Significant enhancement of oleanolic acid accumulation by biotic elicitors in cell suspension cultures of Calendula officinalis L.

The effects of elicitors on cell growth and oleanolic acid (OA) accumulation in shaken cell suspension cultures of Calendula officinalis were investigated. Elicitors were added individually at various concentrations to 5-day-old cell cultures and their effects monitored at 24 h intervals for 4 days. Different effects on OA accumulation were observed depending on the day of treatment. Jasmonic acid was the most efficient elicitor. After 72 h of treatment with 100 μM JA, the intracellular content of OA reached its maximum value (0.84 mg g−1 DW), which was 9.4-fold greater than that recorded in an untreated control cultures. The addition of chitosan at 50 mg l−1 produced a 5-fold enhancement of OA accumulation (0.37 mg g−1 DW) after 48 h of treatment. Treatment with yeast extract at 200 mg l−1 for 96 h or with pectin at 2 mg l−1 for 48 h produced identical cellular levels of OA (0.22 mg g−1 DW). Lastly, 48 h elicitation with homogenate of the fungus Trichoderma viride produced a 1.8-fold increase in oleanolic acid content (0.12 mg g−1 DW). In addition to significantly stimulating OA accumulation and its secretion into the culture medium, the elicitors also caused slight inhibition of cell growth.