Metribuzin metabolism in tomato: Isolation and identification of N-glucoside conjugates

Metribuzin [4-amino-6-tert-butyl-3-(methylthio)-1,2,4-triazin-5(4H)-one] metabolism was studied in tomato (Lycopersicon esculentum Mill. “Sheyenne”). Pulse-treatment studies with seedlings and excised leaves showed that [5-¹⁴C]metribuzin was rapidly absorbed, translocated (acropetal), and metabolized to more polar products. Foliar tissues of 19-day-old seedlings metabolized 96% of the root-absorbed [¹⁴C]metribuzin in 120 hr. Excised mature leaves metabolized 85–90% of the petiole-absorbed [¹⁴C]metrubuzin in 48 hr. Polar metabolites were isolated by solvent partitioning, and purified by adsorption, thin-layer, and high-performance liquid chromatography. A minor intermediate metabolite (I) was identified as the polar β-d-(N-glucoside) conjugate of metribuzin. The biosynthesis of (I) was demonstrated with a partially purified UDP-glucose: metribuzin N-glucosyltransferase from tomato leaves. A possible correlation between foliar UDP-glucose: metribuzin N-glucosyltransferase activity levels and differences in the tolerance of selected tomato seedling cultivars to metribuzin was suggested. The major polar metabolite (II) was identified as the malonyl β-d-(N-glucoside) conjugate of metribuzin.     

Cold storage degradation of the herbicide metribuzin in field soil samples awaiting analysis

In conjunction with a study of herbicide degradation in the field, metribuzin (4-amino-6-tert-butyl-4,5-dihydro-3-methylthio-l,2,4-triazin-5-one) was shown to be degraded at-37±5°C in soil samples awaiting analysis. Products of this degradation process include the known derivatives DADK (6-tert-butyl-2,3,4,5-tetrahydro-l,2,4-triazine-3,5-dione) and DK (4-amino-6-tert-butyl-2,3,4,5-tetrahydro-l,2,4-triazine-3,5-dione) and these products themselves have been shown to be degraded further under the same storage conditions. Loss of the herbicide can be approximated using half order kinetics with respect to metribuzin; however, loss of both previously identified metabolites is most rapid when metabolite concentrations are highest. Assuming half order kinetics, samples of Almasippi very fine sandy loam containing metribuzin (0.5 mg/kg) can be expected to lose 50% of the herbicide in 282 days at - 37°, a period which is short enough to affect residue analysis results significantly should samples be stored for extended lengths of time prior to analysis. Implied in these results is the desirability of immediate analysis following collection of samples; otherwise rates of loss during storage must be determined to allow adjustment of results to compensate for residue losses in samples awaiting analysis.     

Effect of 4-amino-6-methyl-3-phenylamino-l,2,4-triazin-5(4H)-one on the lignification process catalysed by peroxidase from lupin (lupinus albus)

The molecular action of herbicides with a triazine structure, such as atrazine (2-chloro-4-ethylamino-6-isopropylamino-1,3,5-triazine) and metribuzin (4-amino-6-tert-butyl-3-methylthio-1,2,4-triazin-5(4H)-one), has been related to their inhibition of the electron carrier system between chloroplastic photosystems II and I. This report provides evidence that 4-amino-6-methyl-3-phenylamino-1,2,4-triazin-5(4H)-one, a recently synthesised triazine, structurally analogous to metribuzin, causes a powerful inhibition of the cell-wall lignification catalysed by peroxidase from lupin. The two reactions involved in this lignification process are: oxidative polymerisation of coniferyl alcohol and the generation of hydrogen peroxide at the expense of NADH oxidation.     

