AbstractThe nonstructural protein NSm of tomato spotted wilt virus (TSWV) has been identified as the avirulence determinant of the tomato single dominant Sw‐5 resistance gene. Although Sw‐5 effectiveness has been shown for most TSWV isolates, the emergence of resistance‐breaking (RB) isolates has been observed. It is strongly associated with two point mutations (C118Y or T120N) in the NSm viral protein. TSWV‐like symptoms were observed in tomato crop cultivars (+Sw‐5) in the Baja California peninsula, Mexico, and molecular methods confirmed the presence of TSWV. Sequence analysis of the NSm 118–120 motif and three‐dimensional protein modelling exhibited a noncanonical C118F substitution in seven isolates, suggesting that this substitution could emulate the C118Y‐related RB phenotype. Furthermore, phylogenetic and molecular analysis of the full‐length genome (TSWV‐MX) revealed its reassortment‐related evolution and confirmed that putative RB‐related features are restricted to the NSm protein. Biological and mutational NSm 118 residue assays in tomato (+Sw‐5) confirmed the RB nature of TSWV‐MX isolate, and the F118 residue plays a critical role in the RB phenotype. The discovery of a novel TSWV‐RB Mexican isolate with the presence of C118F substitution highlights a not previously described viral adaptation in the genus Orthotospovirus, and hence, the necessity of further crop monitoring to alert the establishment of novel RB isolates in cultivated tomatoes.

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