Identification and characterization of high-molecular-weight secalins from triticale seeds by capillary zone electrophoresis.
A rapid and reliable method for separation and characterization of the variability of high-molecular-weight secalin subunits (HMW-SS) in hexaploid triticale (x 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.
Salmanowicz
Boleslaw P
BP
Institute of Plant Genetics, Polish Academy of Sciences, Poznań, Poland. bsal@igr.poznan.pl
D000097:Acetonitriles / Q000737:chemistry; D000483:Alleles; D002523:Edible Grain / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005983:Glutens / Q000032:analysis* / Q000145:classification; D008970:Molecular Weight; D011092:Polyethylene Glycols / Q000737:chemistry; D012639:Seeds / Q000737:chemistry*
D000097:Acetonitriles; D011092:Polyethylene Glycols; D005983:Glutens; C037302:glutenin; C032159:acetonitrile