Genetic transformation of Vanda lilacina Teijsm. & Binnend. with a chitinase gene

This research was aimed to improve plantlet regeneration and to establish the optimal conditions for transformation of Vanda lilacina Teijsm. and Binnend. using Agrobacterium tumefaciens strain LBA4404 (pCAMBIA 1305.1) harboring a chitinase gene as a fungal resistant one. It was found that seeds successfully germinated, and developed into protocorms and plantlets on solid New Dogashima (ND) medium supplemented with 1 % (w/v) potato juice. Protocorm proliferation was successful when culturing the protocorms on solid ND medium supplemented with 3 mg/l BA. For successful genetic modification, the protocorms were cocultivated with A. tumefaciens for 45 min. Bacterial elimination and selection of putative transformants were implemented using 250 mg/l cefotaxime and 10 mg/l hygromycin, respectively. GUS assay revealed 80% of the protocorms expressing gus gene activity. Availability of 35S and NOS detected by PCR analysis confirmed gene integration.