Figure 5.

Effect of overexpressed thioredoxin h on pullulanase activity in transgenic barley grain during germination and seedling development. A homozygous line, GPdBhssBarWtrx-29–3, and a null segregant, GPdBhssBarWtrx-29–11–10, were used for the pullulanase assays. (Top) Pullulanase was assayed spectrophotometrically by measuring the dye released from red pullulan substrate at 534 nm. (Middle) Pullulanase was separated on native 7.5% polyacrylamide gels containing the red pullulan substrate. Activity, identified by comparison with purified barley pullulanase, is seen as clear areas that developed on incubating the gel in 0.2 M succinate buffer, pH 6.0, for 1 hr at 37°C. (Bottom) The gel in Exp. B was scanned and analyzed by integration of the activity bands.