Characterization of acetylcholinesterases of acaricide-resistant and susceptible strains of the cattle tick Boophilus microplus (Can.): I. Extraction of the critical component and comparison with enzyme from other sources

The critical acetylcholinesterase component, that is responsible for the resistance mechanism of decreased sensitivity to the inhibitor in the cattle tick Boophilus microplus (Can.), has been isolated from the organophosphorus-susceptible Yeerongpilly and resistant Biarra and Ridgelands strains. With the substrate acetylthiocholine this enzyme component gave a pH activity profile similar for all strains. Optimal substrate concentration for Ridgelands was 1.73.10^?2M compared to 2.70.10^?4M for the other strains. Computed Km values were 1.98.10^?5M, 3.65.10^?5M, and 6.43.10^?6M for Yeerongpilly, Biarra, and Ridgelands strains, respectively, while the corresponding V_max figures were 336, 135, and 21 μmoles acetylthiocholine hydrolyzed per gram of larvae per hour. These results are discussed in relation to the resistance mechanism and compared with similar parameters derived for this enzyme from bovine erythrocytes, the sheep blowfly Lucilia cuprina, and the sheep nematode Trichostrongylus colubtiformis.     

Alfalfa metabolism of propham

Root-treated alfalfa absorbs, translocates, and metabolizes [phenyl-¹⁴C]isopropyl carbanilate ([¹⁴C]propham). After 7 days of root treatment, the distribution of radiolabel was 73% for shoots and 27% for roots. Shoots and roots were extracted and separated into the polar, nonpolar, and solid residual components using a mixture of chloroform, methanol and water. The insoluble residues accounted for approximately 40% of the ¹⁴C found in shoots and roots. The nonpolar fraction (6.1% of the radiolabel in shoots and roots) was not characterized, but was shown to be some component other than parent propham. Propham was not found in either shoots or roots. The polar metabolites were partly purified on Amberlite XAD-2. Cellulase-liberated aglycones were derivatized and separated by high-performance liquid and gas-liquid chromatography. The infrared, nuclear magnetic resonance, and mass spectral data showed that the polar metabolites of alfalfa shoots and roots were glycoside conjugates of isopropyl 2-hydroxycarbanilate (2-hydroxypropham) and isopropyl 4-hydroxycarbanilate (4-hydroxypropham). Conjugated 4-hydroxypropham accounted for 45.9% of the ¹⁴C in the shoots and 3.4% of the ¹⁴C in the roots. Conjugated 2-hydroxypropham accounted for 3.4% of the ¹⁴C in the shoots and 1.4% of the ¹⁴C in the roots.     

Mercapturic acid formation in the metabolism of propachlor,CDAA, and fluorodifen in the rat

When [¹⁴C]F₃-fluorodifen (2,4′-dinitro-4-trifluoromethyl diphenylether), carbonyl-[¹⁴C]CDAA (N,N-diallyl-2-chloroacetamide), and carbonyl-¹⁴C-propachlor (2-chloro-N-isopropylacetanilide) were fed to rats, 57 to 86% of the ¹⁴C was excreted via the urine within 48 hr. Although very little radioactivity was excreted in the feces of CDAA-treated rats, 15–22% of the ¹⁴C was excreted in the feces of propachlor- of fluorodifentreated rats and an average of 8% of the ¹⁴C remained in these rats 48 hr after treatment. Oxidation of the ¹⁴C label to [¹⁴C]O₂ was not a major process in the metabolism of these herbicides. The only major radioactive metabolite present in the 24-h urine of fluorodifen-treated rats, 2-nitro-4-trifluoromethylphenyl mercapturic acid, accounted for 41% of the administered dose of ¹⁴C. In the metabolism of CDAA, the corresponding mercapturic acid accounted for 76% of the dose; it was the only major metabolite present in the 24-h urine. In contrast, three major metabolites were detected in the 24-h urine of propachlortreated rats, and the mercapturic acid accounted for only 20% of the dose. The mercapturic acid of each herbicide was identified by mass spectrometry.     

