We cloned the Odontoglossum ringspot virus (ORSV) coat protein (CP) gene from ORSV-infected Epidendrum and constructed a pHPUNOJ E -1 vector by ligating this CP gene, green fluorescence protein (GFP) gene, and hygromycin resistant (Hyg r ) gene into the plasmid pGreenII. This chimeric vector was mediated into A. tumefaciens LBA 4404. By infecting and co-culturing the rhizomes of Cym. niveo-marginatum with A. tumefaciens LBA 4404, ORSV CP gene transgenic shoots were obtained. GFP was observed in 64% of the 81 surviving rhizomes and in 39% of 215 regenerated shoots. The presence of ORSV CP, GFP, and Hyg r genes was confirmed in the transgenic shoots by using polymerase chain reaction (PCR) analysis, whereas that of ORSV CP was confirmed by enzyme-linked immunosorbent assay (ELISA).
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