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.     

Comparison of greenhouse and environmentally controlled growthroom root growth potential testing of 2+0 eastern white pine seedlings

Two-year-old estern white pine (Pinus strobus L.) seedlings were measured for Root Growth Potential (RGP) in a hydroponic system over two years in both a greenhouse and an environmentally controlled growthroom. Analyses of variance showed no significant differences (alpha = 0.05) between testing environments for either year. This study demonstrates the validity of using greenhouse RGP testing for 2+0 eastern white pine.     

Acidification trends in Sweden

A Swedish hydrochemical monitoring program, which was started during the International Hydrological Decade (1965–1975), has been evaluated with respect to trends and regional differences in acidic deposition and stream water quality. Despite the more than twofold decrease in S emissions in Sweden since the early 1970's, there was only a marginal decrease in sulphate level in precipitation, indicating that long-range transport dominates precipitation chemistry. Trend analyses of stream water data gave practically no evidence that surface water quality had been changing since the late 1960's, i.e., no time lagged or accumulating effects of acidic deposition during past decades could be demonstrated. The difference between the wet deposition of S in southern and northern parts of Sweden was found to be smaller than previously reported. In the northern catchment areas, the input of S was considerably larger than the output. The statistical uncertainty of the trend estimates raises questions about the effectivity of present hydrochemical monitoring programs in detecting environmental changes.     

Mungbean [Vigna radiata (L.) Wilczek] globulins: purification and characterization

Vicilin type (8S) and basic 7S globulins and legumin type (11S) globulins were isolated from mungbean [Vigna radiata (L.) Wilczek]. The native molecular weights of the different globulin types were 360000 for legumin, 200000 for vicilin, and 135000 for basic 7S. Some of the 8S globulin apparently complexed and coeluted with the 11S on gel filtration. On SDS-PAGE, 11S was composed of two bands of 40000 and 24000, 8S was composed of 60000, 48000, 32000, and 26000 bands, and basic 7S was composed of 28000 and 16000 bands. The percent composition of total globulins was estimated to be as follow: 8S, 89%; basic 7S, 3.4%; and 11S, 7.6%. The basic 7S and 11S but not the 8S globulins were found to have disulfide bonds. The presence of carbohydrates by conjugated peroxidase reaction was observed in all bands of 8S, the acidic polypeptide of basic 7S, and its complex but not in 11S. The 28000 basic 7S band and its 42000 complex and the first three major bands of 8S cross-reacted with antibodies to all types of soybean conglycinin subunits (alpha, alpha', and beta), whereas the fourth band cross-reacted only with the anti-beta subunit. None of the mungbean globulins cross-reacted with anti-soybean glycinin. Basic 7S was found to be easily extracted with 0.15 M NaCl, 11S was extracted with 0.35 M NaCl,and 8S was extracted over a wide range of NaCl concentrations. The N-terminal sequences of the different subunits/fragments of the globulins were determined and found to have strong homology with storage proteins of other legumes and crops.     

Potassium bentonites reduce radiocaesium availability to plants

After the Chernobyl accident in 1986 the fate of radiocaesium from the fallout became of pressing concern. Specific soil amendments, as K fertilizer and specific clay minerals, promised to mitigate the worst effects. We therefore investigated the influence of bentonite and the K status of the soil on the radiocaesium equilibria in soil and on its availability to ryegrass. A sample of a sandy soil was contaminated with ¹³⁴Cs and amended with K and Ca salts (0–0.97 mmol kg^?1) and K bentonite (0–2%). After 4 weeks' incubation of the soil mixtures, ryegrass was grown for 18 weeks in a pot trial and harvested on seven occasions. No significant treatment effects on ¹³⁴Cs activity concentrations were found at the first and second harvest. From the third harvest onwards, however, ¹³⁴Cs activity concentrations in the grass were reduced up to twofold (P < 0.05) by increasing rates of K bentonite. Adsorption studies with ¹³⁷Cs revealed that the radiocaesium interception potential (RIP) of the soil–bentonite mixtures (> 1% bentonite) increased about 10‐fold during plant growth. The RIP of the K bentonite after plant growth was up to 10 times larger than that of pure illite. The formation of specific Cs sorption sites is ascribed to the in situ illitization of the K bentonite. The increase in RIP during plant growth is reflected in a decrease in exchangeable K⁺ at 2% K bentonite of about 18%. Radiocaesium concentrations in grass could be reliably predicted from the Cs⁺ and K⁺ concentrations in the soil solution. Adding K bentonite to a soil contaminated with radiocaesium is effective in fixing Cs in the soil.     

