Browsing by Author "Slaski, Jan Jacek"
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- Microchip electrophoretic analysis of Phaseolin patterns and Its comparison with currently used SDS-PAGE techniquesPublication . Silva, Emanuel Marques da; Santos, Teresa Maria Marques dos; Ganança, José Filipe Teixeira; Slaski, Jan Jacek; Carvalho, Miguel Â. A. Pinheiro deThe goal of this work was to compare reproducibility of phaseolin patterns of common bean obtained by two electrophoretic protein separation techniques including the conventional SDS-PAGE and an automated chip electrophoresis system. Five standard cultivars of common bean provided by the United States Department of Agriculture (Beltsville, Maryland) that represented five phaseolin types, T (Tendergreen), C (Contender) and S (Sanilac), B (Boyaca) and P (Pampa), were used in this study. Comparison of the phaseolin patterns revealed that the chip-on-a-lab electrophoresis provided a good reproducibility. The phaseolin polymorphism included four to seven polypeptides typical for the pattern composition of the T, C and S types. The polymorphism of the B and P patterns was also established. Phaseolin polypeptides separated by the microchip electrophoresis exhibited differences with respect to the molecular weights and electrophoretic mobility as compared to the SDS-PAGE technique. This phenomenon could be attributed to the absence of a solid separation phase in the microchip electrophoresis. Moreover, this technique has potential to substantially accelerate screening of large bean germplasm collections since it allows for the accurate analysis of the higher number of individual plants within accessions than the conventional, tedious and time consuming SDS-PAGE method.
- Phytogeographical origin of Madeiran common beans based on phaseolin patternsPublication . Silva, Emanuel Marques da; Correia, Anísia Soraia Abreu; Lopes, Nuno Alexandre Amaral; Nóbrega, Humberto Gil Moreira; Ganança, José Filipe Teixeira; Domingues, Ana Maria; Khadem, Manhaz; Slaski, Jan Jacek; Carvalho, Miguel Ângelo Almeida Pinheiro deThe objective of this work was to determine the geographic origin of the Madeiran common bean (Phaseolus vulgaris) gene pool. Phaseolin patterns of 50 accessions representing the diversity of common bean collected in Madeira, Portugal, and conserved in the ISOPlexis Germplasm Bank, were analysed using the Experion automated electrophoresis system, based on lab-on-a-chip technology. Five common bean standard varieties with typical phaseolin patterns were used to determine the phytogeographical origin of the Madeiran common bean accessions. Ninety two percent of the accessions exhibited a phaseolin pattern consistent with the one of common bean types belonging to the Andean gene pool, while the origin of the remaining 8% of the accessions was indistinguishable. The application of a similarity coefficient of 85%, based on Pearson correlations, increases the number of accessions with uncertain pattern. The analytical approach used permitted the determination of the origin of the common bean gene pool, which is Andean in 98% of the cases, and clustering of the observed variability among the Madeiran common beans.