Investigation of spatial and temporal metal atmospheric deposition in France through lichen and moss bioaccumulation over one century.

Lichens and mosses were used as biomonitors to assess the atmospheric deposition of metals in forested ecosystems in various regions of France. The concentrations of 17 metals/metalloids (Al, As, Cd, Co, Cr, Cs, Cu, Fe, Mn, Ni, Pb, Sb, Sn, Sr, Ti, V, and Zn) indicated overall low atmospheric contamination in these forested environments, but a regionalism emerged from local contributions (anthropogenic activities, as well as local lithology). Taking into account the geochemical background and comparing to Italian data, the elements from both natural and anthropogenic activities, such as Cd, Pb, or Zn, did not show any obvious anomalies. However, elements mainly originating from lithogenic dust (e.g., Al, Fe, Ti) were more prevalent in sparse forests and in the Southern regions of France, whereas samples from dense forests showed an accumulation of elements from biological recycling (Mn and Zn). The combination of enrichment factors and Pb isotope ratios between current and herbarium samples indicated the historical evolution of metal atmospheric contamination: the high contribution of coal combustion beginning 150 years ago decreased at the end of the 20th century, and the influence of car traffic during the latter observed period decreased in the last few decades. In the South of France, obvious local influences were well preserved during the last century.

Origin and distribution of rare earth elements in various lichen and moss species over the last century in France.

Rare earth elements (REE) are known to be powerful environmental tracers in natural biogeochemical compartments. In this study, the atmospheric deposition of REE was investigated using various lichens and mosses as well as herbarium samples from 1870 to 1998 from six major forested areas in France. The comparison between the REE distribution patterns in organisms and bedrocks showed a regional uniformity influence from dust particles originating from the bedrock and/or soil weathering that were entrapped by lichens and mosses. These lithological signatures were consistent over the last century. The REE patterns of different organism species allowed minor influence of the species to be highlighted compared to the regional lithology. This was even true where the morphological features played a role in the bioaccumulation levels, which were related to the variable efficiency in trapping atmospheric dust particles. A comparison between REE profiles in the organisms and bark indicated a lack of influence of the substrate on lichen REE content. Lichens and mosses appear to be robust passive monitors of REE atmospheric deposition over decades because the mineral data was preserved in herbarium samples despite organic degradation being shown by carbon isotopes and SEM observations. To overcome the bias of REE concentration that resulted from organic degradation, the use of a normalized method is recommended to interpret the historical samples.

Comparing early twentieth century and present-day atmospheric pollution in SW France: A story of lichens.

Lichens have long been known to be good indicators of air quality and atmospheric deposition. Xanthoria parietina was selected to investigate past (sourced from a herbarium) and present-day trace metal pollution in four sites from South-West France (close to Albi). Enrichment factors, relationships between elements and hierarchical classification indicated that the atmosphere was mainly impacted by coal combustion (as shown by As, Pb or Cd contamination) during the early twentieth century, whereas more recently, another mixture of pollutants (e.g. Sb, Sn, Pb and Cu) from local factories and car traffic has emerged. The Rare Earth Elements (REE) and other lithogenic elements indicated a higher dust content in the atmosphere in the early twentieth century and a specific lithological local signature. In addition to long-range atmospheric transport, local urban emissions had a strong impact on trace element contamination registered in lichens, particularly for contemporary data.

Molecular tools for the identification of Tuber melanosporum in agroindustry.

Tuber melanosporum Vitt., Tuber magnatum Pico, and Tuber uncinatum Chat. can be differentiated by their morphological characters. Fraud problems have arisen recently with the importation to Europe of truffles from China. T. melanosporum is morphologically very close, but distinct from the Chinese species [Tuber indicum (Cooke and Massee) and T. himalayense BC (Zhang and Winter)]. We have optimized molecular tools to unequivocally identify T. melanosporum. DNA extraction from ascocarps of black truffles is not straightforward. Problems to obtain pure DNA are due to high contents of phenolic compounds, melanine, and various polymers (proteins, polysaccharides, etc). These compounds coprecipitate with the DNA during extraction and strongly inhibit the PCR reaction. We have developed an efficient and reliable protocol for DNA extraction from truffle ascocarps. It was used successfully for DNA extraction from mycorrhizal root tips as well as from canned preparations of T. melanosporum. Several approaches to identify T. melanosporum by PCR were developed. Two specific primers for T. melanosporum were designed after comparison of the ITS region of this species with those of three Chinese fungi. They proved to be efficient to specifically detect the presence of T. melanosporum by PCR. The mycorrhizal status of trees inoculated with T. melanosporum but unable to produce truffles was confirmed in a single-step PCR reaction. A multiplex PCR approach was also developed with three sets of primers (including a specific one for Chinese truffles) to detect, in one PCR reaction, the presence of any other Tuber species mixed with T. melanosporum ascocarps. This optimized protocol, in association with the specific primers we designed, is applicable to quality control in the truffle industry from the production stages to final commercial products.

Phylogenetic relationships between European and Chinese truffles based on parsimony and distance analysis of ITS sequences.

Phylogenetic relationships among truffle species from Europe and China were investigated through parsimony analysis of the ITS sequences. Three major clades were obtained among the species analysed. The so-called white truffles appeared polyphyletic since Tuber magnatum was grouped with brown truffles and not with the other white species (T. maculatum, T. borchii, T. dryophilum, T. puberulum). The black truffles investigated in this study, T. brumale, T. melanosporum, T. indicum and T. himalayense, were grouped in an independent clade. The Périgord black truffle T. melanosporum and the Chinese black truffles T. indicum and T. himalayense, were very closely related. The delimitation of these species was estimated by a distance analysis on several isolates collected from different geographic areas. In spite of intraspecific variations of the internal transcribed spacers (ITS) sequences, T. melanosporum and the Chinese black truffles can be unambiguously attributed to distinct taxa.

Purification, Elicitor Activity, and Cell Wall Localization of a Glycoprotein from Phytophthora parasitica var. nicotianae, a Fungal Pathogen of Tobacco.

ABSTRACT A glycoprotein of 34 kDa (GP 34) was solubilized at acidic pH from the mycelium of Phytophthora parasitica var. nicotianae and was purified by ion exchange and gel permeation chromatography. Whole tobacco plants treated with GP 34 through their roots showed an enhanced lipoxygenase activity as well as hydroxyproline-rich glycoprotein accumulation, indicating that this molecule had elicitor properties. An antiserum raised against the pure glycoprotein allowed localization of GP 34 by immunogold-labeling on the cell surface of the mycelium when the fungus was grown in vitro. In the wall-less zoospores, GP 34 was limited to the flagellum surface. It was then abundantly synthesized at the onset of encystment. During infection of tobacco plants, labeling was very faint at early stages of colonization, particularly in the susceptible host cultivar. It appeared earlier in the resistant host cultivar and was restricted to the living fungus, declining with mycelium cell death.

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