Nicotine production by tissue cultures of tobacco as influenced by various culture parameters.

Tissue culture systems of tobacco (Nicotiana tabacum L., Nicotiana rustica L.) are described, in which up to 5% and more nicotine accumulated on a dry weight basis. Maximum amounts of nicotine (5.3 %, yield 920 mg/1) were detected in mixotrophic green cell suspensions of a newly developed flue-cured tobacco cultivar (N. tabacum cv. Delcrest X cv. McNair 133). This cultivar also showed maximum nicotine content compared to other strains when cultured as callus or heterotrophic cell suspension. Nicotine content was increased up to 29-fold (2. 9 %, yield 360 mg/1) compared to the stock cultures, when a two- or three-stage culture was employed by reducing the auxin and cytokinin levels, raising medium sucrose to 40 gil and replacing ammonium nitrate by potassium nitrate in the production medium. Cells in 20 1 batch fermentor cultures accumulated up to 1 % nicotine with a final yield of 28 mg/1, thus producing much less nicotine.