The wild Cymbidium lowianum , a national-level rare and endangered species of Orchid, is an excellent garden plant with ornamental flowers with striking, deep red lips in a V-shaped formation. The objective of this study was to estab lish a micropropagation protocol system via immature seeds of wild Cymbidium lowianum , evaluate the Murashige and Skoog (MS), half-strength Murashige and Skoog ( 1/2 MS) medium, 6-benzylaminopurine (BA), a-napthaleneacetic acid (NAA), organic additions activated charcoal (AC) and banana pulp (BP) effects on the different morphogenesis (seed germination, multiple shoot and rooting) in vitro . The optimal combination for the germination of seed was 1.0 mg ·L - 1 BA with 0.5 mg ·L - 1 NAA in 1/2 MS, and addition 0.3%AC, which resulted in 95% seed germination in 90 days. The best formulation for multiple shoot was 1/2 MS medium containing 2.5 mg·L - 1 BA, 0.5 mg ·L - 1 NAA and addition 8% BP in which produced 19.8 shoots per protocorm in 60 days. Multiple shoots were cut and rooted in 1/2 MS supple mented with 1.5 mg ·L - 1 NAA,0.1 mg·L - 1 BA and 0.3% AC, roots initiated 20 days after culture, the rooting percent ages reached to 100%, in which 4.7 per shoot produced roots in 60 days. The survival rate of plantlet was up to 92% in moss after 30 days. This finding reveals that it is possible to obtain in vitro culture of Cymbidium lowianum using im mature seeds in asymbiotic culture.
J. Y. Chen, “The Top Ten Most Known Flowers of China,” Shanghai Culture Publishing House, Shanghai, 1989.
J. D. Chung, C. K.Chun and S. O. Choi, “Asymbiotic Germination of Cymbidium ensifolium 2; Effects of Several Supplements to the Medium, pH Values and Light and/or Dark Culture Periods on Growth of Rhizome and Organogenesis for Rhizome,” Journal of The Korean Society for Horticultural Science, Vol. 26, No. 2,1985, pp. 186-192.
K. Shimasaki and S. Uemoto, “Micropropagation of a Terrestrial Cymbidium Species Using Rhizomes Developed from Seeds and Pseudo Bulbs,” Plant Cell, Tissue and Organ Culture, Vol. 22, No. 3, 1990, pp. 237-244. doi:10.1007/BF00033642
X. Wang, “Tissue Culture of Cymbidium: Plant and flower Induction in Vitro,” Lindleyana, Vol. 3, 1988, pp. 184-189.
N. R.Nayak, M. Tanaka and J. A. Teixeira da Silva, “Biotechnology of Cymbidium—An Overview of Recent Progress and Future Opportunities,” In: J. A. Teixeira da Silva, Ed., Floriculture, Ornamental and Plant Biotechnology: Advances and Topical Issues, Vol. 4, Global Science Books, London, 2006, pp. 558-562.
R. B. Malabadi, J. A. Teixeira da Silva, K. Nataraja and G. S. Mulgund, “Shoot Tip Transverse Thin Cell Layers and 2,4-Epibrassinolide in the Micropropagation of Cymbidium bicolor Lindl,” Floriculture and Ornamental Biotechnol, Vol. 3, 2008, pp. 44-48.
D. T. Nhut, T. N. T. Tien, M. T. N. Huong, N. T. H. Hien, P. P. X. Huyen, V. Q. Luan, B. V. Le and S. Teixeira da, “Artificial Seeds for Preservation and Propagation of Cymbidium spp,” Prop Ornamental Plants, Vol. 5, 2005, pp. 67-73.
D. Lauzer, M. S. Arnaud and D. Barabe, “Teterazolium Straining and in Vitro Germination of Mature Seeds of Cypripedium acaule (Orchidaceae),” Lindeleyana, Vol. 9, 1994, pp. 197-204.
V. Vujanovic, S. M. Arnaud, D. Barabe and G. Thibeault, “Viability Testing of Orchid Seed and the Promotion of Coloration and Germination,” Annals of Botany, Vol. 86, No. 1, 2000, pp. 79-86. doi:10.1006/anbo.2000.1162
T. R. Johnson, S. L. Stewart, D. Dutra, M. K. Kane and L. Richardson, “A Symbiotic and Symbiotic Seed Germination of Eulophia alta (Orchidaceae)—Preliminary Evidence for the Symbiotic Culture Advantage,” Plant Cell, Tissue and Organ Culture, Vol. 90, No. 3, 2007, pp. 313-323.
J. Arditti, J. D. Michaud and A. P. P. Oliva (1981), “Seed Germination of North American Orchids. I. Native California and Related Species of Calypso, Epipactis, Goodyera, Piperia and Plantantheria,” Botanical Gazette, Vol. 142, No. 4, 1981, pp. 442-453. doi:10.1086/337245
A. Thornhill and H. Koopowitz, “Viability of Disa Uniflora Berg (Orchidaceae) Seeds under Variable Storage Conditions: Is Orchid Gene Banking Possible?” Biological Conservation, Vol. 62, No. 1, 1992, pp. 21-27. doi:10.1016/0006-3207(92)91148-L
L. W. Zettler and T. M. McInnis, “Symbiotic Seed Germination and Development of Spiranthes cernua and Goodyera pubescens (Orchidaceae: Spiranthoideae),” Lindleyana, Vol. 8, 1993, pp. 155-162.
