2003 Volume 20 Issue 2 Pages 145-152
When hypocotyl sections of Raphiolepis umbellata L. were cultured in a woody plant medium containing 8.9μM benzylaminopurine and 1.1μM α-naphthaleneacetic, adventitous shoots were formed. Fully developed plants were obtained in 3 months of the culture. A chimeric plasmid pENiR harboring cDNA of Arabidopsis thaliana nitrite reductase gene (nii) under the control of an enhanced promoter consisting of sequences from both cauliflower mosaic virus 35S promoter and tobacco mosaic virus omega regions and a nopaline synthase terminator was constructed and introduced to the hypocotyl sections by particle bombardment. A plasmid pCH bearing hygromycin phosphotransferase gene (hph) was cointroduced. Transformed shoots were selected for hygromycin resistance. Polymerase chain reaction analysis showed the presence of hph gene in 16 out of 2572 bombarded hypocotyl sections, and two of which were found to bear Arabidopsis nii gene. Translation of this gene in transgenic plants was confirmed by Western blot analysis.
