LAUSR

In 2022, an outbreak of apparent Fusarium wilt raised concern, as the disease has historically been sporadic and infrequent. Previous studies reported that the primary causal agent of this disease in the U.S. is Fusarium oxysporum f. sp. nicotianae, but the identity of other Fusarium spp. responsible for Fusarium wilt and root rot, and their pathogenicity have not been thoroughly investigated. To identify Fusarium species associated with the recent disease outbreak in flue-cured tobacco, 36 symptomatic plants with apparent one-sided wilting were sampled from 14 fields in Virginia and North Carolina. Forty-seven obtained isolates showed colony morphology typical of Fusarium spp. on potato dextrose agar (PDA) medium. Fifteen isolates representing colony variations on PDA were further selected for microscopic examination and molecular identification using the translation elongation factor 1-alpha (tef1-α) gene. Phylogenetic analysis of tef1-α nucleotide sequences identified nine Fusarium species representing five distinct species complexes: Fusarium incarnatum-equiseti (six isolates), F. oxysporum (three isolates), F. solani (three isolates), F. fujikuroi (two isolates), and F. tricinctum (one isolate). The pathogenicity of each isolate was assessed on flue-cured tobacco cultivar 'K326' seedlings at the six-leaf stage in greenhouse inoculation experiments, with or without wounding. All 15 Fusarium isolates were pathogenic, causing root rot, leaf necrosis, and reduced growth parameters compared to the control treatment. The diversity of Fusarium species associated with Fusarium wilt of tobacco and the variability in aggressiveness among Fusarium species within the same species complex or belonging to the different Fusarium species complexes provides a theoretical basis for future research on the epidemiology of Fusarium disease in tobacco and integrated disease management strategies to reduce disease development and severity.