Binding of ioxynil to photosynthetic membranes

Two binding affinities for ioxynil to isolated thylakoids of the alga Bumilleriopsis filiformis are observed. Higher concentrations needed for the “low-affinity” binding site inhibit electron transport, while low ioxynil concentrations saturating the “high-affinity” binding site do not affect electron transport. For higher plant thylakoids (Chenopodium, Spinacia) the high-affinity binding site apparently is also the site to inhibit electron flow. The time dependence known for electron-transport inhibition is also apparent for the binding of phenol-type inhibitors to thylakoids of higher plants. Long incubation times (40 min) with the herbicide are necessary to equilibrate the binding site. Time dependence can be abolished by decreased pH, short trypsin or lysyl-endoprotease digestion of the isolated membranes. These treatments only increase the accessibility of the binding site, but the amount of bound herbicide after equilibration does not change. As a second effect the number of binding sites is reduced by prolonged trypsin digestion. This is not observed by treatment with a lysyl-specific protease. In contrast to herbicides like atrazine or diuron, short trypsin treatment alters the accessibility only (presumably provided by a shielding protein) but not the properties of the binding site of phenol-type herbicides. In contrast, thylakoids from the alga Bumilleriopsis, either from wild-type or a diuron resistant mutant, do not exhibit a time-dependent binding.     

Synthesis and characterization of a group of dihydropyrimido-benzimidazole photosystem II herbicides

A group of 28 dihydropyrimido-benzimidazole analogs and derivatives was tested for herbicidal activities. The 1:1 mixture of 7-fluoro- and 8-fluorodihydropyrimido-benzimidazole had the greatest biological activity. Activity is increased by methylation of nitrogen 2. None of the active compounds was inhibitory to nonphotosynthetic tissue-culture cells in vitro. The I₅₀ of inhibition of growth of green duckweed plants and that of in vitro photosynthesis activity matched the I₅₀ of inhibition of photosystem II activity. Fluorescence analyses of photosystem II activity indicated inhibition at the same step as did many triazine, phenylurea, pyridazinone, and uracil herbicides. As the compounds showed insufficient activity on recently evolved triazine-resistant weed biotypes, their further development was terminated, despite their novel chemistry.     

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.     

Differential effects of phenoxy herbicides on light-dependent 14CO2 fixation in isolated cells of C3 and C4 plants

Cells were isolated from the developing leaves of Ipomoea aquatica and Digitaria sanguinalis. The effects of phenoxy alkanoic acid herbicides on light-dependent ¹⁴CO₂ fixation and oxygen evolution in these leaf cells were studied. (2,4-Dichlorophenoxy)acetic acid and (2,4,5-trichlorophenoxy) acetic acid (2,4,5-T and 2,4-D) caused a 20% stimulation of ¹⁴CO₂ fixation at 0.8 × 10^?5M and an inhibition at 1 × 10^?4M in I. aquatica leaf cells. Temperature seemed to have a marked influence on such action. No effect or very little effect was observed in the leaf cells of D. sanguinalis. The nonactive (2,4,6-Trichlorophenoxy)acetic acid (2,4,6-T) caused a similar stimulation of CO₂ fixation as 2,4-D and 2,4,5-T at low concentrations in I. aquatica leaf cells, but no inhibition was observed at high concentration. Increase of hight intensity increased the rate of CO₂ fixation in both control and 2,4,6-T-treated cells; however, the percentage of stimulation remained the same. At stimulatory concentration, all three compounds did not cause any stimulation in either photosystem I and II or photosystem II-mediated oxygen evolution. At higher concentrations, the differential effects of 2,4-D and 2,4,5-T on the light-induced CO₂ fixation and photosystem II-mediated oxygen evolution in the I. aquatica leaf cells and D. sanguinalis mesophyll (ms) cells may be attributed in part to their selective action against dicotyledonous plants.     

