Phenolic Herbicides Research Articles

One-step hydrothermal synthesis of Bi-TiO2 nanotube/graphene composites: An efficient photocatalyst for spectacular degradation of organic pollutants under visible light irradiation

In the present study, we have adopted a simple one-pot alkaline hydrothermal route to synthesize Bi-doped TiO2NT/graphene composites by using different wt% of Bi with an aim to achieve the excellent photocatalytic activity under visible light source. The nature of GO is changed to deoxygenated graphene with simultaneous embedding of Bi into TiO2 nanotube (TNT), during hydrothermal process. XRD and FTIR analysis confirm the successful conversion of GO to deoxygenated graphene. EPR analysis reveals the co-existence of Ti3+ ion with oxygen vacancy, which is created by the Bi doping. The photocatalytic activity of the prepared samples is measured by the degradation of aqueous suspensions of methylene blue (MB) and Dinoseb (phenolic herbicide), under visible-light irradiation. The prepared TiO2NT/graphene composite with 2-wt% bismuth (2-BTNTG) has shown the improved photocatalytic activity as compared to their counterparts. The improved photocatalytic activity is associated to the synergistic effect of graphene and Bi-TNT, which facilitate the interfacial charge transfer and enhances the efficiency of light harvesting in the visible region. Moreover, the underlying mechanism involving photocatalytic degradation of organic pollutants over 2-BTNTG is explored by using trapping experiments, suggesting that the .OH radicals solely contributed to degradation.

Herbicide and pesticide occurrence in the soils of children's playgrounds in Sarajevo, Bosnia and Herzegovina.

Pesticide pollution in Sarajevo public playgrounds is an important health and environmental issue, and the lack of information about it is causing concerns amongst the general population as well as researchers. Since children are in direct contact with surface soils on children's playgrounds, such soils should be much more carefully examined. Furthermore, herbicides and pesticides get transmitted from soil surfaces brought from outside the urban areas, or they get dispersed following their direct applications in urban areas. Infants' and children's health can be directly affected by polluted soils because of the inherent toxicity and widespread use of the different pesticides in urban environments such as playgrounds. In addition to that, the presence of chromated copper arsenate (CCA) wood preservative pesticide found as soil pollutant in playing equipment was also documented. Soil samples from playgrounds were collected and analyzed for triazines, carbamates, dithiocarbamates, phenolic herbicides and organochlorine pesticides. Samples for the determination of heavy metals Cu, Cr and As were prepared by microwave-assisted acid digestion, and the findings were determined by using an inductively coupled plasma optical emission spectrometer. Triazines, carbamates, dithiocarbamates, chlorphenoxy compounds, phenolic herbicides, organochlorine pesticides and organotin compounds were detected in playground soils and their determined concentrations (mg/kg) were respectively found as follows: <0.005, <0.05, <0.5, < 0.4, <0.1, <0.001 and <0.004. The determined contents (mg/kg) of Cu, Cr and As were in the ranges from 16.77 to 80.21, from 7.14 to 15.45 and from 3.31 to 4.43, respectively. Our preliminary results raise concerns about potential adverse effects of herbicides and pesticides on human health, which strengthens the case for a more preventative and protective approach to the uncontrolled presence of herbicides and pesticides in Sarajevo's playground soils.

Assessment of phenolic herbicide toxicity and mode of action by different assays.

A phytotoxicity assay based on seed germination/root elongation has been optimized and used to evaluate the toxic effects of some phenolic herbicides. The method has been improved by investigating the influence of experimental conditions. Lepidium sativum was chosen as the most suitable species, showing high germinability, good repeatability of root length measurements, and low sensitivity to seed pretreatment. DMSO was the most appropriate solvent carrier for less water-soluble compounds. Three dinitrophenols and three hydroxybenzonitriles were tested: dinoterb, DNOC, 2,4-dinitrophenol, chloroxynil, bromoxynil, and ioxynil. Toxicity was also determined using the Vibrio fischeri Microtox® test, and a highly significant correlation was found between EC50 values obtained by the two assays. Dinoterb was the most toxic compound. The toxicity of hydroxybenzonitriles followed the order: ioxynil >bromoxynil >chloroxynil; L. sativum exhibited a slightly higher sensitivity than V. fischeri to these compounds. A QSAR analysis highlighted the importance of hydrophobic, electronic, and hydrogen-bonding interactions, in accordance with a mechanism of toxic action based on protonophoric uncoupling of oxidative phosphorylation. The results suggest that the seed germination/root elongation assay with L. sativum is a valid tool for the assessment of xenobiotic toxicity and can be recommended as part of a test battery.

