Brazilian Journal of Biological Sciences (ISSN 2358-2731)

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Vol. 2, No. 4, p. 271-286 - Dec. 31, 2015


Asymbiotic seed germination and mass multiplication of Taprobanea spathulata (L.) Christenson (Asparagales: Orchidaceae): a medicinally important epiphytic orchid

Natarajan Parimala Devi, Balamohan Lisipriya and Narmatha Bai

An effective protocol was developed for asymbiotic seed germination and mass multiplication of Taprobanea spathulata (L.) Christenson a medicinally important epiphytic orchid. Different types of media such as MS, B5, Mitra, KCM and LO media was tested for seed germination and protocorm development. Half strength MS medium was found to be best for seed germination (92.73 + 1.18%) and protocorm development. Protocorms were cultured on half strength MS medium with different growth hormones such as BAP, KIN, NAA, IAA (0.5-2.0 mg/L) either individually or in combination. Organic additives such as peptone, coconut water and tomato juice were tested either individually or in combination with KIN for multiple shoot induction. KIN (1.0 mg/L) along with IAA (1.0 mg/L) was found to be best in producing multiple protocorms (22.40 + 1.33) with 1.60 + 0.24 number of roots. The organic additives peptone at 0.5% individually (14.64 + 0.312) and 0.125% in combination with KIN (0.5 mg/L) was effective in producing 16.84 + 0.639 multiple protocorms. Both NAA and IAA at 0.5 mg/L individually were effective for inducing healthy roots. Plantlets with well-developed leaves and roots were transplanted to vermiculite for acclimatization. The survival rate was 67%.

Seed germination, Multiple protocorms, IAA, Peptone, Activated charcoal, Vermiculite.

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Abdel-Rahman; Abdel-Hamd, Y. Nutritional value of some canned tomato juice and concentrates. Food Chemistry, v. 9, p. 303-306, 1982.

Anjum, S.; Zia, M.; Chaudhary, M. F. Investigations of different strategies for high frequency regeneration of Dendrobium malones 'Victory'. African Journal of Biotechnology, v. 5, n. 19, p. 1738-1743, 2006.

Anonymous. Wealth of India Council of Scientific and Industrial Research. New Delhi, 1992.

Arditti, J. Aspects of physiology of orchids. Advances in Botanical Research, v. 7, p. 421-655, 1979.

Arenmongla T.; Deb, C. R. Studies on in vitro morphogenetic potential of nodal segments of Malaxis acuminata D. Don. Applied Biological Research, v. 14, n. 2, p. 156-163, 2013.

Arenmongla, T.; Deb, C. R. Germination of immature embryos and multiplication of Malaxis acuminata D. Don an endangered therapeutically important orchid by asymbiotic culture in vitro. Indian Journal of Biotechnology, v. 11, p. 464-469, 2012.

Basha, S. K. M.; Umamaheswari, P.; Rajyalakshmi, E.; Rambabu, M.; Pullaiah, T. Medicinal flora of Penusila Narasimha Sacred Grove, Eastern Ghats, SPSR Nellore District, Andhra Pradesh, India. Indian Journal of Fundamental and Applied Life Sciences, v. 2, n. 2, p. 334-344, 2012.

Basker, S.; Narmatha Bai, V. In vitro propagation of an epiphytic and rare orchid Eria bambusifolia Lindl. Research in Biotechnology, v. 1, p. 15-20, 2010.

Bektaş, E.; Cuce, M.; Sokmen, A. In vitro germination protocorm formation and plantlet development of Orchis coriophora (Orchidaceae) a naturally growing orchid species in Turkey. Turkish Journal of Botany, v. 37, p. 336-342, 2013.

Bhadra, S. K.; Hossain, M. M. In vitro germination and micropropagation of Geodorum densiflorum Lam. Schltr., an endangered orchid species. Plant Tissue Culture, v. 13, n. 2, p. 165-171, 2003.

Bose, T. K.; Bhattacharjee, S. K.; Das, P.; Basak, U. C. Orchids of India Naya Prakash Bidhan Sarani, Calcutta, India. 1999.