Selectivity of SMY 1500 (4-amino-6-tert-butyl-3-ethylthio-l,2,4,triazin-5(4H)-one) in Triticum durum

Susceptibility to SMY 1500 (4-amino-6-tert-butyl-3-ethylthio-1.2.4-triazin-5(4H)-one) was studied in durum wheat (Triticum durum Desf.) cultivars in growth-chamber assays. Weight reduction, photosynthesis inhibition and herbicide levels were determined at various times after a 24-h herbicide treatment in two cultivars of durum wheat, one cultivar of spring wheat (Triticum aestivum L). and in the weed Bromus diandrus (Roth). Measurements of weight reduction distinguished between plants tolerant and plants susceptible to SMY 1500. Tolerant cultivars showed a lower herbicide content and photosynthesis inhibition than susceptible plants after the 24-h treatment. Thereafter, herbicide levels and photosystem II inhibition decreased faster in tolerant plants. Comparison with previous work showed that plants tolerated doses of SMY 1500 about four times higher than of its analogue, metribuzin. Herbicide content in leaves just after treatment, and rate of detoxification may explain the selectivity of SMY 1500 in these cereals.     

Measurement of simazine residues in chickpeas,Cicer arietinum,by gas–liquid chromatography

A method is described for the measurement of simazine [2-chloro-4,6-bis(ethylamino)-1,3,5-triazine] residues in chickpeas (Cicer arietinum). Ground chickpea samples were extracted with dichloromethane, followed by clean-up on alumina. Gas-liquid chromatography using metribuzin [4-amino- 6-tert-butyl-4,5-dihydro-3-methylthio-1,2,4-triazin-5-one] as internal standard with thermionic detection was used to quantify simazine residues. The limit of detection was 0.02 mg kg^?1 and the recoveries of simazine from chickpea samples (0.1–4 mg kg-1) averaged 92%.     

Phytotoxicity and detoxification of metribuzin in dark-grown suspension cultures of soybean

Differential resistance of soybean to metribuzin (4-amino-6-t-butyl 3-(methylthio)-as-triazin-5(4H)-one) was demonstrated in cell suspensions from resistant and susceptible cultivars. Whereas the herbicide had been reported to inhibit photosynthesis, the observations on achlorophyllous, dark-grown cultures indicated that phytotoxicity was not restricted to photosynthesis. The sites of light-independent herbicide activity were not identified, but the presence of these sites was demonstrated in both the herbicide-susceptible and herbicide-resistant cells. The mechanism to protect these sites was present in all cultures. The mechanism, enzymatic detoxification of the herbicide, was inoperative in the susceptible cultures due to accumulation of a substance which inhibited the enzyme. Resistant cultures metabolized that substance to an ineffectual form. Metribuzin selectivity was therefore determined according to the accessibility of the inhibitor to the enzyme responsible for resistance.     

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.     

Inhibition of chitin biosynthesis by buprofezin analogs in relation to their activity controlling Nilaparvata lugens Stål

Buprofezin (Applaud, 2-tert-butylimino-3-isopropyl-5-phenyl-3,4,5,6-tetrahydro-2H-1,3,5-thiadiazin-4-one) strongly inhibited the [³H]chitin synthesis from N-acetyl-d-[1-³H]glucosamine in the brown rice planthopper, Nilaparvata lugens Stål. No inhibition was observed for [³H]-labeled protein biosynthesis from [5-³H]glucose or l-[3,5-³H]tyrosine but [³H]-labeled nucleic acid synthesis from [5-³H]glucose was weakly reduced by buprofezin. The lethal activity of buprofezin analogs related well to their inhibitory potency against chitin biosynthesis in N. lugens nymphs.     

Effect of 4-amino-6-methyl-3-phenylamino-1,2,4-triazin-5(4H)-one on the integrity of chloroplasts from barley leaves

4-Amino-6-methyl-3-phenylamino-1,2,4-triazin-5(4H)-one alters the integrity of chloroplasts in barley (Hordeum vulgare L. var. Hassan) leaves as judged by their sedimentation profiles in linear gradients of sucrose. This effect may be related to the inhibition caused by 4-amino-6-methyl-3-phenylamino-1,2,4-triazine-5(4H)-one of both ascorbate peroxidase and superoxide dismutase activities, and could thus be explained by an accumulation of toxic reduced-oxygen species in illuminated chloroplasts.     

