Biodegradation of the herbicide bromoxynil and its plant cell wall bound residues in an agricultural soil

The mineralization and formation of metabolites and nonextractable residues of the herbicide [¹⁴C]bromoxyniloctanoate ([¹⁴C]3,5-dibromo-4-octanoylbenzonitrile) and the corresponding agent substance [¹⁴C]bromoxynil ([¹⁴C]3,5-dibromo-4-hydroxybenzonitrile) was investigated in a soil from an agricultural site in a model experiment. The mineralization of maize cell wall bound bromoxynil residues was also investigated in the agricultural soil material. The mineralization of [¹⁴C]bromoxynil and [¹⁴C]bromoxyniloctanoate in soil within 60 days amounted up to 42 and 49%, respectively. After the experiments, 52% of the originally applied [¹⁴C]bromoxynil and 44% of the [¹⁴C]bromoxyniloctanoate formed nonextractable residues in soil. Plant cell wall bound [¹⁴C]bromoxynil residues were also mineralized to an extent of about 21% within 70 days; the main portion of 76% persisted as nonextractable residues in the soil. In bacterial enrichment cultures and in soil two polar metabolites were observed; one of it could be identified as 3,5-dibromo-4-hydroxybenzoate and the other could be described tentatively as 3,5-dibromo-4-hydroxybenzamide.     

Effect of climatic factors on the potency of ioxynil and bromoxynil

Outdoor pot experiments, carried out between 1967 and 1971, mainly with Polygonum lapathifolium L. and Stellaria media (L.) Vill., showed that herbicidal potency of ioxynil and bromoxynil salts and esters was affected by climatic conditions, particularly solar radiation and relative humidity. Multiple regression equations were frequently obtained which accounted for 80% or more of the observed variation in effective dose (ED₉₀). By contrast, experiments in growth cabinets suggested that air temperature was the most important climatic variable; the reasons for this disagreement are discussed. It is concluded that climatic factors affect the potency of ioxynil and bromoxynil via their influence on leaf uptake; ester formulations (emulsions) were much less affected than aqueous salt solutions.     

Degradation of ioxynil to CO2 in soil

Degradation of ioxynil (4-hydroxy-3,5-diiodobenzonitrile) to CO₂ was detected in a clay loam, high organic matter content soil. The majority of radioactivity was recovered as ¹⁴CO₂ from both ring-labeled and cyano-labeled ioxynil; however, ¹⁴CO₂ was always released from cyano-labeled ioxynil at a much faster initial rate. No ¹⁴CO₂ was released in treated sterile soil, either aerobically or anaerobically. Production of ¹⁴CO₂ from cyanolabeled and ring-labeled ioxynil was greatly inhibited by HgCl₂ (10^?5M), and p-chloromercuribenzoate (5 × 10^?5M), but slightly inhibited by ferricyanide (10^?4M). No ¹⁴CO₂ was evolved from ring-labeled ioxynil under anaerobic conditions. These observations indicated that the degradation of ioxynil to CO₂ in soil was a microbial action and was oxygen dependent. This is consistent with the known mechanism of oxygenases in degrading benzene rings. Anaerobically, a small amount of ¹⁴CO₂ was released from cyano-labeled ioxynil. Thin-layer chromatographic analyses of the culture supernatant revealed that 3,5-diiodo-4-hydroxybenzamide and 3,5-diiodo-4-hydroxybenzoic acid were intermediate metabolites.     

Soil persistence studies with [14C]MCPA in combination with other herbicides and pesticides

The persistence of [¹⁴C]MCPA at a rate equivalent to 1 kg ha^?1 was studied under laboratory conditions in a clay loam, heavy clay and sandy loam at 85% of field capacity moisture and 20±1°C both alone and in the presence of tri-allate, trifluralin, tri-allate and trifluralin, malathion, Vitaflow DB, malathion and Vitaflow DB, bromoxynil, bromoxynil and asulam, bromoxynil and difenzoquat, dicamba, dicamba and mecoprop, linuron, MCPB, metribuzin, propanil, TCA, benzoylprop-ethyl, diclofop-methyl, and flamprop-methyl. Except in the soils treated with asulam, the half-lives of [¹⁴C]MCPA in all three soil types were similar, being approximately 13±1 days, thus indicating that none of the other chemicals studied adversely affected the soil degradation of MCPA. In the asulam treated soils, the half-lives of the MCPA were about 3 days longer than in non-asulam treated soils; the effect was most marked in the clay loam.     

