Effect of Different Media and Mannitol Concentrations on Growth and Development of Vandopsis lissochiloides (Gaudich.) Pfitz. under Slow Growth Conditions
In vitro conservation of orchid germplasm provides an effective technique for ex situ conservation of orchid diversity. In this study, an efficient protocol for in vitro conservation of Vandopsis lissochiloides (Gaudich.) Pfitz. plantlet under slow growth conditions was investigated. Plantlets were cultured on different strength of Vacin and Went medium (½VW and ¼VW) supplemented with different concentrations of mannitol (0, 2, 4, 6 and 8%), sucrose (0 and 3%) and 50 g/L potato extract, 150 mL/L coconut water. The cultures were incubated at 25±2 °C and maintained under 20 µmol/m2s light intensity for 24 weeks without subculture. At the end of preservation period, the plantlets were subcultured to fresh medium for growth recovery. The results found that the highest leaf number per plantlet could be observed on ¼VW medium without adding sucrose and mannitol while the highest root number per plantlet was found on ½VW added with 3% sucrose without adding mannitol after 24 weeks of in vitro storage. The results showed that the maximum number of leaves (5.8 leaves) and roots (5.0 roots) of preserved plantlets were produced on ¼VW medium without adding sucrose and mannitol. Therefore, ¼VW medium without adding sucrose and mannitol was the best minimum growth conditions for medium-term storage of V. lissochiloides plantlets.
Digital Object Identifier (DOI): doi.org/10.5281/zenodo.3299541Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 617
 T. Eng-Soon, Orchid of Asia, Marshall Cavendish Editions, Singapore, 2009.
 E. Hágsater, and V. Dumont, “Orchids: Status, Survey and Conservation Action Plan,” Gland, Switzerland and Cambridge, 1996.
 T. Santisuk, K. Chayamarit, R. Pooma, and S. Suddee, Thailand Red Data: Plants. Office of Natural Resources and Environmental Policy and Planning (ONEP), Bangkok,Thailand, 2006.
 F. Engelmann, “In vitro conservation of tropical plant germplasm - a review,” Euphytica, vol. 57, pp. 227−243, 1991.
 B. Grout, “Genetic preservation of plant cells in vitro,” Acta Physiol Plant., vol. 18, no.1, pp. 97−98, 1995.
 I. Theilade, and L. Petri, “Conservation of tropical trees ex situ through storage and use,” Guidelines and Technical Notes No. 65, Danida Forest Seed Centre, Humlebaek, Denmark.
 P.E. Rajasekharan, and L. Sahijram, “In vitro conservation of plant germplasm,” In: BirBahadur et al. (Eds.), Plant biology and biotechnology: Volume II: Plant genomics and biotechnology, doi: 10.1007/978-81-322-2283-5_22, 2003.
 Z. Li, L. Zhao, G. Kai, S. Yu, Y. Cao, Y. Pang, X. Sun, and K. Tang, “Cloning and expression analysis of a water stress-induced gene from Brassica oleracea,” Plant Physiol Bioch., vol. 42, no. 10, pp. 789−94, 2004.
 L. Perales, V. Arbona, A. Gómez-Cadenas, M.J. Cornejo, and A. Sanz, “A relationship between tolerance to dehydration of rice cell lines and ability for ABA synthesis under stress,” Plant Physiol Bioch., vol. 43, no. 8, pp. 786−792, 2005.
 M.R. Morsy, L. Jouve, J.F. Hausman, L. Hoffmann, and J.M. Stewart, “Alteration of oxidative and carbohydrate metabolism under abiotic stress in two rice (Oryza sativa L.) genotypes contrasting in chilling tolerance,” J Plant Physiol., vol. 164, no. 2, pp. 157−67, 2007.
 E.F Vacin, and F. Went, “Some pH changes in nutrient solutions,” Bot Gaz., vol. 110, no. 4, pp. 605−613, 1949.
 F.J.M. Bonnier, and J.M. Van Tuyl, “Long term in vitro storage of lily: Effects of temperature and concentration of nutrients and sucrose,” Plant Cell Tissue Organ Cult., vol. 49, pp. 81−87, 1997.
 P. García-Jiménez, E.P. Navarro, C.H. Santana, A. Luque, and R.R. Robaina, “Anatomical and nutritional requirements for induction and sustained growth in vitro of Cymodocea nodosa (Ucria) Ascherson,” Aquat. Bot., vol. 84, no. 1, pp. 79−84, 2006.