噻二唑基-3-哒嗪酮类化合物的合成及生物活性

将取代的二酰基肼环合后,得到中间体2-芳基-5-氯甲基-1,3,4-噻二唑,然后与2-叔丁基-4-氯-5-羟基-3(2H)-哒嗪酮反应,合成了8个未见文献报道的含噻二唑基哒嗪酮类化合物,其化学结构经1H NMR、高分辩质谱和元素分析确认。生物活性测试结果表明,部分化合物对粘虫P.separate W.有较好的抑制生长活性,其中化合物 3b 的EC50值为21 mg/L。     

Synthesis and insecticidal activity of novel 1,3,4-oxadiazolin-5-one and pyrazolin-5-one derivatives

A series of novel 2-(2,4,6-trisubstituted phenyl)-1,3,4-oxadiazolin-5-one derivatives and 3-(2,4,6-trichlorophenyl)pyrazolin-5-one derivatives were synthesized and evaluated for insecticidal activity. It was found that a moderately bulky alkyl group, such as a tert-butyl group, on the heterocyclic ring, and a trifluoromethyl group on the benzene ring were optimal substituents on the molecule. The oxygen atom in the oxadiazoline ring was essential for insecticidal activity. Of the compounds assayed, 4-tert-butyl-2-(2,6-dichloro-4-trifluoromethylphenyl)-1,3,4-oxadiazolin-5-one gave the highest activity against Nephtotettix cincticeps, with an LC₅₀ value of 0.51 mg litre⁻¹. © 1999 Society of Chemical Industry     

The residual activity of metribuzin in soil

The residual activity of 4-ammo-6-tert-butyl-3-(methylthio)-1,2,4-triazine-5-4(H)one (metribuzin) on indicator plants in Sassafras sandy loam soil was studied. Inhibition of coleoptile. radicle and primary root development were found at 1 ppm in oat (Avena sativa L., var. Beede) and at 5 ppm in cucumber (Cucumis sativus L., var. Pepino). At concentrations below 5 ppm, metribuzin stimulated the elongation of primary root and radicle in cucumber. After 46 days’ incubation. less than 1 ppm of the intact compound remained in soil without affecting the elongation of oat roots while its half life was approximately 6 days at 28deg;C incubation. L'activité résiduelle de la métribuzine dans le sol L'activité résiduelle de la 4-amino-6-t-butyl-3-(méthylthio)-1,2,4-tHazine-S-(4H)-one (métribuzine) a étéétudiée sur des plantes tests dans un sol sablo-limoneux de Sassafras. L'inhibition de la coléoptile, de la radicule et du développement de la racine primaire a été constatéà la dose de 1 ppm pour l'avoine (Avena sativa L. var. Beede) et à 5 ppm pour le concombre (Cucumis sativus L. var. Peplno). A des concentrations inférieures à 5 ppm, la métribuzine a stimulé l'élongation de la racine primaire et de la radicule du concomposé. Après 46 jours d'incubation, moins d'un ppm du composé intact restait dans le sol sans affecter l'élongation des racines d'avoine, alors que sa demi-vie a été approximativement de 6 jours à une température d'incubation de 28°C. Die Residualwirkung von Metribuzin im Boden Es wurde die Rcsidualwirkung von 4-Amino-6-tert,-butyl-3-(methylthio)-1,2,4-triazin-5-(4H)-one (Metribuzin) an Indikalorpflanzen in Sassafras-sandigem-Lehm untersucht. Bei 1 ppm war bei Hafer (Avena sativa L., Sorte Beede) die Entwicklung der Koleoptile, der Keim- und der Primär-wurzel gehemmt und bei 5 ppm desglcichen bei Gurke (Cucumis sativus L., Sorte Pepino). Bei Konzentrationen unter 5 ppm, förderte Metribuzin bei der Gurke die Streckung der Primär- und der Keimwurzel. Nach 46 Tagen Inkubation-szeit war weniger als 1 ppm der unveränderten Verbindung im Boden, was aber das Längenwachstum der Haferwurzeln nicht beeinträchtigte. Die Halbwertzeit betrug bei 28°C ungefahr 6 Tage.     

