N6-methyladenosine RNA methylation is involved in virulence of the rice blast fungusPyricularia oryzae(syn.Magnaporthe oryzae)

N6-methyladenosine (m6A) RNA methylation is a conserved modification of RNA in eukaryotes. Pyricularia oryzae, a filamentous phytopathogenic fungus, is the cause of a destructive rice blast disease that can lead to significant declines in rice production. Here, we characterized the function of m6A RNA methylation in the development and virulence of P. oryzae by studying four genes with functional genomics. We found that PoIme4 is an N6-adenosine-methyltransferase, and deletion of PoIME4 led to decreased levels of m6A RNA methylation. PoYth1 and PoYth2 are two m6A-binding proteins, and deletion of PoYTH2 led to reduced conidiation. Co-localization experiments showed that PoAlkb1 (an mRNA:m6A demethylase) and PoYth1 were co-localized with PoDcp1 in the processing bodies involved in mRNA decay. Virulence tests showed that PoIME4, PoALKB1, PoYTH1 and PoYTH2 were involved in virulence on rice in P. oryzae. Therefore, these experimental evidences provide new and important information about the roles of m6A RNA methylation in fungal asexual reproduction and pathogenicity.