Cu 2 fluorescein cufl fluorescent probe

Nitric oxide detection was performed as in Horchani et al. (2011) using the 4,5‐diaminofluorescein probe (DAF‐2; Sigma‐Aldrich) with the following changes. Either nodules (20–30 mg FW) or root segments (50–100 mg FW) were incubated in 1 ml of detection buffer (10 mM Tris‐HCl pH 7.4, 10 mM KCl) in the presence of 10 μM DAF‐2. As a control, NO production was measured in the same experimental system through the use of the Cu(II) fluorescein (CuFL) fluorescent probe (Strem Chemicals, Bischheim, France) instead of DAF‐2 in the detection buffer as described in Horchani et al. (2011). Similar results were obtained with both probes. The production of NO was measured with a spectrofluorimeter‐luminometer (Xenius, Safas, Monaco).

NO detection was performed as in Horchani et al. (2011) (link) using the 4,5-diaminofluorescein probe (DAF-2, Sigma-Aldrich) with the following changes. Either nodules (20–30 mg fresh weight) or root segments (50–100 mg fresh weight) were incubated in the dark at 23°C in 1 ml of detection buffer (10 mM Tris-HCl pH 7.4, 10 mM KCl) in the presence of 10 μM DAF-2. As a control, NO production was measured in the same experimental system through the use of the Cu(II) fluorescein (CuFL) fluorescent probe (Strem Chemicals) instead of DAF-2 in the detection buffer as described in Horchani et al. (2011) (link). Similar results were obtained with both probes. The production of NO was measured with a spectrofluorimeter-luminometer (Xenius, SAFAS, Monaco). Inhibitors are used as described in Horchani et al. (2011) (link). The inhibitors are added to the reaction medium for the determination of NO at the concentration of 1 mM tungstate (Tg), 1 mM allopurinol, 1 mM propyl gallate, and 300 μM potassium cyanide (KCN). NO production was initiated 1 h after the addition of inhibitors. Three independent biological replicates have been performed with three technical replicates per biological assay.