Callus tissues derived from chilling-tolerant herbaceous plant, Atractylodes lancea, Atropa belladonna, Bupleurum falcatum, Dioscorea tokoro, Lithospermum erythrorhizon and Phytolacca americana could be cold-stored at 4°C for three months or more, whereas those from chilling-sensitive herbaceous plants such as Datura innoxia and Perilla frutescens vat. crispa and a deciduous tree, Mallotus ]aponicus, could not survive after cold storage for two to three months. Tobacco callus cultures could be stored at 4°C for two or four months depending on a callus strain. The effect of cold storage on secondary metabolite production varied. Nicotine and betalain production suffered from cold storage of tobacco and Phytolacca americana callus cultures, respectively. However, production of anthocyanin in cultures of Mallotus ]aponicus and Bupleurum falcatum and shikonin derivatives in Lithospermum erythrorhizon callus was affected very little. Root-forming ability was retained for more than one year in cold-stored callus tissues of Bupleurum falcatum, while the control callus tissues maintained at 25°C completely lost the organogenetic ability six months after the first subculture.
Various characteristics including the saponin content in the root of Bupleurum falcatum plants propagated in vitro through somatic embryogenesis of callus cultures were compared with those of the plants propagated by seeds. The asexually propagated plants had an aerial part of more uniform characteristics than those of sexually propagated ones. However, both the mean and variance of root weight of the former were significantly larger than those of the latter. As for the saponin content of the root on a dry weight basis, there was little difference between the two groups. The amounts of saikosaponins c and d in a root were significantly larger in the asexually propagated plants than in the sexually propagated ones.
Shoot cultures ofAtractylodes lancea DC. (Compositae) have been established by inoculating the flower bud on Linsmaier-Skoog's medium supplemented with 10(-5) M naphthaleneacetic acid and 10(-5) M 6-benzyl-aminopurine. Shoots were multiplied on a medium containing 10(-6) M 6-benzylaminopurine. Propagated shoots rooted on a medium devoid of any plant growth regulators were transferred to potting soil and finally to the field.
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