Expression of foreign genes, GUS and hygromycin resistance, in the halophyteKosteletzkya virginicain response to bombardment with the Particle Inflow Gun

Xianggan Li, John L. Gallagher
1996 Journal of Experimental Botany  
A 'Particle Inflow Gun' was constructed to introduce the glucuronidase (GUS) and hygromycin resistance genes into halophytic suspension cells of Kosteletzkya virginica. The transient expression of the GUS gene was associated with the cell culture conditions, physical parameters during the use of the Particle Inflow Gun, and different promoters coupled to GUS. When the CaMV35S promoter was used, the cells adapted at 85 mM NaCI had a similar gene transfer efficiency to those of the
more » ... the non-salt-adapted control, while expression was less in the 170 mM and 255 mM NaCI-adapted cells. Both elevating bombardment pressure to 1.65 mPa and shortening the distance between the cells and the particle holder from 21 cm to 9 cm enhanced GUS expression in the cells grown in four salinity treatments. An ABA-responsive promoter induced the expression of the GUS gene either with 10 4 M ABA or with salts in the post-bombardment medium in both control and NaCI-adapted cell lines. Stable transgenic callus lines were isolated by using hygromycincontaining medium after bombarding the suspension cells with the Particle Inflow Gun. The presence of the GUS gene in stable transformants was confirmed not only by histochemical and fluorimetric assays for the GUS activity, but also by Southern hybridization of RT-PCR amplified mRNA.
doi:10.1093/jxb/47.9.1437 fatcat:x6rtuuixdrhkrb7h6bvyp5f5ay