Stemphylium botryosum Pathogenic on Spinach Seed Crops in Washington

In September 2000, symptoms typical of leaf spot caused by Cladosporium variabile were observed on a spinach (Spinacea oleracea L.) seed crop in western Washington. Dry, bleached spots (1 to 20 mm) were most abundant on lower leaves. Two isolates of C. variabile and three isolates of Stemphylium were recovered by plating surface-sterilized (0.1% sodium hypochlorite) sections of symptomatic leaf tissue onto water agar and acidified potato dextrose agar (PDA). Transfers of each isolate were made to PDA, and cultures were kept at 24 ± 2°C on a lab bench (natural day/night cycle) for 10 to 14 days. Spore suspensions (10⁵/ml) of the isolates of C. variabile were prepared in a 0.01% solution of Tween 80. Isolates of Stemphylium produced few spores, so mycelial suspensions (10⁵ fragments/ml) were prepared. Five 8-week-old seedlings of each of the cultivars Winter Bloomsdale and Ozarka II were inoculated per fungal isolate by atomizing the inoculum onto each seedling until all leaves were covered with a thin film of droplets (4 to 5 ml of inoculum per seedling). Plants were enclosed in plastic bags on a greenhouse bench (24 ± 3°C) for 72 h (8 h/16 h day/night). Symptoms developed within 80 h of inoculation for both isolates of C. variabile and two isolates of Stemphylium. Small (1 to 2 mm) sunken spots turned white 24 to 48 h later and became dry and bleached. Lesions caused by isolates of Stemphylium enlarged and coalesced more rapidly than lesions caused by C. variabile, and were more irregular and usually not delimited by the thin brown margin typical of lesions caused by C. variabile. The differences in symptoms were consistent on both spinach cultivars. Symptoms were not observed on non-inoculated control plants nor on plants inoculated with the third isolate of Stemphylium. C. variabile and Stemphylium were reisolated from symptomatic leaf tissue. Colony morphology, conidiophores, and conidia of the pathogenic Stemphylium isolates were similar to those of pathogenic isolates of Stemphylium botryosum obtained from spinach plants in California (2). This is the first report of S. botryosum as a foliar pathogen of spinach seed crops in Washington. Although Correll et al. (1) noted Stemphylium to be damaging on mature spinach plants grown for seed production, S. botryosum may not have been diagnosed previously on spinach seed crops in Washington because of the similarity of symptoms caused by S. botryosum and C. variabile. S. botryosum was recently reported as a foliar pathogen of spinach in California (2). References: (1) J. C. Correll et al. Plant Dis. 78:653, 1994. (2) S. T. Koike et al. Plant Dis. 85:126, 2001.