Ioxynil and Tetrabromobisphenol A Suppress Thyroid-Hormone-Induced Activation of Transcriptional Elongation Mediated by Histone Modifications and RNA Polymerase II Phosphorylation

To elucidate molecular mechanisms by which the phenolic herbicide ioxynil (IOX) and the brominated flame retardant tetrabromobisphenol A (TBBPA) exert thyroid hormone (TH) disrupting activity, we investigated the effects of the chemicals on the histone and RNA polymerase II (RNAPII) modifications in Xenopus laevis XL58-TRE-Luc cells in direct TH-response genes encoding TH receptor β (Thrb) and TH-induced basic leucine zipper protein (Thibz) using chromatin immunoprecipitation assays. For both the thrb and thibz genes, 3,3',5-triiodothyronine (T3) enhanced the amounts of gene transcripts and increased the amounts of acetylated histone H4 (H4Ac), trimethylated histone H3 lysine 4 (H3K4me3) and phosphorylated RNAPII serine 5 (RNAPIIS5P), epigenetic markers of gene activation at 5' regulatory regions, and the amounts of trimethylated histone H3 lysine 36 (H3K36me3) and phosphorylated RNAPII serine 2 (RNAPIIS2P), epigenetic markers of activation of transcriptional elongation at protein coding regions. Treatment with IOX and TBBPA reduced the amounts of the thrb transcript and suppressed the T3-induced modifications of H3K4me3, RNAPIIS5P, H3K36me3, and RNAPIIS2P. In the thibz gene, IOX and TBBPA did not suppress the T3-induced histone and RNAPII modifications except for H3K36me3 in the TBBPA treatment, despite both chemicals decreasing the T3-induced transcription. Our results demonstrate that IOX and TBBPA affect TH-induced histone and RNAPII modifications, which are involved in early and progressive stages of RNAPII transcriptional elongation, in direct TH-response genes, in somewhat target gene-dependent and chemical-specific manners. Both IOX and TBBPA are likely to influence epigenetically a cascade of TH receptor-mediated gene regulation.

Salicylic acid may indirectly influence the photosynthetic electron transport

Salicylic acid (SA) is a phenolic phytohormone with important roles in plant development, transpiration, endogenous signaling and defense against pathogens. One of the pathways of SA biosynthesis is located in the chloroplasts. The aim of the present work was to investigate the possible regulatory effects of SA on photosynthetic electron transport processes. Here we show that SA also affects leaf photosynthesis, via inducing stomatal closure and also by slowing down Photosystem II (PS II) electron transport. Photosynthetic CO2 incorporation and stomatal conductivity (measured with an infrared gas analyzer) were much lower in SA-infiltrated tobacco leaves than in untreated or water-infiltrated controls. PS II electron transport (calculated from PAM chlorophyll fluorescence data) was more sensitive to SA than Photosystem I (PS I) (measured with far red absorption). Direct probing of PS II charge separation and stabilization (measured with thermoluminescence), however, showed that these events were less affected in isolated thylakoid membranes than in leaves, suggesting that the effect of SA on PS II is indirect and different from similar effects of phenolic herbicides.

Species-dependent effects of the phenolic herbicide ioxynil with potential thyroid hormone disrupting activity: modulation of its cellular uptake and activity by interaction with serum thyroid hormone-binding proteins

Ioxynil, a phenolic herbicide, is known to exert thyroid hormone (TH) disrupting activity by interfering with TH-binding to plasma proteins and a step of the cellular TH-signaling pathway in restricted animal species. However, comparative studies are still lacking on the TH disruption. We investigated the interaction of [125I]ioxynil with serum proteins from rainbow trout, bullfrog, chicken, pig, rat, and mouse, using native polyacrylamide gel electrophoresis. Candidate ioxynil-binding proteins, which included lipoproteins, albumin and transthyretin (TTR), differed among the vertebrates tested. Rainbow trout and bullfrog tadpole serum had the lowest binding activity for ioxynil, whereas the eutherian serum had the highest binding activity. The cellular uptake of, and response to, ioxynil were suppressed by rat serum greater than by tadpole serum. The cellular uptake of [125I]ioxynil competed strongly with phenols with a single ring, but not with THs. Our results suggested that ioxynil interferes with TH homeostasis in plasma and with a step of cellular TH-signaling pathway other than TH-uptake system, in a species-specific manner.