Chai, M. L.; Xu, C. J.; Senthil, K. K.; Kim, J. Y.; Kim, D. H. Stable transformation of protocorm-like bodies in Phalaenopsis orchid mediated by Agrobacterium tumefaciens. Scientia Horticlturae, v. 96, p. 1-4, 2002.

Chen, J. T.; Chang, W. C. Effects of tissue culture conditions and explants characteristics on direct somatic embryogenesis in Oncidium 'Gower Ramsey'. Plant Cell Tissue and Organ Culture, v. 69, p. 41-44, 2002.

Chen, J. T.; Chang, W. C. Efficient plant regeneration through somatic embryogenesis from callus of Oncidium (Orchidaceae). Plant Science, v. 160, p. 87-93, 2000.

Chen, L. R.; Chen, J. T.; Chang, W. C. Efficient production of protocorm-like bodies and plant regeneration from flower stalk explants of the sympodial orchid Epidendrum radicans. In Vitro Cellular Developmental Biology-Plant, v. 38, p. 441-445, 2002.

Chen, T. Y.; Chang, W. C. Plant regeneration through direct shoot bud formation from leaf cultures of Paphiopedilum orchids. Plant Cell Tissue and Organ Culture, v. 76, p. 11-15, 2004.

Chung, J. D.; Chun, C. K. Asymbiotic germination of Cymbidium ensifolium. I - Effect of basal media and growth regulators on germination of seeds and shoot emergence from rhizomes. J. Korean Society for Horticultural Science, v. 24, p. 236-242, 1983.

Chyuam, Y. N.; Norihan, M. S. In vitro propagation of Paphiopedilum orchid through formation of protocorm-like bodies. Plant Cell Tissue Organ Culture, v. 105, p. 193-202, 2011.

Dasari, R.; Sathyavati, D.; Belide, S. K.; Reddy, P. J.; Abbulu, K. Evaluation of antioxidant activity of two important memory enhancing medicinal plants Celtis timorensis and Vanda Spathulata. Asian Journal of Pharmaceutical and Clinical Research, v. 6, n. 2, p. 153-155, 2013.

Deb, C. R.; Imchen. An efficient in vitro hardening technique of tissue culture raised plants. Biotechnology, v. 9, n. 1, p. 79-83, 2010.

De-Pauw, M. A.; Remphrey, W. R.; Palmer, C. E. The cytokinin preference for in vitro germination and protocorm growth of Cypripedium candidum. Annals of Botany, v. 75, n. 3, p. 267-275, 1995.

Dhiman, K.; Pathak, P.; Vij, S. P. Effect of growth hormones on immature embryo culture of Habenaria edgeworthii Hook. ex collect medicinally important terrestrial orchid. Abstracts Cum Souvenir NRC for Orchids (ICAR). Pakyong, Sikkim, 2013. p. 155-156.

Dohling, S.; Kumaria, S.; Tandon, P. Optimization of nutrient requirements for asymbiotic seed germination of Dendrobium longicornum and D. formosum. Proceedings of the Indian National Science Academy, v. 74, p. 167-171, 2008.

Fay, M. E. In what situations is in vitro appropriate to plant conservation? Biodiversity Conservation, v. 3, p. 176-183, 1994.

Fonnesbech, M. Growth hormones and propagation of Cymbidium in vitro. Physiologia Plantarum, v. 27, p. 310-316, 1972.

Gamborg, O. L. Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res., v. 50, p. 151-158, 1968.

George, E. F.; Hall, M. A.; Jan De Klerk, G. Plant propagation by tissue culture. 3. ed. Dordrecht: Springer, 2008.

Giri, L.; Jugran, A.; Rawat, S.; Dhyani, P.; Andola, H.; Bhatt, I. D.; Rawal, R. S.; Dhar, U. In vitro propagation genetic and phytochemical assessment of Habenaria edgeworthii an important Astavarga plant. Acta Physiologia Plantarum, v. 34, n. 3, p. 869-875. 2012.

Goh, C. J.; Wong, P. F. Micropropagation of the monopodial orchid hybrid Aranda deborah using inflorescence explants. Scientia Horticulturae, v. 44, p. 315-321, 1990.