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.     

Interaction with phospholipids as a possible mode of action of 3-phenylindole on Aspergillus niger

Low concentrations of 3-PI(3-phenylindole) inhibit several uptake and biosynthetic processes in liquid cultures of Aspergillus niger. The incorporation of ³²P_i into phospholipids, [¹⁴C]uridine into nucleic acids, and [¹⁴C]phenylalanine into proteins was inhibited and the uptake of all three precursors was reduced. Studies on an in vitro interaction between 3-phenylindole and phospholipids showed, that at about equimolar concentrations 3-phenylindole prevents phospholipids from swelling in water. At the same ratio a decrease of the absorption intensity of the NH-band of 3-phenylindole was observed in ir spectra. Moreover, the maxima of the uv spectrum of 3-phenylindole shifted after addition of a sonicated phospholipid suspension. It is suggested that binding to phospholipids is the first effect of 3-phenylindole. Apparently this affects, consequently, several membrane-bound reactions, i.e., transport and biosynthetic processes.     

Control of Galium aparine (cleavers) in winter cereals with mecoprop alone or with ioxynil plus bromoxynil

A series of eleven field experiments over two seasons investigated the control of Galium aparine (cleavers) with mecoprop and a proprietary mixture of ioxynil plus bromoxynil. Applications of mecoprop (2?4 kg a.e. ha^?1) between December and March gave variable results but a mixture of ioxynil + bromoxynil (0?6 kg a.i. ha^?1) and mecoprop improved control up to at least 90%, at most sites. Applications of mecoprop alone in April were most effective and frequently achieved more than 90% control. In May, when the G. aparine plants were very large, it was less successful. The reasons for the environmental effects on performance are discussed. Preliminary examination of the effect of soil temperature on mecoprop activity indicates that the reliability of control increases with increasing temperature (0–12°C). An average of 94% control was achieved when soil temperatures were greater than 6°C. La destruction de Galium aparine (Gaillet) dans les cereales d'hiver avec du mecoprop seul ou avec ioxynil plus bromoxynil     

Demonstrating formation of potentially persistent transformation products from the herbicides bromoxynil and ioxynil using liquid chromatography-tandem mass spectrometry (LC-MS/MS)

It is shown that potentially persistent transformation products can be formed from the herbicides bromoxynil (3,5-dibromo-4-hydroxybenzonitrile) and ioxynil (3,5-diiodo-4-hydroxybenzonitrile), and possible leaching to groundwater is discussed. A similar process to the formation of BAM (2,6-dichlorobenzamide) from the herbicide dichlobenil (2,6-dichlorobenzonitrile) can be anticipated as bromoxynil and ioxynil are analogues of dichlobenil and they are degraded by the enzymes nitrilase, nitrile hydratase and amidase. A biodegradation study using cultured Variovorax sp. DSM 11402, a species commonly found in soil, demonstrated that ioxynil and bromoxynil were fully transformed into their corresponding amides in 2-5 days. These amides were not further degraded within 18 days, and formation of other degradation products was not observed. These results are in agreement with biodegradation experiments with dichlobenil. In soil, dichlobenil is transformed into its only observed degradation product BAM, which is persistent and mobile, and has been found in 19% of 5000 samples of Danish groundwater. Variovorax sp. is known to degrade the non-halogenated analogue benzamide, suggesting that degradation of the three amides may be hindered by the halogenated substituents (meta-Br; meta-I; ortho-Cl). This hypothesis is supported by QSAR modelling of fundamental properties. Using a new optimised liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, the sorption and desorption properties of bromoxynil and ioxynil were characterised in sandy topsoil at four concentration levels. The estimated sorption coefficient K(d) was 1.4 L kg(-1) for bromoxynil and 5.4 L kg(-1) for ioxynil, indicating weak to moderate sorption to topsoil. Desorption of the herbicides showed that they were strongly and irreversible bound to the soil (K(des) > K(d)). The amount of herbicide desorbed depended on the initial concentration level. At low levels, K(des) values were higher, indicating stronger binding than at higher levels. The isocratic LC-MS/MS method developed for simultaneous detection of bromoxynil, ioxynil and their main degradation products is described. Using negative electrospray ionisation (ESI-), the detection limits were 0.4-1.0 microg L(-1), with relative standard deviations of 4-10% (n = 10) using direct injection without clean-up steps. The standard curves showed linearity in the range 5-100 microg L(-1) with r(2) > 0.992.     

