Optimization of glufosinate-ammonium (Basta) dose for transgenics selection in banana cv. Grand Naine

Abstract

Plant transformation techniques play an important role in modern agriculture. The dose rate of the selective agent is the primary determinant of the success rate in selecting putative transformants. The survival frequency of transformant and non-transformant cells is determined by selection pressure. In commercial banana cv. Grand Naine, for which we have already developed an effective in-vitro regeneration system, we examined the ideal dosage of a selective agent basta. Different concentrations of basta (1 mg/l, 2 mg/l, 3 mg/l, and 4 mg/l) were initially tested for their efficacy in regeneration media (RM) and germination media (GM). However, no optimal results were obtained within this range. Consequently, the concentration range was refined to lower levels, specifically 0.2 mg/l, 0.4 mg/l, 0.6 mg/l, 0.8 mg/l, and 1 mg/l, for further experiments. For both media, we found optimal results at 0.2 mg/l. Further, the affectivity of the optimal dosages of basta was examined for transgenic selection in banana cv. Grand Naine. After being transformed with the bar gene, the embryogenic cell suspension (ECS) cells were allowed to generate embryos and were transferred to RM and further to GM having 0.2 mg/l of basta. PCR analysis verified the presence of bar (BlpR) gene integration in putative transgenic banana plants.