Two-dimensional electrophoresis and western-blotting analyses with anti Ara h 3 basic subunit IgG evidence the cross-reacting polypeptides of Arachis hypogaea, Glycine max, and Lupinus albus seed proteomes

The allergenicity of seed storage proteins, the major components of edible legume seeds, may cause serious reactions in both children and adult population. Updated methodologies for evaluation of the activity of these proteins are needed. In this paper we used two-dimensional (2D) electrophoretic techniques to investigate the immuno-cross-reactivities of anti Ara h 3 basic subunit IgG to the seed proteomes of three legume species, namely, peanut, soybean, and lupin. The seed proteins, extracted with two different procedures, were separated by 2D electrophoresis, and the electrophoretic maps were analyzed by Western blot. In peanut proteome the antibodies strongly reacted with the 23 kDa polypeptides, corresponding to Ara h 3 basic isoforms, the antigen they were raised to, and three unidentified acidic polypeptides near 45 kDa. Remarkable cross-reactivities with lupin and soybean Ara h 3 homologous polypeptides and nonrelated proteins, namely, lupin conglutin gamma and soybean Bg7S, were detected. Therefore, these proteins may be regarded as new putative allergens. The present findings show the potentiality of 2D electrophoresis in the identification of food allergens and open the way to the traceability of the new cross-reacting proteins in the food chain.     

Genetic variability in cultivated common bean beyond the two major gene pools

It is generally accepted that two major gene pools exist in cultivatedcommon bean (Phaseolus vulgaris L.), a MiddleAmerican and an Andean one. Some evidence, based on unique phaseolin morphotypesand AFLP analysis, suggests that at least one more gene pool exists incultivated common bean. To investigate this hypothesis, 1072 accessions from acommon bean core collection from the primary centres of origin, held at CIAT,were investigated. Various agronomic and morphological attributes (14categorical and 11 quantitative) were measured. Multivariate analyses,consisting of homogeneity analysis and clustering for categorical data,clustering and ordination techniques for quantitative data and nonlinearprincipal component analysis for mixed data, were undertaken. The results ofmost analyses supported the existence of the two major gene pools. However, theanalysis of categorical data of protein types showed an additional minor genepool. The minor gene pool is designated North Andean and includes phaseolintypes CH, S and T; lectin types 312, Pr, B and K; and mostly A5, A6 and A4 types-amylase inhibitor. Analysis of the combined categorical data ofprotein types and some plant categorical data also suggested that some othergermplasm with C type phaseolin are distinguished from the major gene pools.     

Occurrence and distribution of organochlorine pesticides (OCPs) in tomato (Lycopersicon esculentum) crops from organic production

Organochlorine pesticides (OCPs) were quantified by GC-ECD in tomato (Lycopersicon esculentum) during a vegetation period. Plants were harvested at 15, 60, and 151 days after seed germination. Leaves, stem, roots, and fruit (peel and flesh) were analyzed separately. The results showed that tomato plants were able to accumulate OCPs from soils, and a trend to reach the equilibrium among tissues at mature stages was also observed. Endosulfans comprised the main OCP group, probably due to its spray during summer months in the surrounding areas. Banned pesticides such as DDTs, heptachlor, and dieldrin were found. OCPs levels in the fruit were below the maximum residues limits (MRL) considered by the Codex Alimentarius. DDE/DDT and alpha-/gamma-HCH ratios of <1 would indicate recent inputs of DDT and lindane in the environment. The occurrence of OCPs in the study farm, where agrochemicals have never been used, is a result of atmospheric deposition of those pesticides.     

