An Effective Nutrient Media for Asymbiotic Seed Germination and In Vitro Seedling Development of Phalaenopsis ‘Bahia Blanca’

Document Type: Original Article


1 Department of Horticulture, Karaj Branch, Islamic Azad University, Karaj, Iran

2 Department of Horticulture, University of Tehran, Karaj, Iran


Our study was to develop a rapid and successful method for promoting asymbiotic seed germination and growth and development of protocorm like bodies (PLBs) and subsequent plantlets of Phalaenopsis orchid. Four basal media i.e. Murashige and Skoog (MS), half strength MS (½MS), Knudson C (KC) and, Vacin and Went (VW) supplemented with coconut water and peptone were examined for their effectiveness in large-scale multiplication by asymbiotic seed germination. Seed germination, leaf number, root number, root length, seedling length and fresh and dry weight were scored. Well-rooted plantlets 4-cm in height were transferred to a greenhouse in trays containing expanded clay, charcoal, cocopeat and perlite (4:2:1:1) or sphagnum moss. The seed germination frequency and seedling development was found significantly higher in organic growth supplement-enriched MS and ½MS media. The results indicated that the highest seed germination (94 %) was obtained in MS medium supplemented with 100 mg/L coconut water or 1 g/L peptone and ½MS containing 1 or 2 g/L peptone. However, the longest seedling was found in MS or ½MS media supplemented with 100 mg/L coconut water or MS containing 1 g/L peptone. Ninety-seven percent of plantlets survived after 1 month in mixture media expanded clay: charcoal: cocopeat: perlit.

Graphical Abstract

An Effective Nutrient Media for Asymbiotic Seed Germination and In Vitro Seedling Development of Phalaenopsis ‘Bahia Blanca’


  • Simple and low-cost medium for Phalaenopsis in vitro culture.
  • Maximum seed germination and PLB de­velopment on MS or ½MS media.
  • Beneficial effects of coconut water and peptone on asymbiotic seed germination.


Aktar, S., Nasiruddin, K.M. and Hossain, K. 2008. Effects of different media and organic additives interaction on in vitro regeneration of Dendrobium orchid. Journal of Agriculture & Rural Development, 6(1&2): 69-74.

Arditti, J. 1967. Factors affecting the germination of orchid seeds. The Botanical Review, 33(1): 1-97.

Balilashaki, Kh., Gantait, S., Naderi, R. and Vahedi, M. 2015. Capsule formation and asymbiotic seed germination in some hybrids of Phalaenopsis, influenced by pollination season and capsule maturity. Physiology and Molecular Biology of Plants, 21(3): 341-347.

Bejoy, M., Kumar, S.C., Radhika, B.J. and 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.

Chen, Y., Goodale, U.M., Fan, X.L. and Gao, J.Y. 2015. Asymbiotic seed germination and in vitro seedling development of Paphiopedilum spicerianum: An orchid with an extremely small population in China. Global Ecology and Conservation, 3: 367-378.

Erfani, M., Miri, S.M. and Imani, A. 2017. In vitro shoot proliferation and rooting of Garnem rootstock as influenced by basal media, plant growth regulators and carbon sources. Plant Cell Biotechnology and Molecular Biology, 18(3&4): 101-109.

Fallahpour, M., Miri, S.M. and Bouzari, N. 2015. In vitro propagation of Gisela 5 rootstock as affected by media and plant growth regulators. Journal of Horticultural Research, 23(1): 57-64.

Gnasekaran, P., Rathinam, X., Sinniah, U.R. and Subramaniam, S. 2010. A study on the use of organic additives on the protocorm-like bodies (PLBs) growth of Phalaenopsis violacea orchid. Journal of Phytology, 2(1): 29-33.

Griesbach, R.J. 2002. Development of Phalaenopsis orchids for the Mass-Market. p. 458-465. In: J. Janick and A. Whipkey (eds.), Trends in New Crops and New Uses. ASHS Press, Alexandria, VA.

Kaur, S. and Bhutani, K.K. 2012. Organic growth supplement stimulants for in vitro multiplication of Cymbidium pendulum (Roxb.) Sw. Horticultural Science (Prague), 39: 47–52.

Kauth, P.J., Dutra, D., Johnson, T.R., Stewart, S.L., Kane, M.E. and Vendrame, W. 2008. Techniques and applications of in vitro orchid seed germination. p. 375-391. In: J. A. Teixeira da Silva (ed), Floriculture, Ornamental and Plant Biotechnology: Advances and Topical Issues. Global Science Books Ltd, UK.

Knudson, L. 1946. A new nutrient solution for germination of orchid seeds. American Orchid Society Bulletin, 15: 214–217.

Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Plant Physiology, 15: 473–497.

Paul, S., Kumaria, S. and Tandon, P. 2012. An effective nutrient medium for asymbiotic seed germination and large-scale in vitro regeneration of Dendrobium hookerianum, a threatened orchid of northeast India. AoB Plants, 1 (2012): plr032.

Roy, J. and Banerjee, N. 2001. Cultural requirements for in vitro seed germination, protocorm growth and seedling development of Geodorum densiflorum (Lam.) Schltr. Indian Journal of Experimental Biology, 39: 1041-1047.

Shadang, R., Dwivedi, P., Hegde, S.S. and Ahmed, N. 2007. Effects of different culture media on seed germination and subsequent in vitro development of protocorms of Hygrochilus parishii (Veith & Rchb.f.) Pfitz (Orchidaceae). Indian Journal of Biotechnology, 6: 256-261.

Shekarriz, P., Kafi, M., Dianati Deilamy, Sh. and Mirmasoumi, M. 2014. Coconut water and peptone improve seed germination and protocorm like body formation of hybrid Phalaenopsis. Agriculture Science Developments, 3(10): 317-322.

Teixeira da Silva, J.A. 2013. Orchids: Advances in tissue culture, genetics, phytochemistry and transgenic biotechnology. Floriculture and Ornamental Biotechnology, 7(1): 1-52.

Thongpukdee, A., Thepsithar, C. and Rojanawong, T. 2010. Optimum conditions for seed germination of Phalaenopsis ‘Silky Moon’. Acta Horticulturae, 878: 237-242.

Tsai, W.T. and Chu, C.Y. 2001. The industry of Phalaenopsis in Taiwan. Horticulture NCHU, 26(1): 27-41.

Vacin, E. and Went, F. 1949. Some pH changes in nutrient solution. Botanic Gardens Conservation News,110: 605-613.

Yan, T.W. and Arditti, J. 2009. History of orchid propagation A mirror of the history of biotechnology. Plant Biotechnology Reports, 3: 1-56.

Zahara, M., Datta, A., Boonkorkaew, P. and Mishra, A. 2017. The effects of different media, sucrose concentrations and natural additives on plantlet growth of Phalaenopsis hybrid ‘Pink’. Brazilian Archives of Biology and Technology, 60: e17160149.

Zeng, S., Wu, K., Teixeira da Silva, J.A., Zhang, J., Chen, Z., Xia, N. and Duan, J. 2012. Asymbiotic seed germination, seedling development and reintroduction of Paphiopedilum wardii Sumerh., an endangered terrestrial orchid. Scientia Horticulturae, 138: 198–209.