Do anthocyanins function as antioxidants in leaves? Imaging of H2O2 in red and green leaves after mechanical injury


Kevin S. Gould. E-mail:


Purified anthocyanin extracts show strong antioxidant properties in vitro, but it is not known whether they can scavenge reactive oxygen in living cells. The oxidative responses in red and green portions of Pseudowintera colorata leaf laminae were compared by the real-time imaging of H2O2 in cells after mechanical injury. An oxidative burst was elicited almost immediately from chloroplasts in the palisade mesophyll, as evidenced using the fluorochromes dichlorofluorescein and scopoletin. H2O2 accumulated in green lamina regions for 10  min, and then decreased slowly. By contrast, red regions recovered rapidly, and maintained consistently low levels of H2O2. Infusion of cells with N-acetyl-l-cysteine accelerated the depletion of H2O2 from green regions. Wounded leaves ultimately developed a localized necrotic lesion and an intense anthocyanic band. The red regions were enriched in anthocyanins, flavonols, dihydroflavonols, and hydroxycinnamic acids. Only the anthocyanins were suitably located to account for the enhanced rates of H2O2 scavenging. The data support the hypothesis that red cells have elevated antioxidant capabilities in vivo.