Mechanisms of resistance to fungicides in field strains of Botrytis cinerea

Field strains of Botrytis cinerea Pers ex Fr, the causal agent of grey mould diseases, were collected from French vineyards between 1993 and 2000. Several phenotypes have been characterized according to the inhibitory effects of fungicides towards germ-tube elongation and mycelial growth. Two types of benzimidazole-resistant strains (Ben R1 and Ben R2) could be detected; negative cross-resistance to phenylcarbamates (e.g. diethofencarb) was only found in Ben R1. Benzimidazole resistance was related to point mutations at codon 198 (Ben R1) or 200 (Ben R2) of the beta-tubulin gene. Most dicarboximide-resistant strains were also weakly resistant to aromatic hydrocarbon fungicides (e.g. dicloran) but remained sensitive to phenylpyrroles (e.g. fludioxonil). These resistant field strains (Imi R1) contained a single base pair mutation at position 365 in a two-component histidine kinase gene, probably involved in the fungal osmoregulation. Three anilinopyrimidine-resistant phenotypes have been identified. In the most resistant one (Ani R1), resistance was restricted to anilinopyrimidines, but no differences were observed in the amino-acid sequences of cystathionine beta-lyase (the potential target site of these fungicides) from Ani R1 or wild-type strains. In the two other phenotypes (Ani R2 and Ani R3), resistance extended to various other groups of fungicide, including dicarboximides, phenylpyrroles and sterol biosynthesis inhibitors. This multi-drug resistance was probably determined by over-production of ATP-binding cassette transporters. The hydroxyanilide fenhexamid is a novel botryticide whose primary target site is the 3-keto reductase involved in sterol C-4 demethylations. Apart from the multi-drug-resistant strain Ani R3, three other fenhexamid-resistant phenotypes have been recognized. For two of them (Hyd R1 and Hyd R2) fenhexamid-resistance seemed to result from P450-mediated detoxification. Reduced sensitivity of the target site could be the putative resistance mechanism operating in the third resistant phenotype (Hyd R3). Increased sensitivity to inhibitors of sterol 14 alpha-demethylase recorded in Hyd R1 strains was related to two amino-acid changes at positions 15 and 105 of this enzyme.     

Effect of acquisition time and chemical fat suppression on meningeal enhancement on MR imaging in dogs

Our purpose was to characterize meningeal gadolinium enhancement on magnetic resonance (MR) imaging in dogs with inflammatory and neoplastic diseases, and to assess interobserver variability and the impact of delayed acquisition and chemical fat saturation on its conspicuity. Transverse T1-weighted FLAIR images were acquired prior to, and immediately following gadolinium injection (T0), and at 5 (T5) and 15-20 min delay (TD), with and without fat suppression, in 155 consecutive dogs imaged for suspected brain disease. The agreement on meningeal enhancement was globally substantial (kappa = 0.61) and the likelihood of obtaining a definite diagnosis was significantly increased with the use of fat suppression (P < or = 0.004). Meningeal enhancement was judged definitively present by consensus in 46 of 155 (30%) dogs. Of these, meningeal enhancement was characterized qualitatively and quantitatively in 30 dogs with a clinical diagnosis (18 inflammatory, 11 neoplastic, 1 infarct), and image sequences were compared. Meningeal enhancement was more often diffuse and leptomeningeal in animals with inflammation versus neoplasia (50% vs. 42%, and 69% vs. 48%, respectively), but significant associations were not found. Meningeal thickness and contrast ratio were higher with neoplasia (P < or = 0.02), but results did not vary significantly between series for either group. Yet, images with fat suppression were most useful 50% of the time for definite diagnosis and/or characterization of meningeal enhancement. While delayed image acquisition following gadolinium injection does not improve characterization of meningeal enhancement in dogs, fat suppression is beneficial qualitatively.     

