Bioassays using detached tobacco leaves to determine the sensitivity of Peronospora tabacina to fungicides

Two bioassay methods are described which use detached tobacco leaves to measure the sensitivity of Peronospora tabacina to systemic fungicides. Tobacco leaves (13–15 cm²), treated with fungicides before or after detachment from the plant, were inoculated with sporangia in water drops and, after incubation in beakers and Petri plates, the disease severity and/or production of sporangia was determined 4–7 days after treatment with the fungicides. Of 15 systemic fungicides applied to detached leaves, eight N-phenylamides at 0.066?1.0 μg ml^?1 controlled blue mould; metalaxyl was the most effective fungicide. Isolates of P. tabacina, collected in the field from tobacco plants grown in soil treated with metalaxyl, were not resistant to the fungicide applied to detached leaves prior to inoculation. The fungicide, applied to leaves before detachment, was used to measure the efficacy of five systemic N-phenylamide fungicides sprayed on the basal and unsprayed distal portions of the leaves. Blue mould was controlled on the basal portion of the leaf by all the fungicides at 0.66?1.0 μg ml^?1, but it required the application of 3–30 times more chemical on the basal portion to achieve comparable blue mould control on the distal part of the leaf.     

Studies on the systemicity of [14C]metalaxyl in cowpea (vigna unguiculata (L.) walp.)

[¹⁴C]metalaxyl applied to seeds, roots and leaves of cowpea was readily taken up by all plant parts and a major fraction of it was retained within the treated leaves and seeds but not in the roots. The fungicide exhibited ambimobility inside the plant. When applied to the middle of leaflets, metalaxyl mobilised towards proximal and distal parts of the leaflet. Intraleaflet movement seemed to result from translocation through veins and diffusion through cell walls and intercellular spaces. Accumulation of metalaxyl at tips and margins of primary leaves followed application to roots. Metalaxyl was readily extracted from most plant parts except cotyledons and only 0.8% of the fungicide applied as a seed treatment was recovered from 50-day-old plants.     

Studies on the mode of action of asulam,aminotriazole and glyphosate in Equisetum arvense L. (field horsetail). I: The uptake and translocation of [14c]asulam, [14C]aminotriazole and [14C]glyphosate

The uptake and translocation of [¹⁴C]asulam (methyl 4-aminophenyl-sulphonylcarbamate), [¹⁴C]aminotriazole (1-H-1,2,4-triazol-3-ylamine) and [¹⁴C]glyphosate (N-(phosphonomethyl)glycine) were assessed in Equisetum arvense L. (field horsetail), a weed of mainly horticultural situations. Under controlled-environment conditions, 21°C day/18°C night and 70% r. h., the test herbicides were applied to 2-month-old and 2-year-old plants. Seven days following the application of 0.07-0.09 °Ci (1.14mg) of the test herbicides to young E. arvense, the accumulation of ¹⁴C-label (as percentage of applied radioactivity) in the treated shoots, untreated apical and basal shoots was as follows: [¹⁴C]asulam, 13.2, 0.18 and 1.02%; [¹⁴C] aminotriazole, 67.2, 3.65 and 1-91%; [¹⁴C]glyphosate, 35.9, 0.06 and 0.11%. The equivalent mean values for the accumulation of ¹⁴C-label in 2-year-old E. arvense were [¹⁴C]asulam, 12.0, 1-15 and 1.74%; [¹⁴C]aminotriazole, 58.6, 9.44 and 4.12%; [¹⁴C]glyphosate, 33.1, 0.79 and 2.32%. In the latter experiment, test plants received 0.25-0.30 °Ci (4mg) of herbicide, they were assessed after a 14-day period and the experiment was carried out at 3-week intervals between 2 June and 25 August on outdoor-grown plants. Irrespective of test herbicide or time of application, very low levels of ¹⁴C-label accumulated in the rhizome system. Only 0.2% of the applied radioactivity was recovered in 2-year-old plants and 0.4% in 2-month-old plants. In the young plants [¹⁴C]asulam accumulated greater amounts and concentrations of ¹⁴C-label in the rhizome apices and nodes than [¹⁴C]aminotriazole or [¹⁴C]glyphosate treatments. Inadequate control of E. arvense under field conditions may be due to limited basipetal translocation and accumulation of the test herbicides in the rhizome apices and nodes.     