Radiochemical distribution and decline studies with bromoxynil octanoate in wheat

Bromoxynil octanoate labelled with ¹⁴C in the ring or in the cyano-group was applied to wheat seedlings at the two-leaf or fully-tillered stage and at rates equivalent to up to 16 oz a.i./acre. The plants were grown either in environmental chambers under controlled conditions for up to 28 days, or outdoors under field conditions for various periods up to harvest. Initially, elimination of radioactivity occurred more rapidly with bromoxynil-cyano-[¹⁴C]-octanoate than with bromoxynil-ring-[¹⁴C]-octanoate, indicating metabolic attack on the cyano group. Under outdoor conditions with ring-[¹⁴C]-herbicide applied at the two-leaf stage, only 12% of the radioactivity was retained after 28 days, principally in the treated leaves. When application was made at fully-tillered stage, about 33% of the ¹⁴C was retained after 56 days, almost entirely in the treated senescent leaves at the base of the plant. There was very little translocation of the herbicide or of any major metabolite. The level of radioactivity in harvested grain and in straw more than 7.5 cm above the ground was very low, even after very late application of ring-[¹⁴C]-labelled herbicide. The amount of bromoxynil octanoate, together with any metabolite retaining part of the aromatic ring, did not collectively exceed the equivalent of approx. 0.01 parts/million bromoxynil octanoate.     

Bromoxynil degradation in a Mississippi silt loam soil

BACKGROUND: The objectives of these laboratory experiments were: (1) to assess bromoxynil sorption, mineralization, bound residue formation and extractable residue persistence in a Dundee silt loam collected from 0–2 cm and 2–10 cm depths under continuous conventional tillage and no‐tillage; (2) to assess the effects of autoclaving on bromoxynil mineralization and bound residue formation; (3) to determine the partitioning of non‐extractable residues; and (4) to ascertain the effects of bromoxynil concentration on extractable and bound residues and metabolite formation. RESULTS: Bromoxynil K_d values ranged from 0.7 to 1.4 L kg^?1 and were positively correlated with soil organic carbon. Cumulative mineralization (38.5% ± 1.5), bound residue formation (46.5% ± 0.5) and persistence of extractable residues (T_1/2 < 1 day) in non‐autoclaved soils were independent of tillage and depth. Autoclaving decreased mineralization and bound residue formation 257‐fold and 6.0‐fold respectively. Bromoxynil persistence in soil was rate independent (T_1/2 < 1 day), and the majority of non‐extractable residues (87%) were associated with the humic acid fraction of soil organic matter. CONCLUSIONS: Irrespective of tillage or depth, bromoxynil half‐life in native soil is less than 1 day owing to rapid incorporation of the herbicide into non‐extractable residues. Bound residue formation is governed principally by biochemical metabolite formation and primarily associated with soil humic acids that are moderately bioavailable for mineralization. These data indicate that the risk of off‐site transport of bromoxynil residues is low owing to rapid incorporation into non‐extractable residues. Published 2009 by John Wiley & Sons, Ltd     

Effect of some substituted benzonitriles on photosynthetic activity of spinach and wheat in vivo and in vitro

