Enhanced sensitivity of Arabidopsis anthocyanin mutants to photooxidation: a study with fluorescence imaging

Chlorophyll fluorescence imaging and antioxidative capability in detached leaves of the wild-type Arabidopsis thaliana ecotype Landsberg erecta (Ler) and in three mutants deficient in anthocyanin biosynthesis (tt3, tt4 and tt3tt4) were investigated under photooxidation stress induced by methyl viologen (5 μm) in the light. In comparison with the wild-type (WT) plant, photooxidation resulted in significant decreases in the contents of total phenolics and flavonoid, total antioxidative capability and chlorophyll fluorescence parameters (F_v/F_m, qP, Φ_PSII, NPQ and ETR) as determined by chlorophyll fluorescence imaging, and in an increase in cell-membrane leakiness in the three anthocyanin mutants. The sequence of sensitivity to photooxidation in the leaves of the four phenotypes were tt3tt4 (deficient in both chalcone synthase locus (CHS) and dihydroflavonol 4-reductase locus (DFR)) > tt4 (deficient in CHS) > tt3 (deficient in DFR) > WT. The results demonstrate that anthocyanins might, along with other antioxidants, protect the photosynthetic apparatus against photooxidative damage. An interesting phenomenon was observed over the 270 min of the photooxidative treatment, that is, fluorescence imaging revealed that qP, Φ_PSII and ETR appeared in three phases (fall → partial recovery → rapid fall). This was considered to be a modulation of reversible deactivation in PSII to cope with the moderate oxidative stress in the first two stages of short-term treatment (<150 min), followed finally by damage to PSII under severe oxidative stress with prolonged treatment.