Temperature effects on the action of acylurea insecticides against tobacco hornworm (Manduca sexta) larvae

The toxic effects of six acylurea insecticides on larvae of the tobacco hornworm were investigated at each of four environmental temperatures (20, 25, 30 and 35°C). This spans the range of temperatures which the insects can tolerate. For all the acylureas tested, mortality increased with temperature when either newly hatched or fourth-instar larvae were given insecticide in their food. Sub-lethal growth inhibition also became more pronounced at progressively higher environmental temperatures. This temperature dependence of acylurea action was not due to altered uptake of the insecticide, since there was no significant variation with temperature in the amount of [¹⁴C]flufenoxuron taken up by fifth-instar larvae when given a single meal containing labelled insecticide. Additionally, mortality of fourth-instar larvae given a single intra-haemocoelic injection of flufenoxuron was significantly greater at higher temperatures, implying that temperature affects a process that occurs after insecticide uptake. The intrinsic ability of acylureas to inhibit chitin synthesis is temperaturesensitive, since flufenoxuron inhibited the incorporation of [¹⁴C]N-acetylalucosamine into chitin by proleg epidermis in vitro significantly less well at 20°C than at the higher temperatures tested. However, there was no significant variation between the effectiveness of in-vitro chitin synthesis inhibition at 25, 30 and 35°C. These data show that the effectiveness of acylurea insecticides is subject to strong temperature effects in the range of temperatures likely to be experienced in the field.     

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.     

Resistance to acylalanines in Pseudoperonospora cubensis1

Resistance to acylalanine fungicides in Pseudoperonospora cubensis, the downy mildew of cucurbits, is reviewed. Insensitive strains of the fungus predominated during the second year of metalaxyl application in cucumber glasshouses in Greece and Israel. Resistance was detected either on detached cucumber leaves floated on metalaxyl solutions or on potted plants treated (sprayed or drenched) with this fungicide. Metalaxyl-resistant strains of P. cubensis showed good pathogenicity and fitness, and competed favorably with the sensitive strains in the absence of the fungicide. A synergism between these two biotypes resulted in an increased virulence of the resistant forms on metalaxyl-treated plants. Metalaxyl-resistant strains of the pathogen exhibited cross resistance to other acylalanine fungicides. Strains resistant to acylalanines and to the chemically unrelated oomycete-fungicides fosetyl-Al and propamocarb were isolated in Israel, as well. In Greece, however, protective sprays with fosetyl-Al, cymoxanil and mancozeb gave good control of cucumber downy mildew when the pathogen was resistant to metalaxyl. P. cubensis is the first case of fungus which developed resistance to acylalanines in the field.     

Relationship between the Spray Droplet Density of Two Protectant Fungicides and the Germination of Mycosphaerella fijiensis Ascospores on Banana Leaf Surfaces

The effect of fungicide spray droplet density (droplet cm^-2), droplet size, and proximity of the spray droplet deposit to fungal spores was investigated with Mycosphaerella fijiensis ascospores on the banana (Musa AAA) leaf surface for two contact fungicides: chlorothalonil and mancozeb. When droplet size was maintained at a volume median diameter (VMD) of 250 μm while total spray volume per hectare changed, M. fijiensis ascospore germination on the leaf surface fell below 1% for both fungicides at a droplet deposit density of 30 droplet cm^-2. At a droplet deposit density of 50 droplet cm^-2, no ascospores germinated in either fungicide treatment. When both droplet size and droplet cm^-2 varied while spray volume was fixed at 20 litre ha^-1, ascospore germination reached 0% at 10 droplet cm^-2 (VMD=602 μm) for both fungicides. At lower droplet densities (2–5 droplet cm^-2 VMD=989 μm and 804 μm respectively), ascospore germination on the mancozeb-treated leaves was significantly lower than on the chlorothalonil-treated leaves. The zone of inhibition surrounding a fungicide droplet deposit (VMD=250 μm) on the leaf surface was estimated to extend 1·02 mm beyond the visible edge of the spray droplet deposit for chlorothalonil and 1·29 mm for mancozeb. The efficacy of fungicide spray droplet deposit densities which are lower than currently recommended for low-volume, aerial applications of protectant fungicides was confirmed in an analysis of leaf samples recovered after commercial applications in a banana plantation. Calibrating agricultural spray aircraft to deliver fungicide spray droplets with a mean droplet deposit density of 30 droplet cm^-2 and a VMD between 300 and 400 μm will probably reduce spray drift, increase deposition efficiency on crop foliage, and enhance disease control compared to aircraft calibrated to spray finer droplets. © 1997 SCI.     

