MicroRNA-like RNAs in plant pathogenic fungus Fusarium oxysporum f. sp. niveum are involved in toxin gene expression fine tuning

MicroRNA-like RNAs (milRNAs) are short non-coding regulatory sRNAs which play an important role in regulating gene expression at the post-transcriptional level by targeting mRNAs for degradation or inhibiting protein translation. To explore the presence of milRNAs in Fusarium oxysporum f. sp. niveum (Fon) and analyze their expression at different propagules, two categories of sRNAs were identified from Fon hyphae and microconidia using illumina sequencing. A total of 650,960 and 561,114 unique sRNAs were obtained from the hyphae and microconidia samples. With a previously constructed pipeline to search for microRNAs, 74 and 56 milRNA candidates were identified in hyphae and microconidia, respectively, based on the short hairpin structure analysis. Global expression analysis showed an extensively differential expression of sRNAs between the two propagules. Altogether, 78 significantly differently expressed milRNAs were identified in two libraries. Target prediction revealed two interesting genes involved in trichothecene production, necrosis and ethylene-inducing peptide 1 (NEP1) biosynthesis and in silico analysis indicated that they were down-regulated by Fon-miR7696a-3p and Fon-miR6108a. The expression levels of these two milRNAs were further validated by qRT-PCR and the results were consistent. The negative correlation of the expression levels between these two milRNAs and their potential target genes imply that they play a role in trichothecene and NEP1 biosynthesis. And this negative regulation for toxin-related gene expression is more specific in microconidia. The present study provides the first large-scale characterization of milRNAs in Fon and the comparison between hyphae and microconidia propagules gives an insight into how milRNAs are involve in toxin biosynthesis.