Factors controlling degradation of pesticides in soil

Rates of pesticide degradation in soil exhibit a high degree of variability, the sources of which are usually unclear. Combining data from incubations performed using a range of soil properties and environmental conditions has resulted in greater understanding of factors controlling such degradation. The herbicides clomazone, flumetsulam, atrazine, and cloransulam-methyl, as well as the former insecticide naphthalene offer examples of degradation kinetics controlled by coupling competing processes which may in turn be regulated separately by environmental conditions and soil properties. The processes of degradation and volatilization appear to compete for clomazone in solution; sorbed clomazone is degraded only after the solution phase is depleted. Similarly, volatilization of naphthalene is enhanced when degradation has been inhibited by high nutrient levels. Degradation of the herbicide flumetsulam has been shown to be regulated by sorption, even though the compound has a relatively low affinity for the soil. The fate pathway for cloransulam-methyl shifts from mineralization to formation of metabolities, bound residues and physically occluded material as temperature increases. Atrazine degradation in soil may be controlled in part by the presence of inorganic nitrogen, as the herbicide appears to be used as a nitrogen source by micro-organisms. New insight gained from measurement of multiple fate processes is demonstrated by these examples.     

Studies into the action of the diphenyl ether herbicides acifluorfen and oxyfluorfen. Part II: The interaction with photosynthetic electron transport reactions

The effects of acifluorfen and oxyfluorfen on photosynthetic electron transport reactions of pea chloroplasts were compared with those induced by paraquat and monuron. Monuron inhibited electron flow between photosystems I and II, and paraquat acted as an electron acceptor for photosystem I, promoting superoxide formation by illuminated chloroplasts. Neither acifluorfen nor oxyfluorfen at concentrations up to 50 μM affected non-cyclic electron flow or promoted superoxide formation. Both herbicides were shown to repress ferredoxin-dependent NADP⁺ reduction by illuminated chloroplasts. Further experiments showed that, in the presence of ferredoxin-NADP⁺ reductase and chloroplast membranes maintained in the dark, p-nitro diphenyl ether (DPE) herbicides promoted the rate of ferredoxin-dependent oxidation of NADPH, implying that these herbicides can accept electrons from reduced ferredoxin. The interaction between acifluorfen, ferredoxin and chloroplast membranes was examined further by following the effect of this herbicide on the peroxidation of illuminated thylakoids. Lipid peroxidation was promoted by acifluorfen, although this effect was abolished if thylakoids were washed prior to use. The effect of washing could be reversed by adding exogenous ferredoxin. These data demonstrate that interaction of DPE herbicides with photosynthetic electron transport in the vicinity of ferredoxin is necessary for light-dependent herbicide activation.     

The herbicide binding niche of photosystem II-a model

The Q_B binding niche of photosystem II is also the binding site for many different herbicides. In order to understand the mode of binding of the herbicides, a 3-dimensional model of the binding niche was constructed. The model was based upon a comparison of the known structure of the Q_B binding niche in purple bacteria with sequence and mutant data of the D-1 protein of photosystem II. Plastoquinone builds up hydrogen bonds to phenylalanine 265 backbone amide nitrogen, to serine 264 hydroxyl, and to histidine 215 delta-1 nitrogen. In addition to these hydrogen bond donors and acceptors, herbicides can build up hydrogen bonds to backbone carbonyl of alamine 263 and to serine 268 hydroxyl. This is supported by binding data of inhibitors in Chlamydomonas reinhardtii chloroplasts of wild type and of five D-1 protein mutants (Ser264 Ala, Ala251 Val, Phe255Tyr, Val2191le, Leu275Phe).     

psbA mutation (Asn266 to Thr) in Senecio vulgaris L. confers resistance to several PS II-inhibiting herbicides

