Organogenic Potential of Dendrobium Floral Tissues for Stable Transformation Applications
Jorge Sahagun1,2,4, Anupan Kongbangkerd2,3, and Kumrop Ratanasut1,2*
1Department of Agricultural Science, Faculty of Agriculture,
Natural Resources and Environment, Naresuan University, Phitsanulok 65000 Thailand
2Center for Agricultural Biotechnology, Naresuan University, Phitsanulok 65000 Thailand
3Department of Biology, Faculty of Sciences, Naresuan University, Phitsanulok 65000 Thailand
4Department of Science and Technology – Philippine Nuclear Research Institute,
Commonwealth Ave., Diliman, Quezon City 1101 Philippines
*Corresponding author: This email address is being protected from spambots. You need JavaScript enabled to view it.
ABSTRACT
This study aimed to examine a promising procedure for in vitro direct organogenesis from floral tissues of Dendrobium Sonia ‘Earsakul’ as a new potential regeneration system for transformation in orchids. Petal and sepal explants from floral buds (2.0–2.3 cm) survived on ½ MS solid medium supplemented with or without 1-naphthaleneacetic acid (NAA) or/and benzylaminopurine (BA), but only petal explants generated protruding meristemoid tissues when they were cultured on ½ MS medium supplemented with both NAA and BA. A significant increase in the number of protruding meristemoid tissues was observed in the liquid medium. The ½ MS liquid medium supplemented with 0.5 mg L–1 NAA and 1.0 mg L–1 BA was successfully used to induce meristemoids of petal tissues transiently transformed by A. tumefaciens strain EHA105 carrying pCAMBIA–1301 via agroinfiltration and the subsequent use of 20 mg L–1 meropenem was effective in eliminating Agrobacterium from the infected explants. The transformed status of the meristemoid tissues was confirmed by GUS expression analysis, indicating that the T-DNA of the vector used had been successfully incorporated into the genome. For the first time, the researchers have demonstrated the potential methodology for direct organogenesis from agroinfiltration-transformed petal tissues of D. Sonia ‘Earsakul’.
INTRODUCTION
Dendrobium is the second largest genus in the orchid family (Orchidaceae) and contains the greatest diversity of horticulturally interesting specimens, accounting for about 85% of the total orchid cut flower trade worldwide (Puchoaa 2004; Teixeira da Silva et al. 2015). Dendrobium hybrids have dominated international trade due to their floriferous nature, with interesting flower colors, sizes and shapes, year-round availability, and long vase life. D. Sonia ‘Earsakul’ is a common favorite hybrid orchid in Thailand, and it has become popular in many countries due to its spectacular form and color. The high demand for this clone and other varieties of D. Sonia lines have prompted researchers to develop innovative tissue culture techniques dedicated to this hybrid line (Kumari et al. 2013; Puchoaa 2004) – aiming for varietal improvement, high regeneration yield, clonal stability, as well as for transgenic applications (Julkifle et al. 2012). . . . . read more
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