The inhibitory effects of nine nitro and/or bromo-substituted benzonitrile compounds on the photosynthetic electron flow in isolated chloroplasts and on the in vivo CO₂ fixation of spinach (Spinacia oteracea L.) and wheat (Triticum aesticum L. cv. Bezoslaya) were investigated. Bromoxynil and 3-nitro-5-hromo-4-hydroxy-bcnzonitrile were the strongest and equally effective inhibitors of Ihe in vivo CO₂ fixation of spinach, hut in wheat the nitro-bromo-compound is ineffective and 3-nitro-benzonitrile is even more inhibitory than bromoxynil. None of the substances affected DCPIPH → methylviologen reduction. In the inhibition of the DCPIP reduction only the 3,5-disuhstituted 4-hydroxy-derivatives were effective. The fact that these compounds affect only the PS II reaction with both H₂O and DPC as electron donors suggests a site of inhibition on the reducing side of PS II, between Ihe PS II reaction centre and ihe DCPIP Site. It is suggested that in the inhibition of the DCPIP reduction only steric factors are important and the different electron configuration of the sterieally similar molecules may be involved only in the absorption and translocation processes of the compounds.     

DDT and pyrethroids: Receptor binding and mode of action in the house fly

The mode of action of DDT and pyrethroids was investigated in the house fly, Musca domestica L, using drug:receptor binding techniques. Both in vivo and in vitro binding studies demonstrated the existence of membrane receptors which bind specifically to [¹⁴C]DDT and [¹⁴C]cis-permethrin. The receptors show properties to be expected of a critical target site of these insecticides. These include negative temperature correlation with binding, relatively nonsensitivity to DDE, and sensitivity to Ca²⁺. The receptor sites are readily saturated at 45–90 nM [¹⁴C]DDT and have an apparent disassociation constant (K_d) of 12.2 nM. The maximum number of binding sites was estimated to be 17 pmol DDT/mg membrane protein (0.34 pmol/house fly head). Competition studies showed DDT, cis-permethrin, and cypermethrin bind to the same receptor but not at precisely the same site. The addition of Ca²⁺ to the incubation buffer significantly inhibited the binding of both [¹⁴C]DDT and [¹⁴C]cis-permethrin, suggesting the receptor binding is Ca²⁺ sensitive and may have a role in ion conductance.     

Mechanism of resistance to fenarimol in Aspergillus nidulans

Mycelial uptake of [¹⁴C]fenarimol (10 μg/ml) by 20 fenarimol-resistant mutants of Aspergillus nidulans was compared with uptake by wild-type strain 003. Uptake of the fungicide during the initial 10 min of incubation was significantly lower in all mutant strains than in the wild-type strain indicating that resistance is related with reduced uptake. Upon prolonged incubation a gradual decrease of accumulated radioactivity in the wild-type strain was observed. A few mutants displayed resistance to unrelated chemicals such as p-fluorophenylalanine or d-serine; this phenomenon appeared not to be due to a decreased uptake of the corresponding natural amino acids. Incorporation of [³H]adenine and [¹⁴C]leucine by mycelium of mutant M193 was hardly inhibited after 5 hr of incubation with the fungicide, whereas a distinct effect was found with the wild-type strain. At this time also fungitoxicity to the wild-type strain became apparent. Probably, this effect is indirectly caused by inhibition of ergosterol biosynthesis. Mycelium of mutant M193 incorporated [¹⁴C]acetate slightly less effectively than the wild-type strain. After 2 hr of incubation with this radiochemical leakage of [¹⁴C]acetate metabolites from mycelium of the mutant strain was observed. This indicates that resistance might be correlated with increased excretion of fungal metabolites, which in turn may be related with reduced fitness of fenarimol-resistant mutants.     