DNA sequence analysis of the psbA gene encoding the D1 protein of photosystem II (PS II), the target site of PS II-inhibiting herbicides, identified a point mutation (Asn266 to Thr) in a bromoxynil-resistant Senecio vulgaris L. population collected from peppermint fields in Oregon. Although this mutation has been previously reported in Synechocystis, this is the first report of this particular point mutation in a higher plant exhibiting resistance to PS II-inhibiting herbicides. The resistant population displayed high-level resistance to bromoxynil and terbacil (R/S ratio 10.1 and 9.3, respectively) and low-level resistance to metribuzin and hexazinone (R/S ratio 4.2 and 2.6, respectively) when compared with the susceptible population. However, the population was not resistant to the triazine herbicides atrazine and simazine or to the urea herbicide diuron. A chlorophyll fluorescence assay confirmed the resistance levels and patterns of cross-resistance of the whole-plant studies. The resistant S. vulgaris plants produced fewer seeds. Differences in cross-resistance patterns to PS II-inhibiting herbicides and the difference in fitness cost could be exploited in a weed management program.     

Using chlorophyll fluorescence induction for a quantitative detoxification assay with metribuzin and chlorotoluron in excised wheat (Triticum aestivum and Triticum durum) leaves

Chlorophyll fluorescence induction was used as a probe to detect herbicide detoxification in tolerant or susceptible wheat cultivars. Experimental conditions have been carefully examined for establishing detoxification kinetics of chlorotoluron and metribuzin, two photosystem-II-inhibiting herbicides. After a root treatment, leaves were cut, placed in glass tubes and maintained in the dark. The fluorescence induction rise was examined repeatedly and detoxification kinetics were established from these data for the same position on the individual leaves. The herbicide-dependent fluorescence rise decreased within hours in chlorotoluron-tolerant but not in susceptible Triticum aestivum cultivars. In contrast, no significant reversion could be detected after metribuzin application in both tolerant and susceptible cultivars of Triticum durum. Near the fluorescence-determined half-inhibition of photosystem II, linear detoxification kinetics were obtained in individual leaves, thus providing an accurate measurement of relative detoxification rates.     

Inhibition of the photochemical activity of the isolated photosystem ii reaction centre by 2-cyanoacrylates

Certain derivatives of the 2-cyanoacrylates are potent inhibitors of photosystem II (PSII) mediated electron flow. These compounds block electron flow by binding to the D1 polypeptide as do many other types of photosystem II herbicides. In this paper we report the action of these species of 2-cyanoacrylates on the photochemical activity of the isolated PSII reaction centre consisting of the D1 and D2 polypeptides but free of other chlorophyll-binding proteins. The three species tested were 2-ethoxyethyl 2-cyano-3-(4-chlorobenzylamino)-3-isopropylacrylate (cyanoacrylate 1), isopropyl 2-cyano-3-(N-methylanilino)acrylate (cyanoacrylate II) and methoxymethyl 2-cyano-3-(4-chlorobenzylthio)-3-mercapto-acrylate sodium salt (cyanoacrylate III). Unlike the case with thylakoid membranes or PSII enriched membranes, cyanoacrylate III was a more potent inhibitor than cyanoacrylate I of the photoreduction of 2,6-dichlorophenol indophenol (DCPIP) in the isolated PSII reaction centre. This cyanoacrylate was also more effective than diuron (DCMU) in blocking DCPIP reduction in the reaction centre complex. Moreover, unlike DCMU and cyanoacrylates I and II, cyanoacrylate III was more effective in inhibiting the quinone-dependent photoreduction of cytochrome b559 in isolated PSII reaction centres. It is concluded that the hydrophilic nature of cyanoacrylate III gives it greater potency with the isolated reaction centre and that its site of action is likely to be closer to the histidine ligands involved in the binding of non-haem iron rather than in the Q_B binding niche. (See footnote on page 331 for nomenclature of cyanoacrylates).     

Field test of a mathematical model for non-equilibrium transport of pesticides in soil

A model for the transport of pesticides in non-structured arable soil has been tested under field conditions. Three classes of sorption site are distinguished in the model. Sorption at class 1 sites is assumed to be at equilibrium whereas sorption at class 2 and class 3 sites is calculated using rate equations. Class 2 sites equilibrate on a time scale of days and class 3 sites equilibrate on a time scale of hundreds of days. In the model, the liquid phase is assumed to be homogeneous and completely mobile. The model was validated in two field experiments on a loamy sand soil using the herbicides cyanazine and metribuzin and using bromide ion as a tracer of liquid flow in soil. Ignoring sorption at class 3 sites resulted in large discrepancies between calculated and measured concentration profiles. Calculated concentration profiles were sensitive to the desorption rate constant for class 3 sites.     