Uptake and translocation of carpropamid in rice (Oryza sativa L)

Translocation of the antiblast compound, carpropamid, was investigated in rice using [¹⁴C]carpropamid. When applied to the seed, carpropamid was not only readily absorbed but was translocated to different parts of the seedlings emerging from treated seeds. A substantial portion of fungicide appeared to be exuded onto the leaf surface. In 21‐day‐old plants grown from [¹⁴C]carpropamid‐treated seeds, 27.2% of the radioactivity isolated from leaves was present on the surface of lamina. This exuded fraction is probably responsible for its action as a fungal anti‐penetrant compound. Following 30‐min root dipping of 14‐day‐old seedlings, carpropamid was rapidly absorbed and translocated throughout the seedling. Its intra‐laminar distribution was uniform as determined by autoradiography. Only a small fraction (<2%) of fungicide applied to the foliage was translocated beyond the site of application within the treated leaf. Translocation was primarily apoplastic. Approximately 54% of the radioactivity recovered from leaves was in the form of carpropamid. At least seven radiolabelled metabolic products were observed by TLC. Only 8.3% of radioactivity applied through the seeds could be recovered from 21‐day‐old seedlings. © 2001 Society of Chemical Industry     

Estimating benzimidazole residues in thatch and turfgrass by bioassay

Bioassays using pellets of agar, thatch-agar and turfgrass-agar were developed using benzimidazole-sensitive Penicillium expansum Link, to detect the fungicide methyl benzimidazol-2-ylcarbamate (MBC) which is the major fungitoxic degradation product of benomyl [methyl 1-(butylcarbamoyl)benzimidazol-2-ylcarbamate] in thatch and turfgrass clippings. These bioassays were used to estimate the amount of fungicide that was biologically available and hence, by subtraction from that applied, the amount that remained bound and biologically unavailable. The limit of quantitation was 0·5 mg kg⁻¹. From 19·9% to 93·2% of the applied fungicide was bound by thatch and 46·2% to 56·9% was bound to turfgrass clippings depending on the concentrations used. In-vitro degradation studies showed that MBC had a half life of approximately 2·5 weeks at 23°C in non-sterilized thatch.     

Antagonistic effects of 2, 4-D), MCPA and MCPB on uptake and translocation of haloxyfop-ethoxyethyl in oat (Avena sativa L.)

