The DNA isolated from SC50 and SC50×3 was subject to a methylation content-sensitive enzyme and double-digest, restriction-site-associated DNA technique (ddRAD-MCSeEd). The same trend was observed in conidia production. Seedling stem base and head assays revealed a virulence decline of all subcultures, whereas virulence was restored in SC50×3. In vitro conidia production, colonies growth and secondary metabolites production were also determined for SC1, SC23, SC50, and SC50×3. SC50 was also used to re-infect (three times) wheat heads (SC50×3) to restore virulence. A416) were inoculated with subcultures (SC) 1, 23, and 50.
To assess the effect of subculturing on virulence, wheat seedlings and heads (cv. A virulent strain of this fungus was consecutively sub-cultured for 50 times (once a week) on potato dextrose agar. We investigated if a such a strategy is also adopted by Fusarium graminearum in regulating virulence toward its natural hosts. In summary, results from this study indicate that lncRNA transcripts and alternative splicing events are two important mechanisms for regulating the expression of genes important for conidiation, hyphal growth, and pathogenesis, and provide new insights into transcriptomes and gene regulation in the rice blast fungus.ĭNA methylation mediates organisms’ adaptations to environmental changes in a wide range of species. Furthermore, we exemplified that a lncRNA was important for hyphal growth likely by regulating the neighboring protein-coding gene and that alternative splicing of the transcription factor gene CON7 was required for appressorium formation. In addition, 2,358 genes were found to undergo alternative splicing events. Notably, their expressions were concomitant with the transcription of neighboring genes thereof in conidia and hyphae. Interestingly, the vast majority of these novel genes likely transcribed long non-coding RNAs (lncRNAs), and most of them were localized in the intergenic regions. Here, we performed and analyzed RNA-Seq of conidia and hyphae, resulting in the identification of 3,374 novel genes. oryzae is publicly available, its current transcriptomes remain incomplete, lacking the information on non-protein coding genes and alternative splicing. Pyricularia oryzae causes the rice blast, which is one of the most devastating crop diseases worldwide, and is a model fungal pathogen widely used for dissecting the molecular mechanisms underlying fungal virulence/pathogenicity. Our study indicates that TuRLK1 is required for immune response mediated by the unique NLR protein YrU1, and likely plays an important role in disease resistance to other pathogens. Furthermore, overexpression of TuRLK1 could enhance powdery mildew resistance in bread wheat and Arabidopsis thaliana after inoculating with the corresponding pathogens. Silencing of TuRLK1, using barley stripe mosaic virus (BSMV)-induced gene silencing (VIGS) system in PI428309 that contains YrU1, compromised the resistance against stripe rust caused by Pst CY33, indicating that TuRLK1 was required for YrU1-activated plant immunity. Transient expression of TuRLK1 induced hypersensitive cell death response in N. TuRLK1 contained only a small number of LRR motifs, and was localized in the plasma-membrane. In this study, we found that the transcript level of TuRLK1, which encodes a novel leucine-rich repeat receptor-like kinase, was up-regulated after inoculation with Pst in the presence of YrU1, through RNA-seq analysis in T. However, the molecular mechanism and components involved in YrU1-mediated resistance are not clear. YrU1 was positionally cloned in the progenitor species of the A genome of bread wheat, Tricicum urartu, recently.
YrU1 is a nucleotide-binding site (NBS) and leucine-rich repeat (LRR) protein (NLR), with additional ankyrin-repeat and WRKY domains and confers effective resistance to stripe rust fungus Puccinia striiformis f.
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