Effects of physicochemical interactions and microbial activity on the persistence of Cry1Aa Bt (Bacillus thuringiensis) toxin in soil

Genetically modified crops, that produce Cry insecticidal crystal proteins (Cry) from Bacillus thuringiensis (Bt), release these toxins into soils through root exudates and upon decomposition of residues. The fate of these toxins in soil has not yet been clearly elucidated. Persistence can be influenced by biotic (degradation by microorganisms) and abiotic factors (physicochemical interactions with soil components, especially adsorption). The aim of this study was to follow the fate of Cry1Aa Bt toxin in contrasting soils subjected to different treatments to enhance or inhibit microbial activity, in order to establish the importance of biotic and abiotic processes for the fate of Bt toxin. The toxin was efficiently extracted from each soil using an alkaline buffer containing a protein, bovine serum albumin, and a nonionic surfactant, Tween 20. The marked decline of extractable toxin after incubation of weeks to months was soil-dependent. The decrease of extractable toxin with incubation time was not related to microbial degradation but mainly to physicochemical interactions with the surfaces that may decrease immunochemical detectability or enhance protein fixation. Hydrophobic interactions may play an important role in determining the interaction of the toxin with surfaces.     

Exploration of plant genomes in the FLAGdb++ environment

Background

Efficient high throughput screening systems of useful mutants are prerequisite for study of plant functional genomics and lots of application fields. Advance in such screening tools, thanks to the development of analytic instruments. Direct analysis in real-time (DART)-mass spectrometry (MS) by ionization of complex materials at atmospheric pressure is a rapid, simple, high-resolution analytical technique. Here we describe a rapid, simple method for the genetic discrimination of intact Arabidopsis thaliana mutant seeds using metabolic profiling by DART-MS.

Results

To determine whether this DART-MS combined by multivariate analysis can perform genetic discrimination based on global metabolic profiling, intact Arabidopsis thaliana mutant seeds were subjected to DART-MS without any sample preparation. Partial least squares-discriminant analysis (PLS-DA) of DART-MS spectral data from intact seeds classified 14 different lines of seeds into two distinct groups: Columbia (Col-0) and Landsberg erecta (Ler) ecotype backgrounds. A hierarchical dendrogram based on partial least squares-discriminant analysis (PLS-DA) subdivided the Col-0 ecotype into two groups: mutant lines harboring defects in the phenylpropanoid biosynthetic pathway and mutants without these defects. These results indicated that metabolic profiling with DART-MS could discriminate intact Arabidopsis seeds at least ecotype level and metabolic pathway level within same ecotype.

Conclusion

The described DART-MS combined by multivariate analysis allows for rapid screening and metabolic characterization of lots of Arabidopsis mutant seeds without complex metabolic preparation steps. Moreover, potential novel metabolic markers can be detected and used to clarify the genetic relationship between Arabidopsis cultivars. Furthermore this technique can be applied to predict the novel gene function of metabolic mutants regardless of morphological phenotypes.     

Effect of host and inoculum patterns on take-all disease of wheat incidence,severity and disease gradient

The importance of the spatial aspect of epidemics has been recognized from the outset of plant disease epidemiology. The objective of this study was to determine if the host spatial structure influenced the spatio-temporal development of take-all disease of wheat, depending on the inoculum spatial structure. Three sowing patterns of wheat (broadcast sowing, line sowing and sowing in hills) and three patterns of inoculum (uniform, aggregated and natural infestation) were tested in a field experiment, repeated over 2 years. Disease (severity, root disease incidence, plant disease incidence and, when applicable, line and hill incidences) was assessed seven times during the course of each season and the spatial pattern was characterized with incidence-incidence relationships. In the naturally infested plots, disease levels at all measurement scales were significantly higher in plots sown in hills, compared to plots sown in line, which were in turn significantly more diseased than plots with broadcast sowing. Disease aggregation within roots and plants was stronger in line and hill sowing than in broadcast sowing. Analysis of the disease gradient in the artificially infested plots showed that the disease intensified (local increase of disease level) more than it extensified (spatial spread of the disease), the effect of the introduced inoculum was reduced by 95% at a distance of 15 cm away from the point of infestation. Yield was not significantly affected by sowing pattern or artificial infestation.     

