Method for determining reactive oxygen species (ROS) was based on Galsurker et al. (2020) with slight modification [19 (link)]. Five ulvan-treated grapes and 5 DDW-treated grapes were tested on 2 and 4 DPT. The grapes were submerged in 10 μM DCF (2′,7′-Dichlorofluorescein, Sigma-Aldrich, St. Louis, MO, USA) in phosphate-buffered saline (PBS) and placed on an orbital shaker at 1.6 Hz for 15 min in the dark. Followed by de-staining with PBS solution on the orbital shaker for 1 min, and the washing repeated twice.
The fluorescence was measured at 488 nm excitation and 510 nm emission using in- vivo Imaging System (IVIS®, PerkinElmer, Inc., Waltham, MA, USA) Results were calculated using IVIS® Lumina II imaging system software (PerkinElmer, Inc., Waltham, MA, USA) and expressed in units of radiance [p/s/cm²/sr].
The fluorescence was measured at 488 nm excitation and 510 nm emission using in- vivo Imaging System (IVIS®, PerkinElmer, Inc., Waltham, MA, USA) Results were calculated using IVIS® Lumina II imaging system software (PerkinElmer, Inc., Waltham, MA, USA) and expressed in units of radiance [p/s/cm²/sr].
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