Plant cell cultures as a model for the biochemistry of crop selectivity

Plant cell cultures have been used to study the metabolic degradation of 4-amino-5-methyl-2-(tert-butylaminocarbonyl)-1,2,4-triazolin-3-one. The biochemical basis of selectivity was shown to reside in effective metabolic conjugation. The herbicide was eliminated from cell cultures of beet (which is tolerant to it) by conjugation (N-glycolisation), but this occured to only a limited extent with cell cultures of non-target plants such as soybean.     

Differences in the tolerance of metribuzin by varieties of soybeans

Metribuzin [4-amino-6-tert-butyl-3-(methylthio)-as-triazin-5(4/f)-one] has shown promise for weed control in Glycine max (L.) Merr. (soybean), but at rates effective for preemergence weed control soybean tolerance has been narrow and field observations indicated that varieties of soybeans differed in their tolerance. Treatments of 0.0,0.56,1,12 and 1,68 kg/ha an location in the Coastal Plain, Piedmont, and Limestone Valley-area of Georgia indicated that the soybean variety ‘Coker 102’ was Meterially more susceptible to metribuzin injury than were ‘Hardee’, ‘Bragg’, ‘Bienville’, ‘Hampton 266 A’, or ‘Coker 318’. The injury caused smaller soybean stands, heights, and yields. These results confirmed previous observations from field trials using this herbicide. Différences de sensibilités variétales chez le soja vis-à-vis de la métribuzine La métribuzine [4-amino-6-tert-butyl-3-(méthylthio)-as-triazine-5(4H)-one] a donné des résultats prometteurs pour le Désherbage du soja Glycine max (L.) Merr. Aux doses efficacies pour le désherbage en prélevée, la résistance du soja a étéétroite et les observations faites au champ ont mis en évidence des difféarences de sensibilité entre les variétés de soja. Des traitements à raison de 0,0, 0,56, 1,12 et 1,68 kg/ha à des emplacements situfés dans les régions suivantes: Coastal Plain, Piedmont et Limestone Valley, dans l'éatat de Georgie, ont montré que la variété de soja ‘Coker 102’était notablement plus sensible à la métribuzine que les varietés ‘Hardee’, ‘Bragg’, ‘Bienville’, ‘Hampton 266 A’ ou ‘Cocker 318′. Les dégâts se traduisirent par une réduction de Ia vigueur, de lataille et du rendement. Ces résultats confirment les observations antérieures dans des essais au cham pavec cet herbicide. Unterschiede in der Toleranz von Metribuzin gegenüber Sojabohnensorten [4-Amino-6-tert.-butyl-3-(methylthio)-as-triazin-5(4H)-one] ist ein aussichtsreiches Herbizid für die Unkrautbekämpfung in Glycine max. (L.) Merr. (Sojabohne). Bei wirksamen Aufwandraengen im Vorauflaufverfahren war die Sojabohnentoleranz wenig ausgeprägt und Feldbeobachtungen zeigten, dass Sojabohnensorten unterschiedlich tolerant sind. Behandlungen mit 0,0, 0,56, 1,12 und 1,68 kg/ha an Standorten in der Coastal Plain, Piedmont und der Limestone Valley-Gegend in Georgia zeigten, dass die Sojabohnensorte ‘Coker 102’ erheblich empfindlicher gegenuber Metribuzinschaden war als ‘Hardee’, ‘Bragg’, ‘Bleinville’, ‘Hampton 266 A’ oder ‘Coker 318′. Der Schaden äusserte sich in geringeren Bestandesdichten, Pflanzenhöhen und niedrigeren Erträgen. Diese Ergebnisse bestätigen die schon früher mit diesem Herbizid gemachten Beobachtungen in Freilandversuchen.     

