Capillary electrophoresis: a useful tool for the management of plant genetic resources.

Capillary electrophoresis (CE) is a powerful analytical tool that is widely applied to the analysis of biological samples. Proteins, peptides, nonprotein amino acids, phenolic compounds, and ions can be analysed using this electrophoretic methodology. This review summarises some applications of CE to the evaluation and characterisation of plant genetic resources of both Triticum and legume species, as carried out at the Istituto di Genetica Vegetale, National Research Council (IGV-CNR) in Bari (Italy). Different protein fractions as well as nonprotein amino acids were investigated by capillary zone electrophoresis (CZE), the most user-friendly mode of CE application. The described case studies show that CZE can be applied to some institutional activities of gene banks such as the evaluation of genetic diversity within stored collections, the acquisition of new samples, the differentiation of species belonging to the same genus, the identification of misclassified accessions, and the measurement of compounds relevant to nutrition or health.

'Fagiolo a Formella', an Italian lima bean ecotype: biochemical and nutritional characterisation of dry and processed seeds.

BACKGROUND: An ecotype of the lima bean, named 'fagiolo a Formella', which, to the best of our knowledge, is the only example of an Italian lima bean (Phaseolus lunatus L.) ecotype, is cultivated in the Campania region of southern Italy. Physical, nutritional and processing traits of dry seeds were evaluated for two consecutive growing seasons (2009 and 2010). The canning quality was also investigated, but only for the harvest of 2010. RESULTS: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of total seed proteins allowed the attribution of 'fagiolo a Formella' to the Mesoamerican gene pool and Sieva morphotype. Seeds have a trapezoid shape, white coat and 100-seed weight greater than 42 g. Yield, protein, trypsin inhibitor and phytic acid values were found comparable with those reported for lima bean varieties cultivated in sub-tropical areas. Moreover, we found that this ecotype is devoid of lectin. CONCLUSIONS: The good adaptation to growing environment is indicated by the fact that 'fagiolo a Formella' seed quality is comparable to that of lima beans grown in America. Overall the canning quality was found satisfactory and canning significantly destroys the main anti-nutritional compounds present in dry seeds.

Environmental effect on yield, composition and technological seed traits of some Italian ecotypes of grass pea (Lathyrus sativus L.).

BACKGROUND: Grass pea seeds are a good source of vegetable proteins, but the presence of toxic and antinutritional compounds represents a barrier to their large-scale use as food or animal feed. How much growing location and/or seasonal climate might affect the storage of these factors has been little investigated. RESULTS: Fourteen Italian ecotypes of grass pea were cultivated in two locations in southern Italy characterised by different climatic conditions. The seven ecotypes with the best yields and/or seed quality were investigated for a further two growing seasons. From a statistical point of view the physicochemical and nutritional traits among ecotypes were not the same from one year to the next. Moreover, a significant positive correlation was found between ß-oxalyl-diamino-propionic acid and trypsin inhibitor contents. The lowest levels of both these compounds were associated with the highest amount of rainfall during the plant vegetative cycle. CONCLUSION: Principal component analysis of the data showed that the overall seed composition was affected by the growing location. Consequently, each grass pea genotype should also be carefully investigated in relation to different environments before being considered for release as safe for widespread human or animal consumption.

Extraction and separation of water-soluble proteins from different wheat species by acidic capillary electrophoresis.

Optimization of protein extraction and a capillary zone electrophoresis method for water-soluble protein analysis in wheat is described. The optimal separation was obtained with a 50 microm i.d. x 27 cm (20 cm to detector) uncoated capillary filled with 0.1 M phosphoric acid/beta-alanine, pH 2.5, buffer containing urea (1 M), 0.05% (w/v) hydroxypropylmethylcellulose, and 20% (v/v) acetonitrile. Separation was carried out at 15 kV and 35 degrees C for 9 min. Extract stability was also investigated within 2 h from the extraction. Good visual peak parameters and a higher sensitivity can be obtained when 30% ethanol is used as an extraction medium. The method was successfully used to analyze extracts obtained from whole and refined meals of six Triticum spp. Moreover, the described methodology could be applied to the discrimination of species with different ploidy levels and to the detection of durum wheat adulteration, as well as to screen wheat collections for enzymes involved with the quality of wheat derivatives.

Simple and rapid method for the differentiation of Lens culinaris Medik. from false lentil species.

The practice of using both common vetch (Vicia sativa L.) and single-flower vetch (Vicia articulata Hornem.) seeds as food or feed is encouraged by the very high resemblance of their seeds with those of small-seeded lentil cultivars. Among the Vicieae, antinutritional and toxic factors are particularly important, because many species, containing high levels of these compounds, are not safe. A simple and fast capillary electrophoresis (CE) method was proposed for the differentiation of lentil cultivars from false lentil species (i.e., single-flour vetch and common vetch). Proteins were extracted from defatted milled seeds with an alcoholic/saline solution. Extracts were separated in an uncoated fused-silica capillary with iminodiacetic acid (IDA) isolectric buffer containing 0.05% hydroxypropylmethylcellulose (HPMC) and 20% acetonitrile. The presented method is useful also for the detection of contamination of whole or split seeds of lentil by vetch species. With respect to alternative techniques, such as DNA-based markers or thin-layer chromatography (TLC), CE has the advantages of being less expensive, faster, and fully automated.

Specific differentiation in Vicia genus by means of capillary electrophoresis.

Capillary electrophoresis has been applied to the discrimination of 13 Vicia species, belonging to four sections of Vicia genus. The studied species necessitate of plant growing tests or DNA molecular markers to be distinguished being their seeds morphologically very similar. Alcoholic/saline extracts from dry cotyledons were separated in uncoated fused-silica capillary with iminodiacetic acid isolectric buffer containing hydroxypropylmethylcellulose and acetonitrile. The low intra-specific variation observed for 11 species, suggests that this approach is suitable to carry out species discrimination. Species-specific peaks were identified for V. articulata, V. atropurpurea, V. bithynica, V. benghalensis, V. disperma, V. ervilia, V. monantha, V. sativa and V. villosa. Conversely, V. lutea, V. melanops and V. peregrina, showing very similar electrophoregrams, require other methodological approaches to be discriminated. The discussed CE method appears to have a potential to be regarded as an alternative tool to identify some Vicia species being far less expensive and time consuming than plant growing tests and DNA molecular markers.