Genetic manipulation of proline accumulation influences the concentrations of other amino acids in soybean subjected to simultaneous drought and heat stress

The effect of simultaneous drought and heat stress on free amino acid levels was compared in wild type and transgenic soybean (Glycine max (L.) Merr cv Ibis) plants transformed with the cDNA coding for the last enzyme of Pro biosynthesis, l-Delta(1)-pyrroline-5-carboxylate reductase (EC 1.5.1.2), in sense and antisense directions. The most rapid increase in Pro content was found in the sense transformants that exhibited the least water loss, while the slowest elevation of Pro levels was detected in the antisense transformants that exhibited the greatest water loss during stress. Correspondingly, the level of the Pro precursors Glu and Arg was higher in sense transformants and lower in antisense ones compared to the wild type plants during the initial part of the stress. Interestingly, genetic manipulation of Pro levels also affected the stress-induced changes in the concentration of several other amino acids, which indicates the coordinated regulation of their metabolic pathways.     

Volatile components of Loureira, Dona Branca, and Treixadura wines.

White wines experimentally produced from the white grape varieties Loureira, Dona Branca, and Treixadura have been analyzed over four consecutive harvests. The contents of monoterpenes, volatile phenols, alcohols, sulfur components, acetates, fatty acids, and ethyl esters were determined by gas chromatography (FID) and gas chromatography-mass spectrometry. The mean values from four vintages confirmed that these wines have characteristic profiles. Loureira wines are characterized by a high content of free terpenes, 1-hexanol, ethyl acetate, and fatty acids. Dona Branca wines present the highest concentrations of methanol and sulfur constituents, and the lowest concentrations of higher alcohols, acetates, diethyl succinate, and ethyl esters. The levels of monoterpenes in Treixadura wines are very low, but they have the highest concentrations of volatile phenols, principally due to the vanillin, diethyl succinate, ethyl lactate, and ethyl esters. These results were confirmed by principal component and linear discriminant analysis, which show a clear differentiation among these wines as a function of the varietal origin.     

Production, purification, and properties of an endoglucanase produced by the hyphomycete Chalara (Syn. Thielaviopsis) paradoxa CH32

The hyphomycete Chalara (syn. Thielaviopsis) paradoxa produces endoglucanase activity during the late trophophase. The low molecular mass (35 kDa) endoglucanase purified from cultured broths works optimally at 37 degrees C and pH 5.0. The enzyme inactivates at pH below 3.0 and also at temperatures of 50 degrees C or higher, but it is stable at lower temperatures, including refrigeration temperature and freezing. The enzyme is inhibited by detergents, by EDTA, and by the divalent cations Hg(2+) and Ag(2+). It is also inhibited to some extent by 10 mM Zn(2+), Fe(2+), and Mg(2+), but it is stimulated by Mn(2+). Enzyme activity is not affected by reducing agents. In the presence of low concentrations of water miscible organic solvents (20%) endoglucanase activity is inhibited by 7% (for methanol) to 50% (for acetonitrile), and it is totally inhibited at higher solvent concentrations (50%). Enzyme activity is not affected by the water immiscible solvent ethyl acetate. Carboxymethylcellulose is the preferred substrate (K(m(app)) = 8.3 g/L; V(max(app)) = 1.1 microM/min). Hydrolysis of crystalline cellulosic substrates is very limited, but it is greatly enhanced by phosphoric acid swelling. The purified enzyme shows no activity toward disaccharides or aryl-glucosides. Its activity is inhibited by cellobiose.     

Significance of soil properties in the adsorption and mobility of the fungicide metalaxyl in vineyard soils

Adsorption and mobility of the fungicide metalaxyl were studied in 16 vineyard soils from the La Rioja region (Spain), with organic matter (OM) contents in the 0.31--1.37% range, and in 7 natural soils with OM contents in the 3.30--8.24% range. Adsorption isotherms were obtained using the batch equilibrium technique, and mobility was studied by soil thin-layer chromatography (soil-TLC). In all cases, the adsorption isotherms fit the Freundlich equation. The values of the K(f) constants were low in the vineyard soils (0.01--0.64) and increased in the natural soils (1.05--2.83). The n(f) values were in general lower than unity. K(f) constants were significantly correlated (p < 0.001) with the OM content when all of the soils were considered. According to the determination coefficient, r(2), OM would account for 88% of the variance in adsorption. When the vineyard soils alone were considered, a significant correlation was seen between K(f) and the OM and clay contents; both parameters, varying simultaneously, explain 80% of the variance in adsorption. Study of the mobility of metalaxyl with soil-TLC indicated that in vineyard soils the fungicide has the potential for being highly mobile in 19% of the soils and mobile in 81% of them. In natural soils, the fungicide has the potential for being moderately mobile or mobile in 86 and 14% of the soils, respectively. This type of behavior of metalaxyl indicates that in vineyards soils of the La Rioja region (Spain) with low OM contents, where application of the compound is continuous, a leaching of the fungicide from the soil to groundwaters could potentially occur. These results should be borne in mind when metalaxyl is to be used in the soils of this region.     

