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This paper reports a study of the alkaloid content of different parts of Datura stramonium L. (Solanaceae) in various stages of its growth and development, and the location, structure and ultrastructure of alkaloid-biosynthesizing cells (idioblasts) in different parts of naturally grown and cultured plant material. TLC, HPLC, and GC/MS methods were used for alkaloid assays. The results showed that alkaloid production starts from the end of the second week after seed germination, increases in different organs up to the tenth week of growth, and then decreases. Leaves and capsules showed the highest alkaloid content in the vegetative and generative stages, respectively. In leaves the alkaloids decreased rapidly in the generative stage. The highest alkaloid content was recorded in vegetative leaves, followed in descending order by vegetative petioles, generative and vegetative stems, generative petioles, generative roots, generative leaves, vegetative roots and mature seeds. The organs as well as calli derived from different leaf parts were examined for the presence of idioblasts by microscopic and cytochemical methods. Idioblasts were present only in semi-hyaline callus originated from leaf base; they were spherical or oval, with a thick cell wall and large central vacuole. These observations should prove helpful in attempts to produce specific alkaloids in naturally grown plants and cell cultures.
Hairy root culture plays an important role in investigation of alkaloid production in culture in vitro. The complexity of scientific work and the production capacity limitations associated with whole plant systems and unorganized cells can be overcome using culture of hairy roots. This paper presents an example of a strategy to produce secondary metabolites from plants, in which we investigated the alkaloid pathway, modified gene expression, and cultivated and optimized hairy root growth in a bioreactor. Datura innoxia whole plant was transformed by Agrobacterium rhizogenes, and the obtained hairy roots were studied for their tropane alkaloid production. Optimization of medium led to an increase of scopolamine production in hairy root culture of Datura innoxia by increasing the produced biomass, but genetic engineering seems to be the best way to increase the accumulation of scopolamine. These root cultures were not able to stably overaccumulate scopolamine. The involvement of putative enzymatic regulation of the tropane alkaloid pathway is discussed.
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