Utilisation d'anticorps spécifiques d'une fraction obtenue par chromatographie d'exclusion pour une amélioration de la caractérisation sérologique deRhizoctonia solani Kühn AG3

La purification d'un extrait mycélien soluble d'une souche deR. solani AG3 par chromatographie d'exclusion permet de recueillir une fraction antigénique. Une lapine est immunisée par voie intradermique. L'immunsérum titre 0,125 par la technique de double-diffusion en agarose. Différentes techniques sérologiques (immunodiffusion, électrotransfert suivi de révélation des fractions antigéniques, coloration immunoenzymatique du mycélium) sont appliquées à des souches deR. solani AG1, AG2, AG3, AG4, AG5, AG6, AG7, AGBI et deCeratobasidium CAG1, CAG2, CAG3, CAG4 et CAG5. Le sérum se révèle spécifique des souches AG3. A soluble mycelial extract ofR. solani AG3 was purified by size exclusion chromatography. Five peaks were present in the elution profile: the first was well defined and of large volume. It was retained to immunise a female rabbit by intradermic injection. The titre of the antiserum determined by double immuno-diffusion in agarose was 0.125. The antiserum was tested on isolates ofR. solani AG1, AG2, AG3, AG4, AG5, AG6, AG7, AGB1 and ofCeratobasidium CAG1, CAG2, CAG3, CAG4 and CAG5 (Table 1) using three different methods:

Genetically encoded sensors for ions and metabolites

Many of the genes encoding important trans porters and metabolic enzymes have been identified over the last ten years. Moreover it has been possible to study the biochemical properties of the corresponding proteins in great detail. It is expected that by 2,010 biochemical functions will have been assigned to many of the products of the approximately 30,000 Arabidopsis genes. We will get closer to understanding the biological function of the gene products by systematic analysis of mutants using knock-out and TILLING approaches. Metabolomics initiatives complement these approaches by providing insight into the changes in cellular ion and metabolite profiles in the mutants, thus giving information essential for the construction of cellular and whole plant models. However, one important dataset especially relevant to multicellular organisms is lacking: the knowledge of the spatial and temporal profiles of ions and metabolite levels at cellular and subcell ular levels. To address this issue, we have developed protein-based nanosensors for several metabolites, providing a set of tools for the determination of cytosolic and subcellular ion (e.g. iron and zinc) and metabolite levels in real time using fluorescence-based microscopy. The prototypes of these sensors were shown to function in vitro and also in vivo, i.e. in yeast and in mammalian cell cultures. One future goal is to expand the set of sensors to a wider spectrum of targets by using the natural spectrum of periplasmic binding proteins from bacteria and by computational design of proteins with altered binding pockets. Application of nanosensor technology to plant cells and tissues will help to elucidate the special and temporal distribution of ions and metabolites.     

Spatial attributes of fire regime in eastern Canada: influences of regional landscape physiography and climate

The characterization of the fire regime in the boreal forest rarely considers spatial attributes other than fire size. This study investigates the spatial attributes of fires using the physiography of the landscape as a spatial constraint at a regional scale. Using the Canadian National Fire Database, the size, shape, orientation and eccentricity were assessed for 1,136 fires between 1970 and 2010 in Quebec’s boreal forest and were summarized by ecodistrict. These spatial metrics were used to cluster 33 ecodistricts into homogeneous fire zones and then to determine which environmental variables (climate, topography, hydrography, and surficial deposits) influence the spatial attributes of fires. Analyses showed that 28 out of 33 ecodistricts belonging to a given fire zone were spatially contiguous, suggesting that factors driving the spatial attributes of fire are acting at a regional scale. Indeed, the orientation and size of fires vary significantly among the zones and are driven by the spatial orientation of the landscape and the seasonal regional climate. In some zones, prevailing winds during periods conducive to fire events parallel to the orientation of the landscape may favour the occurrence of very large fires (>100,000 ha). Conversely, an orientation of the landscape opposite to the prevailing winds may act as a natural firebreak and limit the fire size and orientation. This study highlights the need to consider the synergistic relationship between the landscape spatial patterns and the climate regime over the spatial attributes of fire at supra-regional scale. Further scale-dependant studies are needed to improve our understanding of the spatial factors controlling the spatial attributes of fire.     

Family Variation in the Morphology and Physiology of White Spruce (Picea glauca) Seedlings in Response to Elevated CO2 and Temperature

Tree improvement programs aim to develop families that are well-adapted to future growing conditions. To gain insight into the stability of the family genetic response to climate change, white spruce (Picea glauca) seedlings from 60 full-sib families were subjected to a combination of two temperature regimes and two levels of CO₂ over two growing seasons. There was positive effect of warmer temperatures and higher CO₂ on some growth variables but no significant family × treatment interactions. Instantaneous water use efficiency was the only physiological trait that was affected positively by the CO₂ treatment, showing a 51% increase that was consistent across families.     

