Sites of action of photosynthetic inhibitor herbicides; Experiments with trypsinated chloroplasts

The effect of four photosynthetic inhibitor herbicides, bromacil, ioxynil, metribuzin and monuron, on chloroplast electron transport was investigated. All four compounds completely inhibited electron flow with tripotassium hexacyanoferrate as oxidant, but the inhibition caused by bromacil, metribuzin and monuron was almost totally reversed by trypsin treatment. With ioxynil, only a partial degree of reversability was shown. With a molybdosilicate as oxidant, electron transport was not completely inhibited by any of the herbicides. Whereas the partial inhibition was reversed by tryptic digestion in the presence of bromacil, metribuzin and monuron, there was virtually no reversal in the presence of ioxynil. The results suggest a common site of action for all four herbicides and an additional site for ioxynil nearer to photosystem II.     

Effect of metribuzin, butachlor and chlorimuron-ethyl on amino acid and protein formation in wheat and maize seedlings

Application of the recommended field dose of metribuzin, butachlor and chlorimuron-ethyl to 10-days-old wheat and maize seedlings differentially reduced shoot fresh and dry weights during the following 16 days. Metribuzin was the most reductive while butachlor was the least. The herbicides slightly affected the activities of nitrate reductase (NR, EC 1.6.6.1) and nitrite reductase (NiR, EC 1.7.7.1) but greatly inhibited glutamine synthetase (GS, EC 6.3.1.2) and glutamate synthase (GOGAT, EC 1.4.7.1) activities. Meanwhile, there were significant accumulations of ammonia and soluble-N accompanied by diminutions in total-N and protein contents; metribuzin exerted the greatest changes. Additionally, aliphatic, aromatic and total amino acids in both species were mostly elevated by the three herbicides; however, valine, leucine and isoleucine were decreased by only chlorimuron-ethyl. These results could conclude that herbicides, particularly metribuzin, cause a shortage in ammonia assimilation and subsequently a decrease in protein formation. Moreover, the elevation of soluble-N and amino acids appeared to result from breakdown of the pre-existing protein, a state that seemed consistent in seedlings treated with metribuzin and, to some extent chlorimuron-ethyl but recovered in those treated with butachlor.     

Removal of the phytotoxicity of uracil herbicides in water by photodecomposition

The efficiency of a photo-oxidation procedure for the detoxication of water containing herbicides of the uracil group was tested by bioassay. Treated herbicide solutions were checked for their inhibiting effect on germination and photosynthesis in sorghum (Sorghum bicolor). The treated solutions, and the main photodegradation products of bromacil and terbacil did not inhibit significantly either germination or seedling development at concentrations of up to 200 and 10 mg litre^?1, respectively. Preliminary tests with industrial waste effluents containing added uracil compounds showed that these compounds were rendered non-phytotoxic by the photo-oxidation procedure.     

Effects of microbial inhibitors on the degradation rates of metamitron,metazachlor and metribuzin in soil

Rates of carbon dioxide evolution and degradation rates of metamitron, metazachlor and metribuzin were measured in two soils in the presence of three microbial inhibitors. The nonselective microbial inhibitor sodium azide reduced both carbon dioxide evolution and the rate of loss of all three herbicides in both soils, although the reduction in degradation rate of metamitron was small. The antibacterial antibiotic novobiocin enhanced carbon dioxide evolution from both soils but had variable effects on the rates of herbicide degradation. It inhibited degradation of metazachlor and metribuzin, and in one of the soils its effects on metazachlor degradation were similar to those of sodium azide. Novobiocin inhibited degradation of metamitron to a small extent in one soil only. The antifungal antibiotic cycloheximide also enhanced carbon dioxide evolution from both soils. In general, its effects on herbicide degradation were similar to those of novobiocin, although the extent of inhibition was usually less pronounced. The results are discussed in terms of the relative involvement of microorganisms in degradation of the three herbicides.     

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.     

