 80% of the bromoxynil had broken down after 7 days. In all three soils degradation was unaffected by the presence of asulam, diclofop-methyl, flamprop-methyl, MCPA, metribuzin or propanil. Propanil underwent rapid degradation in all soil treatments, with over 95% of the applied propanil being dissipated within 7 days. There were no noticeable effects on propanil degradation resulting from applications of asulam, barban, bromoxynil, dicamba, MCPA, MCPB, metribuzin or 2,4-D. The breakdown of [¹⁴C]dicamba in a particular soil was unaffected by being applied alone or in the presence of diclofop-methyl, flampropmethyl, MCPA, metribuzin, propanil or 2,4-D. The times for 50% of the applied dicamba to be degraded were approximately 16 days in both the clay loam and sandy loam, and about 50 days in the heavy clay.