Guha Bakshi Sensarma; Pal, D. C. A lexicon of medicinal plants in India. Naya Prokash Calcutta, India, 2001. v. 2.

Gutierrez, P. M. P. Orchids: a review of uses in traditional medicine its phytochemistry and pharmacology. Journal of Medicinal Plants Resources, v. 4, p. 592-638, 2010.

Hadley, G. The interaction of kinetin auxin and other factors in the development of north temperate orchids. New Phytologist, v. 69, p. 549-555, 1970.

Hajong, S.; Kumaria, S.; Tandon, P. In vitro propagation of the medicinal orchid Dendrobium chrysanthum. Proceedings of the Indian National Science Academy, v. 76, p. 1-6, 2010.

Harvais, G. An improved culture medium for growing the orchid Cypripedium reginae axenically. Canadian Journal of Botany, v. 51, p. 327-332, 1982.

Hossain, M. M. Therapeutic orchids: traditional uses and recent advances - an overview. Fitoterapia, v. 82, p. 102-140, 2011.

Hossain, M. M.; Sharma, M.; Pathak, P. In vitro propagation of Dendrobium aphyllum (Orchidaceae) - seed germination to flowering. Journal of Plant Biochemistry and Biotechnology, v. 22, n. 2, p. 157-167, 2012.

Hossain, M. M.; Sharma, M.; Teixeira da Silva, J. A.; Pathak, P. Seed germination and tissue culture of Cymbidium giganteum Wall ex Lindl. Scientia Horticulturae, v. 123, p. 479-487, 2010.

Ichihashi, S.; Islam, M. O. Effect of complex organic additives on callus growth in three orchid genera Phalaenopsis, Doritaenopsis and Neofinetia. Journal of Japanese Society for Horticultural Science, v. 68, p. 269-274, 1999.

Intuwong, O.; Sagawa, Y. Clonal propagation of sarcanthine orchids by aseptic culture of inflorescences. American Orchid Society Bulletin, v. 42, p. 264-270, 1973.

Jayanthi, P.; Rajendran, A.; Binu Thomas; Aravindhan, V.; Sivalingam, R. Biodiversity of lithophytes in Madukkarai Hills of Southern Western Ghats of Coimbatore District, Tamil Nadu, India. International Journal of Biological Sciences and Technology, v. 2, n. 2, p. 76-82. 2011.

Johnson, T. R.; Kane, M. E. Asymbiotic germination of ornamental Vanda: in vitro germination and development of three hybrids. Plant Cell Tissue and Organ Culture, v. 91, p. 251-261, 2007.

Kano, K. Studies on the media for orchid seed germination. Mam. Fac. Argic. Kagwa Univ., v. 20, p. 1-76, 1965.

Kaur, S.; Bhutani, K. K. In vitro propagation of Vanda testacea Lindl. Reichb f.: a rare orchid of high medicinal value. Plant Tissue Culture and Biotechnology, v. 19, n. 1, p. 1-7, 2009.

Knudson, L. A new nutrient solution for germination of orchid seed. American Orchid Society Bulletin, v. 15, p. 214-217, 1946.

Lauzer, D.; Arnaud, M. S.; Barabe, D. Teterazolium straining and in vitro germination of mature seeds of Cypripedium acaule (Orchidaceae). Lindeleyana, v. 9, p. 197-204, 1994.

Lindemann, E. G. P.; Gunckel, J. E.; Davidson, O. W. Meristem culture of Cattleya. American Orchid Society Bulletin, v. 39, p. 1002-1004, 1970.

Luo, J. P.; Wang, Y.; Zha, X. Q.; Huang, L. Micropropagation of Dendrobium densiflorum through protocorm like bodies: effects of plant growth regulators and lanthanoids. Plant Cell Tissue and Organ Culture, v. 93, n. 3, p. 330-340, 2006.

Machaka-Houri, N.; Al-Zein, M. S.; Westbury, D. B.; Talhouk, S. N. Reproductive success of the rare endemic Orchis galilaea (Orchidaceae) in Lebanon. Turkish Journal of Botany, v. 36, p. 677-682, 2012.

Madhi, R. P.; Chakrabarti, S. Traditional knowledge of NE people on conservation of wild orchids. Indian Journal of Traditional Knowledge, v. 8, n. 1, p. 11-16, 2009.