GISQ, a multifunctional indicator of soil quality

We present here an indicator of soil quality that evaluates soil ecosystem services through a set of 5 subindicators, and further combines them into a single general Indicator of Soil Quality (GISQ). We used information derived from 54 properties commonly used to describe the multifaceted aspects of soil quality. The design and calculation of the indicators were based on sequences of multivariate analyses. Subindicators evaluated the physical quality, chemical fertility, organic matter stocks, aggregation and morphology of the upper 5 cm of soil and the biodiversity of soil macrofauna. A GISQ combined the different subindicators providing a global assessment of soil quality.Research was conducted in two hillside regions of Colombia and Nicaragua, with similar types of land use and socio-economic context. However, soil and climatic conditions differed significantly. In Nicaragua, soil quality was assessed at 61 points regularly distributed 200 m apart on a regular grid across the landscape. In Colombia, 8 plots representing different types of land use were arbitrarily chosen in the landscape and intensively sampled. Indicators that were designed in the Nicaragua site were further applied to the Colombian site to test for their applicability.In Nicaragua, coffee plantations, fallows, pastures and forest had the highest values of GISQ (1.00; 0.80; 0.78 and 0.77, respectively) while maize crops and eroded soils (0.19 and 0.10) had the lowest values. Examination of subindicator values allowed the separate evaluation of different aspects of soil quality: subindicators of organic matter, aggregation and morphology and biodiversity of macrofauna had the maximum values in coffee plantations (0.89; 0.72 and 0.56, respectively on average) while eroded soils had the lowest values for these indicators (0.10; 0.31 and 0.33, respectively).Indicator formulae derived from information gained at the Nicaraguan sites were not applicable to the Colombian situation and site-specific constants were calculated.This indicator allows the evaluation of soil quality and facilitates the identification of problem areas through the individual values of each subindicator. It allows monitoring of change through time and can guide the implementation of soil restoration technologies. Although GISQ formulae computed on a set of data were only valid at a regional scale, the methodology used to create these indices can be applied everywhere.     

Blind inlets: conservation practices to reduce herbicide losses from closed depressional areas

Purpose

In a 6-year study, we investigated the effectiveness of blind inlets as a conservation practice in reducing pesticide losses compared to tile risers from two closed farmed depressional areas (potholes) in the US Midwest under a 4-year cropping rotation.

Materials and methods

In two adjacent potholes within the same farm and having similar soils, a conventional tile riser and blind inlet were installed. Each draining practice could be operated independent of each other in order to drain and monitor each depression with either practice. Sampling events (runoff events) were collected from the potholes from 2008 to 2013 using autosamplers. The samples were analyzed for atrazine, metolachlor, 2,4-D, glyphosate, and deethylatrazine.

Results and discussion

The results of this study demonstrated that the blind inlet reduced analyzed pesticide losses; however, the level of reduction was compound dependent: atrazine (57 %), 2,4-D (58 %), metolachlor (53 %), and glyphosate (11 %).

Conclusions

Results from this study corroborate previous research findings that blind inlets are an effective conservation practice to reduce discharge and pollutants, including pesticides from farmed pothole surface runoff in the US Midwest.

     

Degradation and binding of atrazine in surface and subsurface soils

Understanding the dissipation rates of chemicals in unsaturated and saturated zones of subsurface soils will help determine if reductions of concentrations to acceptable levels will occur. Chemical properties and microbial biomass and activity were determined for the surface (0-15 cm), lower root (50-105 cm), and vadose (175-220 cm) zones in a Huntington silty clay loam (Fluventic Hapludoll) collected from an agricultural field near Piketon, OH. The rates of sorption, mineralization, and transformation (formation of bound residues and metabolites) of atrazine were determined. Microbial activity was estimated from the mineralization of (14)C-benzoate. We observed decreased levels of nutrients (total organic carbon, N, and P) and microbial biomass with depth, while activity as measured with benzoate metabolism was higher in the vadose zone than in either the surface or the root zones. Sorption coefficients (K(f)) declined from 8.17 in the surface to 3.31 in the vadose zone. Sorption was positively correlated with organic C content. Rates of atrazine mineralization and bound residues formation were, respectively, 12-2.3-fold lower in the vadose than in the surface soil. Estimated half-lives of atrazine ranged from 77 to 101 days in the surface soil, but increased to over 900 days in the subsurface soils. The decreased dissipation of atrazine with increasing depth in the profile is the result of decreased microbial activity toward atrazine, measured either as total biomass or as populations of atrazine-degrading microorganisms. The combination of reduced dissipation and low sorption indicates that there is potential for atrazine movement in the subsurface soils.     

