First Report of a Novel Fusarium Species Causing Yellowing Decline of Sugar Beet in Minnesota

In the United States, yellows disease of sugar beet (Beta vulgaris), which causes wilt, early death, and yield reduction, is caused primarily by Fusarium oxysporum f. sp. betae (3,4), but F. graminearum (2) has also been implicated. During the past 3 years, a similar disease causing yellowing and severe decline appeared in some sugar beet fields of central and southwest Minnesota planted with cultivars resistant to yellows. The disease has become a concern to the local sugar beet industry, which produces 56% of sugar beets in the United States. From 2005 to 2007, isolations were made from sugar beets collected in commercial fields and from a Fusarium screening nursery showing symptoms of yellowing, interveinal chlorosis, scorching, stunting, vascular discoloration of the taproot, and early death of plants. Of 96 Fusarium isolates recovered and used in root-dip inoculation trials in the greenhouse, 58 were pathogenic to sugar beets. On the basis of morphology, 12 were identified as F. oxysporum, 6 as F. graminearum, and 40 as a novel Fusarium species. The remaining 38 isolates were nonpathogenic. All three pathogenic Fusarium species were isolated from taproots, but only the novel Fusarium was isolated from petioles. In culture, the novel Fusarium exhibited a bright orange color on the underside of potato dextrose agar medium and produced micro- and macroconidia sparsely. Hyphal tip isolates of all novel Fusarium isolates were pathogenic, causing typical yellowing symptoms and plant death to the Fusarium yellows susceptible sugar beet cv. VDH46177 in replicated greenhouse trials. Isolates were successfully reisolated from the symptomatic plants, fulfilling Koch's postulates. Restriction fragment length polymorphism (RFLP) endonuclease digestion patterns (Alu1, Fnu4HI, HaeIII, and HhaI) of the internal transcribed spacer (ITS) region of 40 pathogenic novel isolates showed a distinct pattern compared with known Fusarium species. Thin layer chromatography analysis of 13 novel isolates detected the type A trichothecenes neosolaniol and 4,15-diacetoxyscirpenol. Partial sequences of the translation elongation factor 1-α (TEF) from 12 single-spored novel Fusarium isolates were generated. BLAST analysis of the TEF sequence against the FUSARIUM-ID (1) and GenBank databases did not match any known Fusarium species. On the basis of pathogenicity, morphology, RFLP patterns, mycotoxin production, and TEF sequence analysis it appears that this is a new species of Fusarium, but additional multilocus phylogenetic analyses are warranted. The natural occurrence of this novel Fusarium pathogen in sugar beet may have implications in breeding for resistance to Fusarium yellows, since yellow decline has been observed in purportedly Fusarium-tolerant cultivars in the Minnesota and North Dakota production regions.