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

This research was aimed to improve plantlet regeneration and to establish the optimal conditionsfor 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 successfullygerminated, and developed into protocorms and plantlets on solid New Dogashima (ND) mediumsupplemented with 1 % (w/v) potato juice. Protocorm proliferation was successful when culturing theprotocorms on solid ND medium supplemented with 3 mg/l BA. For successful genetic modification, theprotocorms were cocultivated with A. tumefaciens for 45 min. Bacterial elimination and selection of putativetransformants were implemented using 250 mg/l cefotaxime and 10 mg/l hygromycin, respectively. GUSassay revealed 80% of the protocorms expressing gus gene activity. Availability of 35S and NOS detected byPCR analysis confirmed gene integration.