Water mobility in multicomponent model media as studied by 2H and 17O NMR

Molecular mobility of water was studied in a microbiological media containing complex and heterogeneous mixtures of cellulose, l-sorbose, and orange serum broth (OSB) using (2)H and (17)O high-resolution NMR. All mixtures showed Lorentian (17)O NMR spectra but complex (2)H NMR line shapes. Sorbose, when solubilized, caused line-narrowing where as cellulose-OSB mixtures showed wide peaks with flat plateaus. Presence of liquid or solvent water had a profound effect on a marked increase in T(2) relaxation time observed in sorbose-containing samples. (17)O NMR data were not composition dependent, while (2)H NMR data were highly sorbose dependent.     

The combined effects of enhanced UV-B radiation and selenium on growth, chlorophyll fluorescence and ultrastructure in strawberry (Fragaria × ananassa) and barley (Hordeum vulgare) treated in the field

The possible ameliorative effects of selenium (Se) addition to soil on the detrimental effects of enhanced UV-B radiation were tested on strawberry and barley during 4 months of field experiment in Kuopio, Central Finland. Control plants were exposed to ambient levels of UV radiation, using arrays of unenergized lamps. A control for UV-A radiation was also included in the experiment. Added Se, applied as H₂SeO₄, at the level of 0.1 mg kg⁻¹ soil (low dosage) and 1 mg kg⁻¹ soil (high dosage) increased Se concentrations in plants more than 10 and 100 times, respectively. After 4 months of exposure, strawberry and barley plants were harvested for biomass analysis. Chlorophyll fluorescence was measured using the Hansatech FMS2 fluorescence monitoring system. Leaf anatomy and ultrastructure were observed by light and transmission electron microscope. Several effects of UV and Se as well as their interaction were found, mostly for strawberry, but not for barley, indicating species-specific responses. Our results provided evidence that the high Se concentration in soil had no ameliorative effect but increased the sensitivity of strawberry to enhanced UV-B radiation in the field. Under ambient radiation, Se did not alter leaf growth of strawberry, whereas under UV-B radiation, the high Se addition significantly decreased leaf growth. Strawberry runner biomass was affected by the interaction of Se and UV. Under ambient radiation Se did not change dry weight of runners, but in combination with UV-A or UV-B radiation the high Se dosage decreased dry weight of runners by about 30%. Although the high Se concentration positively influenced on quantum efficiency of photosystem II (PSII) in strawberry leaves, it reduced runner biomass, leaf number and ratio of starch to chloroplast area. This suggests that the harmful effects of the high Se dosage on photosynthetic processes occurred as a result of changes in activity or/and biosynthesis of enzymes, rather than alteration of PSII. At the low concentration, Se effects were slight and variable.Although barley leaves accumulated higher Se concentrations than strawberry, there were no apparent changes in their growth, biomass or chlorophyll fluorescence due to Se effect either alone or in combination with UV-B. However, at the ultrastructural level, an enlargement in the peroxisome area was found due to combination of UV radiation with Se, suggesting the activation of antioxidative enzymes, possibly catalase. Decrease in mitochondrial density in barley cells in response to Se might be attributed to alteration of mitochondrial division. Increase in the proportion of cells with cytoplasmic lipid bodies due to combined effect of UV-B and Se indicated the alteration of lipid metabolism and the acceleration of cell senescence in barley. Main UV-B effects were found, mostly at the tissue and ultrastructural level in strawberry, but not in barley, indicating species-specific susceptibility to enhanced UV-B radiation. UV-B-treated strawberry plants developed marginally thinner leaves with reduced ratio of starch to chloroplast area in their cells, suggesting negative influence of UV-B on photosynthetic processes.     

Traces of human migrations in Helicobacter pylori populations

Helicobacter pylori, a chronic gastric pathogen of human beings, can be divided into seven populations and subpopulations with distinct geographical distributions. These modern populations derive their gene pools from ancestral populations that arose in Africa, Central Asia, and East Asia. Subsequent spread can be attributed to human migratory fluxes such as the prehistoric colonization of Polynesia and the Americas, the neolithic introduction of farming to Europe, the Bantu expansion within Africa, and the slave trade.