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United States Department of Agriculture

Agricultural Research Service

Research Project: ENHANCED END USE QUALITY AND UTILIZATION OF SORGHUM GRAIN

Location: Grain Quality and Structure Research Unit

Title: Potentials and Method Improvements of Capillary Zone Electrophoresis for Use in Spelt Breeding Programs

Authors
item Schober, Tilman
item Bean, Scott

Submitted to: International Gluten Workshop
Publication Type: Proceedings
Publication Acceptance Date: October 16, 2006
Publication Date: N/A

Technical Abstract: Capillary zone electrophoresis (CZE) in acidic buffer systems is capable of separating cereal storage proteins based on similar separation principles as classical acidic polyacrylamide gel electrophoresis. However, it is faster, its resolution is distinctly higher and data evaluation is much simpler. Applying a 100 mM sodium phosphate buffer system pH 2.5 containing hydroxypropyl methylcellulose (HPMC), and using a 60 cm capillary, CZE was successfully used in a spelt breeding program. Mislabeled samples could be identified, although the differences in the patterns were very small. However, it was not possible to clearly differentiate between pure spelts and wheat-spelt crosses. Crossing spelt with modern wheat may be, but is not necessarily, reflected in the gliadin pattern. The acidic phosphate buffer system was compared to an isoelectric buffer system composed of 50 mM iminodiacetic acid (IDA), acetonitrile, and HPMC, using a short (27 cm) capillary. It was found that the IDA system provided more than 10 times faster separations with almost the same resolution as the sodium phosphate system. It is concluded that CZE, especially with the IDA buffer, is a fast and powerful tool in spelt breeding programs to avoid mislabeling, and gain insight into the relatedness of new lines with unknown pedigrees.

Last Modified: 8/30/2014