Beans in Europe: origin and structure of the European landraces of Phaseolus vulgaris L.

This study focuses on the expansion of Phaseolus vulgaris in Europe. The pathways of distribution of beans into and across Europe were very complex, with several introductions from the New World that were combined with direct exchanges between European and other Mediterranean countries. We have analyzed here six chloroplast microsatellite (cpSSR) loci and two unlinked nuclear loci (for phaseolin types and Pv-shatterproof1). We have assessed the genetic structure and level of diversity of a large collection of European landraces of P. vulgaris (307) in comparison to 94 genotypes from the Americas that are representative of the Andean and Mesoamerican gene pools. First, we show that most of the European common bean landraces (67%) are of Andean origin, and that there are no strong differences across European regions for the proportions of the Andean and Mesoamerican gene pools. Moreover, cytoplasmic diversity is evenly distributed across European regions. Secondly, the cytoplasmic bottleneck that was due to the introduction of P. vulgaris into the Old World was very weak or nearly absent. This is in contrast to evidence from nuclear analyses that have suggested a bottleneck of greater intensity. Finally, we estimate that a relatively high proportion of the European bean germplasm (about 44%) was derived from hybridization between the Andean and Mesoamerican gene pools. Moreover, although hybrids are present everywhere in Europe, they show an uneven distribution, with high frequencies in central Europe, and low frequencies in Spain and Italy. On the basis of these data, we suggest that the entire European continent and not only some of the countries therein can be regarded as a secondary diversification center for P. vulgaris. Finally, we outline the relevance of these inter-gene pool hybrids for plant breeding.

Development and use of chloroplast microsatellites in Phaseolus spp. and other legumes.

Chloroplast microsatellites (cpSSRs) provide a powerful tool to study the genetic variation and evolution of plants. We have investigated the usefulness of 39 primer pairs tagging cpSSR loci on a set of eight different genera of Leguminosae (Papilionoideae subfamily) and five species belonging to the genus Phaseolus. Thirty-six 'universal' primer pairs were retrieved from the literature, one was re-designed and a further two were designed de novo. The cpSSR loci analysed were highly polymorphic across the individuals examined. Twenty-seven primer pairs were polymorphic in the overall sample, 18 within Phaseolus, and 16 in both P. vulgaris and P. coccineus. Analysis of the plastome sequences of four Leguminosae species (obtained from GenBank) showed that in the loci targeted by universal primer pairs: (i) the originally tagged cpSSRs can be lost; (ii) other cpSSRs can be present; and (iii) polymorphism arises not only from differences in the numbers of cpSSR repeats, but often from other insertion/deletion events. Multilocus linkage disequilibrium analysis suggests that homoplasy is not a major problem in our dataset, and principal component analysis indicates intelligible relationships among the species considered. Our study demonstrates that this set of chloroplast markers provides a useful tool to study the diversity and the evolution of several legumes, and particularly P. vulgaris and P. coccineus.

Sorption of pesticides on kaolinite and montmorillonite as a function of hydrophilicity.

Pesticides and other organic species are adsorbed by soil via different mechanisms, with bond strengths that depend on the properties of both the soil and the pesticide. Since the clay fraction in soil is a preferential sorbent for organic matter, reference kaolinite and montmorillonite are useful models for studying the mechanism and the strength of sorption. This paper presents the results of batch experiments to investigate the interactions of kaolinite KGa-1 and montmorillonite SWy-1 with the following pesticides and organic species resulting from the natural degradation of pesticides in the environment: atrazine (1-chloro-3-ethylamino-5-isopropylamino-2,4,6-triazine), simazine (1-chloro-3,5-bisethylamino-2,4,6-triazine), diuron [1,1-dimethyl-3-(3,4-dichlorophenyl)urea], aniline, 4-chlorophenol, 2,4-dichlorophenol, and 2,4,6-trichlorophenol. Each of these chemicals has different hydrophilicity. Systems containing 2.0 g of clay were put in contact with 100.0 mL of solutions of the pesticides at known concentration ranging from 1.0 to 5.0 mg/L, and the amount of solute adsorbed was evaluated through RP-HPLC analysis of the pesticide still present in the aqueous suspension. To test for electrostatic interactions between the clay surface and the pesticides, potentiometric titration was used to determine the permanent surface charge of clays. Experiments were performed at different pH values. The results indicate that, for the chemicals studied, neutral molecules are preferentially retained relative to ionized ones, and that montmorillonite is a more effective sorbent than kaolinite.

Statistical evaluation of recovery of 3,4-dichloroaniline in soil as function of particle size and analyte concentration.

A great mean value of recovery for extraction of 3,4-dichloroaniline from a soil is calculated from individual recovery values evaluated for four different fractions of the soil. Then the uncertainty associated to this great mean recovery is calculated and used to know whether to apply or not the correction in routine analysis performed for the same kind of soil and the same analyte. The most representative fractions that, as a function of particle size, can be identified in a soil are: sand (2.000-0.063mm), coarse silt (0.063-0.020mm), fine silt (0.020-0.002mm) and clay (

Sorption studies of chloroanilines on kaolinite and montmorillonite.

