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

Abstract

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%.

Ultra high performance liquid chromatography coupled with triple quadrupole mass spectrometry and chemometric analysis of licorice based on the simultaneous determination of saponins and flavonoids

“…Consequently, licorice is known as “the king of herbs” or “Guo lao” in Chinese, meaning “national venerable master” . It appears in approximately 60% of TCM prescriptions and 70% of herbal formulas in Japan . In TCM, licorice is usually processed with honey and traditionally used as a decoction , which is considered to change the property of licorice from cool to warm .…”

Development of a Competitive Time‐Resolved Fluoroimmunoassay for Paclitaxel

“…MAb 3A3 we used in this assay was an IgG antibody with a k‐type light chain and showed low cross‐reactivities against the structurally related compounds found in Taxus species, which ensured its high specificity to Tax. Given the rather low Tax content in the shoot bark of Taxus (the highest level being only 0.022% in Taxus samples, as shown in Table ), a highly sensitive and specific assay system is essential for the breeding of medicinal plants like Glycyrrhiza species previously succeeded by us using ELISA (Fujii et al ., ).…”

Quality Control of Natural Products by Fingerprinting of Eastern Blotting

“…Licorice is also widely used as a sweetener in the production of confectioneries, soy sauce and is therefore in high demand worldwide. An anti-GC MAb was prepared in our laboratory [14] and we previously reported on the synergistic effects of GC and other constituents [37], the interfacial behavior of GC [77], and the identification of Glycyrrhiza species with higher GC content [27]. More recently we found that two flavonoids, isoliquiritigenin and formononetin promote in vitro fertilization [78].…”

“…We prepared many MAbs against natural products like forskolin [1], solamargine [2], crocin [3], marihuana compound [4], opium alkaloids [5], ginsenosides [6,7], berberine [8], sennosides [9,10], paeoniflorin [11,12], glycyrrhizin (GC) [13,14], saikosaponin [15][16][17], ginkgolic acid [18], aconitine alkaloid [19] and baicalin [20]. More recently MAbs against capsaicinoids [21], artemisinin [22,23], luteoloside [24], daidzin [25] and puerarin [26] have been prepared and developed individual competitive enzyme-linked immunosorbent assay (ELISA) as a high sensitive, specific, and simple methodology resulting in screening of medicinal plants targeting bioactive component [27], pharmacokinetics for capsaicinoids [21] artemisinin [28], and ginsenoside Rb1 and Rg1 [29].…”