Effect of Culture Origin and Conditions on Duvatrienediol Accumulation in Shoot Cultures of Tobacco

A comparison of morphology, trichome type and duvane accumulationof tobacco shoot cultures originating from seedlings, callusand cell suspension cultures incubated on a range of cytokininconcentrations is presented. A method for the extraction andHPLC analysis of duvatrienediols from cultures is described.Duvatrienediols (DVTs) were not detected in callus or cell suspensioncultures and were only detected in seedling-denved shoot cultures.Shoot development was dependent on explant type probably asa result of pre-exposure of callus and suspension cells to highauxin concentrations. Low cytokinin levels promoted shoot developmentwhilst high cytokinin levels increased the number of shootsregenerated but retarded their development. DVT accumulationoccurred only in ‘normally’ developed shoots withtrichomes. Growth of shoot cultures in liquid medium was inhibitoryto the accumulation of DVTs. Optimum conditions for the accumulationof DVTs were shown to be incubation of seedling-derived shootcultures on media containing 10^–6 M BAP solidified withagar. The yield was 120 µg g^–1 fr. wt. which iscomparable to greenhouse-grown material. Both and ß-DVTwere detected in a ratio of 5: 1. The relationship between tnchomesand DVT accumulation is discussed with reference to the rolesof culture origin, cytokinin, trichome type and environmentaleffects. Key words: Tobacco, duvatrienediol, trichome, shoot culture