Plant regeneration from protocorm-derived callus of Cypripedium formosanum
Totipotent callus of Cypripedium formosanum, an endangered slipper orchid species, was induced from seed-derived protocorm segments on a quarter-strength Murashige and Skoog medium containing 4.52 μM 2,4-dichlorophenoxyacetic acid and 4.54 μM 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (thidiazuron). This callus proliferated well and was maintained by subculturing on the same medium. On average, 13 protocorm-like bodies could be obtained from a piece of 4 mm callus after being transferred to the medium with 4.44 μM N6-benzyladenine after 8 wk of culture. The regenerated protocorm-like bodies formed shoots and roots on medium containing 1 g l−1 activated charcoal and 20 g l−1 potato homogenate. After 24 wk of culture on this medium, well-developed plantlets ready for potting were established.
Key wordsCypripedium formosanum callus induction plant regeneration protocorm-like body
- Arditti, J.; Ernst, R. Paphiopedilum. Micropropagation of orchids. New York: John Wiley & Sons, Inc.; 1993:434–466.Google Scholar
- Stewart, J.; Button, J. Tissue culture studies in Paphiopedilum. Am. Orchid Soc. Bull. 44:591–599; 1975.Google Scholar
- Yeung, E. C. Histological and histochemical staining procedures. In: Vasil, I. K., ed. Cell culture and somatic cell genetics of plants, vol. 1. Laboratory procedures and their applications. Orlando, FL: Academic Press; 1984:689–697.Google Scholar