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This study was performed, using several temperate cymbidiums obtained from asymbiotic seed culture, to assess the change of rhizome poiyphenol contents during culture as well as the process of tissue development during organogenesis. This study was also undertaken to obtain basic data for plant genetics and breeding by comparing the bands of several isoenzymes in rhizomes derived from self- or cross-pollination. The polyphenol contents of rhizomes obtained from cross-polination were higher than those from self-pollination. In this case, the content of ethyl acetate insoluble polyphenol (UP) was higher than that of ethyl acetate soluble polyphenol (FP-I-MAP). Total polyphenol contents were maximum after 30∼60 days in culture but were decreased after 90 days, and also the browning of medium was severe. When the extracts of rhizomes were analysed using TLC (thin-layer chromatography), rhizomes cultured with BAP showed more spots and medium browning than without BAP. The tips of all rhizomes had meristems, and active epidermal and subepidermal cells formed apical meristem. However, the surface area of apical meristem were different. Shoot-forming cells had the large quantity of cytoplasms, and the vacuolization of these cells was delayed. As the result of electrophoresis of rhizomes obtained from self-or cross-pollinaticn, MDH (malate dehydrogenase: E.C.l.l.l.37), 6-PGD (6-phosphogluconate: E.C.1.1.1.44), PGI (phophoglucose isomerase: E.C.4.3.1.9), PGM (phosphoglucomutase: E.C.2.7.5.1), and AcPH (acid phosphatase: E.C. 3.1.3.2) out of 12 isoenzymes used, showed distinctive bands and difference between species in activity. Therefore, these isoenzymes thought to be useful markers for the future genetic analysis.
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