Mahendran, G.; Muniappan, V.; Ashwini, M.; Muthukumar, T.; Narmatha Bai, V. Asymbiotic seed germination of Cymbidium bicolor Lindl. (Orchidaceae) and the influence of mycorrhizal fungus on seedling development. Acta Physiologia Plantarum, v. 35, p. 829-840, 2013.

Mahendran, G.; Narmatha Bai, V. Mass propagation of Satyrium nepalense D. Don: a medicinal orchid via seed culture. Scientia Horticulturae, v. 119, p. 203-207, 2009.

Manners, V.; Kumaria, S.; Tandon, P. Micropropagation of Vanda coerulea Griff ex Lindl.: a study of regeneration competence of roots in vitro. Environmental Engineering and Applications (ICEEA), International Conference. Singapore, 2010. p. 100-102.

Mathews, V. H.; Rao, P. S. In vitro multiplication of Vanda hybrids through tissue culture technique. Plant Science Letters, v. 17, p. 383-389, 1980.

McIntyre, D. K.; Veitch, G. J.; Wrigley, J. W. Australian orchids from seeds. II - Improvement in techniques and further successes. American Orchid Society Bulletin, v. 43, p. 52-53, 1974.

Miria, A.; Khan, A. B.; Rao, B. R. P. Orchids of Talakona Sacred Grove, Andhra Pradesh, India. American-Eurasian Journal of Agriculture & Environment Sci., v. 12, n. 4, p. 469-471, 2012.

Mitra, G. C.; Prasad, R. N.; Roychowdhury, A. Inorganic salts and differentiation of protocorms in seed callus of orchid and correlative changes in its free aminoacid content. Indian Jounal of Experimental Biology, v. 14, p. 350-351, 1976.

Miyazaki, S.; Nagamatsu, T. Studies on the promotion of the early growth in vitro of orchid. Pt. I. Agric. Bull. Saga Univ., v. 21, p. 131-149, 1965.

Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, v. 15, p. 473-497, 1962.

Muthukrishnan, S.; Franklin Benjamin, J. H.; Baradwaj, R. G.; Senbagalakshmi, P.; Senthil Kumar, T.; Rao, M. V. Effect of different organic additives on seed germination of Geodorum densiflorum (Lam.) Schltr.: an endangered orchid. Abstracts Cum Souvenir NRC for Orchids (ICAR), Pakyong, Sikkim, 2013. p. 135-136.

Nadkarni, K. M. The Indian materia medica. Bombay Popular: Prakashana. 1999.

Nasiruddin, K. M.; Begum, R.; Yasmin, S. Protocorm like bodies and plant regeneration from Dendrobium formosum lead callus. Asian Journal of Plant Science, v. 2, n. 13, p. 955-957, 2003.

Ng, C. Y.; Saleh, N. M. In vitro propagation of Paphiopedilum orchid through formation of protocorm-like bodies. Plant Cell Tissue and Organ Culture, v. 105, p. 193-202. 2011.

Pant, B.; Thapa, D. In vitro mass propagation of an epiphytic orchid, Dendrobium primulinum Lindl. through shoot tip culture. African Journal of Biotechnology, v. 11, n. 42, p. 9970-9974, 2012.

Pathak, P.; Mahant, K. C.; Guota, A. In vitro propagation as an aid to conservation and commercialization of Indian orchids: seed culture. In: Pathak, P.; Sehgal, R. N.; Shekar, N. Sharma, M.; Sood, A. (Ed.). Orchid's science and commerce. Dehra Dun Bishen Singh, Mahendra Pal Singh, 2001. p. 319-362.

Pierik, R. L. M.; Steegman, R. H. The effect of 6-benzylaminopurine on growth and development of Cattaleya seedlings growth unripen seeds. Z. Pflanzenphysiol., v. 68, p. 228-234, 1972.

Potshangbam, N.; Nirmala, C. In vitro rapid propagation of Cymbidium aloifolium L. Sw.: a medicinally important orchid via seed culture. Journal of Biological Science, v. 11, n. 3, p. 254-260, 2011.

