Alternaria brassicicola-induced ROS accumulation during black spot disease differentially affects antioxidant efficiency, phenolic content and susceptibility of Brassica species

Early-stage responses of Brassica juncea, B. napus and B. oleracea during black spot disease and the development of Alternaria brassicicola were evaluated. Infection of plant cells by the fungus occurred mainly through direct penetration or by appressoria and, rarely, stomata in B. juncea and B. oleracea; in B. napus, penetration was mostly direct. The process of conidial germination, germ tubes and appressoria formation on a leaf surface was correlated with the post-inoculation time and host species (p < 0.05). Changes in the leaf surface were observed at successful infection sites as bright, wax-deprived areas. Significant differences in progression of infection between Brassica species were observed, with B. oleracea being the most susceptible and B. napus the least. Accumulation of superoxide anion radicals and hydrogen peroxide was not only observed in plant cells, but also in A. brassicicola germ tubes and appressoria at 8 and 12 h post-inoculation (hpi). Enhanced production of reactive oxygen species (ROS) and lipid peroxidation during infection triggered diverse patterns of enzymatic and non-enzymatic antioxidant activities and phenolic compound contents in response to A. brassicicola. In each Brassica species, a different antioxidant was the most active. With their antioxidant properties, phenolic compounds also played an essential role in these interactions. This phenomenon could be related to the disparate levels of susceptibility of the Brassica species to A. brassicicola; for example, the most susceptible, B. oleracea, showed high ROS accumulation with decreasing tendency during disease progression, unchanged phenolics content, and the highest catalase activity, differentiating it from B. juncea and B. napus.