The antagonism of haloxyfop-ethoxyethyl (HE) by selected phenoxy herbicides was evaluated through studies of the foliar absorption and translocation of ¹⁴C]HE in oat (Avena sativa L.). Uptake of [¹⁴C]HE, from simultaneous application in mixture with a phenoxy herbicide, was inhibited by the latter in the order MCPB MCPA2,4-D. In mixtures, the foliar absorption of [¹⁴C]HE was reduced more by salts of the phenoxy herbicides than by the corresponding butyl esters. 2,4-D-butyl enhanced uptake of [¹⁴C]HE. The application rate of phenoxy herbicides (from 0.5 to 1.5 kg a.e. ha^?1) did not affect the uptake of [¹⁴]HE, but did influence translocation. Movement of [¹⁴C]herbicide out of the treated leaf was less than 5% of the total ¹⁴C applied; translocation was significantly reduced by all phenoxy herbicides and was antagonized most by 2,4-D-salt and least by MCPB-butyl. Phenoxy salts invariably reduced [¹⁴C]HE translocation more than the corresponding butyl esters. Prior application of phenoxy salts reduced uptake of [¹⁴C]HE, but this antagonism was reduced as the time interval between spray applications increased. Translocation of ¹⁴C out of the treated leaf was antagonized most by prior application of 2,4-D, and by phenoxy salt formulations. When applied up to 2 days after HE, phenoxy salts reduced uptake, but translocation of ¹⁴C was generally unaffected. Les effets antagonistes du 2,4-D, du MCPA et du MCPB sur la pénétration et la migration de l'haloxyfop-éthoxyéthyl dans l'avoine (Avena sativa L.) L'effet antagoniste de plusieurs herbicides de type phénoxy à l'égard de l'haloxyfopéthoxyéthyl (HE) a étéétudié dans des études de pénétration foliaire et de migration du [¹⁴C]HE chez l'avoine (Avena sativa L.) Lorsqu'il est appliqué en mélange avec un herbicide phénoxy, la pénétration du [¹⁴C]HE est inhibée dans l'ordre suivant: MCPB MCPA 2,4-D. La pénétration foliaire du [¹⁴C]HE était davantage réduite par les sels d'herbicides phénoxys que par les esters butyles correspondants. Le 2,4-D butyle augmentait la pénétration du [¹⁴C]HE. La dose d'herbicides phénoxys (de 0,5 à 1,5 kg m.a. ha^?1) n'affectait pas la pénétration de [¹⁴C]HE mais modifiait sa migration. La migration d'herbicide ¹⁴C hors de la feuille traitée était inférieure à 5 % de la radioactivité appliquée. Elle était significativement réduite par tous les herbicides phénoxys, le plus par le sel de 2,4-D et le moins par le MCPB-butyle. Les phénoxys sous forme de sels diminuaient toujours la migration du [¹⁴C]HE davantage que les esters butyles correspondants. Si l'application de phénoxys sous forme de sel précédait celle de [¹⁴C]HE, sa pénétration était réduite mais cet antagonisme était réduit lorsque l'intervalle de temps entre les deux applications était augmenté. La migration de ¹⁴C hors de la feuille traitée était le plus diminuée par le 2,4-D et par les phénoxys sous forme de sels. Quand ils étaient appliqués jusqu'à deux jours après [¹⁴C]HE, les phénoxys sous forme de sel réduisaient sa pénétration, mais la migration de ¹⁴C n'était généralement pas affectée. Antagonistische Wirkung von 2,4-D, MCPA und MCPB auf die Aufnahme und Translokation von Haloxyfop-ethoxyethyl in Hafer (Avena sativa L.) Die antagonistische Beeinflussung von Haloxyfop-ethoxyethyl (HE) durch ausgewählte Phenoxy-Herbizide wurde anhand der Blattaufnahme und Translokation von [¹⁴C]HE in Hafer (Avena sativa L.) untersucht. Die Aufnahme von [¹⁴C]HE bei gleichzeitiger Anwendung in Mischung mit einem Phenoxy-Herbizid wurde durch die letztgenannten Stoffe in der Reihenfolge MCPB MCPA 2,4-D gehemmt, wobei die Salz-Verbindungen stärker wirkten als die entsprechenden Butylester. 2,4-D-butyl förderte die Aufnahme von [¹⁴C]HE. Die Aufwandmenge der Phenoxy-Herbizide (0,5 bis 1,5 kg AS ha^?1) blieb ohne Einflus auf die Aufnahme von [¹⁴C]HE, beeinflußte aber die Translokation. Aus den behandelten Blättern wurde weniger als 5 % der gesamten [¹⁴C]Menge transloziert; die Translokation wurde durch alle PhenoxyHerbizide signifikant reduziert, am meisten durch 2,4-D-Salz, am wenigsten durch MCPB-butyl. Die Salz-Verbindungen verminderten die [¹⁴C]HE-Translokation mehr als die entsprechenden Butylester. Eine vorausgehende Behandlung mit den Salz-Verbindungen senkte die Aufnahme von [¹⁴C]HE, aber mit zunehmender Zeit zwischen den Anwendungen nahm dieser Antagonis mus ab. Hierbei war der Einfluß von 2,4-D und von den Salz-Verbindungen am stärksten. Wurden diese Stoffe bis zu 2 Tagen nach HE ausgebracht, beeinträchtigten sie die Aufnahme, jedoch im allgemeinen nicht die Translokation von ¹⁴C.     

