Glyphosate tolerance was compared among transgenic tobacco

plants containing gat, G2-aroA, or both genes by assessing germination of T1 transgenic tobacco seeds and by leaf spraying. T1 seeds of transgenic tobacco G2, GAT, and G2-GAT (containing G2-aroA, gat, or G2-aroA/gat, respectively) were germinated after sterilization on MS medium containing different concentrations of glyphosate ( Fig. 5). Glyphosate tolerance was evaluated by seed germination and seedling growth on medium containing glyphosate after 4 weeks. On medium containing 0.2 mmol L− 1 glyphosate, no difference in seed germination was apparent among the 3 types of transgenic tobacco. All transgenic plants germinated and developed normally, and there was selleck little difference in seedling growth vigor compared with the control (plants growing on MS medium without glyphosate). On medium containing 1 mmol L− 1

glyphosate, all of the G2 transgenic plants died. No difference in viability was apparent among controls and GAT or G2-GAT transgenic plants, although the growth vigor of GAT and G2-GAT plants was obviously reduced. On media supplemented with 5 mmol L− 1 glyphosate, a difference in viability was apparent between GAT and G2-GAT transgenic plants, and their growth vigor was reduced compared with the control. On media supplemented with 10 mmol L− 1 glyphosate, all AZD8055 chemical structure GAT transgenic plants died, but 14% of G2-GAT plants survived ( Table 1). The segregation ratio of glyphosate resistant and sensitive plants was 3:1 in selection medium containing 0.2 mmol L− 1 glyphosate. We accordingly postulated that the genes introduced into these transgenic tobacco plants were inserted as single copies. T1 transgenic plants at 6 to 8-leaf-stage were sprayed with a 1.0% (v/v) solution of the herbicide Roundup (isopropylamine glyphosate salt as active ingredient, 41.0%, w/v) at a dose of 0.8 L ha− 1. In non-transgenic plants, the leaves and stem apex began to wilt 1–3 days after treatment. The non-transgenic

control showed severe wilt and chlorosis on all leaves after 5 days and died 7 days after Osimertinib chemical structure treatment. Twenty-four GAT plants grew well with normal morphology for 2 weeks after treatment, and 6 GAT plants begin to wilt 5 days after treatment and died after 2 weeks. Four G2 plants survived, but 3 showed partial leaf chlorosis and bleaching after 6 days. Twenty-six G2-GAT plants grew well with normal morphology for 2 weeks after treatment, and the remaining 4 plants exhibited wilting and bleaching 5 days after treatment and then died. All the three types of transgenic plants, except for 5 G2-GAT plants, died after glyphosate treatment at a dose of 1 L ha− 1 (Table 2 and Fig. 6).