First Report of Fusarium solani on Utah Sweetvetch in the United States.

Utah sweetvetch (Hedysarum boreale Nutt.) is a native American perennial nitrogen fixing legume used mainly in rangeland reclamation, soil rejuvenation, and erosion control. In June 2011, a field of Utah sweetvetch grown for seeds in central Oregon had approximately 15% of the plants exhibiting chlorosis, defoliation, stunting, wilting, and/or death. Dissection of the crown of symptomatic plants revealed discolored pinkish brown vascular tissue. Symptomatic tissues from six random plants were surface sterilized, placed on acidified potato dextrose agar (PDA) medium, and cultured for 7 days at room temperature, which allowed six fungal isolates (SS1 through SS6) to be collected. On PDA, all six isolates had rapid, creamy white colored growth. Based on observations of 1-week-old isolates, microconidia were oval to kidney shaped, single celled, 8 to 10 × 2.5 to 4 μm, and formed at the tips of long unbranched monophialides. Macroconidia were three to four septate, cylindrical to slightly curved, with characteristic foot shaped basal cell and blunt apical cell, 37 to 49 × 4.4 to 5.3 μm. Chlaymydospores observed were 8.5 to 11 × 7.6 to 9 μm. Based on fungal references (1,2,3), the isolates were identified as Fusarium solani (Mart.) Sacc. Identification of the isolates at the molecular level was determined by amplification of the internal transcribed spacer (ITS) region using PCR and amplicon sequencing. Botrytis cinerea and F. graminearum cultures were used as controls for the extraction, amplification, and sequencing steps. Genomic DNA was extracted from mycelia using protocols of the MOBIO Ultraclean Soil DNA Isolation Kit (MO-BIO Laboratories Inc, Carlsbad, CA, USA). PCR was performed using ITS1/ITS4 primers and resulted in 563- to 573-bp amplicons, which were sequenced. Analysis of the ITS sequences (GenBank Accession Nos. JX524018 to JX524023) for the six fungal isolates using BLASTn revealed a 99% sequence identity with F. solani strains (AB470903, AB513851, AJ608989, EF152426, EU029589, and HM214456). Pathogenicity was confirmed on Utah sweetvetch plants in the greenhouse. Seeds of Utah sweetvetch were first plated on acidified PDA for germination; healthy seedlings were then selected and transplanted into pots with sterilized soil after 2 weeks of growth. The plants were kept in a greenhouse at Central Oregon Agricultural Research Center, Madras, Oregon. Ten 40-day-old healthy vetch plants were inoculated by drenching with a mixed conidial suspension (10⁷ conidia/ml) of the six F. solani isolates. Ten plants drenched with sterile distilled water were included as controls. Symptoms of chlorosis and stunting similar to those in the commercial field were observed within 30 days of inoculation on 8 of 10 inoculated plants, while control plants were symptomless. Fungal isolates identical to F. solani were reisolated from the symptomatic plants. To our knowledge, this is the first report of F. solani on Utah sweetvetch plants. References: (1) C. Booth. The Genus Fusarium. CMI, Kew, Surrey, UK, 1971. (2) P. E. Nelson et al. Fusarium species: An illustrated manual for identification. The Pennsylvania State University Press, USA, 1983. (3) H. I. Nirenberg. A simplified method for identifying Fusarium spp. occurring on wheat. Can. J. Bot. 59:1599, 1980.