Intracellular NO and ROS Quantification

According to a previous method, intracellular NO production was measured using the fluorescence probe 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF-FM-DA). Mycelia were stained with DAF-FM-DA for 30 min, and NO was detected with a laser scanning confocal microscope (Carl Zeiss LSM700, Oberkochen, Germany) (14 (link)). The ROS concentration was assessed according to a previously described method, and intracellular ROS production was measured by the fluorescence probe DCFH-DA (52 (link)). The fluorescence was analyzed using Zeiss software (ZEN lite, Zeiss, Göttingen, Germany) (53 (link)).