Agrobacterium-mediated transformation of the wild orchid species Phalaenopsis amabilis

Induction of Somatic Embryogenesis through Overexpression of ATRKD4 Genes in Phalaenopsis “Sogo Vivien”

“…Based on the morphological changes during embryo development of P. "Sogo Vivien", therefore protocorms in phase 4 ( 21 DAS) (Fig.3a) were selected in this study as the target for genetic transformation using Agrobacterium. This phenomenon was very similar to that of embryo development in the wild orchid Phalaenopsis amabilis as reported by Semiarti et al (2007), inwhich 21 DAS protocorms were used as the target for Agrobacterium-mediated transformation. Bunnag and Pilahome (2012) have also used protocorms as the transformation target.…”

“…This compound is highly reactive towards biomolecules, which cause damage of plant tissue followed by browning and necrosis (Kuta and Tripathi, 2005). Several studies of genetic transformation using Agrobacterium with transformation targets in the form of protocorm-like bodies (plb) and orchid protocorm obtained transformation effi ciency of 4.3% in Cattleya (Zhang et al, 2010); 1.91% in Phalaenopsis hybrid (Mishiba et al, 2005); 1.7% in Phalaenopsis amabilis (Semiarti et al, 2007); and 1.3 to 2.7% in Oncidium and Odontoglossum (Raffeiner et al, 2009). To improve the transformation effi ciency, there are many studies using a variety of treatments, such as the provision of antioxidants, desiccation of transformation target before and after infection with Agrobacterium, length of co-cultivation time, density of Agrobacterium tumefaciens used in the transformation, temperature of co-cultivation process, and the addition of surfactant at the time of Agrobacterium tumefaciens infection to the explants (Opabode, 2006).…”

“…The use of the induction system with glucocorticoid is highly efficient, when compared to the other methods. In transgenic technology of plants, generally induction of foreign gene expression was conducted by providing strong promoter(s) such as the Caulifl ower Mosaic Virus-35S promoter (Semiarti et al, 2007), regulatory protein Gal4 from Saccharomyces cerevisiae (Liu and Lehman, 2008), or the herpes simplex virus VP16 as a regulatory switch (Wysocka and Herr, 2003) that positioned upstream of the transcription unit of the foreign gene to be regulated, so as to force the foreign gene can be expressed. But, it could not in this case, Figure 6.…”

“…Agrobacterium tumefaciens-mediated genetic transformation method was used to produce a variety of orchid plants (Semiarti et al, 2007;Sreeramanan and Zuraida, 2010).…”

Agrobacterium-Mediated Transformation of Indonesian Orchids for Micropropagation

“…Therefore, this method is readily adopted for plant genetic engineering. Transformation using Agrobacterium has become common in horticultural plants such as in lettuce (Michelmore et al, 1987), cabbage (Metz et al, 1995;Jin et al, 2000;Lee et al, 2000), kale (Cogan et al, 2001), orchids (Semiarti et al, 2007 and2010;Chin and Mii, 2011), and petunias (Klee et al, 1987). With genes that can modulate auxin concentrations, transgenic techniques have been used to manipulate endogenous levels of auxin in plants.…”