Determination of phytohormones of environmental impact by capillary zone electrophoresis

A test mixture of five phytohormones [naphthaleneacetic acid (NAA), naphthoxyacetic acid (NOA), indoleacetic acid (IAA), indolebutyric acid (IBA), and indolepropionic acid (IPA)] was investigated. These compounds were cleanly separated with good resolution by capillary zone electrophoresis with a UV diode array detector using 20 mM sodium phosphate buffer (pH 7.25). The lowest detection limit was obtained for IPA (0.45 mg L(-)(1) or 0.005 mg kg(-)(1)) and the highest for NAA (1.04 mg L(-)(1) or 0.014 mg kg(-)(1)). The method has been applied for tomato samples fortified with the five phytohormones using a liquid-liquid extraction procedure, obtaining recovery percentages ranging from 91 to 109.0%.     

Separation and characterization of phenolic compounds in fennel (Foeniculum vulgare) using liquid chromatography-negative electrospray ionization tandem mass spectrometry

Liquid chromatography (LC) diode array detection (DAD) coupled to negative electrospray ionization (ESI) tandem mass spectrometry (MS/MS) was used for the rapid and sensitive identification of water-soluble phenolic compounds in fennel waste. The plant material was first extracted and then chromatographed on Sephadex LH-20 to afford seven fractions, each of them being subjected to LC-MS analysis. Identification of the compounds was carried out by interpretation of UV, MS, and MS/MS spectra. Forty-two phenolic substances were identified, 27 of which had not previously been reported in fennel, including hydroxycinnamic acid derivatives, flavonoid glycosides, and flavonoid aglycons.     

Further studies on the analysis of DSP toxin profiles in galician mussels

Further studies on mussel samples from Galicia, Spain, have revealed the presence of okadaic acid (OA), dinophysistoxin 2 (DTX2), and the fatty acid acyl esters of both of these toxins as the "DTX3" complex. Measurements were performed with an improved in situ method for the formation of 9-anthryldiazomethane (ADAM) derivatives followed by liquid chromatography with fluorescence detection. Base hydrolysis of DTX3 toxins gave free OA and DTX2, which were determined following ADAM derivatization. Results were confirmed by liquid chromatography/mass spectrometry analyses, and in most of the samples, free DTX2 was the most abundant toxin. However, the OA/DTX2 ratio in the DTX3 conjugated form was different, with OA being the most abundant in all cases. This difference could be due to different rates of metabolism of OA and DTX2 to the acyl esters or due to contamination of the shellfish by the two toxins at different points in time, resulting in less acyl ester formation for one toxin versus the other. The second possibility would be reasonable if two different source organisms were producing the toxins.     

Relative effects of management and environmental conditions on performance and survival of populations of a terrestrial orchid, Dactylorhiza majalis

For population viability analysis of endangered orchid populations, it is crucial to disentangle the effects of weather and management from intrinsic orchid dynamics. When doing this, typically several months’ average temperatures and/or sums of precipitations are compared with some characteristics of plant performance. Here we tested, whether short averaging intervals (1-2 weeks) are more closely correlated with orchid performance. We used 5 years of data from five Dactylorhiza majalis populations, and have shown that the improvement of prediction by shortening the interval over which the temperatures are averaged or precipitation summed, even if detectable, is only weak and not significant. This, however, may be due to low weather variability during the study. Regarding the second aspect, the effect of management (presence or absence of mowing), we have found that leaf area of D. majalis at the regularly mown site was larger than that at the sites which were mown only once in 2 years, but we did not detect a significant effect of the absence of mowing on the incidence of flowering. Mowing can affect orchid performance in two ways: by reduced shading of orchids and by reducing competition with other species. Therefore, we have determined the co-occurring species associated with presence or absence of mowing and found that shading significantly affected the length of the flower stalk, the ratio of leaf width to leaf length at the end of the season, but did not affect seed weight and probability of flowering the next year. We conclude that the most appropriate management for D. majalis is mowing at least once a year, ideally at the end of June/beginning of July, after its fruiting.     

Reduction of decomposition rates of scots pine needle litter due to heavy-metal pollution

Decomposition of unpolluted Scots pine needle litter was studied in two heavy-metal-pollution gradients in Sweden; one near a brass mill and the other around a primary smelter. In the latter area locally collected polluted Scots pine needle litter was also incubated. Decomposition rates were strongly influenced by the metal pollution and a decrease in the rate of mass-loss occurred. In the brass-mill gradient this occurred until about 1 km from the pollution source which corresponded to about 500 µg Cu and 1 000 µg Zn g^?1 soil. Data are presented to indicate that lignin decomposition was more sensitive to pollution than decomposition of whole litter and affected further away from the pollution sources. At the smelter sites, the metal-polluted needle litter decomposed more slowly than the unpolluted needle litter, and this difference was enhanced close to the smelter. The results indicate that heavy metals accumulated in needles prior to shedding have a long-term impact on the subsequent decomposition of the litter. Both litter quality and soil factors thus contribute to the reduced litter decomposition rate in metal-polluted forests. A new non-linear model with decreasing decay rate was used in the statistical evaluation. The model can be used to characterize the effects of pollution on decomposition rate.