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Henares, B.

Publications and source records attributed to Henares, B..

2 recordsLinked to original sources

Major effector loss reveals compensatory pathogenicity networks in a necrotrophic wheat pathogen

Necrotrophic effectors (NEs) are key determinants of virulence in the necrotrophic fungal pathogen Parastagonospora nodorum that causes septoria nodorum blotch of wheat. However, targeted removal of three important NEs SnToxA, SnTox1, and SnTox3 in the mutant{Delta} toxa13 previously revealed a redundancy mechanism is triggered, whereby pathogenicity on wheat is maintained. In this study, we investigated the gene regulatory profile underpinning this phenomenon and discover that virulence is not dependent on a fixed set of dominant effectors but instead arises from a flexible, epistatic compensatory network. Although host transcriptional responses to the P. nodorum wildtype SN15 and{Delta} toxa13 infection remained largely conserved, consistent with an overlapping disease-susceptibility pathway, a significant upregulation of candidate effector genes was observed in{Delta} toxa13. This included the recently characterised NE SnTox267, and several other candidate effectors able to induce necrosis in the non-host Nicotiana benthamiana, each carrying a predicted structural fold conserved across other pathogens. We therefore provide further direct evidence that virulence is maintained in P. nodorum lacking three NEs by an epistatic and compensatory effector network, underpinned by changes in pathogen gene expression. Targeting conserved effector-mediated virulence mechanisms rather than individual host-specific gene-for-gene interactions may provide a more tractable route to host resistance.

molecular biology↗

A conserved fungal transcription factor domain drives protein-protein interactions and necrotrophic effector-mediated virulence in Parastagonospora nodorum

The fungal pathogen Parastagonospora nodorum utilises necrotrophic effectors (NEs) to cause chlorosis and necrosis on wheat. NE expression is mediated by an assortment of transcription factors (TF), the most well-characterised of which is PnPf2. Orthologues of PnPf2 regulate virulence in phytopathogenic fungi across the Ascomycete fungal lineage, yet their protein architecture remains functionally uncharacterised. These orthologues are characterised by the archetypal N-terminal Zn2Cys6 zinc-finger DNA-binding domain (DBD), a conserved yet poorly characterised middle-homology region (MHR) and a C-terminal disordered region. We investigated the role of each of the three domains through PnPf2 truncation mutants in situ. This revealed the conserved MHR of PnPf2 is required for the development of disease symptoms on wheat, but also that the C-terminal disordered region in-part modulates NE expression. Domain-interaction analysis through yeast-2-hybrid (Y2H) screening reveals PnPf2 forms a homodimer mediated by the MHR, indicating its importance mediating protein-protein interactions. Using the MHR as a bait in library-scale protein-protein interaction Y2H assays, we identified the COP9-signalosome protein PnCsn6 as a key interaction partner. PnCsn6 is essential for disease symptoms during P. nodorum infection on wheat, including NE expression. Our study presents the first domain-level functional investigation this virulence-regulating TF orthologue, the results of which underpin the essential role of the MHR in driving protein-protein interactions and effector regulation. It also reveals an essential protein-signalling pathway with which PnPf2 directly interacts and shares regulatory control of virulence.

molecular biology↗