Regeneration of Plantlets Through PLB (Protocorm-Like Body) Formation in Phalaenopsis ‘Join Angle X Sogo Musadian’
DOI:
https://doi.org/10.5614/j.math.fund.sci.2016.48.3.2Keywords:
growth regulators, in vitro plant regeneration, Phalaenopsis, protocorm-like bodies, thidiazuron.Abstract
Selection and regeneration of specific hybrids of Phalaenopsis typically takes a long time since this plant usually reproduces through seeds. This study was conducted to examine the best medium and explants for regeneration of Phalaenopsis ‘Join Angle X Sogo Musadian’. In vitro and in vivo roots and leaves were used as explants, which were cultured in half-strength Murashige and Skoog medium supplemented with various combinations of plant growth regulators (TDZ, 2,4-D, NAA, BAP and IAA). Theresultsshowed that the in vitro roots produced the highest number of PLBs (49.3 PLBs) when they were cultured in medium containing 0.5 ppm NAA, 5 ppm BAP, and 0.5 ppm IAA. The in vitro leaf explants also regenerated PLBs, however, only two PLBs developed, i.e. when they were cultured in medium containing 1 ppm TDZ and 2ppm 2,4-D. PLB germination into plantlets was performed by culturing each PLB on MS medium without plant growth regulators. Based on the results of the present study, it can be concluded that in vitro roots and half-strength MS medium supplemented with 0.5 ppm NAA, 5 ppm BAP, and 0.5 ppm IAA are the best explant and best medium respectively for plant regeneration through PLBs.
References
Griesbach, R.J., Development of Phalaenopsis Orchids for the Mass-Market, Trends in New Crops and New Uses, Janick, J. & Whipkey, A. Eds, ASHS Press, Alexandria, pp. 458-465, 2002.
Koair, P., A kof, S. & Luthar, Z., Direct Shoot Regeneration from Nodes of Phalaenopsis Orchids, Acta Agriculturae Slovenica, 83, pp. 233-242, 2004.
Kuo, H.L., Chen, J.T. & Chang, W.C., Efficient Plant Regeneration Through Direct Somatic Embryogenesis from Leaf Explants of Phalaenopsis "Little Steve', In Vitro Cell Development Biology-Plant, 41, pp. 453-456, 2004..
Chen, J.T. & Chang, W.C., Direct Somatic Embryogenesis and Plant Regeneration From Leaf Explants of Phalaenopsis amabilis, Biologia Plantarum, 50(2), pp. 169-173, 2006.
Young, P.S., Murthy, H.N. & Yoeup, P.K., Mass Multiplication of Protocorm Like Bodies Using Bioreactor System and Subsequent Plant Regeneration in Phalaenopsis, Plant Cell, Tissue and Organ Culture, 63, pp. 67-72, 2000.
Niknejad, A., Kadir, M.A. & Kadzimin, S.B., In Vitro Plant Regeneration from Protocorms-Like Bodies (PLBs) and Callus of Phalaenopsis gigantea (Epidendroideae: Orchidaceae), African Journal of Biotechnology, 10(56), pp. 11808-11816, 2011.
Tokuhara, K. & Mii, M., Induction Of Embryogenic Callus and Cell Suspension Culture from Shoot Tips Excised from Flower Stalk Buds of Phalaenopsis (Orchidaceae), In Vitro Cell Development Biology-Plant, 37, pp. 457-461, 2001.
Vendrame, W.A. & Maguire, I., In Vitro Propagation and Plantlet Regeneration from Doritaenopsis Purple Gem "Ching Hua' Flower Explant, Hortscience, 42(5), 1256-1258, 2007.
Murashige, T. & Skoog, F. A Revised Medium for Rapid Growth and Bioassay with Tobacco Tissue Culture, Physiologia Plantarum, 15, pp. 473-495, 1962.
Chen, J.T., Chang, C. & Chang, W.C., Direct Somatic Embryogenesis on leaf explants of Oncidium "Gower Ramsey' and Subsequent Plant Regeneration, Plant Cell Reports, 19, pp. 143-149. 1999
Park, S.Y., Yeung, E.C., Chakrabarty, D. & Paek, K.Y., An Efficient Direct Induction of Protocorm-like Bodies from Leaf Subepidermal Cells of Doritaenopsis Hybrid using Thin-section Culture, Plant Cell Reports, 21, pp. 46-51, 2002.
Ishii, Y., Takamura, T., Goi, M. & Tanaka, M., Callus Induction and Somatic Embryogenesis of Phalaenopsis, Plant Cell Reports, 17, pp. 446-450, 1998.
Tokuhara, K. & Mii, M., Highly-efficient Somatic Embryogenesis from Cell Suspension Cultures of Phalaenopsis Orchids by Adjusting Carbohydrate Sources, In Vitro Cellular & Developmental Biology. Plant, 39, pp. 635-639, 2003
Cui, J., Liu, J., Deng, M., Chen, J. & Henny, R.J., Plant Regeneration through Protocorm-like Bodies Induced from Nodal Explants of Syngonium podophyllum "White Butterfly', HortScience, 43, pp. 2129-2133, 2008.
Schulze, J., Improvements in Cereal Tissue Culture by Thidiazuron: A Review, Fruit, Vegetable and Cereal Science and Biotechnology,1, pp. 64-79, 2001.
Murthy, B.N.S., Murch, S.J. & Saxena, P.K., Thidiazuron: A Potent Regulator of In Vitro Plant Morphogenesis, In Vitro Cell Development Biology-Plant, 34, pp. 267-275, 1998.
Vasser, C., Qureshi, J.A., Gill, R. & Saxena, P.K., Morphoregulatory Role of Thidiazuron: Substitution of Auxin and Cytokinin Requirement for the Induction of Somatic Embryogenesis in Geranium Hypocotyl Cultures, Plant Physiology, 99, pp. 1704-1707.1992
Lee, Y-I., Hsu, S-T. & Yeung, E.C., Orchid Protocorm-like Bodies are Somatic Embryos, American Journal of Botany, 100(11), pp. 2121-2131, 2013.
Guo, W.L.G., Chang, Y.C.A. & Kao, C.Y.K., Protocorm-like Bodies Initiation from Root Tips of Cyrtopodium paranaense (Orchidaceae), Hortscience, 45, pp. 1365-1368, 2010.
Pasternak, T., Miskolczi, P., Ayaydin, F., Meszaros, T., Dudits, D. & Feher, A., Exogenous Auxin and Cytokinin Dependent Activation of CDKs and Cell Division in Leaf Protoplast-derived Cells of Alfalfa, Plant Growth Regulation, 32, pp. 129-141, 2000.
Utami, E.S.W., Sumardi, I., Taryono & Semiarti, E., Effect of -Naphtaleneacetic Acid (NAA) on Somatic Embryogenesis of Moth Orchid Phalaenopsis amabilis (L.), Biodiversitas, 8(4), pp. 295-299, 2007. (Text in Indonesian)
