There are no previous reports of Verticillium wilt in fresh and processing spinach (Spinacia oleracea) crops in the United States. In 2002, a hybrid spinach seed crop in the Pacific Northwest developed late-season wilt symptoms. Assays of the harvested seed and stock seed of the male and female parents revealed 59.5, 44.0, and 1.5%, respectively, were infected with Verticillium dahliae. Assays of 13 stock or commercial seed lots grown in 2002 and 62 commercial lots harvested in 2003 in Denmark, Holland, New Zealand, and the United States revealed the prevalence of Verticillium spp. in commercial spinach seed. Sixty-eight lots (89%) were infected with Verticillium spp. at incidences ranging from 0.3 to 84.8%. Five spinach seed isolates of V. dahliae were pathogenic on each of three spinach cultivars by root-dip inoculation. V. dahliae was detected on 26.4% of the seed from 7 of 11 inoculated plants but on none of the seed from 6 control plants, demonstrating systemic movement of V. dahliae. Seed-to-seed transmission was also demonstrated by planting naturally infected seed lots. This is the first report of Verticillium wilt of spinach in the primary region of spinach seed production in the United States.
Cladosporium variabile, Stemphylium botryosum, and Verticillium dahliae are seedborne and seed-transmitted pathogens of spinach. Spinach seed treatments in 1.2% NaOCl for 10 to 60 min, or hot water (40, 45, 50, 55, and 60°C) for 10 to 40 min, were evaluated for eradication of these fungi from seed. C. variabile and V. dahliae were largely eradicated by chlorine treatment for ≥ 10 min. Although chlorine treatment reduced the incidence of S. botryosum, this fungus was not eradicated after 60 min in chlorine. Seed germination was not affected adversely by chlorine treatment, even after 60 min. In contrast, germination was reduced significantly by hot water treatment at 50°C for ≥30 min or 55 or 60°C for ≥10 min. C. variabile was eradicated from seed treated in 40°C water for 10 min. V. dahliaewas eradicated from seed treated at 55°C for ≥30 min or 60 °C for ≥10 min. S. botryosum was eradicated from a lightly infected seed lot (5% incidence) by hot water treatment at 55 or 60 °C for ≥10 min, but could not be eradicated from two heavily infected lots (>65% incidence), even at 60°C for 40 min. Using precisely controlled parameters, chlorine or hot water seed treatments can be used to eradicate C. variabile and reduce the incidence of S. botryosum and V. dahliae in spinach seed without damaging germination.
Assays of 77 spinach (Spinacia oleracea) seed lots produced in the United States, Denmark, the Netherlands, or New Zealand in 2000 to 2003 showed that Stemphylium botryosum, causal agent of Stemphylium leaf spot, was present in every lot, at a mean incidence of 29.1% per lot. Either Cladosporium variabile, causal agent of Cladosporium leaf spot, or the morphologically similar species C. macrocarpum, was present in 37 of the 77 lots, at a mean incidence of 1.8% per lot. Some seed isolates of S. botryosum and C. variabile proved pathogenic on spinach. Nonpathogenic isolates resembling C. variabile were identified as C. macrocarpum by the absence of torulose aerial hyphae. Pathogenic isolates of S. botryosum were also detected in each of 12 seed lots stored for up to 11 years at 4.4°C and 60% relative humidity. C. variabile or C. macrocarpum was detected in only 2 of the 11 lots, which had been stored for 3 and 8 years. Component seed assays demonstrated that S. botryosum and C. variabile (or C. macrocarpum) were internal and external in spinach seed. S. botryosum was detected in 5 to 76% of the embryos of five seed lots, but the two Cladosporium species were detected in only 0 to 1% of the embryos of these lots. This suggests greater potential difficulty at eradicating S. botryosum than C. variabile from infected spinach seed using seed treatments.
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