Genome-wide identification of cassava MeRboh genes and functional analysis in Arabidopsis

Plant respiratory burst oxidase homolog (Rboh) gene family encodes NADPH oxidases, and plays important roles in the production of reactive oxygen species (ROS), plant signaling, growth and stress responses. Cassava is an important starchy crops in tropical region. Environmental stresses, such as drought, pathogen, have caused great yield loss. The mechanisms of stress response are little known in MeRBOH family of cassava. Investigation of Rboh genes response to disease may provide a clue for clarification the disease resistance mechanisms. In this study, eight MeRboh genes were identified from the cassava genome. Comparisons of gene structure, protein motifs, and a phylogenetic tree showed conservation of Rboh gene families in cassava, Arabidopsis and rice. Transcript levels of most MeRboh genes increased following treatment with a pathogen, Xanthomonas axonopodis pv. manihotis, or with phytohormones salicylic acid or jasmonic acid. Analysis of cis-acting elements also indicated that MeRboh genes could response to light, hormone, abiotic and biotic stress. Prediction of miRNA target and post-translation modification sites of MeRboh suggested possible regulations of miRNA and protein phosphorylation; and transient expression of MeRboh in cassava protoplasts confirmed their localization on plasma membrane. Expression of MeRbohB, MeRbohF partially complemented PAMP responses in Arabidopsis rboh mutants, including the expression of PTI marker FRK1, ROS production, peroxide accumulation and callose deposition. It suggesting that MeRbohB and MeRbohF may participate in the PTI pathway and contributed to ROS production triggered by pathogens. Moreover, overexpression of MeRbohB and MeRbohF enhanced the resistance of Arabidopsis against Pseudomonas syringae pv. tomato DC3000. Together, these results suggest the evolutionary conservation of MeRboh gene family and their important role in the immune response and in regulating the plant disease resistance, providing a foundation for revealing molecular mechanisms of cassava disease resistance.