Effects of repeated applications of ten soil-active herbicides on weed population,residue accumulation and nitrification*

Ten herbicides, bromacil, chlorthal-dimethyl, diphenamid, diuron, fluometuron, neburon, prometryne, pyrazon, simazine and trifluralin at two doses were repeatedly sprayed, in autumn and in spring, for 4 consecutive years on non-cultivated, sprinkler-irrigated field plots. Herbicidal effect was assessed at 1–2 month intervals on the natural weed population and after each observation a paraquat + diquat spray destroyed emerged weeds. The response of various weed species to herbicides varied markedly but a herbicide-induced shift in the composition of weed population did not occur, presumably because of the paraquat treatment. The overall phytotoxicity to weeds present was, in decreasing order: diuron, bromacil, simazine, trifluralin, prometryne, neburon, fluometuron, pyrazon, diphenamid, chlorthal-dimethyl. Persistence of herbicides was in decreasing order: diuron = bromacil, simazine, neburon (at higher rate), fluometuron, trifluralin, prometryne. Control produced by pyrazon improved with the number of applications, but that of diphenamid and chlorthal-dimethyl remained weak and short. After repeated applications, the activity of these herbicides increased or remained at similar level, but in no case decreased. Soil samples were taken 5 months after each application and bioassayed. Phytotoxic residues were detected beneath the disturbed top-soil from bromacil, diuron, fluometuron and simazine after the first application, and from neburon after the second application; residues from trifluralin were found in the top soil only after the fifth application. After the seventh spraying, residues of bromacil were found in the 45–60-cm soil layer. Ammonia content in soil samples taken from treated plots after the fourth, sixth and seventh application was generally similar to the untreated control. In these samples, nitrate content appeared to be correlated negatively with remaining weed number; the control thus contained less nitrate than efficient herbicidal treatments. Soil samples taken after the seventh application of bromacil, diuron, fluometuron, neburon and simazine, which contained appreciable residual concentrations, did not show significant differences from control, in an in vitro nitrification test.     

Prognose-Modell zur Erfassung des Rückstandsverhaltens von Metribuzin und Methabenzthiazuron im Boden und deren Auswirkungen auf Folgekulturen

A predictive model for the assessment of metribuzin and methabenzthiazuron residues in soil and their effects on succeeding crops Aus der simulierten Abbaukurve lassen sich somit für jeden beliebigen Zeitraum nach Applikation eines Herbizids die maximalen pflanzenverfügbaren Wirkstoffanteile ableiten, aus denen die Auswirkungen auf mögliche Folgekulturen prognostiziert wurden. Die relative Empfindlichkeit (im Bereich der ED₅₀) der untersuchten Kulturpflanzen in Hydroponik war bei Metribuzin: Möhre < Bohne = Salat < Erbse = Spinat und bei MBT: Bohne = Erbse = Salat < Spinat = Möhre. Weiterhin konnte gezeigt werden, dass unter Labor- und Freilandbedingungen evtl. auftretende Schäden in fast allen Fällen in den gleichen Aktivitäskategorien lagen. The degradation of the herbicides metribuzin and methabenzthiazuron (MBT) was simulated under outdoor conditions in a sandy soil by a computer model based on the dominating influence of temperature and moisture on degradation, which is measured in preceding laboratory experiments. Depending on the conditions of incubation (10–30°C, 20–90% water holding capacity) the half-life of metribuzin was 11–60 days and of MBT 42 > 1200 days. Knowing the plant available soil water, the fraction of a total herbicide residue potentially available to plants can be calculated from the distribution coefficient (K_d-value). In the soil under investigation, for metribuzin 77% and for MBT 16% of the total residue was available to plants. Thus, the maximum residue available to plants can be calculated from the simulated degradation curve for any period after the application of a herbicide and in combination with a predictive model the effect on succeeding crops can be predicted. The specific susceptibility of the crops in question has to be established in preceding hydroponic culture experiments. The relative susceptibility (about ED₅₀) of the plants in hydroponic-culture was earrot相似文献   

