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Deoxynivalenol (DON) is one of the most widespread trichothecene mycotoxins in contaminated cereal products. DON plays a vital role in the pathogenesis of  Fusarium graminearum , but the molecular mechanisms of DON underlying  Fusarium –wheat interactions are not yet well understood. In this study, a novel wheat ADP-ribosylation factor-like protein 6-interacting protein 4 gene,  TaArl6ip4,  was identified from DON-treated wheat suspension cells by suppression subtractive hybridization (SSH). The qRT-PCR result suggested that  TaArl6ip4  expression is specifically activated by DON in both the  Fusarium  intermediate susceptible wheat cultivar Zhengmai9023 and the  F  usarium  resistant cultivar Sumai3. The transient expression results of the TaARL6IP4::GFP fusion protein indicate that  TaArl6ip4  encodes a plasma membrane and nucleus-localized protein. Multiple sequence alignment using microscale thermophoresis showed that TaARL6IP4 comprises a conserved DON binding motif,  67 HXXXG 71 , and exhibits DON affinity with a dissociation constant ( K  D ) of 91 ± 2.6 µM. Moreover, TaARL6IP4 exhibited antifungal activity with IC 50  values of 22 ± 1.5 µM and 25 ± 2.6 µM against  Fusarium graminearum  and  Alternaria alternata , respectively. Furthermore, TaArl6ip4 interacted with the plasma membrane of  Fusarium graminearum  spores, resulting in membrane disruption and the leakage of cytoplasmic materials. The heterologous over-expression of  TaArl6ip4  conferred greater DON tolerance and  Fusarium  resistance in  Arabidopsis.  Finally, we describe a novel DON-induced wheat gene,  TaArl6ip4 , exhibiting antifungal function and DON affinity that may play a key role in  Fusarium– wheat interactions.

journal of fungi

A Novel Deoxynivalenol-Activated Wheat Arl6ip4 Gene Encodes an Antifungal Peptide with Deoxynivalenol Affinity and Protects Plants against Fusarium Pathogens and Mycotoxins