Studies on the effects of herbicides on soil nitrification,II

The effects of selected herbicides have been studied on the following parameters of soil nitrification processes: the rates of nitrate and nitrite formation from ammonia in freshly perfused soils and in soils previously saturated with nitrifying organisms in an improved perfusion apparatus; the rates of oxygen consumption and of oxidation of ammonia and nitrite in washed cell suspensions of Nitrosomonas europaea and Nitrobacter winogradskii, respectively; the rates of growth of those two organisms in newly established cultures; the rates of oxygen uptake by soil enriched in nitrifying organisms; the rates of proliferation of nitrifying populations in freshly perfused soil. ID50 values were computed for all parameters and herbicides studied. On average, the most sensitive parameters were the metabolism and growth of the two organisms in pure culture, while the least sensitive were the corresponding measures in the soil environment. Similarly, herbicides fell into four distinct groups. The most toxic were the formulated octanoates of bromoxynil and ioxynil (NPH1320 and Totril, respectively); next in order of toxicity were chlorbufam, phenmedipham, formulated oxadiazon, formulated legurame, ioxynil, formulated trifluralin, and bromoxynil; low toxicity was shown by terbacil, dicamba, and tricamba, whereas asulam and the related experimental herbicide MB9555 showed activity on some parameters at the very highest concentrations only. Comparisons of soil with pure-culture parameters showed that the relative toxicities of herbicides to Nitrosomonas in culture bore little relationship to those in soil. The inhibitions of Nitrobacter proliferations in soil on the other hand were correlated with the inhibitions of growth and metabolism in pure culture. Within these overall effects, individual herbicides showed marked differential actions on various parameters. Thus, the formulated octanoates of bromoxynil and ioxynil were extremely toxic to the growth of nitrifying organisms in culture, an action probably due to an unknown formulation component. Relative to other herbicides, bromoxynil and ioxynil were more effective on nitrification processes in the soil environment. Both in culture and in the soil, Nitrobacter is more sensitive than Nitrosomonas to these four herbicides. Legurame and oxadiazon are relatively more toxic to Nitrosomonas in culture, but this differential action is not demonstrable in the soil. Dicamba, tricamba, trifluralin, and chlorbufam are more toxic to Nitrobacter than to Nitrosomonas in the soil environment. Formulated trifluralin seems to exert a stimulating action on the growth of nitrifying organisms, but only in the soil; suppression of antagonistic organisms is suggested as a possible cause. Extrapolation of these results to the field situation suggests that the only herbicide which might cause small inhibitions of nitrification at field rates is terbacil, which is disproportionately toxic at low concentrations. At rates somewhat in excess of normal, the formulated octanoates of bromoxynil and ioxynil and possibly dicamba and oxadiazon could also cause small inhibitions.     

Untersuchungen über Sonchus arvensis L. II Transiokation von 14C-MCPA unter verschiedenen Bedingungen