 D.W. Lawlor, and G. Cornic, “Photosynthetic carbon assimilation and associated metabolism in relation to water deficits in higher plants,” Plant Cell Environ., vol. 25, no. 2, pp. 275−294, 2002.
 R.A. Shibli, M.A. Shatnawi, W. Subaih, and M.M. Ajlouni, “In vitro conservation and cryopreservation of plant genetic resources: a review,” World J of Agric Sci., vol. 2, pp. 372−382, 2006.
 M. K. El-Bahr, A. Abd El-Hamid, M. A. Matter, A. Shaltout, S. A. Bekheet, and A. A. El-Ashry, “In vitro conservation of embryogenic cultures of date palm using osmotic mediated growth agents,” Genet Eng Biotechnol J., vol. 14, no. 2, pp. 363−370, 2016.
 E. Zayova, T. Nedev, and L. Dimitrova, “In vitro storage of Stevia rebaudiana Bertoni under slow growth conditions and mass multiplication after storage,” Biol Bull., vol. 3, no. 1, pp. 30−38, 2017.
 S.A. Bekheet, M.A. Matter, H.S. Taha, and A. A. El-Ashry, “In vitro conservation of jojoba (Simmondsia chinensis) shootlet cultures using osmotic stress and low temperature,” 2016, Middle East J. Agric. Res., vol. 5, no. 4, pp. 396−402, 2016.
 X. J. Pan, W.E. Zhang, and L. Xia, “In vitro conservation of native Chinese wild grape (Vitis heyneana Roem. & Schult) by slow growth culture,” Vitis, vol. 53, no. 4, pp. 207–214, 2014.
 R.W. Tahtamouni, and R.A. Shibli, “Preservation at low temperature and cryopreservation in wild pear (Pyrus syriaca),” Adv Hortic Sci., vol. 13, no. 4, pp. 156–160, 1999.
 A. M. M. Gabr, and S. Sawsan Sayed, “Slow growth conservation and molecular characterization of Deutzia scabra Thunb.,” Afr. J. Plant Sci., vol. 4, no. 10, pp. 409–416, 2010.
 M. Ahmed, and M.A. Anjum, “In vitro storage of some pear genotypes with the minimal growth technique,” Turk J Agric For., vol. 34, pp. 25–32, 2010.
 A.R. Reddy, K.V. Chaitanya, and M. Vivekanandan, “Drought-induced responses of photosynthesis and antioxidant metabolism in higher plants,” J. Plant Physiol., vol. 161, pp. 1189–1202, 2004.
 H.B Shao, L.Y. Chu, C.A. Jaleel, and C.X. Zhao, “Water-deficit stress-induced anatomical changes in higher plants,” C R Biol., vol. 331, no. 3, pp. 215–25, 2008.
 M. Ghaheri, D. Kahrizi, and G. Bahrami, “Effect of mannitol on some morphological characteristics of in vitro Stevia rebaudiana Bertoni,” Biharean biol., Oradea, Romania, vol. 11, no. 2, pp. 94–97, 2017.
 A. Blum, “Drought resistance, water-use efficiency, and yield potential—are they compatible, dissonant, or mutually exclusive?,” Aust J Agric Res., vol. 56, pp. 1159–1168, 2005.
 G. Lopez-Puc, “An effective in vitro slow growth protocol for conservation of the orchid Epidendrum chlorocorymbos SCHLTR.,” Trop. Subtrop. Agroecosyst., vol. 16, pp. 61–68, 2013.
 T.L. da Silva, and J.E. Scherwinski-Pereira, “In vitro conservation of Piper aduncum and Piper hispidinervum under slow-growth conditions,” Pesq. Agropec. Bras., vol. 46, no. 4, pp. 383–389, 2011.
 M. M. Rabba, R.A. Shibli, and M.A. Shatnawi, “In vitro medium term conservation of Felty germander (Teucrium polium L.) micro-shoots,” Jordan J Biol Sci., vol. 8, no. 4, pp. 523–535, 2012.
 M.M. El-dawayati, B. EL- I, H.G. Amina, and E.Z. Zeinab, “In vitro conservation of date palm shoot tip explants under minimal growth condition,” Egypt. J. Agric. Res., vol. 91, no. 3, pp. 1043–1062, 2013.
 V. Joshi, and S.K. Jadhav, “Effect of temperature and media supplements on slow growth conservation of medicinal plant Spilanthes acmella,” Botanica Sarbica, vol. 37, 2, pp. 155–160, 2013.