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Abstract
Fusarium head blight (FHB) is a devastating disease of wheat worldwide. Fhb1 is the most consistently reported quantitative trait locus (QTL) for FHB resistance breeding. A pore-forming toxin-like (PFT) gene at Fhb1 was first cloned by map-based cloning and found to confer FHB resistance in wheat. Proteins often interact with each other to execute their functions. Characterization of the proteins interacting with PFT might therefore provide information on the molecular mechanisms of PFT functions. In this study, a high-quality yeast two-hybrid (Y2H) library using RNA extracted from Fusarium graminearum (Fg)-infected wheat spikes of Sumai 3 was constructed. The agglutinin domains of PFT exhibited no self-activation and toxicity to yeast cells and were used as bait to screen the Y2H library. Twenty-three proteins that interact with PFT were obtained, which were mainly involved in the ubiquitination process, clathrin coat assembly, the oxidation-reduction process, and protein phosphorylation. The expression pattern of these interacting genes was analyzed by quantitative real-time PCR. This study clarifies the protein interactions of PFT and raises a regulatory network for PFT regarding FHB resistance in wheat.
Highlights
Fusarium head blight (FHB) is a devastating disease of wheat worldwide
Using the Y2H screening system, we screened some proteins that interact with pore-forming toxin-like (PFT) and provide information on the proteins interacting with PFT that are putatively involved in FHB resistance in wheat
PFT contains a chimeric lectin with two agglutinin domains and an ETX/MTX2 toxin domain
Summary
Fusarium head blight (FHB) is a devastating disease of wheat worldwide. Fhb[1] is the most consistently reported quantitative trait locus (QTL) for FHB resistance breeding. This study clarifies the protein interactions of PFT and raises a regulatory network for PFT regarding FHB resistance in wheat. Functional validation of candidate genes in wheat remained elusive until pore-forming toxin-like (PFT) in the Fhb[1] region was reported to confer FHB resistance by mutation analysis, gene silencing, and transgenic overexpression in wheat[10]. The proteins encoded by the photosynthesis-related genes PSK-I and PsbS1 have been confirmed to interact with the truncated haemoglobin protein trHb in photosynthesis[14] These proteins are good entry points to further elucidate their regulatory network in the development process. To screen proteins that interact with PFT, a high-quality Y2H library using Fusarium graminearum (Fg)-infected wheat spikes of the FHB-resistant cultivar Sumai 3 was constructed. Using the Y2H screening system, we screened some proteins that interact with PFT and provide information on the proteins interacting with PFT that are putatively involved in FHB resistance in wheat
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