In vitro production of phytoecdysteroids from Ajuga turkestanica

Leaf tissue from wild‐harvested A. turkestanica , a source of adaptogenic phytoecdysteroids, was used to initiate callus and cell suspension cultures. Precursors of phytoecdysteroids (mevalonic acid, cholesterol and acetate) or methyl jasmonate were added on day 2 of subculture to elicit accumulation of phytoecdysteroids in cell suspension cultures. In addition, hairy root cultures were developed from in vitro leaf and shoot tissue inoculated with Agrobacterium rhizogenes . Addition of 15 mg L −1 and 150 mg L −1 mevalonic acid increased the phytoecdysteroid 20‐hydroxyedysone from 8.0 mg mg −1 (control) to 40.2 and 29.2 mg mg −1 , respectively, in cell suspension cultures. Turkesterone content was only slightly increased from 4.1 mg mg −1 in control cell cultures to 5.8 and 4.9 mg mg −1 in cultures elicited with 15 mg L −1 and 150 mg L −1 mevalonic acid, respectively. Cholesterol, sodium acetate and methyl jasmonate did not provoke any increased accumulation of 20‐hydroxyedysone but slightly increased turkesterone content. Cyasterone and cyasterone 22‐acetate were detected in trace amounts in both control and elicited cell cultures. Extracts from 28 day old hairy roots contained 7.3 mg mg −1 20‐hydroxyedysone and 2.1 mg mg −1 turkesterone. The production of phytoecdysteroids was demonstrated in hairy root cultures and enhancement of phytoecdysteroid accumulation was demonstrated from cell suspension cultures of A. turkestanica .