手性除草剂研究进展

以几大类已经商品化或具有较好市场前景的手性除草剂为对象,综述了各除草剂的除草活性、对非靶标生物的毒性及其在环境迁移中的对映异构体选择性现象及机理的研究成果。由于对映异构体间的生物学性质迥异,因此,手性除草剂对映异构体的活性、作用机理、代谢转化及其在生物体内的转移等都具有手性选择性差异。同时,手性除草剂对植物的选择性作用还会因作用对象及环境因素的改变而改变,其在环境中的选择性降解情况也十分复杂,不仅与该除草剂自身的理化性质有关,还与其所处的环境因素有关,诸如土壤类型、植物种类、微生物群落结构、温度、pH值等均会改变手性除草剂对映异构体的环境归趋。     

Reversal of the inhibition of photosynthesis by herbicides affecting hydroxyphenylpyruvate dioxygenase by plastoquinone and tocopheryl derivatives in Chlamydomonas reinhardtii

Isoxaflutole or pyrazolate inhibition of tocopherol and plastoquinone biosynthesis in the green alga Chlamydomonas reinhardtii Dang leads to the inactivation of photosystem II and the degradation of its reaction centre D1 protein when exposed to strong light. Cell-permeable short-chain derivatives of plastoquinone and tocopherol were tested in the reversal. Addition of decyl-plastoquinone reverses herbicide-induced inhibition of photosynthesis and inactivation of photosystem II in short-time (1 h) exposure of the algae to high light. In high light longer than 1 h, decyl-plastoquinone alone loses effectiveness, but a synthetic permeable tocopheryl derivative retards the inhibitory effects on photosystem II and on the degradation of the D1 protein. This indicates that tocopherol deficiency induced by the herbicides makes a major contribution to their secondary mode of action in high light stress.     

A modified fluorometric method to quantify the concentration effect (pI50) of photosystem II-inhibiting herbicides

Chlorophyll fluorescence induction curves of isolated thylakoids were measured in the absence and in the presence of various concentrations of photosystem II-inhibiting herbicides. A mathematical program was applied to simulate the curves. Based on these simulated curves a new method is developed to determine the concentration effect (pI₅₀) of the herbicides. The results of the new method correspond well with reported values in the literature. The method is very convenient and time-saving.     

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.     

Influence of Compost Addition to Soil on the Behaviour of Herbicides

The transformations of eight herbicides (atrazine, simazine, terbutryn, pendimethalin, carbetamide, 2,4-D, metsulfuron-methyl and dimefuron) in soil after compost addition were monitored during long-term laboratory incubations. The herbicides were applied to soil, compost and soil-compost mixtures. Herbicide sorption, their kinetics of mineralisation and the extractability of residues were compared in the different treatments. Compost addition to soil generally decreased herbicide mineralisation and favoured the stabilisation of herbicide residues. A fraction of the stabilised residues remained extractable and potentially available. However, most of them were unextractable and formed bound residues. Sorption could be at the origin of a kinetically limited biodegradation, mainly for the most highly-sorbed herbicides (atrazine, simazine, terbutryn, pendimethalin and dimefuron). Compost addition had little effects on the less sorbed herbicides (carbetamide, 2,4-D and metsulfuron- methyl). © 1997 SCI.     

