Plant regeneration from protocorm-derived callus of Cypripedium formosanum

Article

Summary

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 words

Cypripedium formosanum callus induction plant regeneration protocorm-like body 

References

  1. Arditti, J.; Ernst, R. Paphiopedilum. Micropropagation of orchids. New York: John Wiley & Sons, Inc.; 1993:434–466.Google Scholar
  2. Chang, C.; Chang, W. C. Plant regeneration from callus of Cymbidium ensifolium var ‘misericors’. Plant Cell Rep. 17:251–255; 1998.CrossRefGoogle Scholar
  3. Chen, J. T.; Chang, C.; Chang, W. C. Direct somatic embryogenesis on leaf explants of Oncidium ‘Gower Ramsey’ and subsequent plant regeneration. Plant Cell Rep. 19:143–149; 1999.CrossRefGoogle Scholar
  4. Duncan, D. B. Multiple range and multiple F test. Biometrics 11:1–42; 1955.CrossRefGoogle Scholar
  5. Ernst, R. Effects of thidiazuron on in vitro propagation of Phalaenopsis and Doritaenopsis (Orchidaceae). Plant Cell Tiss. Organ Cult. 39:273–275; 1994.CrossRefGoogle Scholar
  6. Ichihashi, S.; Islam, M. O. Effects of complex organic additives on callus growth in three orchid genera, Phalaenopsis, Doritaenopsis and Neofinetia. J. Jap. Soc. Hort. Sci. 68:269–274; 1999.CrossRefGoogle Scholar
  7. Lin, Y. H.; Chang, C.; Chang, W. C. Plant regeneration from callus culture of a Paphiopedilum hybrid. Plant Cell Tiss. Organ Cult. 62:21–25; 2000.CrossRefGoogle Scholar
  8. Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473–497; 1962.CrossRefGoogle Scholar
  9. Spurr, A. R. A low-viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastruct. Res. 26:31–43; 1969.PubMedCrossRefGoogle Scholar
  10. Stewart, J.; Button, J. Tissue culture studies in Paphiopedilum. Am. Orchid Soc. Bull. 44:591–599; 1975.Google Scholar
  11. 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

Copyright information

© Society for In Vitro Biology 2003

Authors and Affiliations

  1. 1.Department of HorticultureNational Taiwan UniversityTaipeiTaiwan

Personalised recommendations