Changes in green coffee protein profiles during roasting

To reveal its flavor, coffee has to be roasted. In fact, the green coffee bean contains all ingredients necessary for the later development of coffee flavor. It is now widely accepted that free amino acids and peptides are required for the generation of coffee aroma. However, the mechanisms leading to defined mixtures of free amino acids and peptides remain unknown. Information pertaining to the identification of precursor proteins is also lacking. To answer some of these questions, two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) was used to follow the fate of green coffee proteins. Two conditions were considered: roasting and incubation of green coffee suspensions at 37 degrees C. Coffee beans were observed to acquire the potential to spontaneously release H(2)O(2) upon polymerization of their proteins during roasting. Fragmentation of proteins was also observed. Conversely, H(2)O(2) was found to control polymerization and fragmentation of green coffee proteins in solution at 37 degrees C. Polymerization and fragmentation patterns under the two conditions were comparable. These observations suggest that the two conditions under study triggered, at least to some extent, similar biochemical mechanisms involving autoxidation. Throughout this study, a unique fragmentation cascade involving the 11S coffee storage protein was identified. Generated fragments shared an atypical staining behavior linked to their sensitivity to redox conditions.     

Ascorbic acid oxidation in sucrose aqueous model systems at subzero temperatures

The reduction of Tempol by ascorbic acid in concentrated sucrose solutions was measured by electron paramagnetic resonance (EPR) at temperatures ranging from 16 to -16 degrees C. This method allowed the determination of the rate constants (k) of this fast reaction, by recording the Tempol reduction as a function of time. The two reactants were initially separated and had to migrate for the reaction to occur. The experimental findings were compared with predicted values according to the equation for diffusion-controlled reaction proposed by Atkins. The experimental reaction rate constants were observed to be lower than the calculated ones. However, the experimental values were found to be controlled by the temperature and viscosity changes of the reaction media, as expected for a diffusion-controlled reaction.     

Experimental and modeling studies showing the effect of lipid type and level on flavor release from milk-based liquid emulsions

The purpose of this work was to study two key parameters of the lipid phase that influence flavor release-lipid level and lipid type-and to relate the results to a mass balance partition coefficient-based mathematical model. Release of 10 volatile compounds from milk-based emulsions at 10, 25, and 50 degrees C was monitored by 1-min headspace sampling with a solid-phase microextraction fiber, followed by GC-MS analysis. As compared to the observations for milk fat, changing to a lipophilic lipid (medium-chain triglycerides, MCT) and adding a monoglyceride-based surfactant did not influence the volatiles release. However, increasing the solid fat content was found to increase the release. At 25 degrees C, and even more so at 10 degrees C, concurrent with an increase in their solid fat content, hydrogenated palm fat emulsions showed increased flavor release over that observed for emulsions made with coconut oil, coconut oil with surfactant, milk fat, and MCT. However, at 50 degrees C, when hydrogenated palm fat emulsions had zero solid fat content, there was no difference in flavor release from that observed for milk fat emulsions. Varying milk fat at nine levels between 0 and 4.5% showed a systematic dependence of the release on the lipid level, dependent on compound lipophilicity. Close correlations were found between the experimental and model predictions with lipid level and percent liquid lipid as variables.     

Analysis of two bentonites of enological interest before and after commercial activation by solid Na(2)CO(3)

Two natural calcium-rich bentonites used for the removal of wine proteins, originating from Greece and Turkey, were studied by X-ray diffraction, thermal analysis, and solid state NMR, before and after activation by solid Na(2)CO(3). Exchange of Ca(2+) by Na(+) mainly occurs for cations located at the edge of layers and only weakly for interlayer cations. This Na(2)CO(3) activation process leads to an increased efficiency in the adsorption process of bovine serum albumin (BSA) used as a model protein for both bentonites. A direct correlation is observed between the extent of Ca(2+)/Na(+) exchange, the strength of adsorption of BSA, and the extent of unfolding of BSA backbone.     