Herbicidal activity and selectivity of new 3-alkyl-4-amino-6-aryl-1,2,4-triazin-5-ones

3-Alkyl-4-amino-6-phenyl-1,2,4-triazin-5-ones are herbicidal compounds which act by inhibition of the electron transport in photosynthesis (Hill reaction). There is some correlation between phytotoxicity and photosynthesis inhibition data. An exception is found with sugar beets which are very tolerant towards triazinones with a methyl group in 3-position and an amino group in 4-position. The best compound from this series, 4-amino-4,5-dihydro-3-methyl-6-phenyl-1,2,4-triazin-5-one, metamitron, shows very promising properties as a selective herbicide in sugar beet crops.     

Effect of different photosystem II inhibitors on chloroplasts isolated from species either susceptible or resistant toward s-triazine herbicides

In chloroplasts isolated from susceptible and atrazine-resistant Amaranthus retroflexus, the inhibition of photosynthetic electron transport by various classes of herbicides has been investigated. Resistance of mutant Amaranthus is not restricted to s-triazines but also extends to uracils, 1,2,4-triazine-5-ones, and ureas. For 1,2,4-triazin-5-ones and chloroplasts of both biotypes, a correlation between inhibition of photosynthetic electron transport and the partition coefficient could be established. In the case of phenolic herbicides only modestly decreased or even higher sensitivity of chloroplasts from the resistant biotype as compared to the susceptible one could be observed. These results are confirmed by binding of radioactively labeled herbicides to chloroplasts of both plants. Specific binding of atrazine or metribuzin to resistant chloroplasts is completely abolished, and that of diuron or phenisopham diminished as compared to susceptible chloroplasts. In contrast, binding of phenolic herbicides generally is enhanced in resistant chloroplasts. Photoaffinity labeling of thylakoids from both biotypes by 2-azido-4-nitro-6-[2′,3′-³H]isobutylphenol yields almost identical labeling patterns. These results are consistent with a recently proposed model (W. Oettmeier, K. Masson, and U. Johanningmeier, Biochim. Biophys. Acta679, 376 (1982) of two different herbicide binding proteins at the reducing side of photosystem II: a 32- to 34-kdalton protein responsible for binding of triazines, triazinones, ureas, and related herbicides and a photosystem II reaction center protein for binding of phenolic herbicides.     

Biocompost from sugar distillery effluent: effect on metribuzin degradation, sorption and mobility

BACKGROUND: Metribuzin (4-amino-6-tert-butyl-4,5-dihydro-3-methylthio-1,2,4-triazin-5-one) is weakly sorbed in soils and therefore leaches easily to lower soil profiles and results in loss of activity. Soil amendments play an important role in the management of runoff and leaching losses of pesticides from agricultural fields. Therefore, the effect of biocompost from sugarcane distillery effluent on metribuzin degradation and mobility was studied in a sandy loam soil.RESULTS: Metribuzin was more persistent in biocompost-unamended (T-0) flooded soil (t(1/2) - 41.2 days) than in non-flooded (t(1/2) - 33.4 days) soil. Biocompost application at the rate of 2.5 and 5.0% (T-1 and T-2) in non-flooded soils increased metribuzin persistence, but no significant effect was observed on persistence in flooded soils. Freundlich adsorption constants (K(f)) for treatments T-0, T-1 and T-2 were 0.43, 0.64 and 1.13 respectively, suggesting that biocompost application caused increased metribuzin sorption. Leaching studies in packed soil columns indicated that biocompost application affected both metribuzin breakthrough time and maximum concentration in the leachate. Leaching losses of metribuzin were drastically reduced from 93% in control soil (T-0) to 65% (T-1) and 31% (T-2) in biocompost-amended soils.CONCLUSION: Biocompost from sugarcane distillery effluent can be used effectively to reduce downward mobility of metribuzin in low-organic-matter sandy loam soil. Copyright (c) 2008 Society of Chemical Industry.     

