Multiple Shoot Formation of Paphiopedilum 'Delrosi'

Authors: Aree Thongpukdee, Ekasit Nisayan, Chockpisit Thepsithar

Abstract:

Shoots, with three leaves, of Paphiopedilum 'Delrosi' were used as explants for multiple shoot induction. Modified Hyponex medium was supplemented with thidiazuron (TDZ), N6- benzyladenine (BA) or kinetin (Kn) alone and in combinations with 2,4-dichlorophenoxyacetic acid (2,4-D). All explants were cultured for 15 weeks. It was found that TDZ alone at the concentration of 0.45μM or in combination with 4.52μM 2,4-D and 8.88μM BA in combination with 13.56μM 2,4-D promoted multiple shoots. The highest shoot sprouting efficiencies (80.0, 90.0 and 80.0%) and new shoot numbers (1.5, 1.3 and 1.1) were obtained, respectively. Fresh weight, height, numbers of leaf and root of new shoots and initial explants were discussed.

Keywords: Paphiopedilum, terrestrial orchids, in vitro culture, micropropagation, multiple shoot induction

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1331287

References:

[1] H. Koopowitz, Tropical slipper orchids: Paphiopedilum and Phragmipedium species and hybrids. TimberPress Inc., Oregon, 2008.
[2] L. Yung, and L. Nean, L. 2001. "Effect of capsule maturation, medium composition and suspension culture on in vitro germination of Paphiopedilum primulinum seed." Journal of the Chinese Society for Horticultural Science, vol. 47, pp. 147-156, 2001.
[3] Y. H. Lin, D. Jun, L. Chen, and C. Z. Lin, C. Z. 2004. "Seed germination in vitro of Paphiopedilum armeniacum and P. micranthum." Acta Horticulturae Sinica, vol. 31, pp. 540-542, 2004.
[4] C. M. Sama, and M. Kalita, "In vitro propagation of a terrestrial endangered orchid Paphiopedilum insigne (Wall. Ex. Lindl) Pfitz." Journal of Phytological Research, vol. 17, pp. 1-5, 2004.
[5] B. Long, A. X. Niemiera, and Z. Y. Chen, "In vitro propagation of four threatened Paphiopedilum species (Orchidaceae)." Plant Cell, Tissue and Organ Culture, vol. 101, pp. 151-162, 2010.
[6] P. I. Hong, J. T. Chen, and W. C. Chang, "Plant regeneration via protocorm-like body formation and shoot multiplication from seedderived callus of a maudiae type slipper orchid." Acta Physiologia Plantarum, vol. 30, pp. 755-759, 2008.
[7] C. Y. Ng, and N. M. Saleh, "In vitro propagation of Paphiopedilum orchid through formation of protocorm-like bodies." Plant Cell, Tissue and Organ Culture, vol. 105, pp. 193-202, 2011.
[8] T. Y. Chen, J. T. Chen, and W. C. Chang, "Multiple shoot formation and plant regeneration from stem nodal explants of Paphiopedilum orchids." In Vitro Cellular & Developmental Biology−Plant, vol. 38, pp. 595-597, 2002.
[9] C. Y. Ng, and N. M. Saleh, "In vitro multiplication of the rare and endangered slipper orchid, Paphiopedilum rothschildianum (orchidaceae). African Journal of Biotechnology, vol. 9, pp. 2062- 2068, 2010.
[10] T. Y. Chen, J. T. Chen, and C. W. Chang, "Plant regeneration through direct shoot bud formation from leaf cultures of Paphiopedilum orchids." Plant Cell, Tissue and Organ Culture, vol. 76, pp. 11-15, 2004.
[11] E. Nisayan, C. Thepsithar, and A. Thongpukdee, "Multiple shoot induction from seed derived shoots of Paphiopedilum." Acta Hortticulturae, vol. 878, pp. 199-204, 2010.
[12] D. B. Duncan, "Multiple range and multiple F test." Biometrics, vol. 11, pp. 1-42, 1955.
[13] T. Murashige, and F. Skoog, "A revised medium for rapid growth and bioassays with tobacco tissue culture." Physiologia Plantarum, vol. 15, pp. 473-497, 1962.
[14] Y. Lee, and N. Lee, "Plant regeneration from protocorm-derived callus of Cyperidium formosanum." In Vitro Cellular & Developmental Biology−Plant, vol. 39, pp. 475-479, 2003.
[15] H. Shimura, and Y. Koda, "Micropropagation of Cyperidium macranthos var. rebunense through protocorm-like bodies derived from mature seeds." Plant Cell, Tissue and Organ Culture, vol. 78, pp. 273--276, 2004.
[16] L. Huang, C. Lin, C. Kuo, B. Huang, and T. Murashige, "Paphiopedilum cloning in vitro." Scientia Horticulturae, vol. 91, pp. 111-121, 2001.