Pullaiah, T. Encyclopedia of world medicinal plants. New Delhi: Regency Publications, 2006. v. 4.

Rahman, A. R. M. M.; Islam, M. O.; Prodhan, A. K. M. A.; Ichihas, S. Effects of complex organic extracts on plantlet regeneration from PLBs and plantlet growth in the Doritaenopsis orchid. Japan Agricultural Research Quarterly, v. 38, p. 55-59, 2004.

Rahman, M. S.; Hasan, M. F.; Das, R.; Hossain, M. S.; Rahman, M. In vitro micropropagation of orchid Vanda tessellata L. from shoot tip explants. Journal of Biological Science, v. 17, p. 139-144, 2009.

Ram Pal, D.; Barman, M.; Dayamma, R.; Medhi, P. Asymbiotic seed germination seedling growth and acclimation of Zygopetalum intermedium for commercial cultivation. Abstracts Cum Souvenir NRC for Orchids (ICAR). Pakyong, Sikkim, 2013. p. 154.

Rao, A. N.; Avadhani, P. N. Some aspects of in vitro culture of Vanda seeds. Current Science, v. 3, p. 467-468, 1963.

Saiprasad, G. V. S.; Raghuveer, P.; Khetarpal, S.; Chandra, R. Effect of various plant growth regulators on the production of protocorm like bodies in three orchid genera. Indian Journal of Plant Physiology, v. 7, p. 35-39, 2002.

Sana, A.; Ahmad, Y.; Yaseen, I. A. H. M. In vitro propagation of orchid Dendrobium nobile var. Emma white. African Journal of Biotechnology, v. 10, n. 16, p. 3097-3103, 2011.

Saranya, B.; Gopalan, R.; Narmatha Bai, V.; Mahendran, G. Rare and threatened orchid species used for various diseases by Irulars Vellingiri Hills, Coimbatore. International Journal of Scientific and Research Publications, v. 2, n. 9, p. 1-2, 2012.

Saxena, S.; Dhawan, B. Regeneration and large-scale production of bamboo Dendrocalamus strictus Nees through somatic embryogenesis. Plant Cell Reports, v. 18, p. 438-443, 1999.

Shanavaskhan, A. E.; Sivadasan, M.; Alfarhan, A. H.; Jacob, T. Ethnomedicinal aspects of angiospermic epiphytes and parasites of Kerala, India. Indian Journal Traditional Knowledge, v. 11, n. 2, p. 250-258, 2012.

Sharma, S. K.; Tandon, P.; Mishra, R. R. Vitamins as related to axenic seed germination and seedling of Cymbidium elegans Lindl. and Coelogyne punctulata Lindl. Journal of the Orchid Society India, v. 5, p. 25-28, 1991.

Sharma, V. Regenerative competence of pseudobulb explants of endangered orchid genera: a study in vitro. Abstracts Cum Souvenir NRC for Orchids ICAR. Pakyong, Sikkim, 2013. p. 158.

Singh, M. K.; Sherpa, A. R.; Hallan, V.; Zaidi, A. A. A potyvirus in Cymbidium spp. in Northern India. Austr. Plant., v. 123, p. 281-287, 2007.

Singh, V.; Pathak, P.; Vij, S. P. In vitro asymbiotic embryo culture in a floriculturally and medicinally important orchid Dendrobium moschatum. Abstracts cum Souvenir, NRC for Orchids (ICAR). Pakyong, Sikkim, 2013. p. 155-156.

Vellupillai, M.; Swaru, P. S.; Goh, C. J. Histological and protein changes during early stage of seed germination in the orchid Dendrobium acurmenatum. Journal of Horticultural Science, v. 76, p. 94-98, 1997.

Vujanovic, V.; Arnaud, S. M.; Barabe, D.; Thibeault, G. Viability testing of orchid seed and the promotion of colouration and germination. Annals of Botany, v. 86, p. 79-86, 2000.

Yakimova, E.; Toteva, V. K.; Groshkoff, I.; Ivanova, G. Effect of BA and CPPU on protease and a-amylase activity of in vitro cultured explants of Rosa hybrida L. Bulg. Journal Plant Physiology, v. 26, p. 39-47, 2000.