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Abstract
Twenty-one hop varieties in Europe and the United States were successfully identified by DNA analysis, based on single nucleotide polymorphisms (SNPs) as identification markers. Several dozen Mb of transcriptome sequencing data were obtained by next generation sequencer (NGS) from three hop varieties, respectively, and compared to search the regions, which contained many SNPs (including indel sequence). Consequently, four SNP-rich regions were selected as candidates for identification markers of hop varieties. Sequence data of these regions in all varieties tested here were obtained by the normal Sanger method and compared in terms of SNPs. Combination of these SNPs could work well for identification of 21 hop varieties. Moreover, the mixture of two varieties could be correctly evaluated by this method. Hop pellet samples of two different varieties were mixed at various ratios and DNA sequencing was carried out. As a result, the coexisting variety at even a 5% level could be detected by examining electropherograms of the SNP positions. A more quantitative method for mixture evaluation can be expected by DNA techniques, such as quantitative real-time PCR. Because this SNP-based identification method utilizes the DNA sequence itself, it could be an accurate and reproducible tool for the identification of hop varieties.