Date Received: Aug 20, 2018
Date Accepted: Aug 20, 2018
Date Published: Aug 20, 2018
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Vu, B., & Sorgan, S. K. T. (2018). Effects of Plant Growth Regulators and Sucrose on the Regeneration of Paphiopedilum micranthum var. North Vietnam. Vietnam Journal of Agricultural Sciences, 1(1), 11–20. https://doi.org/10.31817/vjas.2018.1.1.02
Effects of Plant Growth Regulators and Sucrose on the Regeneration of Paphiopedilum micranthum var. North Vietnam
,
Sorgan S. K. Tai
2
Keywords
Paphiopedilum micranthum var. North Vietnam, regeneration, plant growth regulators (PGRs), half-strength MS medium, sucrose concentration
Abstract
The objective of this study was to determine the effects of 2,4-Dichlorophenoxyacetic acid (2,4-D) combined with 6-Benzylaminopurine (BAP) and sucrose on the shoot and root regeneration of Paph. micranthum var. North Vietnam. Young fresh leaves were cut into small explants and then cultured on media containing 2,4-D (0.5, 1.0, or 2.0 ppm). Within each of the 2,4-D concentration, BAP was added at three different concentrations (0.5, 1.0, or 2.0 ppm) for callus induction after 4 weeks. Shoot tips (2-3 mm in length) from the leaf-derived calluses were then transferred vertically on half-strength MS medium (2,4-D combined with BAP (0.5, 1.0, or 2.0 ppm) for shooting, and rootless shoots with 2 leaves were transferred to the same MS medium for rooting after 4 months of culture. Rootless shoots with 2 leaves were also transferred vertically on half-strength MS medium with different sucrose concentrations (0, 10, 20, 30, or 40 g L-1) to optimize the growth and development of the plantlets. The results indicated that the optimum degree of shoot and root regeneration occurred on half-strength MS medium containing 2.0 ppm 2,4-D combined with 2.0 ppm BAP. Approximately 86.6% of the cultures responded with 3.3 shoots and 3.0 leaves/explant. In addition, under the optimal concentrations, the number and length of roots per explant after 4 months of culture were 2.1 roots per explant and an average root length of 1.1 cm. MS medium supplemented with 30 g L-1 sucrose had the highest number of leaves and roots, and also the longest average length of roots, which gave 3.8 leaves, 3.3 roots/explant, and a root length of 1.8 cm after 4 months.
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31. Smith C., Lea P. and Leegood R. (1995). Carbohydrate chemistry in plant. Biochemistry and Molecular Biology, UK. pp.73-111.
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33. Tehrim S., Mirza M. and Sajid G. (2013). Comparative study of different growth regulators for efficient plant regeneration in grapes. Pakistan Journal Agriculture and Research. Vol 26 (4). pp. 275-289.
34. Teixeira da Silva J. (2013). Orchid: Advances in tissue culture, genetics, photochemistry and transgenic biotechnology. Flori-culture and Ornamental Biotechnology. Vol 7 (1). pp. 1-52.
35. Wu C., Dewir Y., Hahn E. and Paek K. (2006). Optimization of culturing conditions for the production of biomass and phenolic from adventitious roots of Echinacea angustifolia. Plant Biology. Vol 49 (3). pp. 193-199.
36. Yoon S., Joung K., Sang Y., Eui Y., Kee Y. and Sung W. (2016). Effects of altering medium strength and sucrose concentration on in vitro germination and seedling growth of Cypripedium macranthos Sw. Journal Plant Biotechnology. Vol 43 (1). pp. 132-137.
37. Yu L., Yu T., Yung S., Ruey L. and Fang W. (2011). In vitro shoot induction and plant regeneration from flower buds in Paphiopedilum orchids. In Vitro Cellular Developmental Biology-Plant. Vol 47 (6). pp. 702-709.
38. Zha X., Luo J., Jiang S. and Wang J. (2007). Enhancement of polysaccharide production in suspension cultures of protocorm-like bodies from Dendrobium huoshanense by optimization of medium compositions and feeding of sucrose. Process Biochemistry. Vol 42 (3). pp. 344-351.
39. Zhang Y., Wu K., Zhang J., Deng R., Duan J. and Zeng S. (2015). Embryo development in association with a symbiotic seed germination in vitro of P. armeniacum. Scientific Reports. Vol 5. pp. 16356. doi: 10.1038/srep16356.
