ROS generation experiment
Cells were harvested by centrifugation at 1500 g for 10 min the washed 3
times with sterile double distilled water before suspended in TAP
medium. Cultures were then treated with 125 mM of NaCl and with or
without ABA and were illuminated and shaken-incubated for 3 h, as
described above in materials and methods. One-milliner samples were
collected and stained with 20 µM of H2DCFDA for 30 m
(Colle‘n, and Davison, 1997). The fluorescence of the dye was measured
by a Fortessa flow cytometer (Breda, Netherlands). Cell vitality was
carried out by inoculating of 50 µl drops of Chlamydomonascultures after 24 h of treatment on TAP-1.5% Difco agar plates.