Polimorfizam alela glijadina i glutena kod pšenice (Triticum aestivum L.)

Abstract

Glijadini i glutenini se deponuju u endospermu semena i imaju najveći udeo u sadržaju proteina semena. Cilj ovog rada je identifikacija polimorfizama genskih alela koji kodiraju glijadine i glutenine. U istraživanje je uključeno 10 sorti pšenice. Od svakog genotipa je korišćeno 30 semena za ekstrakciju glijadina sa 70% etanolom, a glutenina sa 10% β-merkaptoetanolom. Glijadini su razdvojeni elektroforezom u rastvoru pufera (pH=3,1) na 8,33% poliakrilamidnom gelu, dok su glutenini razdvajani pomoću SDS-PAGE (pH-8,6) na 11,8% gelu. Elektroforegrami su korišćeni za određivanje alela Gli-1 i Gli-2, kao i alela Glu-1. Identifikovano je pet alela (a, b, c, f, m) na Gli-A1, četiri alela (b, g, e, l) na Gli-B1, četiri alela (a, b, g, k) na Gli-D1, pet alela (b, e, g, m, n) na Gli-A2, šest alela (b, c, d, g, k, o) na Gli-B2 i tri alela (a, b , e, h, m) na Gli-D2 lokusu. Za podjedinice glutenina visoke molekularne mase (HMVGS) je identifikovano tri alela (a, b, c) na Glu-A1, sedam alela (a, b, c, d, f, h, i) na Glu-B1 i tri alela (a, c, d) na Glu-D1. Identifikovani aleli na svakom Gli-1, Gli-2 i Glu-1 lokusu ukazuju na visok polimorfizam proteina glutena i genetičku divergentnost analiziranih genotipova pšenice.

Gliadin and glutenins are deposited in endosperm of seed and they have the highest share in the protein content of seeds. The aim of this study is identification of gene allele polymorphisms encoding gliadin and glutenins. Ten varieties of wheat were included in this study and 30 seeds from each genotype were used for the extraction of gliadin by 70% ethanole, and glutenins by 10% β-mercaptoethanol. The gliadins were separated by acid PAGE electrophoresis (pH=3.1) on 8.33% polyacrylamide gel, while glutenins were separated by SDS-PAGE (pH-8.6) on 11.8% gel. Electrophoregrams were used for determining Gli-1 and Gli-2 alleles as well for Glu-1 allele. The five alleles (a, b, c, f, m) at the Gli-A1, four alleles (b, g, e, l) at the Gli-B1, four alleles (a, b, g, k) at the Gli-D1, five alleles (b, e, g, m, n) at the Gli-A2, six alleles (b, c, d, g, k, o) at the Gli-B2 and five alleles (a, b, e, h, m) at the Gli-D2 locus were identified. For high molecular weight glutenin subunists (HMWGS) the three alleles (a, b, c) at the Glu-A1, seven alleles (a,b, c, d, f, h, i) at the Glu-B1 and three alleles (a c, d) at the Glu-D1 were identified. Identified alleles at each Gli-1, Gli-2 and Glu-1 loci indicate high polymorphisms of gluten proteins and genetic divergences of analyzed wheat genotypes.