2. Akinyele A. (2010). Effects of growth hormones, rooting media and leaf size on juvenile stem cuttings of Buchholzia coriacea engler. Annals of Forest Research. Vol 53 (2). pp. 127-133.
3. Arditti J. and Ernst R. (1992). Micro-propagation of orchids. John Wiley and Sons, Inc., New York, USA. pp. 682.
4. Asghar S., Ahmad T., Hafiz I. and Yaseen M. (2011). In vitro propagation of orchid (Dendrobium nobile) var. Emma white. African Journal of Biotechnology. Vol 10 (16). pp. 3097-3103.
5. Bhatt B. and Tomar Y. (2010). Effects of plant growth regulators on rooting performance of Citrus auriantifolia swingle (Kagzi-Lime) in different growing conditions. Nature and Science. Vol 8 (7). pp. 8-11.
6. De Klerk G. (2002). Rooting of micro-cuttings: Theory and practice. Journal of In Vitro cellular and developmental biology-plant. Vol 38 (5). pp. 415-422.
7. Deb C. and Pongener A. (2011). Asymbiotic seed germination and in vitro seedling development of Cymbidium aloifolium (L.) Sw.: a multipurpose orchid. Journal of Plant Biochemistry and Biotechnology. Vol 20 (1). pp. 90-95.
8. Faria R., Rodrigues F., Oliveira L. and Muller C. (2004). In vitro Dendrobium nobile plant growth and rooting in different sucrose concentrations. Horticultural Brasileira. Vol 22 (4). pp. 780-783.
9. Gibson S. (2000). Plant sugar response pathways. Part of a complex regulatory web. Plant Physiology. Vol 124 (4). pp. 1532-1539.
10. Han H., Zhang S. and Sun X. (2009). A Review on the molecular mechanism of plant rooting modulated by auxin. African Journal of Biotechnology. Vol 8 (1). pp. 348-353.
11. Hartmann H., Kester D., Davies F. and Geneve R. (2002). Plant propagation: Principles and practices. 7th ed. Prentice-Hall, Englewood Cliffs. pp. 15-22.
12. Hasbullah N., Saleh A. and Taha R. (2011). Establishment of somatic embryogenesis from Gerbera jamesonii Bolus Ex. Hook F. through suspension culture. African Journal Biotechnology. Vol 10 (63). pp. 13762-13768.
13. Ibanez A., Valero M. and Morte A. (2005). Establishment and in vitro clonal propagation of the Spanish autochthonous table grapevine cultivar Napoleon: an improved system where proliferating cultures alternate with rooting ones. Anales de Biologia. Vol 27. pp. 211-220.
14. Johnson T., Kane M. and Perez H. (2011). Examining the interaction of light, nutrients and carbohydrates on seed germination and early seedling development of Bletia purpurea (Orchidaceae). Plant Growth Regulation. Vol 63 (1). pp. 89-99.
15. Khatun H., Khatun M., Biswas M., Kabir M. and Al-Amin M. (2010). In vitro growth and development of dendrobium hybrid orchid. Bangladesh Journal Agricultural Research. Vol 35 (3). pp. 507-514.
16. Khatun M. and Al-Amin M. (2006). In vitro germination and micro-propagation from F1 seeds of Dendrobium orchid. Bangladesh Journal of P1ant Breeding and Genetics. Vol 19 (2). pp. 37-42.
17. Klaus T. (2007). Influence of 2,4-D and BAP on callus growth and the subsequent regeneration of somatic embryos in long-term cultures of King’s Ruby. Journal of Plant Biotechnology. Vol 10 (1). pp. 25-33.
18. Konate S., Kon M., Kouakou H., Kouadio J. and Zouzou M. (2013). Callus induction and proliferation from cotyledon explants in Bambara groundnut. African Crop Science Journal. Vol 21 (3). pp. 255-263.
19. Kulaeva O. (1980). Cytokinin action on enzyme activities in plants, Proceedings of the 10th international conference on plant growth substances. Tokyo, Japan, August, 1980. pp.119-128
20. Kumar V., Rashmi D. and Banerjee M. (2014). Callus induction and plant regeneration in Solanum tuberosum L. cultivars (Kufri Chipsona 3 and MP-97/644) via leaf explants. International Research Journal of Biological Sciences. Vol 3 (6). pp. 66-72.
