Abstract
An efficient method of Coelogyne cristata mass propagation was developed using segment of protocorm-like bodies (PLBs) (3 mm2 in size). It was observed that ½ MS medium showed to be more effective to induce shoots through PLBs segment. The explants when cultured on ½ MS media containing TDZ and CP showed relatively superior effect on shoot regeneration as compared to the media containing TDZ alone or in combination with BP. Addition of BP and CP to the medium containing NAA and BA combinations proved distinctly better for shoot multiplication than that of the medium with NAA and BA combinations alone. The highest percentage of explants producing shoots, with a maximum average of 8.1 per explant, was induced on the medium supplemented with 1.0 mg l−1 NAA and 0.5 mg l−1 BA with CP. Shoots produced an average of 15 roots per explant on ½ MS medium supplemented with 2.0 mg l−1 IBA and BP. The 4 cm height plantlets with well-developed roots were successfully acclimatized. The results suggest that CP and BP can be used effectively to initiate shooting and rooting of Coelogyne cristata. Ploidy analysis of regenerated plants using flow cytometry revealed the same ploidy level (diploid). This efficient and reliable protocol could be useful for mass multiplication and germplasm conservation of the wild medicinal orchid.
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Abbreviations
- AC:
-
Activated charcoal
- BA:
-
6-Benzylaminopurine
- BP:
-
Banana powder
- CP:
-
Coconut powder
- HP:
-
Hyponex basal medium
- IBA:
-
Indole butyric acid
- MS:
-
Murashige and Skoog
- ½ MS:
-
Half strength Murashige and Skoog
- NAA:
-
Naphthalene acetic acid
- PGRs:
-
Plant growth regulators
- PLBs:
-
Protocorm-like bodies
- TDZ:
-
Thidiazuron
- VW:
-
Vacin and Went medium
References
Aktar S, Nasiruddin KM, Huq H (2007) In vitro root formation in Dendrobium orchid plantlets with IBA. J Agric Rural Dev 5(1&2):48–51
Aktar S, Nasiruddin KM, Hossain K (2008) Effects of different media and organic additives interaction on in vitro regeneration of Dendrobium orchid. J. Agric Rural Dev 6(1&2):69–74
AmoMarco JB, Lledo MD (1996) In vitro propagation of Salix tarraconensis Pau ex Font Quer, an endemic and threatened plant. In Vitro Cell Dev Biol Plant 32:42–46. doi:10.1007/BF02823012
Arditti J (1967) Orchid biology: reviews and perspective. Cornel University Press, Ithaea, New York, pp 114–1255
Arditti J, Ernst R (1993) Micropropagation of orchids. Wiley, New York
Begum AA, Tamaki M, Kako S (1994) Formation of protocorm-like bodies (PLB) and shoot development through in vitro couture of outer tissue of cymbidium PLB. J Jpn Soc Hortic Sci 63(3):663–673
Chen LR, Chen JT, Chang WC (2002) Efficient production of protocorm-like bodies and plant regeneration from flower stalk explants of the sympodial orchid Epidendrum radicans. In Vitro Cell Dev Biol Plant 38:441–445. doi:10.1079/IVP2002315
Dhar U, Upreti J, Bhatt ID (2000) Micropropagation of Pittosporum napaulensis (DC.) Rehder & Wilson—a rare, endemic Himalayan medicinal tree. Plant Cell Tissue Organ Cult 63:231–235. doi:10.1023/A1010610603893
Lakshmanan P, Lob CS, Gob CJ (1995) An in vitro method for rapid regeneration of a monopodial orchid hybrid Aranda Deborah using thin section culture. Plant Cell Rep 14:510–514. doi:10.1007/BF00232785
Luo JP, Wang Y, Zha XQ, Haung L (2008) Micropropagation of Dendrobium densiflorum Lindl. ex Wall.through protocorm-like bodies: effects of plant growth regulators and lanthanoid. Plant Cell Tissue Organ Cult 93:333–340. doi:10.1007/s11240-008-9381-1