Metabolism and toxicity of metribuzin in mouse liver

Metribuzin was hepatotoxic in mice when administered intraperitoneally (ip) at sublethal doses of 150 to 250 mg/kg. Four dose-dependent abnormalities were evident. Histopathological examination revealed a fulminant centrilobular hepatic necrosis. The serum glutamic-pyruvic transaminase (GPT) activity was elevated. The liver glutathione (GSH) content was almost completely depleted. There was extensive covalent binding of radiocarbon from [carbonyl-¹⁴C]metribuzin to liver proteins and also high blood levels of metribuzin fragments. Each of these four effects of metribuzin on the liver or blood was alleviated or blocked in mice pretreated with piperonyl butoxide (PB), which inhibits the cytochrome P-450-dependent monooxygenase. PB also reduced the lethality of metribuzin by three-fold. In contrast, pretreatment with diethyl maleate to suppress the liver GSH content increased the lethality of metribuzin by twofold. The hepatotoxicity and acute lethality of metribuzin were probably due to reactive intermediates which are normally detoxified by GSH conjugation. The principal urinary metabolites of metribuzin in mice and rats are mercapturic acids, which arise via metribuzin sulfoxide or deaminometribuzin sulfoxide reacting with GSH. Sulfoxidation therefore appears to activate metribuzin to an electrophilic metabolite which, in the absence of GSH, binds to tissue proteins producing hepatotoxicity.     

A rapid quantitative bioassay for the determination of biologically available bromacil in soils

A simple rapid bioassay is described for the determination of biologically available bromacil residues in soils. A clear aqueous extract was made from a soil fortified with a known amount of the herbicide, and similar extracts were made from samples of soil taken from plots that had been sprayed with a bromacil formulation at a rate of 4 kg ha⁻¹. Samples of these extracts were added to a suspension of the unicellular green alga Selenastrum capricornutum. The net photosynthetic oxygen production by the alga was then measured using an oxygen electrode. The results were expressed as a percentage of the oxygen production by a control suspension. The concentration of biologically available bromacil was determined by reference to a previously established dose-response curve of the percentage reduction in oxygen production against bromacil concentration. The accuracy of this bioassay was determined by comparing the results with those obtained using capillary gas-liquid chromatography. The results obtained by the two different methods showed good agreement.     

The possible role of quinate in the mode of action of glyphosate and acetolactate synthase inhibitors

BACKGROUND: The herbicide glyphosate inhibits the biosynthesis of aromatic amino acids by blocking the shikimate pathway. Imazethapyr and chlorsulfuron are two herbicides that act by inhibiting branched‐chain amino acid biosynthesis. These herbicides stimulate secondary metabolism derived from the aromatic amino acids. The aim of this study was to test if they cause any cross‐effect in the amino acid content and if they have similar effects on the shikimate pathway. RESULTS: The herbicides inhibiting two different amino acid biosynthesis pathways showed a common pattern in general content of free amino acids. There was a general increase in total free amino acid content, with a transient decrease in the proportion of amino acids whose pathways were specifically inhibited. Afterwards, an increase in these inhibited amino acids was detected; this was probably related to proteolysis. All herbicides caused quinate accumulation. Exogenous application of quinate arrested growth, decreased net photosynthesis and stomatal conductance and was ultimately lethal, similarly to glyphosate and imazethapyr. CONCLUSIONS: Quinate accumulation was a common effect of the two different classes of herbicide. Moreover, exogenous quinate application had phytotoxic effects, showing that this plant metabolite can trigger the toxic effects of the herbicides. This ability to mimic the herbicide effects suggests a possible link between the mode of action of these herbicides and the potential role of quinate as a natural herbicide. Copyright © 2009 Society of Chemical Industry     

The phytotoxicity of various herbicides in two sandy loam soils and the effect of liming

The phytotoxicity of five soil-acting herbicides was investigated in two sandy loam soils and that of a further five herbicides in one of these soils. The effect of liming on phytotoxicity was also examined. Metoxuron, fenuron, metribuzin and fluometuron were more active in Boddington Barn soil than in Bledington soil. Prometryne was similar in effect in both soils. Liming significantly increased the effect of metoxuron in Boddington Barn soil and metoxuron, atraton, difenoxuron and metribuzin in Bledington soil. Monolinuron was less toxic in limed Bledington soil. Reasons for these differences have not been determined.     

