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Genetic analysis of β-amylase thermostability to develop a DNA marker for malt fermentability improvement in barley, Hordeum vulgare
Authors:T. Kaneko  M. Kihara   K. Ito
Affiliation:Plant Bioengineering Research Laboratories, Sapporo Breweries Ltd, 37-1, Kizaki, Nitta-machi, Nitta-gun, Gunma 370-0393, Japan
Abstract:
β‐Amylase thermostability is one of the major factors affecting fermentability in the brewing process; consequently, it could be used as a selection marker for the trait. In order to clarify what controls its thermostability, the linkage analysis of β‐amylase thermostability and its genotype as restriction fragment length polymorphism patterns was performed in three cross populations. Then, β‐amylase cDNAs cloned from the three varieties which had a different thermostability type were expressed in Escherichia coli. According to the results of the linkage analysis and gene expression test, it was concluded that β‐amylase thermostability resulted from a difference in its structural gene. Furthermore, to construct an STS marker for the gene, the gDNA sequences of β‐amylase were compared among the three varieties, which had different thermostabilities. Although there were many differences in the intron sequence, few nucleotides differed in the exon region. Based on the variation in the intron region, a sequence‐tagged‐site marker was constructed to detect β‐amylase genotypes in breeding material.
Keywords:Hordeum vulgare    AAL    β-amylase    malting quality    STS marker    thermostability    variation
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