Transverse shrinkage in G-fibers as a function of cell wall layering and growth strain

Transverse drying shrinkage was measured at microscopic and mesoscopic levels in poplar wood characterised by an increasing growth strain (GS), from normal to tension wood. Results show that: (a) the drying shrinkage, measured as a relative thickness decrease, was significantly higher for G-layer (GL) than for the other layers (OL), GL shrinkage was not significantly correlated with GS, and OL shrinkage was negatively correlated with GS. (b) In gelatinous fibre (G-fibre), lumen size increased during drying and this increase was positively related with GS, but in normal wood fibre, lumen size decreased during drying. These findings suggest that GL shrank outwards (i.e., its internal perimeter increases), so that its shrinkage weakly affected the total cell shrinkage and the mesoscopic shrinkage was controlled by the OL shrinkage which shrank inwards (i.e., its external perimeter decreases). (c) Measurements done on 7 × 7 mm2 thin sections evidenced a negative correlation between transverse shrinkage and GS, significant in T direction but weak in R direction. These observations at both levels allow to discuss the contribution of GL to the mesoscopic shrinkage of tension wood.     

Urodynamic and morphologic changes in the lower portion of the urogenital tract after administration of estriol alone and in combination with phenylpropanolamine in sexually intact and spayed female dogs

OBJECTIVE: To compare the urodynamic and morphologic effects of the administration of estriol alone and in combination with phenylpropanolamine on the lower portion of the urogenital tract in female dogs. ANIMALS: 3 sexually intact and 3 spayed female Beagles without urinary incontinence. PROCEDURE: Dogs received estriol (2 mg, PO) once daily for 7 days followed by estriol (2 mg, PO) and phenylpropanolamine (1.5 mg/kg, PO) once daily for 7 days. Urethral pressure profilometry, diuresis cystometry, and vaginourethrography were performed before treatment (day 0) and at days 7 and 14. The maximum urethral pressure (MUP) and closure pressure (MUCP), urethral functional and anatomic profile lengths, integrated pressure (IP), plateau, distance before MUP, maximum meatus pressure, threshold pressure, threshold volume, compliance, urethral length, and vaginal length and width were measured. RESULTS: Before treatment, no urodynamic differences were observed between the 2 groups; however, vaginal length and width were significantly shorter in spayed dogs. Compared with day 0 values, estriol treatment significantly increased MUP, MUCP, and IP values at day 7, but at day 14, this effect decreased despite phenylpropanolamine administration. No morphologic changes from baseline were detected after either treatment in any dog. CONCLUSIONS AND CLINICAL RELEVANCE: Data suggest that estriol mainly acts on the urethral sphincter mechanism by increasing urethral resistance in sexually intact and spayed female dogs without urinary incontinence. Administration of estriol and phenylpropanolamine did not increase the urethral resistance more than estriol alone. The urodynamic effects of estriol in female dogs with urinary incontinence remain to be elucidated.     

Can light intensity influence the tolerance of Synedrellopsis grisebachii to glyphosate?

Plant susceptibility to herbicides is related to several factors, including the environmental conditions under which the plants develop. Two experiments were carried out using Synedrellopsis grisebachii plants in two different developmental stages (vegetative and reproductive), with the goal of studying plant susceptibility to the herbicide, glyphosate, and the dependence of this susceptibility on light intensity (full sunlight and 70% shading), correlated with leaf anatomy. The experimental design for both experiments was completely randomized, with a 2 × 7 factorial scheme, with two light intensities and seven different doses of glyphosate (0D, 1/4D, 1/2D, D, 2D, 4D and 6D, where D is the recommended dose of 1800 g ae ha^?1) as the factors and four replicates per treatment. The leaf anatomy was characterized with optical and scanning electron microscopy. The plants that were grown in full sunlight were more tolerant of glyphosate because of thickening of the adaxial epidermis, parenchyma and main vein structures, which required higher glyphosate doses for effective weed control. The plants that were in the reproductive stage were more tolerant of glyphosate, probably because of lower glyphosate absorption and translocation to the reproductive organs.     

