Study On ELISA For Glycyrrhizinic Acid And Regulation Of The Yield And Quality Of Licorice (Glycyrrhiza Uralensis Fisch.)

The enzyme-linked immunosorbent assay(ELISA) for glycyrrhizinic acid in licorice(Glycyrrhiza uralensis Fisch.) was developed. The method developed was used for evaluating the quality of licorice treated by plant growth substance HKL-4. The effects of HKL-4 on yield, component, leaf physiological activities, leaf photosynthesis and root physiological function in licorice was also investigated.1.It is the first time that haptens and glycyrrhizinic acid(GA) were covalently conjugated to bovine serum albumin(BSA) or ovalbumin(OVA) to prepare antigens(immunogens) and coating antigens by carbodiimide(CDI) method. The conjugates were identified and their molecular ratio of hapten to carrier proten were determined by UV spectrophotometry. Conjugates GA-BSA were used to immunize Balb/c mouse to prepare polyclone antibodies(PcAbs) for GA. The liters of antisera were higher than 3.2 × 106 . Powdered licorice root samples were extracted. The contents glycyrrhizinic acid determined by ELISA agreed well with those determined by HPLC. The ELISA results were good correlation with HPLC, the correlation coefficient is r2=0.9918. The establishment of the enzyme-linked immunosorbent assay (ELISA) for glycyrrhizinic acid is reported first time in China.2.The contents of glycyrrhizinic acid in licorice roots were determined by ELISA. It is the cooperational time of the contents of glycyrrhizinic acid and yield in the later autumn. The contents of glycyrrhizinic acid tended to be higher in the younger parts of the root and in the older parts of the rhizome.3. HKL-4 regulated the hormonal systems, then improved the cooperational expression of yield and quality of roots and rhizomes. The yield of roots and rhizomes increased by 13.8% through HKL-4 treatment. The contents of glycyrrhizinic acid and flavornes in roots increased by 32.4% and 35.3-47.5% respectively. Using of the fingerprints of extracted ion chromatograms(EIC),the safety of chemical components in the licorice roots treated by HKL-4 was evaluated. No new compounds was found.4. Biochemcial control increased the physiological activities in leaves. HKL-4 treatments significantly increased the activity of SOD and CAT, while the activity of POD and the content of MDA was reduced . Treated by HKL-4, the submicroscopic structure of leaf changed. Chloroplast was most sensitive, then mitochondria. We supposed that the appropriate dosage of plant growth substance can improve the ability of biosynthesis and energy metabolism in leaf, so that photochemical efficiencies(Fv/Fm ) increased.5. The physiological activity of root was improved by HKL-4 treatment. HKL-4 treatments significantly increased the activity of SOD and CAT, while the activity of POD and the content of MDA reduced. HKL-4 treatments increased the biomass of roots and rhizomes of licorice.6. The contents of IAA, GA4 and CTKs in leaves, stems and rhizome significant increased by HKL-4 treatment, while ABA contents decreased. HKL-4 treatments significantly increased the contents of IAA, GA4 and CTKs in roots, while the contents of ABA in later growth stage are higher than that in the control.