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Genetics, genomics, and transcriptomics reveal candidate avirulence/virulence effectors in Pyrenophora teres f. teres
Jonathan Richards: North Dakota State University; Vaidehi Koladia: North Dakota State University; Robert Brueggeman: North Dakota State University; Timothy Friesen: USDA ARS
<div><i>Pyrenophora teres </i>f. <i>teres</i>, a necrotrophic fungal pathogen of barley and causal agent of net form net blotch, utilizes a repertoire of effector molecules to facilitate disease. Previous research identified 19 QTL associated with virulence/avirulence on 10 barley lines in a fungal mapping population derived from a cross between isolates FGOH04Ptt-21 (North Dakota) and BB25 (Denmark), of which, three QTL contributed a large effect to the phenotypic variation (R<sup>2 </sup>> 30%). Reference quality genomes of each parental isolate were developed via Pac-Bio sequencing and RNAseq analyses conducted to identify candidate effectors underlying the major QTL. Two loci, designated <i>Pin1</i> and <i>Bee1 </i>are hypothesized to be virulence loci, as analysis of QTL effects in progeny isolates appear additive. RNAseq analysis revealed single polymorphic, small secreted proteins underlying <i>Pin1 </i>and <i>Bee1</i>, exhibiting ~4.5x and ~4x upregulation post-inoculation, respectively. A third locus, <i>Tif1</i>, appears to condition avirulence in a qualitative manner. The genomic region underlying <i>Tif1 </i>in the avirulent isolate BB25 contains an ~100 kb expansion, containing 23 genes with low or no homology to annotated genes in the FGOH04Ptt-21 genome. A candidate gene within this region containing a predicted transmembrane domain was identified to be upregulated ~34x at 48 hours post-inoculation. <i>Pin1</i>, <i>Bee1</i>, and <i>Tif1 </i>candidate genes are currently being targeted for functional validation.</div>

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