PRODUCTION OF TERPENOID FROM ARTEMISIA-ANNUA L PLANTLET CULTURES IN BIOREACTOR

Seeds of Artemisia annua obtained from different locations in Europe, Michigan, Washington, Hoechst (India), and Lucknow (India) were grown at the Experimental Field Station at Trombay, Bombay. The plants synthesized various levels of terpenoids of which camphor, 1,8-cineol and beta-caryophyllene were the major constituents. The Lucknow variety synthesized the highest level 1.99 mg% fresh weight (FW) of camphor, the Michigan variety produced 1.08 mg% FW of 1,8-cineol, and the Washington variety contained 0.38 mg% FW of -caryophyllene. Callus cultures interspersed with shoot buds were obtained from all the plant varieties on Murashige and Skoog's medium containing 1 mg l(-1) 6-benzyladenine and 1 mg l(-1) napthaleneacetic acid (Murashige and Skoog, 1962). Multiple shoot cultures were established from the Europe variety on MS medium supplemented with 0.1 mg l(-1) 6-benzyladenine and 1 mg l(-1) indole-3-acetic acid. Plantlet formation occurred on medium containing 1 mg l(-1) napthaleneacetic acid and 0.1 mg l(-1) kinetin. The plantlets produced camphor (0.81 mg% FW), 1,8-cineol (0.07 mg% FW) and beta-caryophyllene (0.025 mg% FW). Multiple shoot and plantlets were grown in 1-1 capacity bioreactors. The plantlet cultures from the bioreactors produced 1.08 mg% FW camphor followed by 0.14 mg% FW 1,8-cineol and 0.65 mg% FW beta-caryophyllene. Addition of gibberellic acid and ethephone to the culture medium in the bioreactor stimulated the synthesis of terpenoids. The cultures produced 3.41 mg% FW camphor after 15 d of cultivation in the bioreactor with ethephone addition.