21. Laboney U., Biswas G. and Abdullah-Al-Shoeb M. (2013). Callus induction and regeneration of orchid shoot tip culture. International Journal Agricultural Sciences. Vol 3 (2). pp. 40-45.
22. Mongomake K., Tchoa K., Nakpalo S., Andre B. and Tanoh H. (2015). In Vitro seeds germination and seedling growth of Bambara Groundnut (Vigna subterranea (L.) Verdc. (Fabaceae)). Hindawi Publishing Corporation, the Scientific World Journal, Article ID 595073, 8 pages.
23. Murashige T. and Skoog F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum. Vol 15 (3). pp. 473-497.
24. Nasrin S., Hossain M. and Anjumarana K. (2003). Induction and evaluation of soma clonal variation in potato (Solanum tuberosum L.). Journal of Biological Sciences. Vol 3 (2). pp. 183-190.
25. Nipawan J., Kanchit T. and Keiko I. (2013). Efficient adventitious shoot regeneration from shoot tip culture of Vanda coerulea, a Thai orchid. Science Asia. Vol 13 (9). pp. 449-455.
26. Rasmussen H. N. (1995). Terrestrial orchids from seed to mycotrophic plant. Cambridge University Press, New York. pp. 444.
27. Sagawa Y. and Kunisaki J. (1982). Clonal propagation of orchids by tissue culture. In: Fujiwara A. (Ed.), Plant tissue culture, Maruzen, Tokyo. pp. 683-684.
28. Shim S., Hahn E. and Paek K. (2003). In vitro and ex vitro growth of grapevine rootstock ‘5BB’ as influenced by number of air exchanges and the presence or absence of sucrose in culture media. Plant Cell, Tissue and Organ Culture. Vol 75 (1). pp. 57-62.
29. Shohael A., Chakrabarty D., Ali M., Yu K., Hahn E., Lee H. and Paek K. (2006). Enhancement of eleuthero sides production in embryogenic cultures of eleuthero coccus sessiliflorus in response to sucrose-induced osmotic stress. Process Biochemistry. Vol 41 (3). pp. 512-518.
30. Singh A., Rajesh S., Mittal A., Singh Y. and Shiva J. (2003). Effect of plant growth regulators on survival rooting and growth characters in Long Pepper (Piper longum L.). Progressive Horticulture. Vol 35. pp. 208-211.
31. Smith C., Lea P. and Leegood R. (1995). Carbohydrate chemistry in plant. Biochemistry and Molecular Biology, UK. pp.73-111.
32. Taiz L. and Zeiger E. (2006). The growth hormone. In: Plant Physiology, 4th Edition, Sinauer Associates, Inc., Sunderland. pp. 467-504.
33. Tehrim S., Mirza M. and Sajid G. (2013). Comparative study of different growth regulators for efficient plant regeneration in grapes. Pakistan Journal Agriculture and Research. Vol 26 (4). pp. 275-289.
34. Teixeira da Silva J. (2013). Orchid: Advances in tissue culture, genetics, photochemistry and transgenic biotechnology. Flori-culture and Ornamental Biotechnology. Vol 7 (1). pp. 1-52.
35. Wu C., Dewir Y., Hahn E. and Paek K. (2006). Optimization of culturing conditions for the production of biomass and phenolic from adventitious roots of Echinacea angustifolia. Plant Biology. Vol 49 (3). pp. 193-199.
36. Yoon S., Joung K., Sang Y., Eui Y., Kee Y. and Sung W. (2016). Effects of altering medium strength and sucrose concentration on in vitro germination and seedling growth of Cypripedium macranthos Sw. Journal Plant Biotechnology. Vol 43 (1). pp. 132-137.
37. Yu L., Yu T., Yung S., Ruey L. and Fang W. (2011). In vitro shoot induction and plant regeneration from flower buds in Paphiopedilum orchids. In Vitro Cellular Developmental Biology-Plant. Vol 47 (6). pp. 702-709.
38. Zha X., Luo J., Jiang S. and Wang J. (2007). Enhancement of polysaccharide production in suspension cultures of protocorm-like bodies from Dendrobium huoshanense by optimization of medium compositions and feeding of sucrose. Process Biochemistry. Vol 42 (3). pp. 344-351.
39. Zhang Y., Wu K., Zhang J., Deng R., Duan J. and Zeng S. (2015). Embryo development in association with a symbiotic seed germination in vitro of P. armeniacum. Scientific Reports. Vol 5. pp. 16356. doi: 10.1038/srep16356.