Es wird über Untersuchungen zur Transiokation von ¹⁴C-MCPA unter verschiedenen Bedingungen in Sonchus arvensis L. berichtet. Die Pflanzen wurden von Wurzetausläufern bestimmter Längen angezogen und nur ein Blatt je Pflanze mit markiertem MCPA behandelt. Die Versuche wurden im Gewächaus und im Freiland durchgeführt. Bereits bei kleinen Pflanzen (Rosettenblätter nicht uber 3 cm) wurde ¹⁴C-MCPA innerhalb von 24 h aus dem behandelten Blätt in die anderen Blätter der Rosette transloziert. In die Wurzein wurde dagegen nur wenig Aktivität eingelagert. Die grössten Mengen an ¹⁴C-MCPA in der Wurzelzone wurden am Ende einer fünftägigen Versuchsperiode bei Pflanzen gefunden, die zur Zeit der Behandlung 5–7 bzw. 12–15 cm lange Rosettenblätter entwickelt hatten. Bei einer Sprosshöhe von 15–20 cm wurde ¹⁴C-MCPA aus einem Rosettenblatt und einem nahe dem Blutenstande inserierten Blatt in ungefahr gleichen Mengen in die Wurzein geleitet. Kurz vor der Blüte war dagegen die basipetale Transiokation aus diesem Blatt erheblich reduziert. Vom Rosettenblatt wurde noch ¹⁴C-MCPA in die Wurzein geleitet. Pflanzen im Rosettenstadium, die nach der Ernte in der Stoppel stehen geblieben waren, zeigten eine erhebliche Transiokation von ¹⁴C-MCPA in den Wurzelauslaufer, von dem sie ausgetrieben waren. Die Verlagerung von markiertem MCPA wurde durch Spritzen mit 1 kg/ha nichtradioaktivem MCPA gehemmt—im Gewachshaus mehr als im Feld. Die Kontaktherbizide Dinoseb, Ioxynil und Bromoxynil hemmten in praxisublichen Aufwandmengen die Transiokation des ¹⁴C-MCPA noch starker. Geringe Mengen dieser Herbizide beeinflussten jedoch die Transiokation weniger, forderten sie aber nie. Im Freiland waren gegen Ioxynil und Bromoxynil kleine Pflanzen weniger empflndlich als grossere. Im Gewachshaus aber verhielten sich die Pflanzen umgekehrt. ¹⁴C-MCPA wurde viel langsamer als Assimilate in S. arvensis transloziert und im Laufe von 10 Tagen nahm die Menge an ¹⁴C-MCPA in den Wurzein ständig zu. Ahnlich wie die Assimilate vermochte auch ¹⁴C-MCPA die Insertionsstellen von Nebensprossen an der Mutterwurzel nur in geringem Ausmass zu durchdringen. Research on Sonchus arvensis. L. II. Translocation of¹⁴C-MCPA under various conditions Trials are reported on the translocation of ¹⁴C-MCPA in Sonchus arvensis under various conditions. Plants were grown from creeping root pieces of a prescribed length and only one leaf on each plant was treated with labelled MCPA. Experiments were conducted in the glasshouse and in the open. Even in small plants (rosette leaves not more than 3 cm) ¹⁴C-MCPA was translocated within 24 h from the treated leaf into other leaves in the rosette. In the roots, on the other hand, only slight activity was observed. The largest quantities of ¹⁴C-MCPA in the root zone were found at the end of a 5-day experimental period in plants, which had developed rosette leaves 5–7 and 12–15 cm long respectively at the time of treatment. When shoots were 15–20 cm tall, ¹⁴C-MCPA was carried to the roots in almost equal quantities from a rosette leaf and from a leaf near the flower head. Plants at the rosette stage which had been left in stubble after the harvest showed a considerable translocation of ¹⁴C-MCPA into the creeping roots from which they had grown. The displacement of labelled MCPA was checked by spraying with non-radioactive MCPA at 1kg/ha—this was more marked in the glasshouse than in the open. Contact herbicides dinoseb, ioxynil and bromoxynil at normal rates checked the translocation of ¹⁴C-MCPA to an even greater extent. Low rates of these herbicides had less effect on translocation but they never accelerated it. In the open, small plants were less sensitive to ioxynil and bromoxynil than larger ones, but in the glasshouse the opposite was true. ¹⁴C-MCPA was translocated much more slowly than assimilates in Sonchus arvensis, and in the course of 10 days the quantity of ¹⁴C-MCPA in the roots steadily increased. Like the assimilates, ¹⁴C-MCPA was able to penetrate beyond the insertion of an additional shoot on the main root to only a slight extent. Reeherches sur Sonchus arvensis. II. Migration du¹⁴C-MCPA dans diverses conditions L'auteur expose les essais efl'ectués sur la migration du ¹⁴C-MCPA chez Sonchus arvensis dans diverses conditions. Les plantes ont été obtenues à partir de stolons d'ume longueur fixée à l'avance et une seule feuille de chaque plante a été traitée avee le MCPA marqué. Les expériences ont été réalisées en serre et en plein air. Même chez de petites plantes (dont les feuilles dela rosette n'avaient pas plus de 3 cm), le¹⁴C-MCPA a migré dans les 24 heures depuis la feuille traitée jusque dans les autres feuilles de la rosette. En revanche, une faible activity seulement a été induite dans les racines. Les plus grandes quantités de ¹⁴C-MCPA dans Ia zone des racines, ont été trouves à la fin d'une pèriode experimental de 5 jours, dans les plantes qui avaient développé des feuilles de rosette de 5 à 7 cm et de 12 à 15 cm de haut. Lonsque les pousses élàient hautes de 15 à 20 cm, le ¹⁴C-MCPA a été transporté dans les racines en quantités presque égales à celles qui ont migré d'une feuille de la rosette à une fleur voisine du bouton floral. Dans les plantes au stade rosette qui ont subsisté dans les chaumes après la moisson, la migration du ¹⁴C-MCPA a été considérable dans le stolon à partir duquel elles s'étaient développées. Le déplacement du MCPA marqué a été empêché par une pulvérisation avec du MCPA non radio-actif à 1 kg/ha; cet effet a été plus marqué en serre qu'en plein air. Les herbicides de contact: dinosèbe, ioxynil et bromoxynil aux doses nor-males d'utilisation ont, dans une large mesure, arrêté la migration du ¹⁴C-MCPA. A doses faibles, ces herbicides ont eu moins d'effet sur la migration, mais ils ne l'ont jamais accél-érée. En plein air, les petites plantes ont été moins sensibles que les grandes à l'ioxynil et au bromoxynil, mais en serre, ce fut l'inverse. Le ¹⁴C-MCPA a migré beaucoup plus lentement que les métabolites dans le Sonchus arvensis et pendant une période de 10 jours la quantité de ¹⁴C-MCPA dans les racines a augmenté régulierement. Comme les metabolites, le ¹⁴C-MCPA n'a été capable de pénétrer dans les noeuds des racines adventices sur la tige principale que dans une faible mesure.     

