Hort. Sci. (Prague), 2012, 39(1):47-52 | DOI: 10.17221/52/2011-HORTSCI
Organic growth supplement stimulants for in vitro multiplication of Cymbidium pendulum (Roxb.) Sw.
- Plant Tissue Culture Laboratory, Department of Natural Products, National Institute of Pharmaceutical Education and Research, Mohali, India
The aim of this study was to establish protocol for in vitro regeneration and multiplication of Cymbidium pendulum through protocorms, as explants using organic growth supplements such as banana homogenate, coconut water, and peptone. The effect of growth supplements was tested on multiplication (neo-formation of secondary protocorms from primary protocorms) of protocorms, development of maximum number of shoots and early plantlets in M medium (Mitra medium). Though the explants regenerated in the absence or presence of growth adjuncts in the medium; the regeneration frequency was found significantly higher in organic growth supplement-enriched medium than control. The protocorm segments (primary) regenerated into protocorm-like bodies (secondary protocorms) which eventually differentiated into shoots; no intervening callus stage was observed. Among the treatments, the highest regeneration frequency, robust shoots and root formation was recorded in banana homogenate (50 g/l); the roots were lengthy, thick, and covered all over the surface by root hairs. Higher concentration of banana homogenate (75 g/l) proved detrimental for the survival of cultures; the protocorm-like bodies showed necrosis and they perished soon after. Coconut water (10%) and peptone (2 g/l) supplemented medium proved optimum for profuse multiplication of protocorm-like bodies (PLBs) their early growth into plantlets. The treatments with organic growth supplements gave better and early plantlets as compared to control. Among these tested organic growth supplements, peptone (2 g/l) and coconut water (10%) proved beneficial for multiplication of regenerants, maximum number of shoots formation and early plantlet development; whereas banana homogenate (50 g/l) favoured the highest regeneration frequency and healthy growth of plantlets.
Keywords: banana homogenate; coconut water; orchids; peptone; protocorms; regeneration
Published: March 31, 2012 Show citation
References
- Aktar S., Nasiruddin K.M., Hossain K., 2008. Effects of different media and organic additives interaction on in vitro regeneration of Dendrobium orchid. Journal of Agricultural and Rural Development, 6: 69-74.
Go to original source...
- Arditti J., 1967. Factors affecting the germination of orchid seeds. Botanical Review, 3: 1-97.
Go to original source...
- Arditti J., 1979. Aspects of physiology of orchids. Advances in Botanical Research, 7: 421-655.
Go to original source...
- Arora R., Bhojwani S.S., 1989. In vitro propagation and low temperature storage of Saussurea lappa C.B. Clarke - an endangered medicinal plant. Plant Cell Reports, 8: 44-47.
Go to original source...
Go to PubMed...
- Bejoy M., Kumar S.C., Radhika B.J., Joemon J., 2004. Asymbiotic seed germination and early seedling development of the dove orchid Peristeria elata Hook. The Journal of the Orchid Society of India, 17: 75-79.
- Bhadra S.K., Hossain M.M., 2003. In vitro germination and micropropagation of Geodorum densiflorum (Lam.) Schltr. an endangered orchid species. Plant Tissue Culture and Biotechnology, 13: 165-171.
- Bhasker J., 1996. Micropropagation of Phalaenopsis. [Ph.D. Thesis.] Thrissur, Kerala Agricultural University.
- Chen J.T., Chang W.C., 2002. Effects of tissue culture conditions and explants characteristics on direct somatic embryogenesis in Oncidium 'Gower Ramsey'. Plant Cell Tissue and Organ Culture, 69: 41-44.
Go to original source...
- Chen S.C., Tang T., 1982. A general review of the orchid flora of China. In: Arditti J. (ed.), Orchid Biology, Reviews and Perspectives II. Ithaca, Cornell University Press: 39-81.
- Chowdhury I., Rahman A.R.M., Islam M.O., Matsui S., 2003. Effect of plant growth regulators on callus proliferation, plantlet regeneration and growth of plantlets of Doritaenopsis orchid. Biotechnology, 2: 214-221.
Go to original source...
- Curtis J.T., 1947. Studies on nitrogen nutrition of orchid embryos. I. Complex nitrogen sources. American Orchid Society Bulletin, 16: 654-660.
- Flamee M., 1978. Influence of selected media on the germination and growth of Paphiopedilum seedlings. American Orchid Society Bulletin, 47: 419-423.
- Fonnesbech M., 1972. Growth hormones and propagation of Cymbidium in vitro. Physiologia Plantarum, 27: 310-316.
Go to original source...
- Goh C., Wang P.F., 1990. Micropropagation of the monopodial orchid hybrid Aranda 'Deborah' using inflorescence explants. Scientia Horticulturae, 44: 315-321.
Go to original source...