Antifungal mode of action of triforine

Rapidly growing mycelia of Aspergillus fumigatus treated with 10 μg/ml triforine (N,N′-bis-(1-formamido-2,2,2-trichloroethyl)-piperazine) showed little or no inhibition in dry weight increase prior to 2 h. By 2.5–3 h, triforine inhibited dry weight increase by 85%. The effects of triforine on protein, DNA, and RNA syntheses corresponded to the effect on dry weight increase both in time of onset and magnitude. Neither glucose nor acetate oxidation were inhibited by triforine.Ergosterol synthesis was almost completely inhibited by triforine even in the first hour after treatment. Inhibition of ergosterol synthesis was accompanied by an accumulation of the ergosterol precursors 24-methylenedihydrolanosterol, obtusifoliol, and 14α-methyl-Δ^{8, 24 (28)}-ergostadienol. Mycelia treated with 5 μg/ml of triarimol (α-(2,4-dichlorophenyl)-α-phenyl-5-pyrimidinemethanol) also accumulated the same sterols as well as a fourth sterol believed to be Δ^{5, 7}-ergostadienol.Identification of 4,4-dimethyl-Δ^{8, 24 (28)}-ergostadienol in untreated mycelia indicates that the C-14 methyl group is the first methyl group removed in the biosynthesis of ergosterol by A. fumigatus. The lack of detectable quantities of 4,4-dimethyl-Δ^{8, 24 (28)}-ergostadienol in triforine or triarimol-treated mycelia and the accumulation of C-14 methylated sterols in treated mycelia suggests that both fungicides inhibit sterol C-14 demethylation. The accumulation of Δ^{5, 7}-ergostadienol in triarimol-treated mycelia further implies that triarimol also inhibits the introduction of the sterol C-22(23) double bond.Two strains of Cladosporium cucumerinum tolerant to triforine and triarimol were also tolerant to the fungicide S-1358 (N-3-pyridyl-S-n-butyl-S′-p-t-butylbenzyl imidodithiocarbonate).     

Etude des phénomènes de sorption entre deux triazines herbicides et des acides humiques

A study of sorption phenomena between two triazine herbicides and humic acids Terbutryne is very readily adsorbed by humic acids while atrazine is only slightly adsorbed and this only in an acid environment. The influence of pH on adsorption and the competitive effect of the cations Ca²⁺, Al³⁺ and Fe³⁺ shows that the proton form of the molecules of the two herbicides can be adsorbed by an ion exchange-type mechanism; the neutral form of terbutryne molecules could be adsorbed by other mechanisms. Desorption of terbutryne is accompanied by a more marked hysteresis phenomenon in the case of neutral molecules, and, in an acid environment, calcium shows a weak capacity for displacement in relation lo the adsorbed herbicide.     

Trichoderma harzianum produces nonanoic acid, an inhibitor of spore germination and mycelial growth of two cacao pathogens

An isolate of Trichoderma harzianum Rifai from an infected cacao pod produces and secretes nonanoic (pelargonic) acid into a liquid culture medium. Nonanoic acid (NA) was very inhibitory to spore germination and mycelial growth of two cacao pathogens, Crinipellis perniciosa Stahel and Moniliophthora roreri Cif. H.C. Evans. It was highly active causing 75% inhibition of spore germination in an in vitro assay at a rate as low as 0.09 μM for M. roreri and 0.92 μM for C. perniciosa. Mycelial growth was comparatively less sensitive to inhibition, but still there was a 75% reduction in growth with 0.62 μM in M. roreri and 151 μM NA in C. perniciosa. In contrast, NA did not affect Trichoderma mycelial growth or spore germination at concentrations that were inhibitory to the pathogens. 6-pentyl-α-pyrone was also produced and secreted into the medium by T. harzianum, however; it was not antagonistic to the cacao pathogens. Although a number of metabolites produced by Trichoderma spp. have been identified in the past, this is the first report of NA production and secretion by any Trichoderma. The results suggest that NA may play a role in the successful use of some Trichoderma spp. isolates in the biocontrol of fungal diseases of plants.     

