Fig 3. Confirmation of TaEXPA2 transgenic tobacco plants.

(A) The relative expression level of TaEXPA2 was assessed by quantitative real-time PCR. The mRNA levels in the WT plants and five different TaEXPA2 transgenic lines (OE-6, OE-9, OE-15, OE-16, and OE-24). (B) TaEXPA2 protein abundance in the cell walls of WT and transgenic tobacco leaves by immunoblot analysis. Cell wall protein extracts were prepared from growing leaves of transgenic and WT tobacco plants. RuBisCO large subunit (Agrisera, AS03037) was used as a loading control. (C) Expansin activity in different transgenic lines and WT plants. Expansin activity was assayed by measuring the increase in the extension rate of wheat coleoptiles after the addition of the cell wall extract from leaves of different tobacco lines. Data represent the mean values for the three independent biological replicates. Standard errors are indicated by vertical bars. * and ** indicate significant differences from the WT values at P < 0.05 and P < 0.01, respectively, according to Duncan’s multiple range test.