- Hong P.I., Chen J., Chang W., 2010. Shoot development and plant regeneration from protocorm-like bodies of Zygopetalum mackayi. In Vitro Cellular and Developmental Biology - Plant, 46: 306-311.
Go to original source...
- Ichihashi S., Islam M.O., 1999. Effect of complex organic additives on callus growth in three orchid genera, Phalaenopsis, Doritaenopsis and Neofinetia. Journal of Japanese Society of Horticultural Science, 68: 269-274.
Go to original source...
- Intuwong O., Sagawa Y., 1973. Clonal propagation of sarcanthine orchids by aseptic culture of inflorescences. American Orchid Society Bulletin, 42: 264-270.
- Jawahar M., Karthikeyan A.V.P., Vijai D., Mahajan M., Ravipaul S., Jeyseelan M., 2008. In vitro plant regeneration from different explants of Cardiospermum helicacabum L. International Journal of Biological and Chemical Sciences, 2: 14-20.
Go to original source...
- Karuppusamy S., Pullaiah T., 2007. In vitro shoot multiplication of Bupleurum distichophyllum Wight. A native medicinal plant of southern India. Plant Tissue Culture, 17: 115-124.
Go to original source...
- Krishnamohan P.T., Jorapur S.M., 1986. In vitro seed culture of Acampe praemorsa (Roxb.) Blatt. and McC. In: Vij S.P. (ed.), Biology, Conservation and Culture of Orchids. New Delhi, East-West Press: 437-439.
- Kusumoto M., Furukuwa J., 1977. Effect of organic matter on growth of Cymbidium protocorms cultured in vitro. Journal of Japanese Society of Horticultural Science, 45: 421-426.
Go to original source...
- Lee Y.L., Lee N., 2003. Plant regeneration from protocorm derived callus of Cypripedium formosanum. In Vitro Cellular and Developmental Biology - Plant, 39: 475-479.
Go to original source...
- Lekha Rani C., Vidya C., Rajmohan K., Mercy S.T., 2005. Protocorm differentiation and seedling growth in Dendrobium hybrid seed cultures as influenced by organic additives. The Journal of the Orchid Society of India, 19: 67-70.
- Mathews V.H., Rao P.S., 1980. In vitro multiplication of Vanda hybrids through tissue culture technique. Plant Science Letters, 17: 383-389.
Go to original source...
- McIntyre D.K., Veitch G.J., Wrigley J.W., 1974. Australian orchids from seeds II. Improvement in techniques and further successes. American Orchid Society Bulletin, 43: 52-53.
- Minea M., Piluek C., Menakani T.A., Tantiwiwat S., 2004. A study on seed germination and seedling development of Spathoglottis kimballiani. Journal of National Academy of Sciences, 38: 141-156.
- Mitra G.C., Prasad R.N., Chowdhary A.R., 1976. Inorganic salts and differentiation of protocorms in seed callus of an orchid and correlated changes in its free amino acid content. Indian Journal of Experimental Biology, 14: 350-351.
- Murdad R., Hwa S.K., Seng C.K., Latip A.M., Aziz A.Z., Ripin R., 2006. High frequency multiplication of Phalaenopsis gigantea using trimmed bases protocorms technique. Scientia Horticulturae, 111: 73-79.
Go to original source...
- Sahoo Y., Chand P.K., 1998. Micropropagation of Vitex negundo L., a woody aromatic medicinal shrub through high-frequency of axillary shoot proliferation. Plant Cell Reports, 18: 301-307.
Go to original source...
Go to PubMed...
- Sharma N., Chandel K.P.S., Srivastava V.K., 1991. In vitro propagation of Coleus forskohlii Briq. a threatened medicinal plant. Plant Cell Reports, 10: 67-70.
Go to original source...
Go to PubMed...
- Sinha P., Roy S.K., 2004. Regeneration of an indigenous orchid, Vanda teres (Roxb.) Lindl. through in vitro culture. Plant Tissue Culture, 14: 55-61.
- Sudha G.C., Seeni S., 1994. In vitro multiplication and field establishment of Adhatoda beddomei C.B. Clarke, a rare medicinal plant. Plant Cell Reports, 13: 203-207.
Go to original source...
Go to PubMed...
- Sudeep R., Rajeevan P.K., Valasalakumari P.K., Geetha C.K., 1997. Influence of organic supplements on shoot proliferation in Dendrobium. Journal of Horticulture, 3: 38-44.
- Villolobus L., Munoz A.M., 1994 Tissue culture of orchids Cattleya, Encyclia, Oncidium and Stanhopea. Orchid Review, 102: 58-62.
- Znaniecka J., Królocka A., Sidwa-Gorycka M., Rybczynski J.J., Szlachetko L.D., Lojkowska E., 2005. Asymbiotic germination and seedling development and plantlet propagation of Encyclia aff. oncidioides - an endangered orchid. Acta Societatis Botanicorum Poloniae, 74: 193-198.
Go to original source...
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