Early detection of green-fluorescent protein in plantain banana (Musa spp.) via particle bombardment
2000
Deswina P. | Marziah M. | Maheran Abd. Aziz | Nor' Aini Mohd. Fadzillah
Genetic transformation of Musa spp. cv. Nangka using the green fluorescent protein (GFP) gene as a marker gene was developed, the explants used were callus and rhizome slices. Bombardment parameters were optimized for high levels of transient expression of the gfp gene and exhibiting the results were recorded 72 hours after bombardment. Efficiency of the transformation was based on the gfp-fluorescent spots in callus and rhizome slices. The results obtained showed that 90% of culture exhibited gfp-fluorescence spots with parameter of 1100 rupture disk pressure. The target distance utilised was 12 cm for callus and 6 cm for rhizome slices. Using 1350 psi disk pressure and 12 cm target distance the highest efficiency was 80% for rhizome slices and 60% for callus. The gfp-expressing spots in callus could be detected after 3 months of bombardment, however, from the rhizome slices, it could be achieved after regeneration (2 months). Visualisation of GFP fluorescent in plant tissues was achieved under the Leica MZFLIII Stereo Microscope and also from photographs taken using a Nikon F60D automatic exposure system.
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