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

ABSTRACT: 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.

SUBMITTER: Jiang X

PROVIDER: S-EPMC5617829 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature

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

Jiang Xuefei X Qiao Fei F Long Yali Y Cong Hanqing H Sun Huapeng H

3 Biotech 20170927 5

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 ...[more]

PMID: 29062675

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Project description:Successive cultivation of fungi on artificial media has been reported to cause the sectorization, which leads to degeneration of developmental phenotype, and virulence. Fusarium oxysporum f. sp. niveum (Fon), the causal agent of watermelon Fusarium wilt, forms degenerated sectors after successive cultivation. In the present research, we demonstrated that subculture with aged mycelia increased the incidence of degenerations. To further investigate the differences between the Fon wild type (sporodochial type, ST) and variants (MT: mycelial type and PT: pionnotal type), developmental phenotypes and pathogenicity to watermelon were examined. Results in variants (PT2, PT3, PT11, and MT6) were different from ST with mycelia growth, conidia production and chlamydospore formation. Virulence of degenerated variants on susceptible watermelon Grand Baby (GB) cultivar was determined after inoculation with Fon variants and Fon ST. In root dipping methods, Fon variants showed no significant differences in disease progress compared with ST. Fon variants showed a significant decrease in disease progression compared with ST through infested soil inoculation. The contrasting results of two inoculation methods suggest that the degenerative changes due to repeated successive cultivation may lead to the loss of pathogen virulence-related factors of the early stage of Fon infection process. Therefore, cell wall-degrading enzymes (CWDEs; cellulase, pectinase, and xylanase) activities of different variants were analyzed. All Fon degenerated variants demonstrated significant decreases of CWDEs activities compared with ST. Additionally, transcript levels of 9 virulence-related genes (fmk1, fgb1, pacC, xlnR, pl1, rho1, gas1, wc1, and fow1) were assessed in normal state. The degenerated variants demonstrated a significantly low level of tested virulence-related gene transcripts except for fmk1, xlnR, and fow1. In summary, the degeneration of Fon is triggered with successive subculture through aged mycelia. The degeneration showed significant impacts on virulence to watermelon, which was correlated with the reduction of CWDEs activities and declining expression of a set of virulence-related genes.

Dataset Information

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

Publications

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

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