PRODUCTION OF TOBACCO ALKALOIDS BY IMMOBILIZED PLANT CELLS (NICOTIANA TABACUM)

Available from UMI in association with The British Library. Requires signed TDF.; The production of tobacco alkaloids by callus, suspension and immobilized cultured cells of Nicotiana spp. was investigated. Cultures of several species of Nicotiana demonstrated a capacity for nicotine production throughout prolonged periods in culture.; In undifferentiated callus cultures nicotine production was observed to be stimulated by addition of low concentrations of auxin, but suppressed by high levels. The other growth hormone, kinetin, was observed to have a similar effect on nicotine production. This was frequently associated with the differentiation of buds and shoots.; With suspension and immobilized cultures of Nicotiana tabacum var. Havana several factors including the pretreatment of cultures during the generation prior to culture on a nicotine production medium were identified as having a substantial effect on the amount of nicotine produced. Conditions of immobilization also affected the viability and productivity of the culture.; Light, which is one important environmental factor for plant growth, was found to suppress nicotine biosynthesis in callus and suspension cultures. Callus and suspension cultures cultured in the dark produced up to three times the nicotine content of cultures grown in continuous low intensity light.; The design and application of a small scale airlift fermenter in both batch and fed-batch modes of culture with immobilized N. tabacum var. Havana cells was investigated. A calcium-alginate gel entrapment system was used for plant cell immobilization. During airlift fermentation, the elicitation of nicotine by the addition of the oligosaccharin, ML-5(a trirhamnoside), and nicotine release induced by adding various concentrations of DMSO were also investigated. Elicitation did not occur in this system, but the use of DMSO resulted in almost all the accumulated nicotine in the cells being released into the medium within 48 hours of treatment.; Studies of immobilized plant cells culture using the airlift fermenter suggested it had very good prospects in plant biotechnology.