Inhibition of Botrytis cinerea growth and suppression of botrytis bunch rot in grapes using chitosan

Chitosan inhibited growth of Botrytis cinerea in liquid culture and suppressed grey mould on detached grapevine leaves and bunch rot in commercial winegrapes. Germination of B. cinerea was completely inhibited in malt extract broth containing chitosan at concentrations greater than 0·125 g L^?1. However, treated conidia were able to infect detached Chardonnay leaves and pathogenicity was not affected, even after incubation for 24 h in chitosan at 10 g L^?1. When added after conidial germination, chitosan inhibited B. cinerea growth and induced morphological changes suggestive of possible curative activity. The effective concentration of chitosan that reduced mycelial growth by 50% (EC₅₀) was 0·06 g L^?1. As a foliar treatment, chitosan protected detached Chardonnay leaves against B. cinerea and reduced lesion diameter by up to 85% compared with untreated controls. Peroxidase and phenylalanine ammonia‐lyase activities were also induced in treated leaves. In vineyard studies, Chardonnay winegrapes exhibited 7·4% botrytis bunch rot severity at harvest in 2007 after treatment with an integrated programme that included chitosan sprays from bunch closure until 2 weeks preharvest, compared with 15·5% in untreated controls and 5·9% with fungicide treatment. In the following season, botrytis bunch rot severity was 44% in untreated Chardonnay at harvest and the integrated programme (21%) was less effective than fungicides (13·8%). However, in Sauvignon blanc winegrapes, the integrated and the fungicide programme each reduced botrytis bunch rot severity to 4% and were significantly different from the untreated control (11·5%). This study provides evidence that suppression of botrytis in winegrapes by chitosan involves direct and indirect modes of action.     

Some physicochemical factors influencing foliar uptake enhancement of glyphosatemono(isopropylammonium) by polyoxyethylene surfactants

Structure-concentration–foliar uptake enhancement relationships between commercial polyoxyethylene primary aliphatic alcohol (A), nonylphenol (NP), primary aliphatic amine (AM) surfactants and the herbicide glyphosatemono(isopropylammonium) were studied in experiments with wheat (Triticum aestivum L.) and field bean (Vicia faba L.) plants growing under controlled-environment conditions. Candidate surfactants had mean molar ethylene oxide (EO) contents ranging from 5 to 20 and were added at concentrations varying from 0·2 to 10 g litre^?-1 to [¹⁴C]glyphosate formulations in acetone–water. Rates and total amounts of herbicide uptake from c. 0·2–μl droplet applications of formulations to leaves were influenced by surfactant EO content, surfactant hydrophobe composition, surfactant concentration, glyphosate concentration and plant species, in a complex manner. Surfactant effects were most pronounced at 0·5 g acid equivalent (a.e.) glyphosate litre^?-1 where, for both target species, surfactants of high EO content (15–20) were most effective at enhancing herbicide uptake: surfactants of lower EO content (5–10) frequently reduced, or failed to improve, glyphosate absorption. Whereas, at optimal EO content, AM surfactants caused greatest uptake enhancement on wheat, A surfactants gave the best overall performance on field bean; NP surfactants were generally the least efficient class of adjuvants on both species. Threshold concentrations of surfactants needed to increase glyphosate uptake were much higher in field bean than wheat (c. 2 g litre^?-1 and < 1 g litre^?-1, respectively); less herbicide was taken up by both species at high AM surfactant concentrations. At 5 and 10 g a.e. glyphosate litre^?-1, there were substantial increases in herbicide absorption and surfactant addition could cause effects on uptake that were different from those observed at lower herbicide doses. In particular, the influence of EO content on glyphosate uptake was now much less marked in both species, especially with AM surfactants. The fundamental importance of glyphosate concentration for its uptake was further emphasised by experiments using formulations with constant a.i./surfactant weight ratios. Any increased foliar penetration resulting from inclusion of surfactants in 0·5 g litre^?-1 [¹⁴C]glyphosate formulations gave concomitant increases in the amounts of radiolabel that were translocated away from the site of application. At these low herbicide doses, translocation of absorbed [¹⁴C]glyphosate in wheat was c. twice that in field bean; surfactant addition to the formulation did not increase the proportion transported in wheat but substantially enhanced it in field bean.     