Acquired resistance triggered by elicitins in tobacco and other plants

Elicitins are a family of proteins excreted byPhytophthora spp. They exhibit high sequence homology but large net charge differences. They induce necrosis in tobacco plants which then become resistant to the tobacco pathogenPhytophthora parasitica var.nicotianae. In stem-treated plants, resistance was not restricted to the site of elicitin application, but could be demonstrated by petiole inoculation at all levels on the stem. Resistance was already maximum after two days and lasted for at least two weeks. It was effective not only towardsP. p. var.nicotianae infection, but also against the unrelated pathogenSclerotinia sclerotiorum. In contrast to dichloroisonicotinic acid, an artificial inducer of systemic acquired resistance, which was increasingly effective with doses ranging from 0.25 to 5mole per plant, the basic elicitin cryptogein exhibited a threshold effect, inducing near total resistance and extensive leaf necrosis above 0.1 nmole per plant. Between 1 and 5 nmole, acidic elicitins (capsicein and parasiticein) protected tobacco plants with hardly any necrotic symptom. Elicitins exhibited similar effects in various tobacco cultivars andNicotiana species, although with quantitative differences, but induced neither necrosis nor protection in other SolanaceÆ (tomato, petunia and pepper). Among 24 additional species tested belonging to 18 botanical families, only some BrassicaceÆ, noticeably rape, exhibited symptoms in response to elicitins, in a cultivar-specific manner. Elicitins appear to be natural specific triggers for systemic acquired resistance and provide a tool for unraveling the mechanisms leading to its establishment.Abbreviations AR acquired resistance - HR hypersensitive response - INA 2,6-dichloroisonicotinic acid - Ppn Phytophthora parasitica var.nicotianae - SAR systemic acquired resistance     

West Nile virus and its emergence in the United States of America

Zoonotic West Nile virus (WNV) circulates in natural transmission cycles involving certain mosquitoes and birds, horses, humans, and a range of other vertebrates are incidental hosts. Clinical infections in humans can range in severity from uncomplicated WNV fever to fatal meningoencephalitis. Since its introduction to the Western Hemisphere in 1999, WNV had spread across North America, Central and South America and the Caribbean, although the vast majority of severe human cases have occurred in the United States of America (USA) and Canada. By 2002-2003, the WNV outbreaks have involved thousands of patients causing severe neurologic disease (meningoencephalitis and poliomyelitis-like syndrome) and hundreds of associated fatalities in USA. The purpose of this review is to present recent information on the epidemiology and pathogenicity of WNV since its emergence in North America.     

Modeling the egg and larval transport pathways of the Antarctic toothfish (Dissostichus mawsoni) in the East Antarctic region: New insights into successful transport connections

The Antarctic toothfish (Dissostichus mawsoni) is an important fishery species widely distributed in the Southern Ocean, especially in areas covered by sea ice. Understanding fish distributions and life cycles, including the transport and survival of eggs and larvae, is essential for the assessment and sustainable management of the fishery. However, owing to difficulties with in situ winter observations, information on the early life stages of D. mawsoni is lacking. Here, we investigated the transport pathways of fish eggs and larvae through a particle tracking study, using satellite-derived ocean surface velocities in the East Antarctic region, which includes important D. mawsoni habitats and exploratory fisheries. Our results indicate that particles released from continental slopes are more likely to be successfully transported to suitable settlement grounds than those released from the BANZARE Bank (the southern region of the Kerguelen Plateau), which is situated further north and has been hypothesized to be a potential spawning ground for D. mawsoni. This study demonstrates successful source–settlement connections in relation to ocean recirculation and suggests important settlement regions for D. mawsoni larvae in the East Antarctic region.