Aminoaciduria Caused by Fanconi Syndrome in a Heifer

A case study of renal tubular dysfunction consistent with idiopathic Fanconi syndrome is reported in an 18‐month‐old Holstein heifer. The clinical, biochemical, and histopathological features are described. The heifer had clinical signs of growth retardation, wasting, and persistent diarrhea. Biochemical blood analysis identified hypokalemia, hyponatremia, and hypochloremia. Urinalysis identified glycosuria, proteinuria, and acidic pH. Histological examination of the kidney disclosed mild tubular necrosis with proteinaceous casts in the lumina of renal tubules. We performed LC‐HRMS on urine to confirm Fanconi syndrome. Using this technique, we identified severe generalized aminoaciduria suggestive of idiopathic renal Fanconi syndrome in this heifer.     

Identification and Characterisation of Bacteria Causing Soft-rot in Agave tequilana

Agave tequilana is the raw material for the production of the alcoholic beverage tequila. A bacterial disease has affected the A. tequilana crop in recent years. Previous reports based on colony and cell morphology, Gram stain and potato rot indicated that Erwinia sp. is the main pathogen. We isolated a several bacterial isolates capable of producing soft-rot symptoms in greenhouse pathogenicity assays. An extensive characterisation involving pathogenicity tests, fatty acid profile, metabolic and physiological properties, ribosomal DNA sequence and intergenic transcribed spacer amplification (ITS-PCR) and restriction banding pattern (ITS-RFLP) was made of each isolate. Three different species: Erwinia cacticida, Pantoea agglomerans and Pseudomonas sp. were identified. Fatty acid and metabolic profiles gave low similarity values of identification but 16S rDNA sequence, ITS-PCR and ITS-RFLP confirmed the identification of E. cacticida. In the phylogenetic tree, E. cacticida from blue agave was grouped neither with E. cacticida type strains nor with Erwinia carotovora. This is the first report that associates E. cacticida with A. tequilana soft-rot symptoms.     

Seasonal effect on infection and development of lesions caused by Cryphonectria parasitica in Castanea sativa

Seasonal variation in the development of chestnut blight, caused by Cryphonectria parasitica, was investigated by inoculating in situ chestnut trees and in vitro excised chestnut segments, at either monthly or 3‐monthly intervals throughout 30 months. Inoculations were made with conidia and mycelium of a virulent isolate and with mycelium of a hypovirulent isolate. Conidial inoculations of living sprouts or excised segments between May and July resulted in the greatest incidence of infection whereas inoculations in autumn and winter, in vitro as well as in situ, did not reveal any visible disease. However, from these symptomless inoculated stems, C. parasitica was isolated 3 months after inoculation. Inoculations with the mycelium of the virulent isolate always resulted in lesions, except in January 1999, and the greatest rate of lesion development occurred for inoculations made in the spring and summer. There was a significant seasonal effect on lesion development. Lesions caused by the hypovirulent isolate, smaller than those caused by the virulent isolate, followed a similar seasonal pattern. The same seasonal variations were observed for inoculations in vitro of excised segments. Relative water content (RWC) of chestnut bark significantly varied with bark sampling date. The rate of lesion development in sprouts significantly correlated with average minimum (ATn) and maximum (ATx) temperatures and the sum of rainfall during inoculation period, with the rate of lesion development measured in excised segments 10 days after inoculation (R10d) and with RWC measured on the day of inoculation. In multiple regression models, variables ATx and R10d best explained variation in lesion development.     

Genetic variation for tolerance or resistance to barley yellow dwarf virus in durum wheat

Summary One durum wheat line (Triticum durum), cv. 82PCD476, with useful BYDV tolerance or resistance, was singled out of 5 152 lines evaluated between 1979 and 1986. A few other lines such as cv. Boohai and cv. 12th IDSN 227, slightly inferior to cv. 82PCD476, also showed some value. With an hybrid of cv. 12th IDSN227 with the susceptible cv. 84PCY-S531, broad-sense heritability values of 0.37–0.41 were obtained for symptoms and a heritability value of 0.55 was obtained for the total weight of spikes. The weight of spikes was considered as a good indicator of wheat tolerance to BYDV. Although BYDV resistance or tolerance genes are not very common in durum wheat, sources of heritable resistance could be found. However, the resistance ofT. aestivum to BYDV was superior to the one found inT. durum.Cintribution no. 323