In Vitro Cellular & Developmental Biology - Plant

, Volume 41, Issue 6, pp 765–769 | Cite as

Cytokinins induce direc somatic embryogenesis of Dendrobium chiengmai pink and subsequent plant regeneration

  • Hsiao-Hang Chung
  • Jen-Tsung Chen
  • Wei-Chin ChangEmail author


Four auxins (2,4-dichlorophenoxyacetic acid [2,4-D], indole-3-acetic acid [IAA], indole-3-butyric acid [IBA], and naphthaleneacetic acid [NAA]), and five cytokinins (N 6-[2-isopentenyl]-adenine [2iP], N 6-benzyladenine [BA], 6-furfurylaminopurine [kinetin], 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea [TDZ], and 6-[4-hydroxy-3-methylbut-2-enylamino]purine [zeatin]) were examined for their effects on direct embryo induction from leaf explants of Dendrobium cv. Chiengmai Pink cultured on 1/2 Murashige and Skoog (MS) medium. Whether in light or darkness, explants easily became necrotic and no embryos were obtained on growth regulator-free or auxin-containing media after 60 d of culture. By contrast, five cytokinins tested induced direct embryo formation from leaf explants, and explants cultured in light had a higher embryogenic response compared with those cultured in darkness. The best condition for direct embryo induction was at 18.16 μM TDZ cultured in light for 60 d, where 33% of explants formed a mean number of 33.6 embryos per explant. During subculture on growth regulator-free 1/2 MS medium, embryos gradually developed into plantlets. Secondary embryogenesis was occasionally found on sheath leaves of embryos. Regenerated plantlets were successfully transplanted and grown in a greenhouse environment.

Key words

cytokinin direct somatic embryogenesis leaf explant 


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  1. Arditti, J.; Ernst, R. Micropropagation of orchids. New York: John Wiley; 1993:311–365.Google Scholar
  2. Chang, C.; Chang, W. C. Plant regeneration from callus culture of Cymbidium ensifolium var. misericors. Plant Cell Rep. 17:251–255; 1998.CrossRefGoogle Scholar
  3. Chen, J. T.; Chang, W. C. Efficient plant regeneration through somatic embryogenesis from callus cultures of Oncidium (Orchidaceae). Plant Sci. 160:87–93; 2000.PubMedCrossRefGoogle Scholar
  4. Chen, J. T.; Chang, W. C. Effects of auxins and cytokinins on direct somatic embryogenesis from leaf explants of Oncidium ‘Gower Ramsey’. Plant Growth Regul. 34:229–232; 2001.CrossRefGoogle Scholar
  5. Chen, J. T.; Chang, W. C. Direct somatic embryogenesis and plant regeneration from leaf explants of Phalaenopsis amabilis var. formosa Shimadzu. Bio. Plant. (in press); 2005.Google Scholar
  6. Chen, J. T.; Chang, C.; Chang, W. C. Direct somatic embryogenesis from leaf explants of Oncidium ‘Gower Ramsey’ and subsequent plant regeneration. Plant Cell Rep. 19:143–149; 1999.CrossRefGoogle Scholar
  7. Dawns, C. J. Biological techniques in electron microscopy. New York: Barnes and Noble; 1971.Google Scholar
  8. Duncan, D. B. Multiple range and multiple F test, Biometrics 11:1–42; 1955.CrossRefGoogle Scholar
  9. Huan, L. V. T.; Takamura, T.; Tanaka, M. Callus formation and plant regeneration from callus through somatic embryo structures in Cymbidium orchid. Plant Sci. 166:1443–1449; 2004.CrossRefGoogle Scholar
  10. Ishii, Y.; Takamura, T.; Goi, M.; Tanaka, M. Callus induction and somatic embryogenesis of Phalaenopsis. Plant Cell Rep. 17:446–450; 1998.CrossRefGoogle Scholar
  11. Manorama, P.; Rao, A. N.; Goh, C. J.; Loh, C. S. Leaf callus development in Aranda and Dendrobium. In: Fifth ASEAN Orchid Congress, Singapore, 1984:102–109.Google Scholar
  12. Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:495–497; 1962.CrossRefGoogle Scholar
  13. Nayak, N. R.; Rath, S. P.; Patnaik, S. In vitro propagation of three epiphytic orchids, Cymbidium aloifolium (L.) Sw., Dendrobium aphyllum (Roxb.) Fisch and Dendrobium moschatum (Buch-Ham) Sw. through thidiazuron-induced high frequency shoot proliferation. Sci. Hortic. 71:243–250; 1997.CrossRefGoogle Scholar
  14. Nayak, N. R.; Sahoo, S.; Patnaik, S.; Rath, S. P. Establishment of thin cross section (TCS) culture method for rapid micropropagation of Cymbidium aloifolium (L.) Sw. and Dendrobium nobile Lindl. var. oculatum Hk. f. Sci. Hortic. 94:107–116; 2002.CrossRefGoogle Scholar
  15. Tokuhara, K.; Mii, M. Induction of embryogenic callus and cell suspension culture from shoot tips excised from flower stalk buds of Phalaenopsis (Orchidaceae). In Vitro Cell. Dev. Biol. Plant 37:457–461; 2001.CrossRefGoogle Scholar
  16. Zimmerman, J. L. Somatic embryogenesis: a model for early development in higher plants. Plant Cell 5:1411–1423; 1993.PubMedCrossRefGoogle Scholar

Copyright information

© Society for In Vitro Biology 2005

Authors and Affiliations

  • Hsiao-Hang Chung
    • 1
  • Jen-Tsung Chen
    • 1
  • Wei-Chin Chang
    • 1
    Email author
  1. 1.Institute of Plant and Microbial Biology BotanyAcademia SinicaTaipeiTaiwan, Republic of China

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