Malabadi RB, Mulgund GS, Nataraja K (2004) Efficient regeneration of Vanda coerulea, an endangered orchid using thidiazuron. Plant Cell Tissue Organ Cult 76:289–293. doi:10.1023/B:TICU.0000009255.69476.b7
Martin KP, Madassery J (2005) Rapid in vitro propagation of Dendrobium hybrids through direct shoots formation from foliar inoculums, and protocorm-like bodies. Sci Hortic 108:95–99
Nayak NR, Rath SP, Patnaik S (1997) In vitro propagation of three ephytic orchids, Cymbidium aloifolium (L) Sw. Dendrobium aphyllum (Roxb.) Fisch. and Dendrobium moschatum (Buch-Ham) Sw. through thidiazuron-induced high frequency shoot proliferation. Sci Hortic 71:243–250. doi:10.1016/S0304-4238(97)00075-7
Park SY, Murthy HN, Peak KY (2000) Mass multiplication of protocorm-like bodies using bioreactor system and subsequent plant regeneration in Phalaenopsis. Plant Cell Tissue Organ Cult 63:67–72. doi:10.1023/A1006420116883
Park SY, Murthy HN, Peak KY (2003) Protocorm-like-body induction and subsequent plant regeneration from root tip cultures of Doritaenopsis. Plant Sci 164:919–923. doi:10.1016/S0168-9452(03)00019-0
Pierik RLM (1987) In vitro culture of higher plants. In: Vegetative propagation of orchids. Martinus Nijhoff Publishers, Dordrecht, The Netherlands
Ravindra M, Gangadhar M, Nataraja K (2004) Efficient regeneration of Vanda coerulea, an endangered orchid using thidiazuron. Plant Cell Tissue Organ Cult 76:289–293
Razden MK (2003) Introduction to plant tissue culture, 2nd edn. Science Publisher, Inc., Enfield. Printed in India
Robinson JP, Balakrishnan V, Britto J (2009) In vitro seed germination and protocorm development of Dendrobium aqueum Lindl. a rare orchid species from eastern ghats of Tamil Nadu. Botany Research International 2(2):99–102
Roy J, Naha S, Majumdar M, Banerjee N (2007) Direct and callus-mediated protocorm-like body induction from shoot-tips of Dendrobium chrysotoxum Lindl. (Orchidaceae). Plant Cell Tissue Organ Cult 90:31–33. doi:10.1007/s11240-007-9244-1
Saiprasad GVS, Polisetty R (2003) Propagation of three orchid genera using encapsulated protocorm-like bodies. In Vitro Cell. Dev. Biol. Plant. 39:42–48. doi:10.1079/IVP2002360
Satake M, Ijung L (2004) Flowers in Myanmar (Part II): wild orchid and medicinal orchid. Aroma res 5:83–89
Sheelavanthmath SS, Murthy HN, Hema BP, Hahn EJ, Paek KY (2005) High frequency of protocorm like bodies (PLBs) induction and plant regeneration from protocorm and leaf sections of Aerides crispum. Sci Hortic 106:395–401. doi:10.1016/j.scienta.2005.04.012
Shimura H, Koda Y (2004) Micropropagation of Cypripedium macranthos var. rebunense through protocorm- like bodies derived from mature seeds. Plant Cell Tissue Organ Cult 78:273–276
Yi-xun Y, Ling L, Juan-xu L, Jing W (2009) Plant regeneration by callus-mediated protocorm-like body induction of Anthurium andraeanum hort. Agric Sci China 8(5):572–577. doi:10.1016/S1671-2927(08)60248-5
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The authors acknowledge the financial support from Bio Green 21 Program (code # 20070301034033), Rural Development Administration, Republic of Korea.
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Communicated by E. Lojkowska.
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Naing, A.H., Chung, J.D., Park, I.S. et al. Efficient plant regeneration of the endangered medicinal orchid, Coelogyne cristata using protocorm-like bodies. Acta Physiol Plant 33, 659–666 (2011). https://doi.org/10.1007/s11738-010-0586-7
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DOI: https://doi.org/10.1007/s11738-010-0586-7