An effective rapid propagation method was established through in vitro cultures of the medicinal plant, Cudrania tricuspidata. In vitro plantlets were obtained from in vitro germinated seeds. The various levels of cytokinins (BAP, Kinetin and TDZ) were tested on multiple shoot formation from plantlets. BAP (1.0 mg/l) treatment induced highest number of multiple shoots. Single shoot cultures gave higher initial shoot numbers than 5 shoots per culture. Among the various culture media, the shoot elongation was optimal on 2 MS basal medium without growth regulators. The IAA (2.0 mg/l) treatment induced highest number of roots. IBA (2.0 mg/l) treatment more promoted in vitro root growth than other concentrations. Rooted shoots were transferred directly to small pots with an artificial soil and successfully acclimatized.

Scopolamine and hyoscyamine are important anticholinergic compounds obtained from Hyoscyamus niger. Adventitious roots induced from rhizome of H. niger and roots were cultured in SH medium supplemented with 3% (w/v) sucrose and 0.5 mg/L IBA. Roots were grown rapidly after 10 days of cultures. Scopolamine production was increased 7 times and hyoscyamine production was increased 12 times after 10 day of cultures. SH medium was best in root growth. But, highest scopolamine productivity was observed in WPM medium, followed White medium and best hyoscyamine productivity was resulted in MS medium. Sucrose was increased scopolamine and hyoscyamine production were increased the medium supplemented by sucrose comparing to than those by other carbon sources.

Extracts from sawdust of Pinus densiflora were showed antifungal activity against Lentinula edodes. It was extracted by hot water and then successively extracted by n-hexane, ethyl acetate, and methanol. The yields of the n-hexane-soluble, ethyl acetatesoluble, methanol-soluble and methanol-insoluble fractions of water extracts were 8.2%, 10.6%, 32.0%, and 49.2%, respectively. The ethyl acetate-soluble fraction showed the greatest antifungal activity against L. edodes: 41.5% inhibition at 1,000 ppm. However, there were not significant differences of antifungal activities between n-hexane-soluble fraction and methanol-soluble fraction at a concentration of 1,000 ppm. The hot water extracts showed 23.5% of antifungal activity against L. edodes at a concentration of 1000 ppm. The four antifungal compounds were separated from ethyl acetate fraction by thin layer chromatography.

Optimal culture conditions for efficient in vitro propagation and polysaccharide production of Orostachys japonicus were established. O. japonicus was cultured in media containing various growth regulators and carbon sources. The highest regeneration rate was achieved in 1.0 and 3.0 mg l -1 of 2,4-D concentration, while the lowest was obtained in 10.0 mg l -1 2,4-D concentration. When different carbone sources were added in the culture medium, plant growth was high in 3% sucrose treatment. The micropropagated shoots were successfully acclimatized in artificial soils and produced comparable amont of polysaccharide compred to parent cultivated plants.

An in vtro nucellar polyembryo propagation method was established with mature seed of the Citrus junos Sieb. 7-8 nucellar polyembryos per seed were induced on MS basal medium without plant growth regulators. The polyembryos developed to complete plantlets on teatment with IBA. These shoots grew further in MS medium without plant growth regulators. Rooting of shoots occurred on MS medium supplemented with IBA. These plantlets were successfully transplanted to small plastic pot containing soil mixture. Somatic embryos were induced from nucellar polyembryo and maturation occurred spontaneously from proliferating cultures on MS medium without growth regulators. Random Amplified Polymorphic DNA (RAPD) marker analysis of in vitro and in vivo grown junos orange showed identical polymorphism indicative of their genetic stability. The RAPD polymorphism produced revealed same banding pattern in each regenerant. Hence, propagaton of junos orange by nucellalr polyembryos was efficient and produced in genetically stable plants under in vitro conditions.

Scopolamine and hyoscyamine are important anticholinergic compounds. To increase the productivity, we have selected various elicitors and developed culture system using a bubble column bioreactor (BCB). As the same manner of elicitation in flask cultures, the elicitors were introduced into BCB cultures and the productivity was investigated. Except the bacterial elicitor of Staphyllococcus aureus, the elicitors inhibited hyoscyamine production. In scopolamine production, the elicitors revealed different responses from the results obtained in flask cultures. The elicitors of KCl and Candida albicans less increased the production than flask cultures. However, methyl jasmonate and S. aureus showed stronger positive effects on tropane alkaloid production. In particular, S. aureus was the most effective elicitor on scopolamine production and the elicitor resulted in the highly increased production, approximately 10 times higher than the control culture.

Scopolia parviflora Nakai, a rare and endangered species, is the sole plant producing tropane alkaloids (TA) among the Korean native species. In order to enhance TA productivity the SP72 root line was selected by screening 100 of root line, and the optimal culture media for root growth and TA production were investigated with the SP72 roots. Based on the several media, SH and 2B5 medium were determined as growth medium and White and NN medium as production medium. Among the four combinations of two-stage culture, 2BN (2B5 as growth medium plus NN as production medium) showed more enhanced root growth and TA production as compared with production media of White and NN medium and growth media of SH and 2B5 medium, respectively. However, bubble column bioreactor (BCB) cultures applying two-stage culture did not reveal the effective results despite of the each successful operation of two-stage culture in conical flasks and BCB cultures.

Total RNAs were isolated from cultured roots of Scopolia parviflora, poly(A)+ RNA was obtained through the mRNA purification, cDNA library of Hnh6h was constructed. Recombinant baculoviruses in Spodoptera frugiperda (Sf) cells were constructed by use of the transfer vector pBacPAK, which has the AcNPV sequence under the polyhedrin promoter. The expression vector carrying Hnh6h gene was transferred to S. parviflora and obtained transgenic hairy root lines. Our results confirmed the over expression of the H6H protein was used by anti-pBacPAK about cDNAs of S. parviflora. This study will served for production of tropane alkaloids by metabolic engineering.

Optimal culture conditions for efficient in vitro propagation and polysaccharide production of Orostachys japonicus were established. The highest growth yield was achieved in 1/2 MS medium, while the lowest growth yield was obtained in 4 MS medium. The patterns of polysaccharide formation were a little similar in all cases, but on MB5 medium, the po]ysaccharide contents of plant were higher than others. Among the modified nitrate levels, effective growth level were obtained in 1/4 N and 1/2 N. High contents of polysaccharide were obtained in 4 N. Different concentration of potassium and calcium did not improve the growth and polysaccharide production. The micropropagated shoots were successfully acclimatized artificial soils.

Hyoscyamine and scopolamine are two most common tropane alkaloids found in the Solanaceae. The pEB expression vector carrying Nspmt gene was transformed to Agrobacterium rhizogenes. The growth of transgenic hairy roots were approximately to 80% of the wild type root. Transgenic hairy roots are less developed on only their branch roots than those of the wild type root. The extracts from Nspmt transgenic hairy root lines, 3 and 5 contained between 3.52 and 4.23 mg/g dry weight as hyoscyamine and did between 5.23 to 6.40 mg/g dry weight as scopolamine. These results showed that the overexpression of the pmt gene enhanced tropane alkaloids production of S. parviflora transformed roots and this improvement affected both hyoscyamine and scopolamine production.