Figure 2

Flowchart showing the preparation of WGE (top panel) and BYL (bottom panel). WGE (top panel). The endosperm is removed from wheat seeds and embryo particles are washed extensively to remove translation inhibitors from the endosperm. To obtain the extract, the washed embryo particles are ground with a mortar under liquid nitrogen or in a Waring blender. BYL (bottom panel). BY-2 cells are cultivated in a 5-L fermenter, harvested in the exponential growth phase (at least 5 days post-inoculation) and converted into protoplasts. The vacuoles are separated from the protoplasts by centrifugation over a stepwise Percoll gradient. The resulting evacuolated protoplasts are lysed using the nitrogen decompression method.