Proliferation of Protocorm-Like Bodies of Dendrobium mannii and Dendrobium mirbelianum in Chitosan-containing Media In Vitro

  • Syifaur Rahmah Bogor Agricultural University
  • Ni Made Armini Wiendi Bogor Agricultural University
  • Willy B. Suwarno Bogor Agricultural University
  • Krisantini Krisantini Journal of Tropical Crop Science


Dendrobium mannii and Dendrobium mirbelianum were rare and endangered orchid species. In vitro micro propagation could be used to conserve and proliferate these species. The aims of this study were to determine the effects of chitosan on proliferation of the protocorm-like-bodies (PLBs) of in vitro grown Dendrobium mannii and D. mirbelianum. Chitosan has been reported as an important component to promote orchid growth in vitro. In this study chitosan was included in the in-vitro media at the concentrations of 0, 1, 2, 3, and 4 mg.L-1. The highest PLBs formation, clump diameter, and fresh weight on D. mannii were obtained from explants cultured on medium supplemented with 1 mg.L-1 chitosan. After 14 weeks of culture, fresh weight, clump diameter and average number of PLBs of D.mannii were 29.52 %, 33.00 % and 35.88% greater at chitosan (1 mg.L-1) compared to control media.  D. mirbelianum treated with low chitosan concentrations (1 and 2 mg.L-1) had 65.71% more leaves than those without chitosan treatment. Chitosan, however, had an adverse effects on growth of both species at  4 mg.L-1. Medium contain 4 mg.L-1 chitosan induced PLBs proliferation of D.mannii, but inhibited PLBs proliferation of D.mirbelianum.  These results suggested that chitosan had important roles in PLB proliferation and could be used for the propagation of D. mannii PLBs in vitro.


Keywords: rare orchid species, in-vitro


Anonim. 2014. [July 2014].

Australian Department of the Environment. (2008). Approved Conservation Advice (s266B of the Environment Protection and Biodiversity Conservation Act 1999). 3 pp.

Barka, E.A., Dullaffroy, P., Clement, C., and Vernet, G. (2004). Chitosan improved the development and protects Vitis vinivera L. againts Botrytis cinerea. Plant Cell Report 22, 608-614.

Chandrkrachang, S., Sompongchaikul, P., and Sangtain, S. (2005). Profitable spin-off from using chitosan in orchid farming in Thailand. Journal of Metals, Materials, and Minerals 15, 45-48.

Chang, C. and Chang, W. (2000). Effect of thidiazuron on bud development of Cymbidium sinensis Wild in vitro. Plant Growth Regulation 30, 171-175.

Changdrkrachang, S. (2006). Chitosan as a growth stimulator in orchid tissue culture and agriculture in Thailand. Plant Science 170, 1185-1190.

CITES Plants .(2015). ENVIS centre on floral diversity. [February 24, 2015].

Deflieghere, F.A., Vermeulen, and Debereve, J. (2004). Chitosan: antimicrobial activity, interaction with food components and applicability as a coating on fruit and vegetables. Food Microbiology 26, 703-714.

Dewanty, R. (2011). Aplikasi penggunaan chitosan terhadap pembentukan protocorm like body (PLB) pada anggrek Phalaenopsis sp L. 62 pp. Skripsi. Budidaya pertanian, Fakultas pertanian. Universitas Jember.

Hargono and Djaeni, M. (2003). Utilization of chitosan prepared from shrimp shell as fat diluent. Journal of Coastal Development 7, 31-37.

Hasegawa A., Kanechika R., and Oguni, S. (2005). Effect of low temperature and chitosan on dormancy breaking and growth of young corms of three Arisaema species. Acta Horticulturae 673, 603-609.

Kanika, G. and Vij, S.P. (2004). Micropropagation of Vanda coerulea (Orchidaceae) through shoot tip culture. Journal of Horticultural Science 33, 227-228.

Latha, P.G. and Seeni, S. (1994). Multiplication of the endangered Indian pitcher plant (Nepenthes khasiana) through enhanced axillary branching in vitro. Plant Cell, Tissue, and Organ Culture 38, 69-71.

Limpanavech, P., Pichyangkura, R., Khunwasi, C., Chadchawan, S., Lotrakul, P., Bunjongrat, P., Chaidee, A., and Akaraeakpanya, T. (2003). The effect of polymer type, concentration and % DD of biocatalyst modified chitosan on flora production of Dendrobium ‘Eiskul’. 4 pp. Chulalongkorn University. Bangkok.

Lokho, A. (2013). Diversity of Dendrobium Sw. its distributional patterns and present status in the Northeast India. International Journal of Scientific and Research Publication (IJSRP) 3, 5.

Luo, J.P., Wawrosch, C., and Kopp, B. (2009). Enhanced micropropagation of Dendrobium huoshanense C.Z. Tang et S.J. Cheng through protocorm-like bodies: The effect of cytokinins, carbohydrate sources and cold pretreatment. Scientia Horticulturae 123, 258–262.