The in vitro degradation of [14C]malathion by enzymatic extracts from resistant and susceptible strains of Drosophila melanogaster

Enzyme preparations from Drosophila melanogaster flies degraded [¹⁴C]malathion to α- and β-malathion monoacids and, hence, were considered to contain malathion carboxylesterase (ME) activity. Although ME- activity was stable during preincubation in the absence of malathion, it decreased dramatically during the course of the reaction, and could not be completely recovered by Sephadex G-25 chromatography. Furthermore, the protein fraction after chromatography still contained ¹⁴C, suggesting that the enzyme had become inhibited by a bound, ¹⁴C-labeled derivative. Extracts from a resistant (malathion-selected), an intermediate control, and the susceptible Canton S strains of D. melanogaster differed in the lability of ME activity during the reaction. This difference was partly attributed to the production of small amounts of malaoxon (2–8%) by the extracts from the more resistant strains. No consistent strain differences were found when the rate of malathion degradation was measured during the first minute of reaction, either with or without a microsomal oxidase inhibitor (metyrapone) present. These results, together with the cross-resistance of the malathion-selected strain to other insecticides and the lack of a synergistic effect of two carboxylesterase inhibitors (triphenyl phosphate and S,S,S-tributylphosphorotrithioate) suggested that malathion carboxylesterase does not contribute significantly to the observed differences in malathion resistance between strains.     

Biochemical identification of putative GABA/benzodiazepine receptors in house fly thorax muscles

[³H]Flunitrazepam ([³H]Flu) was used to identify benzodiazepine binding sites in house fly thorax muscle membranes using a filter assay. [³H]Flu bound to a finite number of sites in a concentration- and time-dependent manner, reaching equilibrium in 10 min. Scatchard plots of the binding indicated a high-affinity site at 0.2 pmol/mg protein (K_d 24.3 nM) and a low-affinity site at 8.2 pmol/mg protein (K_d994nM). Binding of [³H]Flu to the high-affinity binding site was inhibited by several benzodiazepine analogs, with Flu, diazepam, and Ro 5-4864 being more potent than β-CCE, Ro 5-3027, and Ro 5-2180. Clonazepam was least potent in inhibiting [³H]Flu binding. Thus, the drug specificity of these insect muscle benzodiazepine binding sites was quite different from both the mammalian central and peripheral benzodiazepine receptor sites, though closer to the peripheral ones. GABA (γ-aminobutyric acid) and its agonists enhanced the specific binding of [³H]Flu in a dose-dependent manner, and this effect was inhibited with the GABA antagonist bicuculline. The effect was biphasic since at high GABA concentrations this stimulation was reduced. The data suggest that house fly muscles have benzodiazepine receptors, which are coupled allosterically to GABA receptors, analogous to the GABA/benzodiazepine receptors of vertebrates, but with some differences in their drug specificities.     