Structures and Binding Sites of Phenolic Herbicides in the QB Pocket of Photosystem II

Herbicides targeting photosystem II (PSII) block the electron transfer beyond Q(A) by binding to the Q(B) site. Upon binding, the redox potential of Q(A) shifts differently depending on the types of herbicides. In this study, we have investigated the structures, interactions, and locations of phenolic herbicides in the Q(B) site to clarify the molecular mechanism of the Q(A) potential shifts by herbicides. Fourier transform infrared (FTIR) difference spectra upon photoreduction of the preoxidized non-heme iron (Fe(2+)/Fe(3+) difference) were measured with PSII membranes in the presence of bromoxynil or ioxynil. The CN and CO stretching vibrations of these phenolic herbicides were identified at 2215-2200 and 1516-1505 cm(-1), respectively, in the Fe(2+)/Fe(3+) difference spectra. Comparison with the spectra of bromoxynil in ethanol solutions along with density functional theory analysis strongly suggests that the phenolic herbicides take a deprotonated form in the binding pocket. In addition, the CN stretching, NH bending, and NH stretching vibrations of a His side chain, which were found at 1109-1101, 1187-1185, and 3000-2500 cm(-1), respectively, in the Fe(2+)/Fe(3+) difference spectra, showed characteristic features in the presence of phenolic herbicides. These signals are probably attributed to D1-His215, one of the ligands to the non-heme iron. Docking calculations for herbicides to the Q(B) pocket confirmed the binding of deprotonated bromoxynil to D1-His215 at the CO group, whereas the protonated form of bromoxynil and DCMU were found to bind to the opposite side of the pocket without an interaction with D1-His215. From these results, it is proposed that a strong hydrogen bond of the phenolate CO group with D1-His215 induces the change in the hydrogen bond strength of the Q(A) CO group through the Q(A)-His-Fe-His-phenolate bridge causing the downshift of the Q(A) redox potential.

Herbicide effect on the hydrogen-bonding interaction of the primary quinone electron acceptor QA in photosystem II as studied by Fourier transform infrared spectroscopy

The redox potential of Q(A) in photosystem II (PSII) is known to be lower by approximately 100 mV in the presence of phenolic herbicides compared with the presence of DCMU-type herbicides. In this study, the structural basis underlying the herbicide effects on the Q(A) redox potential was studied using Fourier transform infrared (FTIR) spectroscopy. Light-induced Q(A)(-)/Q(A) FTIR difference spectra of Mn-depleted PSII membranes in the presence of DCMU, atrazine, terbutryn, and bromacil showed a strong CO stretching peak of Q(A)(-) at 1,479 cm(-1), while binding of phenolic herbicides, bromoxynil and ioxynil, induced a small but clear downshift by approximately 1 cm(-1). The CO peak positions and the small frequency difference were reproduced in the S(2)Q(A)(-)/S(1)Q(A) spectra of oxygen-evolving PSII membranes with DCMU and bromoxynil. The relationship of the CO frequency with herbicide species correlated well with that of the peak temperatures of thermoluminescence due to S(2)Q(A)(-) recombination. Density functional theory calculations of model hydrogen-bonded complexes of plastoquinone radical anion showed that the small shift of the CO frequency is consistent with a change in the hydrogen-bond structure most likely as a change in its strength. The Q(A)(-)/Q(A) spectra in the presence of bromoxynil, and ioxynil, which bear a nitrile group in the phenolic ring, also showed CN stretching bands around 2,210 cm(-1). Comparison with the CN frequencies of bromoxynil in solutions suggested that the phenolic herbicides take a phenotate anion form in the Q(B) pocket. It was proposed that interaction of the phenolic C-O(-) with D1-His215 changes the strength of the hydrogen bond between the CO of Q(A) with D2-His214 via the iron-histidine bridge, causing the decrease in the Q(A) redox potential.