Batch experiments have been performed in order to evaluate the ability of the two reference clays kaolinite (KGa-1) and Na-montmorillonite (SWy-1) to retain three representative chloroanilines: 3-chloroaniline, 3,4-dichloroaniline and 2,4,6-trichloroaniline. Systems containing the clay mineral and the pollutant solution (at concentration levels ranging between 1.0 and 10.0mg/L) were considered and RP-HPLC methods were employed to follow the sorption processes as a function of time. The results indicate that montmorillonite shows a general higher sorption capacity with respect to kaolinite and that for both the reference clays, in the concentration range investigated, the amount of pollutant sorbed increases with concentration. The sorption coefficient K(d) ranges between 0.0030 L/g for the system 3-chloroaniline-kaolinite and 0.0488L/g for the system 2,4,6-trichloroaniline-montmorrillonite. The most lipophilic trichloroaniline shows the greater sorption. X-ray analyses suggest for kaolinite a preferential sorption onto the mineral surface, while for montmorillonite a progressive swelling of the structure is observed, likely due to sorption processes that also take place in the interlayer.

Oxidative degradation of food dye E133 Brilliant Blue FCF Liquid chromatography-electrospray mass spectrometry identification of the degradation pathway.

This paper is devoted to the evaluation of the degradation pathway of the E133 Brilliant Blue FCF (C.I. 42090) that is largely used in the food industry. The degradation is studied in oxidation conditions obtained by addition of potassium persulfate at different persulfate to dye molar ratios under natural sunlight irradiation. The degradation pathway of the dye passes through a species coloured in dark blue and then gives rise to uncoloured species. Due to the low volatility and the poor thermal stability of the dye, reversed-phase liquid chromatography associated to mass spectrometry and tandom mass spectrometry was employed to follow the kinetics of degradation and identify some intermediates. The identification of organic species still present in the decoloured dye and the value of COD obtained in these conditions show evidence that complete decolorization does not correspond to complete mineralisation. No direct information of toxicity is available for the uncoloured degradation products but the further formation of aromatic amines can not be excluded.

Optimization by experimental design and artificial neural networks of the ion-interaction reversed-phase liquid chromatographic separation of twenty cosmetic preservatives.

Particular attention are recently receiving antimicrobial agents added as preservatives in hygiene and cosmetics commercial products, since some of them are suspected to be harmful to the human health. The preservatives used belong to different classes of chemical species and are generally used in their mixtures. Multi-component methods able to simultaneously determinate species with different chemical structure are therefore highly required in quality control analysis. This paper presents an ion interaction RP-HPLC method for the simultaneous separation of the 20 typical antimicrobial agents most used in cosmetics and hygiene products, that are: benzoic acid, salicylic acid, 4-hydroxybenzoic acid, methyl-, ethyl-, propyl-, butyl-, benzyl-benzoate, methyl-, ethyl-, propyl-, butyl-, benzyl-paraben, o-phenyl-phenol, 4-chloro-m-cresol, triclocarban, dehydroacetic acid, bronopol, sodium pyrithione and chlorhexidine. For the development of the method and the optimization of the chromatographic conditions, an experimental design was planned and models were built by the use of artificial neural network to correlate the retention time of each analyte to the variables and their interactions. The neuronal models developed showed good predictive ability and were used, by a grid search algorithm, to optimize the chromatographic conditions for the separation of the mixture.

Intercalibration of chromatographic methods for auxino phytodrugs in Solanaceae.

Three chromatographic methods are considered for the determination in Solanaceae of auxino-similar phytodrugs, so called because their structure resembles an auxine plant hormone. The phytodrugs studied were: 2,4-dichlorophenoxyacetic acid, 2,4-dichlorophenoxypropionic acid, 2,4-dichlorophenoxybutyric acid, 2-methyl-4-chlorophenoxyacetic acid, 2,4,5-trichlorophenoxyacetic acid, naphthylacetic acid and 2-naphthyloxyacetic acid. Three chromatographic methods, respectively based on ion-interaction HPLC, GC-MS with intra-injector derivatisation and CC-MS with pre-injection derivatisation, were developed, optimised and validated. A comparative discussion of the advantages/disadvantages of the methods suggests a strategy for their preferential use, that is essentially a function of the matrix complexity.

Simultaneous reversed-phase high-performance liquid chromatographic separation of mono-, di-and trichloroanilines through a gradient elution optimised by experimental design.

A new RP-HPLC method for the simultaneous determination of the 13 mono-, di- and trichloroanilines has been developed. In order to obtain the analyte resolution within an acceptable analysis time, a gradient elution program has been optimised through the use of an experimental design and a grid search algorithm. The optimized conditions provided the resolution of all the analytes in less than 80 min. The primary validation of the analytical method gave limit of detection values ranging between 0.02 and 0.06 mg/l and very good linearity of the calibration curves.