DETOXIFICATION OF FLUOMETURON BY CITRUSTISSUES*

Summary. The metabolism of [¹⁴C]fluometuron in Citrus was studied by feeding the herbicide to either young seedlings or to excised organs. Most of the uptake of fluometuron occurred via the roots during the first 24 h and radioactivity was found after 16 days to be in the rootlets (36·5%), mainroot (34·5%), stem (13·7)% and leaves (15·2%). By feeding [¹⁴C]fluometuron to excised organs it was established that although most of the fluometuron breakdown occurred in the rootlets, other plant parts were also capable of metabolizing the herbicide. Therefore, the presence of metabolites in the upper plant organs was not entirely due to translocation from the rootlets. These results suggest that the resistance of Citrus to fluometuron is due to its breakdown in the tissues, probably induced by an N-demethylase enzyme system, similar to that reported for cotton (Frear, Swanson & Tanaka, 1969), in which harmless metabolites arc formed. Détoxification du fluométuron par les tissus de Citrus     

Surfactant and salt affect glyphosate retention and absorption

The influence of nonylphenoxy surfactants and glyphosate salt formulation on spray retention, phytotoxicity and [¹⁴C]glyphosate uptake was investigated in wheat (Triticum aestivum L). and Kochia scoparia L. The amount of spray retained, and uptake of [¹⁴C]glyphosate increased with increasing hydrophilic-lipophilic balance (HLB) value of surfactants. The volume of spray delivered to the plant treatment area and retained by wheat and K. scoparia plants increased with increasing surfactant HLB values, but this only partly accounted for the higher spray retention. Spray retention by leaves of plants was not affected by calcium chloride, either alone or with ammonium sulphate in the glyphosate spray solution. [¹⁴C]Glyphosate absorption by wheat and K. scoparia was reduced by calcium chloride alone, but not in mixtures with ammonium sulphate, regardless of surfactant. Phytotoxicity and uptake of glyphosate salt formulations for wheat was: isopropylamine > ammonium > sodium > calcium; these results indicate that the surfactant selected is important to maintain glyphosate efficacy and that sodium and calcium cations antagonize glyphosate by forming salts that are absorbed less than commercial isopropylamine formulations.     

Development of an effective screening method for partial resistance to Alternaria brassicicola (dark leaf spot) in Brassica rapa

In order to develop a method to measure resistance to Alternaria brassicicola (cause of dark leaf spot disease) in Brassica rapa, the effects of inoculum concentration, leaf stage, leaf age and incubation temperature of inoculation on infection were studied under controlled conditions using several B. rapa genotypes. Three inoculation methods (cotyledon, detached leaf and seedling inoculation) were evaluated for this purpose. The detached leaf inoculation test was the most suitable for screening B. rapa genotypes because clear symptoms were observed on the leaves in less than 24 h, and there was a significant positive correlation between the results from the detached leaf inoculation test and the seedling inoculation test, an established method considered to yield reliable results. In addition, it was very easy to screen plants for resistance on a large scale and to maintain standard physical conditions using detached leaves. For successful infection, inoculum concentration should be adjusted to 5 × 10⁴ conidia ml⁻¹, and incubation temperature should be between 20 °C and 25 °C. The 3rd/4th true leaves from 30 day-old plants were optimal for inoculation. In a screening test using 52 cultivars of B. rapa, the detached leaf test effectively discriminated between various levels of partial resistance among cultivars. As a result, we identified two cultivars, viz Saori and Edononatsu, as highly resistant and five cultivars, viz Tokinashi Taisai, Yajima Kabu, Purara, Norin-F₁-Bekana and Tateiwa Kabu, as having borderline resistance.     