P. P. J. Kauth, W. A. Vendrame and M. E. Kane, “In Vitro Seed Culture and Seedling Development of Calopogon tuberosus,” Plant Cell, Tissue and Organ Culture, Vol. 85, No. 1, 2006, pp. 91-102. doi:10.1007/s11240-005-9055-1
S. L. Stewart and M. K. Kane, “A Symbiotic Seed Germination and in Vitro Seedling Development of Habenaria macroceratitis (Orchidaceae) a Rare Florida Terrestrial Orchid,” Plant Cell, Tissue and Organ Culture, Vol. 86, No. 2, 2006, pp. 147-158. doi:10.1007/s11240-006-9098-y
G. Mahendran and V. Narmatha Bai, “Mass Propagation of Satyrium nepalense D. Don.—A Medicinal Orchid via Seed Culture,” Scientia Horticulturae, Vol. 119, No. 2, 2009, pp. 203-207. doi:10.1016/j.scienta.2008.07.029
R. Mitchell, “Growing Hardy Orchids from Seeds at Kew,” Plantsman, Vol. 1, 1989, pp. 152-169.
A. B. Anderson, “Symbiotic and Asymbiotic Germination and Growth of Spiranthes magnicamporum (Orchidaceae),” Lindleyana, Vol. 6, 1991, pp. 183-186.
Z. L. Li, C. X. Yu, Y. Y. Wang and B. Q. Wang, “Research on Aseptic Budding and Fast Propagation of the Cymbidium tracyanum Seed,” Chinese Agricultural Science Bulletin, Vol. 21, No. 8, 2005, pp. 269-281.
V. L. Sheela, S. Sarada and S. Anita, “Micropropagation of Orchids,” Journal of Ornamental Horticulture, Vol. 9, No. 1, 2006, pp. 1-11.
P. P. D. Davies, “The Plant Hormones: Their Nature, Occurrence and Functions,” Plant Hormones, Springer, 2010, pp. 1-15.
K. A. Marrs, W. R. Alfenito, A. M. Lloyd and V. Walbot, “A Glutathione S-Glucosyltransferase Involved in Vacuolar Transfer Encoded by the Maize Gene Bronze-2,” Nature, Vol. 375, No. 6530, 1995, pp. 397-400. doi:10.1038/375397a0
J. Ross and Damian O’Neill (2001), “New Interactions between Classical Plant Hormones,” Plant Science, Vol. 6, No. 1, 2001, pp. 2-4. doi:10.1016/S1360-1385(00)01795-7
E. V. Soniya and M. Sujitha, “An Efficient in Vitro Propagation of Aristolochia indica,” Biologicl Plantarum, Vol. 50, No. 2, 2006, pp. 272-274. doi:10.1007/s10535-006-0018-0
S. Manjula, A. Thomas, B. Daniel and G. M. Nair, “In Vitro Plant Regeneration of Aristolochia Indica through Axillary Shoot Multiplication and Organogenesis,” Plant Cell Tissue Organ Culture, Vol. 51, No. 2, 1997, pp. 145-148. doi:10.1023/A:1005978125424
G. Birmeta and M. Welander, “Efficient Micropropagation of Ensete Ventricosum Applying Meristem Wounding: A Three-Step Protocol,” Plant Cell Reports, Vol. 23, No. 5, 2004, pp. 277-283. doi:10.1007/s00299-004-0832-9
T. V. Nguyen, T. T. Thu, M. Claeys and G. Angenon, “Agro Bacterium Mediated Transformation of Sorghum (Sorghum bicolor (L.) Moench) Using an Improved in Vitro Regeneration System,” Plant Cell, Tissue and Organ Culture, Vol. 91, No. 2, 2007, pp. 64-155.
J. C. Jang and F. H. Tainter, “Micropropagation of Short Leaf, Virginia and Loblolly Short Leaf Pine Hybrids via Organogenesis,” Plant Cell, Tissue and Organ Culture, Vol. 25, No. 1, 1991, pp. 57-61. doi:10.1007/BF00033914
M. O. Islam, S. Matsui and S. Ichihashi, “Effects of Complex Organic Additives on Seed Germination and Carotenoid Content in Cattleya Seedlings,” Lindleyana, Vol. 15, 2000, pp. 81-88.
J. VanStaden and J. Stewart, “Cytokinins in Banana Fruit,” Zeitschrift für Pflanzenphysiologie, Vol. 76, No. 3, 1975, pp. 280-283.
T. D. Thomas and A. Michael, “High-Frequency Plantlet Regeneration and Multiple Shoot Induction from Cultured Immature Seeds of Rhynchostylis retusa Blume, an Exquisite Orchid,” Plant Biotechnology Reports, Vol. 1, No. 1, 2007, pp. 243-249. doi:10.1007/s11816-007-0038-z