生物肥料磷的利用效率：生物有效性和组分

Although to date some technologies producing bio-based phosphorus (P) fertilizers have been proposed and implemented, the efficient use of the recovered products is still limited due to legislative constraints and lack of insights in the P release with time and in the corresponding mechanisms. The aim of this work was to evaluate the fertilizer performance in terms of P release and use efficiency of recovered struvite, FePO₄-sludge, digestate, and animal manure as compared to fossil reserve-based mineral triple superphosphate (TSP). First, product physicochemical characteristics and P fractions in the context of European fertilizer legislation were assessed. Next, a controlled greenhouse experiment was set up to evaluate plant reactions as well as changes of P availability in a nutrient-rich sandy soil with high P status and a nutrient-poor Rheinsand soil with low P status. Soil P fractions were determined in the extracts with water, ammonium lactate and CaCl₂, and in soil solution sampled with Rhizon soil moisture samplers. Based on all results, it is worth conducting long-term field trials to evaluate the P release effect of struvite and digestate as compared to animal manure and TSP on different soil types with varying P status. These products showed promise as sustainable substitutes for conventional P fertilizers and could contribute to a more efficient use of P in agriculture. A refined classification of P application standards/recommendations in terms of soil P status, soil texture, and fertilizer characteristics, next to the crop P demand, is recommended. Moreover, the additional use of Rhizon samplers for determination of direct available P, including dissolved organic P, is proposed for better understanding and categorization of different P fertilizers in environmental and fertilizer legislations.     

Diversity among melon (Cucumis melo L.) landraces from the Indo-Gangetic plains of India and their genetic relationship with USA melon cultivars

We report here the first broad genetic characterization of farmer-developed landraces of melon (Cucumis melo L.) from the Indo-Gangetic plains of India, an area overlooked in previous melon genetic diversity analyses of Indian melon germplasm. Eighty-eight landraces from three melon Groups in two subspecies (C. melo subsp. agrestis Momordica Group, and C. melo subsp. melo Cantalupensis Group and Reticulatus Group) were collected from the four agro-ecological regions (six sub-regions) of two northern states of the Indo-Gangetic plains of India, Uttar Pradesh and Uttarakhand. Significant differences were found among the landraces and eight USA Reticulatus Group reference cultivars for 18 plant and fruit traits: no. of primary branches per plant, days to marketable maturity, sex expression, fruit shape, flesh colour, netting, no. of fruit per plant, fruit weight, shelf life, total soluble solids (°Bx), ascorbic acid (mg/100 g), titratable acidity (%), fruit length and diameter, seed cavity length and diameter, flesh thickness, and resistance to Cucumber mosaic virus. The three melon groups differed significantly for 10 of the plant and fruit traits. Cantalupensis Group and Reticulatus Group accessions were andromonoecious, and the Momordica Group was monoecious. Neighbour-joining (NJ) tree and factorial correspondence analysis (FCA) of simple sequence repeat loci also revealed a high level of genetic variability in this germplasm. The 96 melon genotypes clustered into five groups in the NJ tree analysis: the 16 Indian Reticulatus Group accessions and eight USA reference cultivars formed a distinct group; and the 60 Cantalupensis Group accessions clustered in four other groups with the 12 Momordica Group accessions in a distinct subgroup of one of the Cantalupensis groups. The FCA plot largely confirmed the NJ tree with three distinct groups, one for each melon group. The close affinity of the Indian and USA Reticulatus melons was not unexpected, but it is not clear whether it was inherent in the group and maintained as Reticulatus melons moved from India through Central Asia and Europe to North America, or the result of recent intercrossing of Indian landraces with the USA-derived cultivars and selection for a broad range of Reticulatus type melons.     

Postharvest storage of white asparagus (Asparagus officinalis l.): changes in dietary fiber (Nonstarch polysaccharides).

Changes occurring in the content and composition of the dietary fiber of white asparagus during storage in different conditions were studied (2 degrees C; 2 degrees C in polyethylene bags with air; 2 degrees C in polyethylene bags with a selected gas mixture). The neutral sugars and uronic acid composition of dietary fiber was determined by gas chromatography and by a spectrophotometric method. The modifications observed in the dietary fiber of the asparagus stored at 2 degrees C were more rapid and pronounced than those in polyethylene bags. The most important changes corresponded to xylose and glucose from insoluble dietary fiber and galactose from soluble dietary fiber. Statistical analysis indicated that the modifications were significantly affected by the type of storage and time.     

Authentication of natural vanilla flavorings: isotopic characterization using degradation of vanillin into guaiacol

The isotopic investigation of vanillin has been extended to the new sources of natural precursors of vanillin recently introduced with a view of obtaining natural vanillin by biotechnological processes. To check the consistency of the isotopic composition of vanillin with that of the corresponding aromatic fragment, a selective degradation reaction into guaiacol was carried out. The reaction was shown to proceed without significant isotopic fractionation at the sites of interest, and an optimized procedure was defined from the results of an experimental design involving the quantity of reagent and the temperature and duration of the experiment. Guaiacol, which can be easily obtained in a reasonable time, is an interesting isotopic probe for carbon- and oxygen-isotope ratio mass spectrometry (IRMS) determinations. It provides (13)C information specific to the aromatic fragment and, combined with delta(13)C values measured on vanillin itself, it improves the authentication potential of carbon-IRMS. Thus, the natural status of ferulic acid may be characterized by significant (13)C depletion at the formyl site. Similarly, the oxygen-18 content of guaiacol is a better authentication tool than delta(18)O of vanillin because it does not suffer the drawback of being altered by chemical exchange of the sp(2) oxygen atom with water in industrial or laboratory procedures. Although collaborative studies are still necessary to improve the interlaboratory reproducibility of the delta(18)O parameters, consistent results can be obtained in an intralaboratory context. It is shown in particular that chemical oxidation of ferulic acid is characterized by a relative enrichment of the aromatic moiety of vanillin.