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SDS-PAGE 및 PCR 을 활용한 국산 밀 품종의 글루텐닌 조성 분석

Analysis of Glutenin Compositions in Korean Wheat Cultivar Using SDS-PAGE and PCR

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키워드

주제분류농학

초록

PCR-based assays with co-dominant or dominant allele-specific STS (sequence tagged sites) markers and sodiumdodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) have been used to discriminate Glu-1 and Glu-3 alleles in Korean wheat cultivars. Three alleles were identified at each high molecular weight glutenin (Glu-1) homoeologous locus and 10 alleles were identified at low molecular weight glutenin (Glu-3) homoeologous locus in Korean wheat cultivars. At Glu-1 loci,wheat cultivars contained Glu-A1a, b and c alleles amplified three fragments of 1093, 1063 and 920bp, and Glu-B1b, c and f alleles discriminated with 707bp (Glu-B1b and f) and 662bp (Glu-B1c) fragments and Glu-D1a, d and f alleles amplified two fragments576bp (Glu-D1d) and 612bp (Glu-D1a and f), respectively. At Glu-3 loci, wheat cultivars contained Glu-A3c, d and e alleles amplified a 573bp, 967bp and 158bp fragments, respectively, Glu-B3d, g, h and i alleles gave a 662bp, 853bp, 1022bp and 621bp, respectively and c) bands and Glu-D3a, b and c alleles amplified a 884bp (Glu-D3a and b) and 338bp (Glu-D3c)fragments. A multiplex-PCR assay was also established which permitted the discrimination of the major Glu-1 and Glu-3 to determine glutenin compositions in a single PCR reaction and agarose gel assay because these systems could be useful to select DNA-based identification of wheat lines carrying good bread making performance in Korean wheat breeding programs. Three multiplex-PCRs for Glu-1 and Glu-3, Glu-A1c/ Glu-B1acf/Glu-D1d, Glu-A3ac/Glu-A3d/Glu-A3e and Glu-B3d/Glu-B3fg /Glu-B3h were established in this study.

Abstract

PCR-based assays with co-dominant or dominant allele-specific STS (sequence tagged sites) markers and sodiumdodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) have been used to discriminate Glu-1 and Glu-3 alleles in Korean wheat cultivars. Three alleles were identified at each high molecular weight glutenin (Glu-1) homoeologous locus and 10 alleles were identified at low molecular weight glutenin (Glu-3) homoeologous locus in Korean wheat cultivars. At Glu-1 loci,wheat cultivars contained Glu-A1a, b and c alleles amplified three fragments of 1093, 1063 and 920bp, and Glu-B1b, c and f alleles discriminated with 707bp (Glu-B1b and f) and 662bp (Glu-B1c) fragments and Glu-D1a, d and f alleles amplified two fragments576bp (Glu-D1d) and 612bp (Glu-D1a and f), respectively. At Glu-3 loci, wheat cultivars contained Glu-A3c, d and e alleles amplified a 573bp, 967bp and 158bp fragments, respectively, Glu-B3d, g, h and i alleles gave a 662bp, 853bp, 1022bp and 621bp, respectively and c) bands and Glu-D3a, b and c alleles amplified a 884bp (Glu-D3a and b) and 338bp (Glu-D3c)fragments. A multiplex-PCR assay was also established which permitted the discrimination of the major Glu-1 and Glu-3 to determine glutenin compositions in a single PCR reaction and agarose gel assay because these systems could be useful to select DNA-based identification of wheat lines carrying good bread making performance in Korean wheat breeding programs. Three multiplex-PCRs for Glu-1 and Glu-3, Glu-A1c/ Glu-B1acf/Glu-D1d, Glu-A3ac/Glu-A3d/Glu-A3e and Glu-B3d/Glu-B3fg /Glu-B3h were established in this study.

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