Transformation of orchid (Dendrobium sp.) via Agrobacterium coated microparticle bombardment

Abstract

MS medium with Vitamin B5 and coconut water with different concentration were used for investigating the suitable medium of shoots. After six weeks, MS medium supplemented with 20% coconut water was concerned that the optimal medium for shoots growth with a large number and strength. These shoots also developed strongly and healthy in the medium which contained MS medium, 1g l-1 activated carbon and 0.3 mg l-1 growth hormone NAA. Agrobacterium coated particle bombardment was seemed to be as a new method which enhanced the transgenic efficiency of Dendrobium sp. Its was used to introduce ipt gene into Dendrobium sp. in order to inhibit leaf senescence of transgenic orchid, based on micro-wounding of PLBs by particle bombardment prior to inoculation with an Agrobacterium suspension. This method combined the advantages of somatic embryo-genesis and gene transfer through an integrated transformation system. This was a first report of application of this technique to transformation of orchid.