In Vitro Propagation of Cephalanthera rubra (L.) Rich., an Endangered Orchid, Using 2,4-D, NAA and BA

الموضوعات : مجله گیاهان زینتی

Mahdi Zargar Azad ¹ , Behzad Kaviani ² , Shahram Sedaghathoor ³

1 - Ph.D. Candidate, Department of Horticultural Science, Rasht Branch, Islamic Azad University, Rasht, Iran
2 - Department of Horticultural Science, Rasht Branch, Islamic Azad University, Rasht, Iran
3 - Department of Horticultural Science, Rasht Branch, Islamic Azad University, Rasht, Iran

تاريخ الإرسال : 17 الخميس , شعبان, 1444 تاريخ التأكيد : 11 الأربعاء , ذو القعدة, 1444 تاريخ الإصدار : 16 الجمعة , صفر, 1445

الکلمات المفتاحية: tissue culture, plant growth regulators, Micropropagation, Orchidaceae, Threatened ornamental plants,

ملخص المقالة :

Orchids are universally popular due to the wide range of colors, sizes, shapes, and scents in their flowers. The demand for orchids as cut flowers and pot plants is increasing in the flower market. Seed germination of orchids is comparatively low in the wild as they typically require fungal symbionts during their germination. Micropropagation of orchids is a major approach to orchid conservation and commercial production. Cephalanthera rubra (L.) Rich., one of Iran’s endangered orchid species, has been cloned using leaf as explant, Murashige and Skoog (MS) as culture medium, and α-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D) (both at the concentrations of 0, 0.1, 0.2, 0.3 and 0.4 mg L–1) as well 6-benzyladenine (BA) (at 0, 1, 2, 3 and 4 mg L–1) as plant growth regulators (PGRs). In order to initiate an axenic culture, the disinfection of leaf explants was performed with sodium hypochlorite and mercury chloride. The highest number of shoots (4.33) was obtained in medium enriched with 4 mg L–1 BA. Maximum stem length (4.73 cm), leaf number (5.33) and node number (2.86) was obtained in medium supplemented with 3 mg L–1 BA. The largest number of root (5) and the highest length of root (4.83 cm) was produced on medium augmented with 0.3 mg L–1 NAA. Rooted plantlets were transferred to pots filled with peat and perlite in 1:1 proportion and acclimatized to ambient greenhouse conditions with an average of 90% survival rate. This is the first report on the micropropagation of C. rubra (L.) Rich.

المصادر:

Amoo, S.O., Aremu, A.O., Moyo, M., Szučova, L., Dolžal, K. and van Staden, J. 2014. Physiological effects of a novel aromatic cytokinin analogue in micropropagated Aloe arborescens and Harpagophytum procumbens. Plant Cell, Tissue and Organ Culture, 116: 17-26.

Baker, A., Kaviani, B., Nematzadeh, Gh. and Negahdar, N. 2014. Micropropagation of Orchis catasetum ‐ A rare and endangered orchid. Acta Scientiarum Polonorum Hortorum Cultus, 13 (2): 197-205.

Bhattacharyya, P., Kumaria, S. and Tandon, P. 2016. High frequency regeneration protocol for Dendrobium nobile: A model tissue culture approach for propagation of medicinally important orchid species. South African Journal of Botany, 104: 232-243.

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 Conservation, 3: 367-378.

Dohling, S., Kumaria, S. and Tandon, P. 2012. Multiple shoot induction from axillary bud cultures of the medicinal orchid, Dendrobium longicornu. AoB Plants, 1–7. http://dx.doi.org/10.1093/aobpla/pls032

Engelmann, F. 2011. Use of biotechnologies for the conservation of plant biodiversity. In vitro Cellular Developmental Biology - Plant, 47: 5-16.

Huang, L.C., Lin, C.J., Kuo, C.I., Huang, B.L. and Murashige, T. 2001 Paphiopedilum cloning in vitro. Scientia Horticulturae, 91: 111-121.

Kalimuthu, K., Senthikumar, R. and Vijayakumar, S. 2007. In vitro micropropagation of orchid, Oncidium sp. (Dancing Dolls). African Journal of Biotechnology, 6 (10): 1171-1174.

Kaviani, B., Negahdar, N., Baker, A. and Mosafer, N. 2017. In vitro micropropagation of an endangered orchid species (Orchis catasetum) through protocorms: The effect of plant growth regulators and iron nanochelate. Plant Research Journal, 30 (1): 215-225. (In Persian with English abstract)

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.

Mahendran, G. and Narmatha Bai, V. 2009. Mass propagation of Satyrium nepalense D.Don.‐A medicinal orchid via seed culture. Scientia Horticulturae, 119: 203-207.

Mohammadi, M., Kaviani, B. and Sedaghathoor, Sh. 2019. Micropropagation of two near threatened orchid. Part 2: Phalaenopsis amabilis Blume var. Grandiflora. Advances in Horticultural Sciences, 33 (4): 485-493.

Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiology Plant, 15: 473–479.

Murthy, H.N., Paek, K.Y. and Park, S.Y. 2018. Micropropagation of orchids by using bioreactor technology. In: Lee, Y.I.  and Yeung, E.C-T (eds.), Orchid propagation: From Laboratories to Greenhouses—Methods and Protocols, Springer Protocols Handbooks, https://doi.org/10.1007/978-1-4939-7771-09

Panwar, D., Ram, K. and Shekhawat, H.N. 2012. In vitro propagation of Eulophia nuda Lindl., an endangered orchid. Scientia Horticulturae, 139: 46-52.

Park, S.Y., Huh, Y.S. and Paek, K.Y. 2018. Common protocols in orchid micropropagation. pp. 179-194. In: Lee, Y.I.  and Yeung, E.C-T (eds.), Orchid propagation: From laboratories to greenhouses—methods and protocols. Springer Protocols Handbooks, https://doi.org/10.1007/978-1-4939-7771-0_8

Roy, A.R., Patel, R.S., Patel, V.V., Sajeev, S. and Deka, B.C. 2011. Asymbiotic seed germination, mass propagation and seedling development of Vanda coerulea Griff ex. Lindl. (Blue Vanda): An in vitro protocol for an endangered orchid. Scientia Horticulturae, 128: 325-331.

Seeni, S. and Latha, P.G. 2000. In vitro multiplication and ecorehabilitation of the endangered Blue Vanda. Plant Cell, Tissue and Organ Culture, 61: 1-8.

Yam, T.W. and Arditti, J. 2018. Orchid micropropagation: An overview of approaches and methodologies. In: Lee, Y.I. and Yeung, E.C.T (eds.), Orchid propagation: From laboratories to greenhouses—methods and protocols. Springer Protocols Handbooks, https://doi.org/10.1007/978-1-4939-7771-0_9

Yeung, E.C., Li, Y.Y. and Lee, Y.I. 2018. Understanding seed and protocorm development in orchids. In: Lee, Y.I. and Yeung, E.C.T (eds.), Orchid propagation: From laboratories to greenhouses—methods and protocols. Springer Protocols Handbooks, https://doi.org/10.1007/978-1-4939-7771-0_9

Zakizadeh, Z., Kaviani, B. and Hashemabadi, D. 2019. Micropropagation of two near threatened orchid. Part 1. Catasetum pileatum cv. Alba. Advances in Horticultural Sciences, 33 (4): 475-483.

Zeng, S., Wua K., Teixeira da Silva, J.A., Zhanga, J., Chena, Z., Xiaa, 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.