Differential Pathogenicity and Secreted in Xylem Gene Patterns in Fusarium spp. Causing Onion Basal Rot

Fusarium basal rot (FBR) poses a serious threat to onion (Allium cepa L.) production worldwide. In South Korea, FBR is primarily associated with Fusarium oxysporum, F. commune, and F. proliferatum. To investigate the relationship between effector gene profiles and virulence, we screened 34 isolates collected from FBR-affected fields for 14 Secreted in Xylem (SIX) genes and three additional effector candidates (CRX1, CRX2, and C5). F. oxysporum isolates carrying the effector suite SIX3, SIX5, SIX7, SIX9, SIX10, SIX12, SIX14, together with CRX1, CRX2 and C5, exhibited significantly higher aggressiveness on onion seedlings and bulbs than effector-negative strains. Among F. commune isolates lacking SIX genes, those carrying both CRX1 and CRX2 tended to show greater pathogenicity than CRX-negative strains. Nevertheless, SIX-negative strains still caused substantial seedling loss and bulb-rot, indicating the involvement of SIX-independent virulence factors. All F. proliferatum isolates were comparably pathogenic to SIX-negative F. oxysporum and F. commune strains, and uniformly carried SIX2-1 and CRX2, with a subset also harboring the SIX2-2 homologue. Across all isolates, SIX9 was the most frequently detected SIX gene and was markedly enriched in strains exhibiting strong pathogenicity. We developed and validated a SIX9-targeted quantitative PCR (qPCR) assay that specifically detects SIX9-positive Fusarium isolates (mainly F. oxysporum and F. commune), with detection limits of 1 pg of DNA or 10⁴ conidia/g soil. These findings enhance our understanding of effector repertoires contributing to Fusarium pathogenicity on onion and provide a molecular tool to support FBR diagnosis.