Effect of herbicide clomazone on photosynthetic processes in primary barley (Hordeum vulgare L.) leaves

The effect of pre-emergently applied herbicide clomazone on the photosynthetic apparatus of primary barley leaves (Hordeum vulgare L.) was studied. Clomazone application caused a reduction in chlorophyll (a+b) and carotenoid levels that was accompanied by a decline in the content of light harvesting complexes as judged from the increasing chlorophyll a/b ratio. The pigment reduction also resulted in changes in 77 K chlorophyll fluorescence emission spectra indicating lower chlorophyll (Chl) fluorescence reabsorption and absence of the long-wavelength emission forms of photosystem I. The maximal photochemical yield of photosystem II (PSII) and the reoxidation kinetics of the primary quinone acceptor Q_A⁻ were not significantly influenced by clomazone. A higher initial slope of Chl fluorescence rise in the Chl fluorescence induction kinetic indicated an increased delivery of excitations to PSII. Simultaneously, analysis of the Chl fluorescence quenching revealed that clomazone reduced function of the electron transport chain behind PSII. The decrease in the saturation rates of CO₂ assimilation paralleled the decrease of the Chl content and has been suggested to be caused by a suppressed number of the electron transport chains in the thylakoid membranes or by their decreased functionality. The obtained results are discussed in view of physiological similarity of the clomazone effect with changes of photosynthetic apparatus during photoadaptation.     

Sorption of low levels of nonionic and anionic surfactants on soil: effects on sorption of triticonazole fungicide

The sorption of two anionic surfactants and a series of seven nonionic alkylphenolethoxylate surfactants of increasing hydrophilic/lipophilic balance (HLB) in a loamy clay soil was evaluated. The effect of low doses of these surfactants on the sorption characteristics of the fungicide triticonazole was investigated. The critical micellar concentration (CMC) of the surfactants in pure water and soil–water systems, and surfactant sorption were estimated by surface tension measurements using a batch equilibration technique. Triticonazole sorption, alone and in the presence of low doses of surfactants, was also measured by batch equilibration. CMC of the alkylphenol surfactants increased with their HLB. The sorption of surfactants increased with their lipophilicity. CMC in the soil–water systems were considerably higher than in pure water. Sorption of the most lipophilic alkylphenol surfactants at the higher doses significantly increased triticonazole sorption. Proposed mechanisms are modifications of soil surface properties, and increase of soil organic carbon content. Sorption of the other nonionic and anionic surfactants only resulted in monomeric surfactant concentrations in pore water, and did not affect triticonazole sorption. © 1998 Society of Chemical Industry     

Inhibition of photosystem II in spinach chloroplasts by trachyloban-19-oic acid

The effect of trachyloban-19-oic acid isolated from Iostephane heterophylla (Asteraceae), was investigated on several photosynthetic activities in spinach thylakoids. The results indicated that this compound inhibited ATP synthesis and uncoupled electron transport, as well as basal and phosphorylating electron flow. Therefore, trachyloban-19-oic acid acts as Hill reaction inhibitor. This compound did not affect photosystem I activity but inhibited uncoupled photosystem II electron flow from H₂O to DCPIP, and has not effect on electron flow from H₂O to SiMo, indicating that the site of inhibition of this compound is at the level of Q_A-Q_B. Chlorophyll a fluorescence measurements confirm the behavior of this diterpene as Q_A-Q_B inhibitor, and in the other hand, this results indicate that a perturbation in the thylakoid membranes at the level of LHC II occurs.     

Effect of a nonhost-selective toxin from Alternaria alternata on chloroplast-electron transfer activity in Eupatorium adenophorum

AAC-toxin, a putative nonhost-selective phytotoxin, was obtained from Alternaria alternata causing a brown leaf spot disease of Crofton weed ( Eupatorium adenophorum ). The effect of AAC-toxin on the electron transfer reaction of chloroplasts showed that the activity of photosystem II, but not photosystem I, was completely inhibited by the toxin. AAC-toxin affected the following chlorophyll fluorescence parameters: coefficient of photochemical quenching ( q _P), the half-time value of fluorescence rise, and the O–J–I–P fluorescence induction kinetics curve, but not the ratio values of F _v/ F _m (the quantum yield of photosystem II) and the half-time value of fluorescence quenching. It was concluded that the toxin inhibited electron transfer from Q_A to Q_B (primary and secondary quinine acceptors of photosystem II) in photosystem II by competing with Q_B for the binding site in D1 protein on the thylakoid membrane.