Screening of Glycyrrhiza uralensis Fisch. ex DC. containing high concentrations of glycyrrhizin by Eastern blotting and enzyme-linked immunosorbent assay using anti-glycyrrhizin monoclonal antibody for selective breeding of licorice

A rapid selection system was used to screen Glycyrrhiza uralensis plants containing high concentrations of glycyrrhizin (GC) by Eastern blotting using anti-GC monoclonal antibody (MAb). Chromatographic fingerprinting by Eastern blotting correlated with the GC concentration analyzed by enzyme-linked immunosorbent assay (ELISA). The roots of wild G. uralensis growing in Mongolia were analyzed by Eastern blotting to identify plants containing high concentrations of GC, and the GC concentration was confirmed by ELISA. G. uralensis plants cultivated in the greenhouse were also analyzed in the same manner. GC concentrations in wild G. uralensis roots and cultivated plants varied widely: between 0.06 and 9.36 percent dry weight (dw%). To confirm the homogeneity of GC concentrations in the cultivated plants, we monitored GC concentrations in the plants over 2 years. Although GC concentrations changed in two plants, they remained comparatively constant in the other five plants, suggesting that GC concentrations are genetically determined. To identify high GC-producing plants, 1025 plants were analyzed, and the highest concentration of GC was 5.36 dw%.