Shoot-Forming Cultures. The shoot-forming cafli of D. purpurea L., which had been established in the previous study (5), were transferred into the liquid basal medium, supplemented with 1 mg/L BA and 1 mg/L IAA, and cultured in the light. Thus, green, shoot-forming cultures without root were established and subcultured every 3 weeks. White, shoot-forming cultures were obtained by sub-culturing a portion of the green, shoot-forming cultures in the dark.Culture Condition. Murashige and Skoog medium (9)-with 1 mg/L thiamin-HCl and without agar, edamin, IAA, and kinetinwas used as the basal medium. Approximately 1.5 g fresh weight of cells were inoculated into a 500-ml Erlenmeyer flask containing 100 ml of liquid medium and cultured in continuous light (fluorescent lamp: about 4 x 105 erg/s.cm2) or in the dark at 28°C on a reciprocal shaker (100 strokes/min; 2.0 cm in length).Measurement of Growth. Fresh weight was measured after removing culture medium by suction filtration. The harvested fresh culture was lyophilized, and its dry weight was determined.Assay for Digitoxin. Lyophilized cells (0.1-2 g) were homogenized with 50 ml ethanol in a glass homogenizer. The homogenate was heated at 74°C for 4 h and filtered. The filtrate was dried in vacuo, and the residue was taken up in 2 ml ethanol and diluted with 18 ml H20. When necessary, the extract was further diluted with H20 to a desirable level. Determination of digitoxin concentration of the extract was done by radioimmunoassays, as described (5).Assay for Chi. Chl content was determined spectrophotometric-
Muchattention has been paid to the aroma compounds formed by degradation of carotenoids in plants.:) In the previous paper, the usefulness of the microbial transformation for the preparation of these aromacompounds was demonstrated; jS-ionone was converted to more than 13 compounds containing (S)-2hydroxy-/?-ionone and (7?)-4-hydroxy-/?
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