The glutathione peroxidase homologous gene Gpxh in Chlamydomonas reinhardtii is upregulated by singlet oxygen produced in photosystem II

The expression of the glutathione peroxidase homologous gene Gpxh, known to be specifically induced by the formation of singlet oxygen (1O2), was analyzed in cells of Chlamydomonas reinhardtii exposed to environmental conditions causing photoinhibition. Illumination with high light intensities, leading to an increased formation of 1O2 in photosystem II, continuously induced the expression of Gpxh in cell for at least 2 h. Phenolic herbicides like dinoterb, raise the rate of 1O2 formation by increasing the probability of charge recombination in photosystem II via the formation of the primary radical pair and thereby 3P680 formation (Fufezan C et al. 2002, FEBS Letters 532, 407-410). In the presence of dinoterb the light-induced loss of the D1 protein in C. reinhardtii was increased and the high light-induced Gpxh expression was further stimulated. DCMU, a urea-type herbicide, causing reduced 1O2 generation in photosystem II, protected the D1 protein slightly against degradation and downregulated the expression of the Gpxh gene compared to untreated cells exposed to high light intensities. This indicates that the Gpxh expression is induced by 1O2 under environment conditions causing photoinhibition.

Study on the toxicity of phenolic and phenoxy herbicides using the submitochondrial particle assay

A simple and rapid in vitro toxicological assay, utilizing submitochondrial particles (SMP), has been used to evaluate the toxic effects of fifteen herbicides belonging to the phenol and phenoxyalkanoic acid chemical classes. The SMP assay allows the quantitative evaluation of the toxicity of compounds with different mechanisms of action: uncouplers, inhibitors of the enzyme complexes involved in reverse electron transfer and in oxidative phosphorylation and chemicals that alter the membrane structure. The two groups of herbicides showed different levels of toxicity. For phenol derivatives, EC 50 values ranged from 0.16 μM (ioxynil) to 6.7 μM (2,4-dinitrophenol), whereas for phenoxy herbicides EC 50 values ranged from 21 μM (2,4,5-trichlorophenoxyacetic acid, 2,4,5-T) to 110 μM (4-chloro-2-methylphenoxyacetic acid, MCPA). On the average, the toxicity of phenolic compounds is greater than that of phenoxyalkanoic acids by two orders of magnitude. Quantitative structure–activity relationships (QSAR) were developed between EC 50 values and various molecular descriptors. The results suggest the existence of different mechanisms of action for the two classes of compounds. The findings obtained for phenolic herbicides are consistent with a protonophoric uncoupling mechanism, whereas for phenoxy herbicides a non-specific mode of action at membrane level can be hypothesized.

Herbicide-Induced Changes in Charge Recombination and Redox Potential of QAin the T4 Mutant ofBlastochloris viridis

To gain new insights into the function of photosystem II (PSII) herbicides DCMU (a urea herbicide) and bromoxynil (a phenolic herbicide), we have studied their effects in a better understood system, the bacterial photosynthetic reaction center of the terbutryn-resistant mutant T4 of Blastochloris (Bl.) viridis. This mutant is uniquely sensitive to these herbicides. We have used redox potentiometry and time-resolved absorption spectroscopy in the nanosecond and microsecond time scale. At room temperature the P(+)(*)Q(A)(-)(*) charge recombination in the presence of bromoxynil was faster than in the presence of DCMU. Two phases of P(+)(*)Q(A)(-)(*) recombination were observed. In accordance with the literature, the two phases were attributed to two different populations of reaction centers. Although the herbicides did induce small differences in the activation barriers of the charge recombination reactions, these did not explain the large herbicide-induced differences in the kinetics at ambient temperature. Instead, these were attributed to a change in the relative amplitude of the phases, with the fast:slow ratio being approximately 3:1 with bromoxynil and approximately 1:2 with DCMU at 300 K. Redox titrations of Q(A) were performed with and without herbicides at pH 6.5. The E(m) was shifted by approximately -75 mV by bromoxynil and by approximately +55 mV by DCMU. As the titrations were done over a time range that is assumed to be much longer than that for the transition between the two different populations, the potentials measured are considered to be a weighted average of two potentials for Q(A). The influence of the herbicides can thus be considered to be on the equilibrium of the two reaction center forms. This may also be the case in photosystem II.

