Summary
An in vitro culture procedure was established for repetitive embryogenesis and plant regeneration from seed-derived protocorms of Phalaenopsis amabilis var. formosa Shimadzu (Orchidaceae). Seed-derived protocorms were cultured on modified half-strength Murashige and Skoog (1962) basal medium (1/2MS) devoid of plant growth regulators. After 45 d, 28.1% of protocorms formed embryos from their posterior regions. 1-Phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (TDZ; 0.45, 4.54, and 13.62 μM) promoted direct embryo formation. The best response was at 13.62 μM TDZ, and 100% of the protocorms formed a mean number of 13.5 embryos after 45d of culture. By contrast, naphthaleneacetic acid (NAA) at 0.54 and 5.37 μM inhibited direct embryo formation. On basal medium devoid of plant growth regulators, 18.8% of primary proliferating embryos could form more embryos. TDZ (0.45, 4.54, and 13.62 μM) also promoted this process. Proliferating embryos/protocorms were transferred to basal medium devoid of plant growth regulators for plantlet formation. Plantlets were successfully obtained from the embryos after 4–6 wk. Following subculture every 6 wk for three passages, the plantlets were transferred to sphagnum moss in a container for acclimatization in the greenhouse. The survival rate was 100%.
Similar content being viewed by others
References
Arditti, J.; Ernst, R. Micropropagation of orchids. New York: John Wiley & Son; 1993:467–520.
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.
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.
Chen, Y.; Piluek, C. Effects of thidiazuron and N6-benzylaminopurine on shoot regeneration of Phalaenopsis. Plant Growth Regul. 16:99–101; 1995.
Chen, Y. C.; Chang, C.; Chang, W. C. A reliable protocol for plant regeneration from callus culture of Phalaenopsis. In Vitro Cell. Dev. Biol. Plant 36:420–423; 2000.
Dawns, C. J. Biological techniques in electron microscopy. New York: Barnes and Noble; 1971.
Duan, J. X.; Chen, H.; Yazawa, S. In vitro propagation of Phalaenopsis via culture of cytokinin-induced modes. J. Plant Growth Regul. 15:133–137; 1996.
Duncan, D. B. Multiple range and multiple F test. Biometrics 11:1–42; 1955.
Ernst, R. Effects of thidiazuron on in vitro propagation of Phalaenopsis and Doritaenopsis (Orchidaceae). Plant Cell Tiss. Organ Cult. 39:273–275; 1994.
Ishii, Y.; Takamura, T.; Goi, M.; Tanaka, M. Callus induction and somatic embryogenesis of Phalaenopsis. Plant Cell Rep. 17:446–450; 1998.
Islam, M. O.; Ichihashi, S. Effects of sucrose, maltose and sorbitol on callus growth of Phalaenopsis, Doritaenopsis and Neofinetia. J. Jap. Soc. Hort. Sci. 68:1124–1131; 1999.
Jensen, W. A. Botanical histochemistry. San Francisco: Freeman; 1962.
Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473–497; 1962.
Tanaka, M.; Hasegawa, A.; Goi, M. Studies on the clonal propagation of monopodial orchids by tissue culture. I. Formation of protocorm-like bodies from leaf tissues in Phalaenopsis and Vanda. J. Jap. Soc. Hort. Sci. 44:47–58; 1975.
Teob, E. S. Orchids of Asia. Singapore: Times Books International; 1989:125–134.
Tokuhara, K.; Mii, M. Micropropagation of Phalaenopsis and Doritaenopsis by culturing shoot tips of flower stalk buds. Plant Cell Rep. 13:7–11; 1993.
Young, P. S.; Murthy, H. N.; Yoeup, P. K. Mass multiplication of protocorm-like bodies using bioreactor system and subsequent plant regeneration in Phalaenopsis. Plant Cell Tiss. Organ Cult. 63:67–72; 2000.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Chen, JT., Chang, WC. Induction of repetitive embryogenesis from seed-derived protocorms of Phalaenopsis amabilis var. Formosa shimadzu. In Vitro Cell.Dev.Biol.-Plant 40, 290–293 (2004). https://doi.org/10.1079/IVP2003527
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1079/IVP2003527