Abstract
The 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.
Highlights
Common bean (Phaseolus vulgaris L.) is an important staple food providing protein, fiber, minerals and vitamins [1]
The electrophoretic results, including composition, number and molecular weight of polypeptides in the phaseolin patterns calculated with assistance of the Experion and Fingerprinting software, as well as the phaseolin type classified according to [6, 9] are summarized (Table 1)
The individual phaseolin polypeptides detected by the microchip electrophoresis were validated by the elution profiles and peak areas generated by the Experion software (Fig. 2)
Summary
Common bean (Phaseolus vulgaris L.) is an important staple food providing protein, fiber, minerals and vitamins [1]. This crop has gained increasing attention as functional food [2]. The distinct geographical distribution of beans wild relatives raise the questions of where common bean domestication occurred and what is the gene pool origin of modern crop cultivars. Two major gene pools are commonly distinguished with the use of morphological and biochemical markers [5]. These pools can be detected at different ratios in modern crop cultivars, depending on their place of origin. The H and B types have been linked to Pampa and Boyaca cultivars [9]
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