Unusual Glycosaminoglycans from a Deep Sea Hydrothermal Bacterium Improve Fibrillar Collagen Structuring and Fibroblast Activities in Engineered Connective Tissues

Biopolymers produced by marine organisms can offer useful tools for regenerative medicine. Particularly, HE800 exopolysaccharide (HE800 EPS) secreted by a deep-sea hydrothermal bacterium displays an interesting glycosaminoglycan-like feature resembling hyaluronan. Previous studies demonstrated its effectiveness to enhance in vivo bone regeneration and to support osteoblastic cell metabolism in culture. Thus, in order to assess the usefulness of this high-molecular weight polymer in tissue engineering and tissue repair, in vitro reconstructed connective tissues containing HE800 EPS were performed. We showed that this polysaccharide promotes both collagen structuring and extracellular matrix settle by dermal fibroblasts. Furthermore, from the native HE800 EPS, a low-molecular weight sulfated derivative (HE800 DROS) displaying chemical analogy with heparan-sulfate, was designed. Thus, it was demonstrated that HE800 DROS mimics some properties of heparan-sulfate, such as promotion of fibroblast proliferation and inhibition of matrix metalloproteinase (MMP) secretion. Therefore, we suggest that the HE800EPS family can be considered as an innovative biotechnological source of glycosaminoglycan-like compounds useful to design biomaterials and drugs for tissue engineering and repair.     

Evaluation of sample pre-treatment and potential contamination of soluble carbon in soil extracts during lyophilisation

Lyophilisation of K₂SO₄ soil extracts has been proposed as a sample preparation technique before elemental analysis of carbon or nitrogen. However, previous measurements, based on wet oxidation or catalytic combustion, indicated that C measurements in lyophilised samples not always proved to be accurate. To determine whether the C analysis was affected by the lyophilisation process, an exploratory study was conducted to investigate the potential effects of the sample pre-treatment and of the lyophilisation process itself. This paper puts forward that the use of soil extracts, previously stored at −20 °C, may affect the recovery of salt in the samples and that contamination of the soluble carbon with exogenous C during lyophilisation is feasible. Therefore we recommend to use freshly prepared soil extracts for lyophilisation and always to include an internal standard among the unknown samples to account for a possible contamination.     

Simultaneous and selective detection of two major soft rot pathogens of potato: Pectobacterium atrosepticum (Erwinia carotovora subsp. atrosepticum) and Dickeya spp. (Erwinia chrysanthemi)

Dickeya spp. and Pectobacterium atrosepticum are major pathogens of potato. Current methods to detect these soft-rotting bacteria require separate identification steps. Here we describe a simple method allowing simultaneous detection of both pathogens based on multiplex PCR. The sensitivity of the primer sets was first examined on purified genomic DNA of the type strains Dickeya chrysanthemi 2048^T and P. atrosepticum 1526^T. The specificity and detection limits of the primer sets were successfully tested on 61 strains belonging to various Dickeya and Pectobacterium species, on artificially inoculated and on naturally contaminated potato plants. This new method provides a gain in time and materials, the main advantages for large-scale processes such as pathogen-free seed certification.     

Fishing impact and environmental status in European seas: a diagnosis from stock assessments and ecosystem indicators

Stock‐based and ecosystem‐based indicators are used to provide a new diagnosis of the fishing impact and environmental status of European seas. In the seven European marine ecosystems covering the Baltic and the North‐east Atlantic, (i) trends in landings since 1950 were examined; (ii) syntheses of the status and trends in fish stocks were consolidated at the ecosystem level; and (iii) trends in ecosystem indicators based on landings and surveys were analysed. We show that yields began to decrease everywhere (except in the Baltic) from the mid‐1970s, as a result of the over‐exploitation of some major stocks. Fishermen adapted by increasing fishing effort and exploiting a wider part of the ecosystems. This was insufficient to compensate for the decrease in abundance of many stocks, and total landings have halved over the last 30 years. The highest fishing impact took place in the late 1990s, with a clear decrease in stock‐based and ecosystem indicators. In particular, trophic‐based indicators exhibited a continuous decreasing trend in almost all ecosystems. Over the past decade, a decrease in fishing pressure has been observed, the mean fishing mortality rate of assessed stocks being almost halved in all the considered ecosystems, but no clear recovery in the biomass and ecosystem indicators is yet apparent. In addition, the mean recruitment index was shown to decrease by around 50% in all ecosystems (except the Baltic). We conclude that building this kind of diagnosis is a key step on the path to implementing an ecosystem approach to fisheries management.     

Assessing the agronomic potential of linseed genotypes by multivariate analyses and association mapping of agronomic traits

High prices of fish oil make linseed attractive for aquaculture and animal feed. To ensure a constant supply of linseed, the development of stable cultivars is of strategic importance. In this study, 35 linseed genotypes were evaluated in five Chilean environments (E) from 2009 to 2012. The additive main effect and multiplicative interaction analysis (AMMI), genotype (G) plus genotype by environment (GE) interaction (GGE) biplot analysis and three stability parameters were tested with the aim of identifying adapted genotypes for the development of linseed cultivars. An association mapping (AM) analysis was also conducted for four agronomic traits and the stability of the associated markers was evaluated using the QQE (QTL main effect and QTL by environment interaction) approach. Combined analysis of variance for yield, seeds per boll (SPB), plant height (PH) and days to flowering (DTF) were significant for G, E and GE (P < 0.001). The combined stability analysis identified some Canadian, Argentinean and Chilean accessions to be the best adapted and highest yielding genotypes. Coancestry analysis indicated that crossing Canadian and Chilean genotypes could maximize transgressive segregation for yield. Significant associations for DTF, PH and SPB explained up to 59 % of the phenotypic variation for these traits. The QQE and AM analyses were consistent in identifying marker LGM27B as the most stable and significant across all environments with the largest effect in reducing DTF. The combined application of the stability, AM and QQE analyses could accelerate the development of marketable linseed cultivars adapted to Southern Chile.