Aerobic soil metabolism of metsulfuron-methyl

A laboratory study was conducted to determine the degradation rates and identify major metabolites of the herbicide metsulfuron-methyl in sterile and non-sterile aerobic soils in the dark at 20°C. Both [phenyl-U-¹⁴C]- and [triazine-2-¹⁴C]metsulfuron-methyl were used. The soil was treated with [¹⁴C]metsulfuron-methyl (0.1 mg kg⁻¹) and incubated in flow-through systems for one year. The degradation rate constants, DT₅₀, and DT₉₀ were obtained based on the first-order and biphasic models. The DT₅₀ (time required for 50% of applied chemical to degrade) for metsulfuron-methyl, estimated using a biphasic model, was approximately 10 days (9–11 days, 95% confidence limits) in the non-sterile soil and 20 days (12–32 days, 95% confidence limits) in the sterile soil. One-year cumulative carbon dioxide accounted for approximately 48% and 23% of the applied radioactivity in the [phenyl-U-¹⁴C] and [triazine-2-¹⁴C]metsulfuron-methyl systems, respectively. Seven metabolites were identified by HPLC or LC/MS with synthetic standards. The degradation pathways included O-demethylation, cleavage of the sulfonylurea bridge, and triazine ring opening. The triazine ring-opened products were methyl 2-[[[[[[[(acetylamino)carbohyl]amino]carbonyl]amino] carbonyl]-amino]sulfonyl]benzoate in the sterile soil and methyl 2-[[[[[amino[(aminocarbonyl)imino]methyl] amino]carbonyl]amino]sulfonyl]benzoate in the non-sterile soil, indicating that different pathways were operable. © 1999 Society of Chemical Industry     

Uptake and translocation of benomyl and carbendazim (methyl benzimidazol-2-yl carbamate) in the symplast

Cytoplasmic uptake of carbendazim (methyl benzimidazol-2-yl carbamate; MBC) from an aqueous solution was demonstrated with isolated mesophyll cells. About 2.5% of the labelled MBC (ring-2-[¹⁴C]) in the treatment solution (1.85 μg/ml) was taken up in 44 h. When cotyledons of cucumber seedling were treated with either 347 or 36 μg [¹⁴C]-MBC/plant 1.11 and 0.13% were extracted, respectively, from the remainder of the plant, 5 days after treatment. Greatest amounts were detected in shoot apices. Likewise, when MBC and benomyl were applied at the dose of 2 μmol, 0.34 and 0.57% were detected in the untreated part of the plant with a bioassay procedure. Foliar application with 347 or 36 μg[¹⁴C]-MBC/leaf resulted in the translocation of 1.68 and 0.11% out of the treated area. By scalding the living cells of the petiole translocation was prevented suggesting that long distance movement occurred in the symplast. During a period of 14 days 1.56% of [¹⁴C]-MBC applied to cucumber leaves was metabolised and respired as CO₂. This degradation was assumed to occur enzymically within the symplast.     

Studies on the mode of action of asulam in bracken (Pteridium aquilinum L. Kuhn) I. Absorption and translocation of [14C]asulam

