2003 Volume 49 Issue 1 Pages 33-37
We have improved our previously published technique for plant regeneration from seed-derived embryogenic callus of bahiagrass. Embryogenic callus was initiated from mature seeds on Murashige and Skoog (MS) liquid medium and solid medium (0.3% Gellan Gum) supplemented with 2mg/1 2,4-dichlorophenoxyacetic acid (2,4-D) and 3% sucrose. This modified culture can be kept small size through regular partition and be more efficient for callus induction and remains regencrable for at least 18 months. Also, in transformation experiments, a greater surface is available for gene delivery and relatively more cells can potentially regenerate. Using a simple self-built particle inflow gun with improved technical features, we have optimized delivery conditions and bombarded the small calli with pDBI, a construct carrying both the GUS reporter gene and a bialaphos resistance gene (bar). Numerous calli have been selected on bialaphos-supplemented medium and two calli displayed GUS activity. One of these calli regenerated many albino plants, but the green plants could not be recovered. However, theses plants displayed GUS activity and integration of the transgenes were confirmed by PCR amplification analysis.
