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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: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: June 5, 2006
Publication Date: September 14, 2006
Citation: Schober, T.J., Bean, S. 2006. Potentials and method improvements of capillary zone electrophoresis for use in spelt breeding programs [abstract]. International Gluten Workshop. Poster Paper No. 19.

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. Several examples are given: mislabeled samples could be identified, although the differences in the patterns were very small. Relatedness between different spelt cultivars could be shown. 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. This latter finding is in agreement with several studies, showing that one single protein class (LMW glutenin subunits, gliadins) did not always reflect purity of spelt. The acidic phosphate buffer system was compared to an isoelectric buffer system composed of 50 mM iminodiacetic acid (IDA), HPMC and acetonitrile, 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: 9/10/2014