Secondary metabolite production is greatly influenced by other culture conditions that need to be identified and optimized for maximum production. Intensity of light and wavelength used has an influence over secondary metabolite production. In cultures of Dioscorea deltoidea, while blue light stimulated alkaloid production, red light decreased it. Similar reports are associated with cell culture accumulation of cardenolides, flavonoids, and betacyanins. Catharanthus roseus cultures when grown under dark conditions accumulated more of serpentine and ajmalicine than in light. When calli were grown under white light, first ajmalicine got accumulated, but when they were exposed to red or blue light there was a constant ajmalicine content and serpentine content was lower.

Presence of gases, oxygen and carbon dioxide in the culture environment has an important role in secondary metabolite production. A higher dissolved oxygen in the media stimulated greater serpentine accumulation in C. roseus cultures. Likewise, formed carbon dioxide and accumulated ethylene, if not quickly removed from the culture head space, inhibited ajmalicine production in these cultures.

UV irradiation induces production of important dimeric alkaloids in C. roseus shoot cultures.