Functional study of Csrbohs in defence response against Xanthomonas citri ssp. citri

NADPH oxidases, encoded by rbohs (respiratory burst oxidase homologues), transfer electrons from NADPH to molecular oxygen (O-2) to generate superoxide anion (O-2(center dot-)), which is the first step in the formation of hydrogen peroxide (H2O2) in the plant-pathogen interaction system. In the present work, six citrus rbohs (Csrbohs) genes were identified in citrus, and their possible involvement in resistance to Xanthomonas citri ssp. citri (Xcc) was examined. Inoculation with Xcc promoted the H2O2 production and induced expression of the Csrbohs, especially CsrbohD. Results showed that CsrbohD was markedly induced in the resistant genotype kumquat Luofu' [Fortunella margarita (Lour.) Swingle] compared with grapefruit Duncan' [Citrus paradisi (Linn.) Macf.]. Virus-induced gene silencing (VIGS) of CsrbohD resulted in reduced resistance to Xcc in grapefruit, but not in kumquat. Compared with non-silenced plants, canker-like symptoms were observed earlier, and they were more extensive in the CsrbohD-silenced grapefruit. Silencing of CsrbohD also suppressed the Xcc induced reactive oxygen species (ROS) burst, and resulted in accumulation of more Xcc bacterial colonies. Taken together, these data indicate that CsrbohD promotes resistance to Xcc, especially in grapefruit.