Studies on the modes of action of n-alkylguanidines and triorganotins on photosynthetic energy conservation in the pea and the unicellular alga Chlamydomonas reinhardi Dangeard

The pea (Pisum sativum L.) and the CW 15 (cell wall-less) strain of the unicellular alga Chlamydomonas reinhardi Dangeard were used to investigate the modes of action of two n-alkylguanidines and several triorganotin salts on reactions associated with photosynthetic energy conservation. The two n-alkylguanidines, n-octylguanidine and n-dodecylguanidine, affected state 3 (ADP stimulated) noncyclic electron transport of pea chloroplasts in a concentration-dependent manner, viz, energy transfer inhibition at low concentrations, uncoupling of electron transport from photophosphorylation at intermediate concentrations, and direct inhibition of electron transport at relatively high concentrations. The Ca²⁺-dependent ATPase (EC 3.6.1.5) activity of C. reinhardi chloroplast fragments was inhibited by both n-alkylguanidines over concentration ranges similar to those required to affect state 3 electron transport in pea chloroplasts. Dodecylguanidine required lower concentrations than octylguanidine to produce similar effects in both assays. The triorganotins produced uncoupler-reversible inhibition of the state 3 electron transport of pea chloroplasts in a sucrose medium and uncoupled the electron transport in a medium containing high chloride concentrations, as shown by their stimulation of both the light-induced transmission changes and the decay of the light-induced pH gradients of chloroplast suspensions. The inhibition of the Ca²⁺-dependent ATPase activity of C. reinhardi by most of the triorganotins was correlated to their inhibition of the growth of the alga on agar medium under different trophic conditions. The efficacies of the tri-n-alkyltins in all the assays used generally increased along the homologous series.     

N-Dealkylation of atrazine and simazine in Senecio vulgaris biotypes: A major degradation pathway

Separate populations of Senecio vulgaris were found that evolved partial tolerance to s-triazine herbicides and others that were totally resistant (plastid resistance). In plants from the susceptible, tolerant, and resistant populations, about one half of applied [¹⁴C]atrazine (2-chloro-4-ethylamino-6-isopropylamino-s-triazine) was rapidly N-dealkylated to the des-ethyl and des-isopropyl products. These products were relatively inactive in inhibiting photosystem II and did not compete with atrazine. After 6 days, less than 25% of the applied [¹⁴C]atrazine was metabolized to water-soluble degradation products but they were not 2-hydroxy derivatives. Less than 2% of the applied atrazine was incorporated into methanol-insoluble residues. The results on metabolism do not explain the tolerance of Senecio to atrazine. However, our results show that N-dealkylation of the s-triazines is more active than previously reported.     

Penetration and metabolism of topically applied permethrin and cypermethrin in pyrethroid-tolerant Wiseana cervinata larvae

The penetration, excretion, and metabolism of topically applied [¹⁴C]permethrin and [¹⁴C]cypermethrin have been examined in larvae of the porina moth Wiseana cervinata to determine the factors which affect body levels of unchanged pyrethroids. Metabolism was by hydrolysis and to a lesser extent oxidation and the primary metabolites were quickly conjugated to water-soluble products. Little excretion occurred and body levels of unchanged pyrethroids were dependent on the interaction of penetration and metabolism. cis-Cypermethrin was more resistant to metabolism than trans-cypermethrin and cis- and trans-permethrin. trans-Permethrin most readily penetrated into larvae. The body levels of unchanged permethrin were enhanced by pretreatment of larvae with the metabolic inhibitors carbaryl or piperonyl butoxide. Tolerance of the pasture pest porina to the synthetic pyrethroids is discussed in relation to these findings.