H2O2 degradation is suppressed in kumquat leaves infected with Xanthomonas axonopodis pv. citri

We found in a previous study that after leaves of kumquat [Fortunella margarita (Lour.) Swingle] cv ‘Nagami’ were inoculated with Xanthomonas axonopodis pv. citri (Xac), total superoxide dismutase activity (SOD) increased to promote higher H2O2 concentrations that coincided with a 4-fold decline in Xac populations ( Kumar et al., 2011a). The objective of the current study was to determine how activities and isoforms of important enzymes that catabolize H2O2, specifically catalase (CAT), ascorbate peroxidase (APOD), and the Class III peroxidases (POD) that are located in the apoplast, change in infected kumquat leaves to affect concentration and compartmentalization of H2O2. DAB (3,3-diaminobenzidine) staining of the Xac-infected leaves confirmed higher overall concentration of H2O2 as in our earlier study. One day after inoculation (dai), APOD activity declined below the controls and declines steadily up to 10 dai when the experiment was terminated. CAT activity was similar to the controls until 4 dai then declined rapidly to about 60% the activity of the controls by 6 dai, after which it remained fairly constant until 10 dai. There were 4 CAT isoforms in control leaves and 5 isoforms in infected leaves. The CAT-1 isoform band was much smaller in infected plants than the control at all sampling times. The CAT-3 isoform band disappeared at 10 dai. The CAT-5 isoform band, which was not observed in control leaves, appeared only at 4 dai in infected leaves. POD activity of infected leaves increased above the controls starting 1 dai and reached a maximum of about 3-fold higher than the controls 8 dai after which it declined. Two POD isoforms were detected in control and infected plants. This study demonstrated that the higher accumulation of H2O2 in kumquat leaves infected with Xac was promoted during pathogenesis first by the suppression of APOD activity and later by suppression of CAT activity. We propose that the higher SOD and lower APOD and CAT activities in the symplast contributed H2O2 substrate for the higher POD activity in the apoplast, which is known to be involved in plant defense against pathogens.

• Citrus canker is an economically important disease worldwide. • H2O2 degradation was suppressed in canker infected ‘kumquat’ leaves. • Activities of catalase and ascorbate peroxidase were decreased during Xac–‘kumquat’ interaction.