Evaluation of microsatellite detection using autoradiography and capillary electrophoresis in hops

Resumo

Two sequence tagged sites (STS) detection techniques were used in the identification of 25 hop cultivars. In cultivar identification, capillary electrophoresis is more efficient than autoradiography. First, it was more sensitive in detection of fragments that allowed observation of the instability in the locus 7a82 due to the small size differences of its alleles. Additionally, the fluorescent detection of fragments was a nonsubjective technique suitable of automation that increased the flexibility of work because no radioactive labeling or long vertical polyacrylamide electrophoresis was needed. Only two cultivars, Hersbrucker and Hallertauer, were indistinguishable, but further molecular analysis revealed that both were included in a separate cluster within the European hop cultivars. The statistical analysis of fluorescence and radioactive detection data including allelic frequencies, heterozygosity, and probability of identity are displayed in this study, which corroborates the following advantages of the capillary electrophoresis technique: high sensitivity, amenable to automation, and its nonsubjectivity. Therefore, fluorescent-labeled primer microsatellite detection by capillary electrophoresis is a powerful tool in cultivar identification for hop breeders, merchants, and brewers. © 2005 American Society of Brewing Chemists, Inc.