Cereal Chem 66:186-192   |  VIEW ARTICLE

Fast Horizontal Sodium Dodecyl Sulfate Gradient Polyacrylamide Gel Electrophoresis for Rapid Wheat Cultivar Identification and Analysis of High Molecular Weight Glutenin Subunits.

B. A. Marchylo, K. A. Handel, and V. J. Mellish. Copyright 1989 by the American Association of Cereal Chemists, Inc. 

Fast horizontal sodium dodecyl sulfate gradient polyacrylamide gel electrophoresis (SDSGPAGE) with silver staining in conjunction with the PhastSystem was used to separate wheat storage proteins (gliadin and glutenin). Resolution achieved with miniature 8-25% gradient PhastGels (50 x 43 x 0.45 mm) was almost equal to that of conventional SDSGPAGE and was superior to that of 10-15% gradient PhastGels. Wheat cultivars distinguishable by conventional electrophoresis also were distinguishable by fast horizontal SDSGPAGE using either 8-25% or 10-15% gradient PhastGels. The 10-15% gradient format had the shortest analysis time (45 min electrophoresis and 44 min staining) and was used for the analysis of 70 wheat cultivars. The majority of cultivars could be distinguished by qualitative differences in protein patterns with the exception of three groups of cultivars comprised of seven red spring, two white winter, and two red winter wheats. Identification of high molecular weight glutenin subunits by fast horizontal SDSGPAGE also was possible with the limitation that subunits 2* and 5 were not resolved and 1 and 2 were partially resolved. An overall evaluation of the advantages of fast horizontal SDSGPAGE and silver staining in conjunction with the PhastSystem suggests that this technique could be used effectively in situations where smaller numbers of samples would be analyzed on an intermittent basis as contrasted to daily analysis of large numbers of samples.