Liquid Culture Systems for in vitro Plant Propagation

564 S. M. Nalawade et al.
roots (Figure 2 G). Converted embryos developed tubers on MS medium
without phytohormones when incubated for four months (Figure 2 H). The
plantlets were hardened for two months in growth chambers (Figure 2 I).
HPLC analysis revealed presence of both D,L-tetrahydropalmatine and
D-corydaline in the tubers. A marked variation in the content of D,Ltetrahydropalmatine
and D-corydaline among different treatments was
observed. However, in most of the treatments, D,L-tetrahydropalmatine and
D-corydaline content in the tubers was greater than the marketed crude drug
and varied depending on the treatment (Table 1). The low amount of the
compounds in the crude drug may be due to their partial loss during
processing which usually involves either boiling in water, vinegar or wine
(Anonymous, 1999).
4. Conclusions
Using the plant regeneration protocol described above, somatic embryos
were induced in tuber-derived callus with subsequent recurrent somatic
embryos from epidermal cells of the converted primary embryos of C.
yanhusuo. Tuber formation in vitro has been achieved in the converted
somatic embryos. By using appropriate growth regulators and prolonged
incubation under controlled conditions it is possible to produce good quality
tubers with uniform high contents of the two protoberberine-type alkaloids
D,L-tetrahydropalmatine and D-corydaline. These protocols can be used for
mass propagation, biochemical or genetic transformation studies.
Acknowledgements
This research was supported by a grant from the National Science
Council (NSC 88-2317-B-055-001), Taiwan.
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