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.     

Mode of action,crop selectivity,and soil relations of the sulfonylurea herbicides

The sulfonylurea herbicides are characterized by broad-spectrum weed control at very low use rates (c. 2–75 g ha^?1), good crop selectivity, and very low acute and chronic animal toxicity. This class of herbicides acts through inhibition of acetolactate synthase (EC 4.1.3.18; also known as acetohydroxyacid synthase), thereby blocking the biosynthesis of the branched-chain amino acids valine, leucine and isoleucine. This inhibition leads to the rapid cessation of plant cell division and growth. Crop-selective sulfonylurea herbicides have been commercialized for use in wheat, barley, rice, corn, soybeans and oilseed rape, with additional crop-selective compounds in cotton, potatoes, and sugarbeet having been noted. Crop selectivity results from rapid metabolic inactivation of the herbicide in the tolerant crop. Under growth-room conditions, metabolic half-lives in tolerant crop plants range from 1–5 h, while sensitive plant species metabolize these herbicides much more slowly, with half-lives > 20 h. Pathways by which sulfonylurea herbicides are inactivated among these plants include aryl and aliphatic hydroxylation followed by glucose conjugation, sulfonylurea bridge hydrolysis and sulfonamide bond cleavage, oxidative O-demethylation and direct conjugation with (homo)glutathione. Sulfonylurea herbicides degrade in soil through a combination of bridge hydrolysis and microbial degradation. Hydrolysis is significantly faster under acidic (pH 5) than alkaline (pH 8) conditions, allowing the use of soil pH as a predictor of soil residual activity. Chemical and microbial processes combine to give typical field dissipation half-lives of 1–6 weeks, depending on the soil type, location and compound. Very short residual sulfonylurea herbicides with enhanced susceptibility to hydrolysis (DPX-L5300) and microbial degradation (thifensulfuron-methyl) have been developed.     

Recovery of non-target plants affected by airborne bromoxynil-octanoate and metribuzin

The present study was conducted to evaluate the recovery potential of non‐target plants affected by two airborne herbicides. Sunflower at the two‐leaf stage was used as a test plant and exposed for 24 h in a wind tunnel to a range of concentrations of airborne bromoxynil‐octanoate and metribuzin. Quantum yield (φ_PSII) of exposed leaves and of the second leaf pair developed after exposure was determined at a particular time up to 16 days following exposure. Maximum depression in quantum yield of exposed leaves from which a complete recovery occurred within 16 days was 63% for bromoxynil‐octanoate and 60% for metribuzin respectively. The corresponding maximum concentrations were 1.310 and 0.390 μg m^?3 respectively. The second leaf pair was also affected and showed a similar recovery potential. From the results it can be concluded that the significance of airborne bromoxynil‐octanoate and metribuzin must not be overestimated, as sunflower and non‐target plants with a similar sensitivity are likely to recover from air concentrations of both herbicides reported under field conditions.     

Herbicide Effects on Sugarcane Growth, Pythium Root Rot, and Pythium arrhenomanes

ABSTRACT Six herbicides were evaluated for their effects on Pythium root rot and growth of sugarcane in greenhouse experiments and on in vitro mycelial growth rate of Pythium arrhenomanes. Pendimethalin and atrazine were most inhibitory to mycelial growth, but neither reduced root rot severity. Asulam, atrazine, and metribuzin were not phytotoxic to sugarcane and did not affect root rot symptom severity in clay loam or silt loam field soils. Atrazine and metribuzin increased shoot number, and atrazine increased total shoot weight for treated plants in silt loam soil. Glyphosate, pendimethalin, and terbacil were phytotoxic to sugarcane. These herbicides increased root rot severity, but the extent to which growth reductions resulted from increased disease severity or from direct herbicide injury was not clear. Adverse effects on plant growth and root rot severity were greater in clay loam than in silt loam soil. The results suggest that sugarcane injury from some herbicides is compounded by increased severity of root rot.     

