In Vitro Organogenesis of a Slipper Orchid, Paphiopedilum ‘Alma Gavaert’
Keywords:callus; heterogeneity; organogenesis; shoot formation; slipper orchid
The aim of the present study was to improve the regeneration efficiency of callus lines in a slipper orchid, Paphiopedilum ‘Alma Gavaert’. Three kinds of vegetative tissues, root, stem and leaf segments, were used as explants to induce callogenesis; out of these, only root explants formed callus and was subcultured in the presence of 5 mg/L dicamba and 5 mg/L 2,4-D combined with 1 or 2 mg/L TDZ. The resulting four callus lines, assigned as 5Di1T, 5Di2T, 5D1T and 5D2T, respectively, were used to test the effect of NAA to BA ratios on re-differentiation, wherein the highest number of shoots (approximately 2 shoots/0.1 g callus clump) were obtained in callus line 5D2T at ratios of 0.001 and 0.002. A largely improvement of shoot regeneration efficiency was obtained by continuous selection of callus lines which derived from different explant positions. Eventually, six callus lines, including 5D2T-T6-G5 to 5D2T-T6-G10, were able to produce approximately 10 times of shoots per callus clump when compared with the parental callus line 5D2T.
Chang C, Chang WC (1998). Plant regeneration from callus culture of Cymbidium ensifolium var. misericors. Plant Cell Reports 17(4):251-255.
Chen JT (2012). Induction of petal-bearing embryos from root-derived callus of Oncidium ‘Gower Ramsey’. Acta Physiologiae Plantarum 34(4):1337-1343.
Chen JT, Chang WC (2000). Efficient plant regeneration through somatic embryogenesis from callus cultures of Oncidium (Orchidaceae). Plant Science 160(1):87-93.
Chen TY, Chen JT, Chang WC (2002). Multiple shoot formation and plant regeneration from stem nodal explants of Paphiopedilum orchids. In Vitro Cellular Developmental Biology - Plant 38(6): 595-597.
Chen TY, Chen JT, Chang WC (2004). Plant regeneration through direct shoot bud formation from leaf cultures of Paphiopedilum orchids. Plant Cell Tissue and Organ Culture 76(1)):11-15.
Chen Y, Goodale UM, Fan XL, Gao JY (2015). Asymbiotic seed germination and in vitro seedling development of Paphiopedilum spicerianum: An orchid with an extremely small population in China. Global Ecology and Conservation 3:367-378.
Chen YC, Chang C, Chang WC (2000). A reliable protocol for plant regeneration from callus culture of Phalaenopsis. In Vitro Cellular & Developmental Biology -Plant 36(5):420-423.
Compton M (2005). Plant regeneration from protocorm-derived callus of five Paphiopedilum hybrids. HortScience 40:1101.
Compton ME (1994). Statistical methods suitable for the analysis of plant tissue culture data. Plant Cell Tissue and Organ Culture 37:217-242.
Dawns CJ (1971). Biological techniques in electron microscopy. Barnes and Noble, New York.
Duncan DB (1955). Multiple range and multiple F test. Biometrics 11:1-42.
Guo B, Abbasi BH, Zeb A, Xu LL, Wei YH (2011). Thidiazuron: a multi-dimensional plant growth regulator. African Journal of Biotechnology 10(45): 8984-9000.
Hong PI, Chen JT, Chang WC (2008). Plant regeneration via protocorm-like body formation and shoot multiplication from seed-derived callus of a maudiae type slipper orchid. Acta Physiologiae Plantarum 30(5):755-759.
Huang LC, Lin CJ, Kuo CI, Huang BL, Murashige T (2001). Paphiopedilum cloning in vitro. Scientia Horticulturae 91(1-2):111-121.
Immonen AST (1996). Influence of media and growth regulators on somatic embryogenesis and plant regeneration for production of primary triticales. Plant Cell Tissue and Organ Culture 44(1): 45-52.
Lee PL, Chen JT (2014). Plant regeneration via callus culture and subsequent in vitro flowering of Dendrobium huoshanense. Acta Physiologiae Plantarum 36(10): 2619-2625.
Lin YH, Chang C, Chang WC (2000). Plant regeneration from callus culture of a Paphiopedilum hybrid. Plant Cell Tissue and Organ Culture 62(1):21-25.
Long B, Niemiera AX, Cheng ZY, Long CL (2010). In vitro propagation of four threatened Paphiopedilum species (Orchidaceae). Plant Cell Tissue and Organ Culture 101(2):151-162.
Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15(3):495-497.
Murthy BNS, Murch SJ, Saxena PK (1998). Thidiazuron: a potent regulator of in vitro plant morphogenesis. In Vitro Cellular Developmental Biology - Plant 34(4):267-275.
Ng CY, Saleh NM, Zaman FQ (2010). In vitro multiplication of the rare and endangered slipper orchid, Paphiopedilum rothschildianum (Orchidaceae). African Journal of Biotechnology 9(14):2062-2068.
Ng CY, Saleh NM (2011). In vitro propagation of Paphiopedilum orchid through formation of protocorm-like bodies. Plant Cell Tissue and Organ Culture 105(2):193-202.
Osuna P, Barrow JR (2004). Regeneration of black grama (Bouteloua eriopoda Torr. Torr) plants via somatic embryogenesis. In Vitro Cellular developmental Biology - Plant 40(3): 299-302.
Płażek A, Filek M, WÄ™dzony M (1999). Improvement of regeneration ability in Phleum pratense L. in vitro culture by dicamba. Acta Physiologiae Plantarum 21(4):397-403.
Rout GR, Samantaray S, Das P (1998). In vitro selection and characterization of Ni-tolerant callus lines of Setaria italica L. Acta Physiologiae Plantarum 20(3):269-275.
Sheehan TJ, Sheehan M (1994). An illustrated survey of orchid genera. Timber Press Inc, Portland.
Skoog F, Miller CO (1957). Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Symposia of the Society for Experimental Biology 11:118-131.
Smigocki AC, Owens LD (1989). Cytokinin-to-auxin ratios and morphology of shoots and tissues transformed by a chimeric isopentenyl transferase gene. Plant Physiology 91(3):808-811.
Steinmacher DA, Clement CR, Guerra MP (2007). Somatic embryogenesis from immature peach palm inflorescence explants: towards development of an efficient protocol. Plant Cell Tissue and Organ Culture 89(1):15-22.
Thorpe TA (1994). Morphogenesis and regeneration. In: Vasil IK, Thorpe, TA (Eds). Plant Cell and Tissue Culture, Kluwer Academic Publishers, Dordrecht pp 17-36.
Tsai KL, Chen EG, Chen JT (2016). Thidiazuron-induced efficient propagation of Salvia miltiorrhiza through in vitro organogenesis and medicinal constituents of regenerated plants. Acta Physiologiae Plantarum 38(1): 29.
Udomdee W, Wen PJ, Chin SW, Chen FC (2012). Shoot multiplication of Paphiopedilum orchid through in vitro cutting methods. African Journal of Biotechnology 11(76):14077-14082.
Wang HC, Chen JT, Chang WC (2010). Morphogenetic routes of long-term embryogenic callus culture of Areca catechu. Biologia Plantarum 54(1):1-5.
Wu IF, Chen JT, Chang WC (2004). Effects of auxins and cytokinins on embryo formation from root-derived callus of Oncidium ‘Gower Ramsey’. Plant Cell Tissue and Organ Culture 77(1):107-109.
Zeng SJ, Huang WC, Wu KL, Zhang JX, da Silva JAT, Duan J (2015). In vitro propagation of Paphiopedilum orchids. Critical Reviews in Biotechnology 36(3):521-534.
Zeng SJ, Wang J, Wu KL, da Silva JAT, Zhang JX, Duan J (2013). In vitro propagation of Paphiopedilum hangianum Perner & Gruss. Scientia Horticulturae 151:147-156.
Zhang YY, Wu KL, Zhang JX, Deng RF, Duan J, da Silva JAT, Huang WC, Zeng SJ (2015). Embryo development in association with asymbiotic seed germination in vitro of Paphiopedilum armeniacum SC Chen et FY Liu. Scientific Reports 5:16356.
How to Cite
Papers published in Notulae Scientia Biologicae are Open-Access, distributed under the terms and conditions of the Creative Commons Attribution License.
© Articles by the authors; licensee SMTCT, Cluj-Napoca, Romania. The journal allows the author(s) to hold the copyright/to retain publishing rights without restriction.
Open Access Journal - the journal offers free, immediate, and unrestricted access to peer-reviewed research and scholarly work, due SMTCT supports to increase the visibility, accessibility and reputation of the researchers, regardless of geography and their budgets. Users are allowed to read, download, copy, distribute, print, search, or link to the full texts of the articles, or use them for any other lawful purpose, without asking prior permission from the publisher or the author.