BODY MASS INDEX AND BROMOXYNIL EXPOSURE IN A SAMPLE OF RURAL RESIDENTS DURING SPRING HERBICIDE APPLICATION

Bromoxynil (3,5-dibromo-4-hydroxybenzonitrile), a phenolic herbicide, is widely used in production of cereals and other crops. Little is known, however, about bromoxynil exposure in humans. Results of previous research suggest a longer residence time in the body for bromoxynil compared to phenoxy herbicides [e.g., (2,4-dichlorophenoxy)acetic acid (2,4-D), 4-chloro-2-methylphenoxyacetic acid (MCPA)] and that bromoxynil would tend to partition into fatty tissue more so than 2,4-D. In previous research, body mass index (BMI) was found to be an independent predictor of plasma concentrations of 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE), the persistent lipophilic metabolite of the chlorinated pesticide bis(4-chlorophenyl)-1,1,1-trichloroethane (DDT). As part of the Prairie Ecosystem Study, gas chromatography/mass spectrometry analysis was used to measure concentrations of bromoxynil and seven other herbicides (2,4-D, dicamba, fenoxaprop, MCPA, ethalfluralin, triallate, and trifluralin) in plasma from residents (104 men, 88 women, 24 youths age 12–17yr) of a cereal-producing region in Saskatchewan, Canada, during spring herbicide application, 1996. Multiple logistic regression analysis was used to explore whether BMI predicted detection of bromoxynil in plasma from the adults. The prevalence of detection (detection limits: 2–50μg/L) was markedly higher for bromoxynil (men, 44.2%; women, 14.8%; youths, 20.8%) compared to each of the other herbicides including 2,4-D (men, 16.5%; women, 3.4%; youths, 12.5%) and MCPA (men, 6.8%; women, 1.1%; youths, 4.2%), although bromoxynil is commonly formulated or tank mixed with these herbicides. In the multiple logistic regression analysis, the variables BMI, exposure group [bromoxynil applicators, non-applicator family members of bromoxynil applicators, all others (reference group)], and days elapsed since the last use of bromoxynil were found to be independent predictors of detection of bromoxynil, while age, gender, and farm residency were not statistically significant. With adjustment for exposure group [bromoxynil applicators: odds ratio (OR=24.30, 95% confidence interval (CI)=9.59–61.58; nonapplicator family members of bromoxynil applicators: OR=3.53, 95% CI=1.19–10.44; all others (reference group)], the OR for detection of bromoxynil was 2.35 (95% CI=0.87–6.33) for participants in the middle (25.53–29.00kg/m2) tertile (men: OR=2.85, 95% CI=0.75–10.82; women: OR=1.63, 95% CI=0.36–7.40) of BMI and 4.01 (95% CI=1.46–11.03) for participants in the highest (>29.00kg/m2) tertile (men: OR=4.67, 95% CI=1.17–18.58; women: OR=2.20, 95% CI=0.44–10.99) with participants in the lowest (<25.53kg/m2) tertile as the reference group. Similar ORs were observed for BMI with adjustment for days elapsed since the last use of bromoxynil. In conclusion, further research is needed to investigate whether adiposity is an important modifying factor for persistence of bromoxynil in the body.

Derivatisation in gas chromatographic determination of acidic herbicides in aqueous environmental samples.

This paper reviews derivatisation processes applied in chromatographic determination of acidic herbicides (with carboxyl and phenol groups), mainly in aqueous environmental samples. The discussion focuses on the basic derivatisation reactions used to convert herbicides to derivatives to make them analysable by gas chromatography, and possibly to reduce detection limits and/or increase extraction recovery from aqueous samples. The reactions are transesterification, esterification, silylation, alkylation, and extractive and pyrolytic alkylation. The reagents used to conduct the reactions are numerous. Diazomethane is a very efficient methylation reagent but explosive and toxic. Methyl iodide also ensures rapid and efficient methylation. Benzyl bromide can be used directly in water but derivatisation yield is low and reproducibility is poor. Butyl chloroformate and dimethyl sulfite can also be used for derivatisation in water, and acetic anhydride can be used for in-situ derivatisation of phenolic herbicides. For increasing selectivity of GC detection pentafluorobenzyl bromide (for ECD) and (2-cyanethyl)dimethyl(diethylamino)silane (for NPD) have been applied. Very characteristic ions are produced in mass spectra if silyl groups are introduced, e.g. by using bis(trimethylsilyl)trifluoroacetamide. Tetramethylammonium, trimethylphenylammonium, tetraalkylammonium, and trimethylsulfonium hydroxides and salts can by used for derivatisation at elevated temperature in the GC injection port. Extractive alkylation is relatively efficient if tetraalkylammonium salts with long chain alkyl groups are used.

