Abstract
High-resolution melting (HRM) analysis is a closed-tube method for detecting nucleotide polymorphisms, enabling rapid genotyping. In this study, we have developed a rapid strain-typing method for Kinko 115, a premium shiitake cultivar, by HRM analysis. When HRM analyses of tyrosinase (tyr) genes of 64 shiitake cultivars were carried out, 3 cultivars including Kinko 115 could be distinguished from another 61 cultivars. After that, HRM analyses of the ribosomal intergenic spacer (IGS) region in these 3 cultivars revealed that Kinko 115 was distinguished from the other 2 cultivars. Consequently, Kinko 115 could be distinguished from other cultivars by HRM analyses of the tyr gene and the IGS region. Furthermore, Kinko 115 could be distinguished from the others by HRM analyses using not only purified DNA, but also crude DNA extracts from mycelia or gills of dried fruiting bodies. Thus, the HRM analyses developed in this study would be useful for rapidly and easily identifying Kinko 115.
