As. Pac. J. Mol. Biol. & Biotech., June 2000 Vol. 8(1) :85-94
Potential selective agents for orchid transformation
A. Janna Ong1, M.Marziah 1*, G.K.A. Ahmad Parveez2
>1Department of Biochemistry and Microbiology, Faculty of Science and Environmental
Studies,
University Putra Malaysia, 43400 UPM, Serdang, Selangor Darn 1 Ehsan, Malaysia
2Palm Oil Research Institute of Malaysia, Biology Division, 50720 Kuala Lumpur,
Malaysia
Received 5 February 2000 / Accepted 17 July 2000
Abstract.
Protocorm like bodies (PLBs) of Dendrobium Sonia 17 were bombarded with 1.0 mm gold microparticles at 1100 psi using the PDS 1000/helium powered biolistic gun. At two weeks post bombardment the PLBs were subjected to different concentrations of antibiotic selections. The five selective agents tested were hygromycin, geneticin, Basta, paromomycin, and kanamycin each at 0, 25, 50, 100, 150, 200, and 300 mg/L. For hygromycin, the experiment was further carried out at lower concentrations of 0, 5, 10, 15, 20, and 25 mg/L. After one month into selection, fresh weights of the PLBs were taken and the number of surviving PLBs counted. It was observed that hygromycin was the best selective agent as it gave a total kill off (all PLBs died) at 25 mg/L. Geneticin was second with 100 % dead at 150 mg/L. For Basta, 10 % of the PLBs still survived at the highest concentration, 300 mg/L, tested. As for paromomycin and kanamycin, all the PLBs survived at 300 mg/L.
Key Words: Dendrobium orchid, transformation, selective agent