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The necrotrophic fungus  Stagonospora nodorum  produces multiple proteinaceous host-selective toxins (HSTs) which act in effector triggered susceptibility. Here, we report the molecular cloning and functional characterization of the SnTox3-encoding gene, designated  SnTox3 , as well as the initial characterization of the SnTox3 protein.  SnTox3  is a 693 bp intron-free gene with little obvious homology to other known genes. The predicted immature SnTox3 protein is 25.8 kDa in size. A 20 amino acid signal sequence as well as a possible pro sequence are predicted. Six cysteine residues are predicted to form disulfide bonds and are shown to be important for SnTox3 activity. Using heterologous expression in  Pichia pastoris  and transformation into an avirulent  S. nodorum  isolate, we show that  SnTox3  encodes the SnTox3 protein and that  SnTox3  interacts with the wheat susceptibility gene  Snn3 . In addition, the avirulent  S. nodorum  isolate transformed with  SnTox3  was virulent on host lines expressing the  Snn3  gene.  SnTox3 -disrupted mutants were deficient in the production of SnTox3 and avirulent on the  Snn3  differential wheat line BG220. An analysis of genetic diversity revealed that  SnTox3  is present in 60.1% of a worldwide collection of 923 isolates and occurs as eleven nucleotide haplotypes resulting in four amino acid haplotypes. The cloning of  SnTox3  provides a fundamental tool for the investigation of the  S. nodorum –wheat interaction, as well as vital information for the general characterization of necrotroph–plant interactions.

SnTox3 Acts in Effector Triggered Susceptibility to Induce Disease on Wheat Carrying the Snn3 Gene