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Identification and characterization of high‐molecular‐weight secalins from triticale seeds by capillary zone electrophoresis
Author(s) -
Salmanowicz Boleslaw P.
Publication year - 2010
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200900691
Subject(s) - glutenin , triticale , chemistry , chromatography , capillary electrophoresis , electrophoresis , isoelectric focusing , linum , polyacrylamide gel electrophoresis , common wheat , protein subunit , biochemistry , biology , botany , enzyme , chromosome , gene
A rapid and reliable method for separation and characterization of the variability of high‐molecular‐weight secalin subunits (HMW‐SS) in hexaploid triticale (× Triticosecale Wittmack) by CZE has been developed. In this method, a mixture of two poly(ethylene oxide) polymers differing in molecular weight and a high concentration of ACN in isoelectric buffer was applied as the running electrolyte. For dynamic coating of the capillary inner wall, a low‐concentration mixture of poly(vinylpyrrolidone) and hydroxypropylmethylcellulose was employed. Wide allelic variations in rye HMW‐SS composition, including some novel x‐ and y‐type HMW‐SS, were detected by CZE. The CZE electropherograms of HMW‐SS showed two groups of peaks in accordance with y‐ and x‐type subunits, with migration times of 8.0–8.8 and 11.0–13.3 min, respectively. HMW‐SS differed in migration times from the simultaneously resolved HMW glutenin subunits, but frequently had very similar electrophoretic mobilities during separation by SDS‐PAGE. Each of the two rye subunits 2r and 6.5r detected by SDS‐PAGE represents in fact two subunits (5.1r or 5.2r, and 6.4r or 6.5r, respectively). After analyzing 106 European triticale cultivars, 12 HMW‐SS were identified (six x‐type and six y‐type). They form six allelic variants of these subunits. The simultaneous separation and identification of triticale HMW glutenin and secalin subunits by CZE is an efficient alternative to SDS‐PAGE and should facilitate breeding of valuable cultivars.