Gesunde Pflanzen unter zukünftigem Klima

Predicted changes in average values of global climate variables (increased temperatures, altered precipitation patterns, increased concentrations of atmospheric CO₂) and changes in the frequency, duration, and degree of extremes (frost, heat, drought, hail, storms, floods, etc.) will affect agricultural crops, agroecosystems, and agricultural productivity. Although forecasts of regional climate changes are still imprecise, mean temperature increases in Europe are expected to be greater in the north (2.5–4.5°C) than in the south (1.5–4.5°C). Regional forecasts for precipitation changes are also very far from precise; however, problems with drought are expected to increase, especially in Mediterranean countries. Overall, shortage of water will be the predominant factor affecting plant growth. As higher temperatures are known to enhance plant development and especially the grain-filling duration of cereals, grain yield losses are possible in a warmer climate. On the other hand, elevated atmospheric CO₂ concentrations are known to stimulate photosynthesis and enhance growth and yield (“CO₂ fertilization”); concomitantly, leaf transpiration is reduced, resulting in improved water use efficiency. Total biomass and yield were enhanced by 20–30% in experiments with elevated CO₂ exposure (550–700 ppm) under more or less ideal growth conditions. Elucidating the interactions between positive and negative effects of climate change is of crucial importance for any prediction of future crop yields. The present paper is a brief summary mainly of the potential effects of elevated temperatures and atmospheric CO₂ on crop growth, quality, and yield. Also, adaptation measures, possible interactive effects of different climate variables, and interactions of climate change components with other growth variables (pathogens, air pollutants) are briefly described.     

Kohlfliegenauftreten im Raps

A new pest in oilseed rape the cabbage root fly (Delia radicum L.) has succeded to establish – at least regional –during the recent years. It is not easy to monitor the cabbage root fly. Although the usual yellow water trap works quite well, it is still difficult for the farmers to determine Delia radicum exactly. The amount of infestation can be assessed best in the middle of November by analysing the damages of the roots. In this article a classification scheme is proposed. It is also difficult to control this pest. While applications of Pyrethroids against the imagines and the use of Dimethoat against the larva mostly showed only little effect in the tests, some new dressing-substances proved better against the standard “Chinook” (Imidacloprid+beta-Cyfluthrin). To forward this research and to clarify factors, which influence the infestation, a project titled “Possibilities of defence and control of cabbage root fly in winter oilseed rape” has been started in November 2003. Eight crop-protection services at German Federal State level and the BBA are involved in this project, which is supported by UFOP.     

Development of an EU protocol for the detection and diagnosis of Potato spindle tuber pospiviroid*

The work described here formed part of the EU SMT DIAGPRO project, to develop diagnostic protocols for 18 regulated pests. The Potato spindle tuber pospiviroid (PSTVd) protocol was developed primarily for testing in vitro‐ and glasshouse‐grown potato plants for the purposes of post‐entry quarantine and the production of pathogen‐tested nuclear stock. After a performance audit of methods used by 12 laboratories in Europe and America by ring testing, four methods were chosen for multilaboratory validation. For most laboratories, the detection limits were 10–20 mg of PSTVd‐infective tissue for R‐PAGE; 0.25–0.5 mg for DIG‐probe; 0.062 mg for RT‐PCR; and 0.0155 mg for TaqMan (this was the lowest weight of infective tissue tested). Some laboratories were able to extend the detection limit to 0.0155 mg for DIG‐probe and RT‐PCR. The DIG‐probe and R‐PAGE are recommended as primary detection methods, with confirmation of viroid presence by any of the four validated detection methods. Specific diagnosis requires the viroid to be sequenced. Other methods may be used for primary detection, providing that they preferably detect all PSTVd isolates and other Pospiviroids that have the potential to infect potato, and detect viroid in at least 1/10 of the tissue weight normally tested per plant.