Acceptor and donor-side interactions of phenolic inhibitors in Photosystem II

Certain phenolic compounds represent a distinct class of Photosystem (PS) II Q B site inhibitors. In this paper, we report a detailed study of the effects of 2,4,6-trinitrophenol (TNP) and other phenolic inhibitors, bromoxynil and dinoseb, on PS II energetics. In intact PS II, phenolic inhibitors bound to only 90–95% of Q B sites even at saturating concentrations. The remaining PS II reaction centers (5–10%) showed modified Q A to Q B electron transfer but were sensitive to urea/triazine inhibitors. The binding of phenolic inhibitors was 30- to 300-fold slower than the urea/triazine class of Q B site inhibitors, DCMU and atrazine. In the sensitive centers, the S 2Q A − state was 10-fold less stable in the presence of phenolic inhibitors than the urea/triazine herbicides. In addition, the binding affinity of phenolic herbicides was decreased 10-fold in the S 2Q A − state than the S 1Q A state. However, removal of the oxygen-evolving complex (OEC) and associated extrinsic polypeptides by hydroxylamine (HA) washing abolished the slow binding kinetics as well as the destabilizing effects on the charge-separated state. The S 2-multiline electron paramagnetic resonance (EPR) signal and the ‘split’ EPR signal, originating from the S 2Y Z state showed no significant changes upon binding of phenolic inhibitors at the Q B site. We thus propose a working model where Q A redox potential is lowered by short-range conformational changes induced by phenolic inhibitor binding at the Q B niche. Long-range effects of HA-washing eliminate this interaction, possibly by allowing more flexibility in the Q B site.

Singlet oxygen production in herbicide-treated photosystem II

Photo-generated reactive oxygen species in herbicide-treated photosystem II were investigated by spin-trapping. While the production of OH and O2− was herbicide-independent, 1O2 with a phenolic was twice that with a urea herbicide. This correlates with the reported influence of these herbicides on the redox properties of the semiquinone QA− and fits with the hypothesis that 1O2 is produced by charge recombination reactions that are stimulated by herbicide binding and modulated by the nature of the herbicide. When phenolic herbicides are bound, charge recombination at the level of P+Pheo− is thermodynamically favoured forming a chlorophyll triplet and hence 1O2. With urea herbicides this pathway is less favourable.

Herbicide Resistance and Supersensitivity in Ala250 or Ala251 Mutants of the D1 Protein in Chlamydomonas reinhardtii

Various site-directed mutants at position Ala250 or Ala251 within the herbicide-binding niche of the D1 subunit of photosystem II were generated by transforming a Chlamydomonas reinhardtii strain specifically engineered to lack a psbA gene fragment coding for the binding region. The following mutants were obtained: Asp250, Phe250, His250, Ile250, Asn250, Arg250, Tyr250, Cys251, Gly251, and the double mutant Ser250/Ile255. A variety of herbicides and inhibitors, such as s-triazines, phenolic herbicides, p-benzo- and naphthoquinones, acridones, a NH-thiazol, and phenmedipham, were assayed for their inhibitory activity in these mutants. In addition, in the Ser250/Ile255 mutant 32 different triazinones were tested for biological activity. In all mutants, either resistance and/or supersensitivity against the herbicides and inhibitors was observed. The resistance and/or supersensitivity against inhibitors in the Ala250 mutants demonstrated for the first time the involvement of this amino acid in herbicide binding.

A Multi‐Year Study of Four Herbicides in Air and Precipitation from a Small Prairie Watershed

AbstractTemporal trends of three phenoxyacid herbicides: 2,4‐D, dichlorprop and MCPA and the phenolic herbicide: bromoxynil, were determined in ambient air and precipitation over a 4‐yr period within a well‐defined watershed in southern Manitoba. Air concentrations ranged from below detection limits to 3.5, 1.0, 13, and 2.0 ng/m3, for 2,4‐D; dichlorprop; MCPA; and bromoxynil, respectively. The herbicides under investigation were four of the highest use herbicides in the South Tobacco Creek Watershed and were present in both air and precipitation samples at maximum concentrations during periods of local use. Concentration patterns for each herbicide in precipitation were similar each year of the study. Of the four herbicides, precipitation fluxes were greatest for MCPA in the watershed and the surrounding region. Although bromoxynil and dichlorprop were less widely used than 2,4‐D, bromoxynil precipitation fluxes exceeded those of 2,4‐D in 1993 and 1994, and dichlorprop precipitation fluxes were greater than 2,4‐D in 1994. Elevated levels of these herbicides in creek water samples were observed during periods of high concentration in both atmospheric compartments, despite the lack of surface runoff events within the watershed over that period.

