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KMID : 0613820090190030378
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Journal of Life Science
2009 Volume.19 No. 3 p.378 ~ p.383
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Agrobacterium-Mediated Transformation of Phalaenopsis by Using Protocorm-Like Body
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Hur Yeon-Jae
Nam Jae-Sung
Chung Young-Soo
Kim Doh-Hoon
Lee Jai-Heon
Lee Young-Byung
Yun Dae-Jin
Kim Eun-Young
Yang Won-Tae
Yi Ki-Hwan
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Abstract
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Agrobacterium tumefaciens-mediated transformation procedure for the phalaenopsis orchid, established by using Protocormlike bodies (PLBs), was aimed at the introduction of target genes into individuals with divergent genetic backgrounds. PLBs obtained from the axillary bud of a peduncle were maintained on a hyponex medium supplemented with 1 g/l of activated charcoal, 30 g/l of sucrose and 0.1 mg/l thiamine. The multiplication rate of PLBs was about 90% in case of subculture PLBs to be cut transversely into 1/3 part from top position. The PLBs were inoculated with Agrobacterium strain EHA105 harboring both β-glucuronidase (GUS) and hygromycin-resistant genes for 20 minutes after dipping treatment. Transformation efficiency was the highest with a Agrobacterium culture medium and dipping treatment of O.D. 0.8. Newly induced PLBs were put on selection medium containing 1 ㎎/l hygromycin for 2 months. Hygromycin-resistant phalaenopsis plants that regenerated after the selection culture of PLBs showed histochemical blue staining due to GUS. Transgene integration of the hygromycin-resistant plants was confirmed by PCR and Southern blot using GUS specific primers and probe.
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KEYWORD
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Phalaenopsis, transformation, Agrobacterium, protocorm-like body
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