Detection of GUS activity was performed using 5-bromo-4-chloro-3-indoyl-β-D-glucuronide (X-Gluc) as described in67 (link), with modifications68 (link). Briefly, 4, 7 or 12 dps seedlings were infiltrated into fresh GUS substrate for 5 min (30,000 Pa) and then incubated for 72 h at 37 °C. Seedlings were, washed twice with water and preserved in 1x PBS-50% glycerol at 4 °C until observation under an Axioskop2 plus microscope (Zeiss) or a MZ9.5 stereomicroscope (Leica).
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