Influence of herbicide binding on the redox potential of the quinone acceptor in photosystem II: relevance to photodamage and phytotoxicity.

Here we show that herbicide binding influences the redox potential (Em) of the plastoquinone QA/QA- redox couple in Photosystem II (PSII). Phenolic herbicides lower the Em by approximately 45 mV, while DCMU raises the Em by 50 mV. These shifts are reflected in changes in the peak temperature of thermoluminescence bands arising from the recombination of charge pairs involving QA-. The herbicide-induced changes in the Em of QA/QA- correlate with earlier work showing that phenolic herbicides increase the sensitivity of PSII to light, while DCMU protects against photodamage. This correlation is explained in terms of the following hypothesis which is based on reactions occurring in the bacterial reaction center. The back-reaction pathway for P680+QA- is assumed to be modulated by the free-energy gap between the P680+QA- and the P680+Ph- radical pairs. When this gap is small (i.e., when the Em of QA/QA- is lowered), a true back-reaction is favored in which P680+Ph- is formed, a state which decays forming a significant yield of P680 triplet. This triplet state of chlorophyll reacts with oxygen, forming singlet oxygen, a species likely to be responsible for photodamage. When the free-energy gap is increased (i.e., when the Em of QA/QA- is raised), the yield of the P680+Ph- is diminished and a greater proportion of the P680+QA- radical pair decays by an alternative, less damaging, route. We propose that at least some of the phytotoxic properties of phenolic herbicides may be explained by the fact that they render PSII ultrasensitive to light due to this mechanism.

Reduced turnover of the D1 polypeptide and photoactivation of electron transfer in novel herbicide resistant mutants of Synechocystis sp. PCC 6803.

Two missense mutants, A263P and S264P, and a deletion mutant des-Ala263, Ser264, have been constructed in the D1 protein of the cyanobacterium Synechocystis sp PCC 6803. All were expected to induce a significant conformational change in the QB-binding region of photosystem II (PSII). Although the des-Ala263, Ser264-D1 mutant accumulated some D1 protein in the thylakoid membrane it was unable to grow photoautotrophically or evolve oxygen. Thermoluminescence and chlorophyll fluorescence studies confirmed that this deletion mutant did not show any functional PSII activity. In contrast, [S264P]D1 was able to grow photoautotrophically and give light-saturated rates of oxygen evolution at 60% of the rate of the wild-type control strain, TC31. The A263P missense mutant was also able to evolve oxygen at 50% of TC31 rates although it did not readily grow photoautotrophically. Thermoluminescence, flash oxygen yield and chlorophyll fluorescence measurements indicated that in both missense mutants electron transfer from QA to QB was significantly impaired in dark adapted cells. However, QA to QB electron transfer could be photoactivated in the mutants by background illumination. Both the A263P and S264P mutants also showed an increase in resistance to the s-triazine family of herbicides although this feature did not hold for the phenolic herbicide, ioxynil. Of particular interest was that the two missense mutants, especially S264P, possessed a slower rate of turnover of the D1 protein compared with TC31 and in vivo contained detectable levels of a 41-kDa adduct consisting of D1 and the alpha subunit of cytochrome b559. When protein synthesis was blocked by the addition of lincomycin, D1 degradation was again slower in S264P than TC31. The results are discussed in terms of structural changes in the QB-binding region which affect herbicide and plastoquinone binding and perturb the normal regulatory factors that control the degradation of the D1 protein and its synchronisation with the synthesis of a replacement D1 protein.

Structure and Site of Action of an Algicide from a Cyanobacterium, Oscillatoria late-virens

Summary An algicidal (herbicidal) metabolite from cyanobacterium Oscillatoria late-virens was purified and the mechanism of action determined. The algicide had a molecular weight of 444 Da, an emperical formula of C 29 H 48 O 3 , and contained phenol, C 20 alkane and an α, β-unsaturated carbonyl residue. With K 3 Fe(CN) 6 and benzoquinones, 50% inhibition of photosynthetic electron flow (I 50 ) was attained by 1.25-2.35 μM of algicide. Chlorophyll fluorescence data indicate that the algicide interacted at the acceptor site of photosystem (PS)II, although the mechanism may be different from that of another herbicide, 3-(3,4-dichlorophenyl)-1, 1-dimethyl urea (DCMU). Structural resemblance with phenolic herbicides indicates their common mode of interaction.