Plant pathogens elicit host transcriptome changes that facilitate hyphal infection and proliferation throughout host tissues. Host genes targeted by the pathogen to facilitate infection have been identified as susceptibility genes (S-genes). As biotrophic pathogens, rust fungi must infect the host to obtain nutrients while suppressing defense responses and likely target host-derived genes to aid in this process. To further understand the host-pathogen interaction RNA-seq analyses were performed on three compatible host-rust pathogen systems: barley cv. Morex was inoculated with Pgt race TTKSK (Ug99), soybean cv. Williams 82 was inoculated with a field collection of Phakopsora pachyrhizi, and maize cv. B73 was inoculated with an isolate of P. sorghi collected at St. Paul. Differential expression was characterized at 6, 12, 24, and 72 h post inoculation. Homologous and differentially expressed host genes were identified and compared for similarities in patterns of differential expression. In each system, a transmembrane nitrate transporter was identified as significantly upregulated early in the interaction compared to mock inoculated controls. Transporters with this molecular function were previously correlated with altering the host-pathogen interaction by knockdown and knockout mutants that were associated with resistance and heightened expression was associated with enhanced susceptibility. The transporter identified in barley was chosen for analysis to understand its effect on the host-pathogen interaction. Current objectives include site directed mutagenesis utilizing multiplexed CRISPR/Cas9 constructs to induce knockout host alleles.
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