Changes in free amino acid pools can predict the mode of action of herbicides

Studies were conducted to determine the short-term changes in free amino acid levels in the meristematic zone of maize after treatment with various herbicides with different modes of action. These herbicides included inhibitors of various amino acid biosynthetic pathways, photosynthesis, and fatty acids biosynthesis. Inhibitors of various amino acid biosynthetic pathways caused specific reduction in the pools of amino acids being produced by the particular pathway. Inhibitors of other metabolic pathways also caused significant changes in pools of various amino acids. Very similar changes in the pools of amino acids were seen in plants treated with different chemical classes of inhibitors affecting a certain metabolic pathway. However, the changes were quite different between inhibitors of different metabolic processes. The data generated from these studies could be used as a diagnostic tool to determine the mode of action of novel herbicides.     

Metribuzin mobility in soil columns as affected by urea fertiliser

Application of urea fertilisers to soils influences the soil solution characteristics and thus may affect the sorption of soil-applied herbicides. The present investigation reports the influence of urea co-application on sorption and leaching of metribuzin, a triazine herbicide. Urea application at 60 and 120 kg N ha(-1) increased metribuzin sorption in soils over that in untreated natural soil. The Kf (Freundlich adsorption coefficient) values of metribuzin for natural, 60 and 120 kg N ha(-1) treatments were 0.43, 0.46 and 0.84 microg(1 - 1/n) g(-1) ml1/n respectively. Downward mobility of metribuzin was studied in packed soil columns (300 mm length x 59 mm i.d.) at two irrigation intensities, 720 m3 ha(-1) (72 mm) and 3600 m3 ha(-1) (360 mm). After 720 m3 ha(-1) irrigation, metribuzin did not leach out of any column and was not detected in the leachate. Urea amendment slowed the leaching of metribuzin by 20 and 40% in 60 and 120 kg N ha(-1) urea-treated columns respectively. Also, following urea application, greater amounts of metribuzin were retained in the application zone. Upon increasing the irrigation intensity fivefold, urea application did not have any effect on metribuzin mobility, and its breakthrough from both natural and urea-amended columns occurred after 126 mm irrigation. However, there was a marked difference in the maximum concentration of metribuzin in the breakthrough curves obtained from natural and urea-amended columns. The study indicated that co-application of metribuzin and urea fertiliser is a safe practice as far as leaching of herbicide is concerned.     

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.     

Use of activated carbon to protect tomato against metribuzin

Protection of tomato (Lycopersicon esculentum Mill., cv. UC 134) against metribuzin was increased by placing a small quantity of activated carbon near each seed position before pre-emergent application of the herbicide. The most protective location of the carbon was the surface of the soil, immediately above the seed. In this location, 0.10 g of activated carbon protected tomatoes from the phytotoxic effects of 0.700 kg metribuzin ha^?1 whereas, without the activated carbon, 0.175 kg metribuzin ha^?1 killed both tomato and blackberry nightshade (Solanum nigrum L.). Placing plastic discs in this location did not reduce the phytotoxicity; however, a barrier that prevented lateral invasion by the herbicide, of the zone of soil through which the plumule emerged, partially protected the plains from 0.350 kg metribuzin ha^?1, showing that metribuzin entered the plants in substantial quantities via the plumule as well as the roots. This result suggests that activated carbon maintained a lower metribuzin concentration in the plumule/root zone than the bulk concentration. It also explains way the soil surface was the most protective carbon location. These findings provide the basis for extending the use of preemergent herbicides to situations in which previously they could not be used safely or economically.