新型2-酰基氰基乙酸类衍生物SYP-10898的研究

以对叔丁基氯苄为起始原料,经氰化、酯化、酯交换和酰基化4步反应合成了新化合物2-酰基氰基乙酸酯类衍生物 SYP-10898 ,其化学结构经核磁共振氢谱和元素分析确证。室内生物活性测试结果表明, SYP-10898 对朱砂叶螨Tetranychus cinnabarinus成螨的LC50值为3.67 mg/L,优于对照药剂丁氟螨酯(cyflumetofen)。田间试验结果表明,在50 mg/L剂量下, SYP-10898 对柑桔红蜘蛛Panonychus citri表现了良好的速效性和持效性,3 d后的防效在93%以上,30 d后的防效在79%以上。     

Monitoring of the photochemical degradation of metamitron and imidacloprid by micellar electrokinetic chromatography and differential-pulse polarography

The photochemical degradation of metamitron (4-amino-4,5-dihydro-3-methyl-6-phenyl-1,2,4-triazin-5-one) and imidacloprid (1-(6-chloro-3-pyridylmethyl)-N-nitroimidazolidin-2-ylideneamine) has been investigated by differential-pulse polarography (DPP) and micellar electrokinetic chromatography (MEKC); the degradation pathways of these pesticides were elucidated and their degradation products proposed. The electrochemical study of imidacloprid by DPP at different pH values demonstrated the occurrence of two different reduction processes; at pH 6.8, two peaks at −0.90 V and −1.38 V, respectively, were obtained, which are related to the photochemical reduction processes. The photochemical degradation of imidacloprid caused by sunlight was polarographically monitored and its degradation products elucidated. The polarographic reduction of deaminometamitron (obtained by photochemical reduction of metamitron) yielded two peaks at −0.62 and −1.37 V, which are related to the reduction of the CN bonds. The effect of sunlight on the reduction of metamitron was monitored by DPP, and an increase of the concentration of the degradation products was observed with time. MEKC with UV-visible detection was used to separate the pesticides and the products of their photochemical degradation, which were identified in combination with DPP. © 1999 Society of Chemical Industry     

Effects of various classes of herbicides on four species of algae

Chlorella pyrenoidosa, Chlorococcum sp., Lyngbya sp., and Anabaena variabilis were cultured in Bold's basal medium. They were treated with 0.1, 1.0, and 10 μM concentrations of 2-chloro-2′, 6′-diethyl-N-(methoxymethyl)acetanilide (alachlor), 2-chloro-4-(ethylamino)-6-(tert-butyl-amino)-s-triazine (terbuthylazine), 2-sec-butyl-4,6-dinitrophenol (dinoseb), 1,1-dimethyl-3-(α,α,α-trifluoro-2,6-dinitro-N-propyl-p-toluidine) (profluralin), 2, 4-bis(isopropylamino)-6-(methylthio)-s-triazine (prometryne), and (2,4-dichlorophenoxy)acetic acid (2,4-D). Growth of all algal species tested was markedly reduced by the triazines. Alachlor, dinoseb, and fluometuron inhibited growth of some algae at higher concentrations while 2,4-D and profluralin did not inhibit growth at the concentrations tested. Photosynthesis was greatly inhibited by the triazines, even at the 0.1 μM concentration. Fluometuron was very toxic to the blue-green algae but had less effect on the green algae tested. Lyngbya was most susceptible to photosynthesis reduction by the herbicides. The concentrations of herbicides tested had little effect on respiration of the algae species. It appears that effects on algal growth were due primarily to inhibition of photosynthesis rather than to other metabolic processes.