Studies of the absorption and translocation of foliage-applied ring-labelled [¹⁴C]asulam [methyl (4-aminobenzenesulphonyl) carbamate] were carried out using glasshouse and field-grown bracken plants. Translocation of ¹⁴C from the treated frond was primarily according to a 'source to sink’pattern with intense accumulation of radioactivity in the metabolically active sinks viz. rhizome apices, frond buds, root tips and young frond tissue. In the case of field bracken, translocation and distribution of ¹⁴C was extensive in the rhizome system, accumulation occurring in the active as well as dormant buds situated on the non-frond-bearing and storage rhizome branches. Treatment of fully expanded fronds with 100μl of [¹⁴C]asulam (1 mg, 1.0–1.5 μCi) as 2 μl droplets resulted in a rapid initial uptake during the first week, followed by progressive entry and distribution with time. Basipetal translocation to the rhizome system was positively correlated with total uptake. High humidity (95%) and high temperature (30°C) stimulated uptake and subsequent basipetal translocation to a considerable degree. Uptake was greater through the stomatal-bearing abaxial than through the adaxial cuticle. Incorporation of a surfactant (Tergitol-7, 0.1%) increased penetration by up to 30%. Uptake declined markedly as the frond aged, while translocation was predominantly acropetal in young treated fronds, becoming exclusively basipetal when the fronds matured. Optimum uptake and maximum distribution of [¹⁴C]asulam in the rhizome and its associated buds was achieved when treatments were applied to almost fully expanded fronds. The translocated ¹⁴C (asulam and possibly some of its metabolites) showed a considerable degree of persistence in the rhizome system, 8% of the applied activity still remaining in the rhizome 40 weeks after treatment.     

The metabolism of fenvalerate in plants: The conjugation of the acid moiety

The metabolism of the pyrethroid insecticide fenvalerate [(RS)-α-cyano-3-phenoxybenzyl (RS)-2-(4-chlorophenyl)-3-methylbutyrate] ( I ), and of its most insecticidal (αS,2S) isomer ( II ), has been examined in cabbage plants grown and treated under laboratory conditions with [¹⁴C]chlorophenyl- and [ring-¹⁴C]benzyllabelled preparations of the two compounds. Both insecticides disappeared from the treated leaves with similar half-lives of approximately 12–14 days; they underwent ester cleavage to a significant extent, together with some hydroxylation at the 2- or 4-position of the phenoxy ring, and hydrolysis of the nitrile group to amide and carboxyl groups. Most of the carboxylic acids and phenols thus produced occurred as glycoside conjugates. In separate experiments, the uptake and metabolism of 2-(4-chlorophenyl)-3-methylbutyric acid ( X ), the acidic half of the molecule, were examined in the laboratory, using abscised leaves of kidney bean, cabbage, cotton, cucumber and tomato plants. The acid X was found to be readily converted, mainly into glucose and 6-O-malonylglucose esters in kidney bean, cabbage and cucumber plants, into glucosylxylose, sophorose and gentiobiose esters in cotton, and into two types of triglucose esters with differing isomerism in tomato. One of the acetyl derivatives of the trisaccharide conjugates was identical with the synthetic deca-acetyl derivative of the [1 → 6]-triglucose ester.     

A study of ethylene and CO2 evolution from ethephon in tobacco

Buffers and leaf discs of mature tobacco (Nicotiana tabacum L.) were utilized to study [¹⁴C]-ethylene and ¹⁴CO₂ evolution from radiolabeled ethephon, (2-chloroethyl)phosphonic acid. Metabolic fate of [¹⁴C]ethephon in leaf discs was investigated by use of thin-layer chromatography, high-voltage paper electrophoresis, autoradiography, and liquid scintillation spectroscopy. The evolution of labeled ethylene generally increased with increasing buffer pH, buffer volume, and dosage of [¹⁴C]ethephon. [¹⁴C]Ethylene was evolved, increasingly with time, from [¹⁴C]ethephon either added to the buffer or applied to leaf discs. The rate of [¹⁴C]ethylene evolution was maximum during the first day and leveled off on the fourth day. More than 50% of the total [¹⁴C]ethylene evolution over a 96-hr period was recovered during the first 24 hr after [¹⁴C]ethephon application. No ¹⁴CO₂ was evolved when [¹⁴C]ethephon was degraded in the presence of buffer or leaf discs. Only ethephon itself, and no detectable metabolite thereof, was discovered in the methanolic extract of the leaf disc tissue. An insignificant amount of ¹⁴C activity (approximately 2% of the extracted ¹⁴C) was detected in the residue. By means of gas chromatography, it was confirmed that in buffers and tobacco leaf tissue ethephon breaks down to release ethylene but not CO₂.