Mitra, G.C., Prasad, R.N., and Chowdhury, R.A. (1976). Inorganic salts and differentiation of protocorms in seed callus of orchid and correlative changes in its free amino acid content. Indian Journal of Experimental Biology 14, 350-351.

Nahar, S.J., Kazuhiko, S., and Haque, S.M. (2012). Effect of polysaccharides including elicitor on organogenesis in protocorm-like body (PLB) of Cymbidium insigne in vitro. Journal of Agricultural Science and Technology B2, 1029-1033. David Publishing.

Nge, K.L., Nwe, N., Chandrkrachang, S., and Stevens, S. (2005). Chitosan as a growth stimulator in orchid tissue culture. Plant Science 170, 1185-1190.

Novak, S.D., Luna, L.J., and Gamage, R.N. (2014). Mini-review: role of auxin in orchids development. Plant Signaling and Behaviour 9, e972277.

Nurhasanah, E., and Wiendi, N.M.A. (2009). Perbanyakan anggrek Gramatophyllum scriptum melalui proliferasi tunas adventif secara in vitro. 61 pp. Skripsi. Fakultas Pertanian. Institut Pertanian Bogor. Bogor.

Pornpeanpakdee, P., Pichyangkura, R., Chadchawan, S., and Limpanavech, P. (2006). Chitosan effects on Dendrobium ‘Eiskul’ protocorm-like body production In “31st Congress on Science and Technology of Thailand October18-20 2005”, pp 1-3, Suranaree University of Technology, Thailand.

Prasertsongskun, S., and Chaipakdee, W. (2011). Effect chitosan on growth and development of Phalaenopsis cornucervi (Breda) Blume & Rchb.f. Khon Kaen University (KKU) Science Journal 39,113-119.

Rao, N.A. (1990). Dendrobium mannii. Ridl. A rare orchid new to Arunachal Pradesh, India. Journal of Economic and Taxonomic Botany 14, 459-470.

Rinaudo, M. (2006). Chitin and chitosan: properties and application. Progress in Polymer Science 31, 603-632.

Samarfard, S., Kadir, M.A., Kadzimin, S.B., Ravanfar, S., and Saud, H.M. (2013). Genetic stability of in vitro multiplied Phalaenopsis gigantean protocorm-like bodies as affected by chitosan. Notulae Botanicae Horti Agrobotanici 41, 177-183.

Seidenfaden, G., and Wood, J.J. (1992). The orchids of Peninsular Malaysia and Singapore. 779 pp. Olsen and Olsen Publisher.

Shimura, H., and Koda, Y. (2004). Micropropagation of Cypripedium macranthos var. Rebunerse through protocorm-like bodies derived from mature seed. Plant Cell, Tissue and Organ Culture 78, 273–276.

Sopalun, K., Thammasiri, K., and Ishikawa, K. (2010). Effect of chitosan as growth stimulator for Grammatophyllum speciosum in vitro culture. World Academy of Science, Engineering and Technology 4, 381-383.

Souter, M. and Lindsay, K. (2000). Polarity and signallling in plant embryogenesis. Journal of Experimental Botany 51, 971-983.

Sulistiana, E. and Sukma, D. (2014). Growth of Phalaenopsis amabilis orchids on chitosan and salicylic acid treatment. Buletin Agrohorti 2, 75-85.

Suningsih, T. (2012). Pengaruh bahan organic chitosan terhadap pertumbuhan dan perkembangan anggrek Dendrobium ‘Woxinia’. 46 pp. Skripsi. Fakultas Pertanian. Institut Pertanian Bogor.

Teoh, E.S. (2005). “Orchids of Asia”. 367 pp. Times Edition - Marshall Cavendish, Singapore.

Tirta, I.G. (2013). Diversity and Collection of Orchids in the Forest Waifoi Raja Ampat and Sorong Nature Park-West Papua In “Proceedings of 4th International Conference on Global Resource Conservation and 10th Indonesian Society for Plant Taxonomy Congress”. Pp 69-76.

Uthairatanakij, A., Da Silva, J.A.T., and Obsuwan, A. (2007) Chitosan for improving orchid production and quality. Orchid Science and Biotechnology 1, 1-5.

Vogler, H. and Kuhlemeier, C. (2003). Simple hormones but complex signaling. Current Opinion in Plant Biology 6, 51-56.

Young, D. H., and Kauss, H. (1983). Release of calcium from suspension cultured Glycine max cells by chitosan, other polycations, and polyamines in relation to effects on membrane permeability. Plant Physiology 73, 698–702.

Zhou, Y. G., Yang, Y. D., Qi, Y. G., Hang, Z. M. Z., Wang, X .J., and Hu, X. J. (2002). Effects of chitosan on some physiological activity in germinating seed of peanut. Journal of Peanut Science 31, 22–25.