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.

Carrot (Daucus carota subsp. sativus) seed crops in Oregon and Washington were surveyed in 2001-02 and 2002-03 for development of Xanthomonas campestris pv. carotae, causal agent of bacterial blight. For each state and season, 20 plants were sampled from each of 7 to 12 direct-seeded crops twice in the fall or winter and three times from spring to summer; and from each of 2 to 4 steckling (root-to-seed) crops three times from spring to summer. X. campestris pv. carotae was detected in 1 of 15 and 6 of 32 stock seed lots planted in the fall in Oregon and Washington, respectively, and in 2 of 6 steckling shipments planted in each state in the spring. The pathogen was detected at 102 to 108 CFU/g foliage in 1 of 22 and 10 of 14 direct-seeded crops within 2 months of planting in 2001 and 2002, respectively. The prevalence of X. campestris pv. carotae then increased through the season in most seed crops, although bacterial blight symptoms were not observed until April in Oregon and July in Washington in both seasons. In August 2002 and 2003, X. campestris pv. carotae was detected in all 10 and 13 crops surveyed in Oregon, respectively; and in 11 of 12 and 7 of 10 crops in Washington, respectively. The pathogen was typically less prevalent in steckling versus direct-seeded crops. X. campestris pv. carotae was detected in 20 of 22 and 19 of 23 harvested seed lots in Oregon and Washington, respectively, at populations ranging from 1.3 × 101 to 1.4 × 108 CFU/g seed. Airborne X. campestris pv. carotae, detected ≤1,600 m downwind of crops being threshed in Oregon in September of 2003 and 2004, may provide a source of inoculum for newly planted seed crops between overlapping biennial seasons for carrot seed production. Despite the prevalence of this pathogen in the Pacific Northwest, carrot seed lots free of X. campestris pv. carotae were detected, demonstrating the ability to produce clean seed in this region by adhering to recommended practices for management of bacterial blight.

Molecular assays to detect and quantify DNA from viable cells of the seedborne pathogen Xanthomonas hortorum pv. carotae in carrot seed were developed and evaluated for use on nontreated and hot-water-treated seed lots. Both a TaqMan real-time polymerase chain reaction (PCR) assay and a loop-mediated isothermal amplification (LAMP) dilution endpoint assay detected and quantified DNA from viable pathogen cells after treatment of carrot seed washes with the live-dead discriminating dye propidium monoazide (PMA). The detection limits of the assays were approximately 101 CFU for pure cultures of X. hortorum pv. carotae, and 102 to 103 CFU/g seed from naturally infested carrot seed lots. X. hortorum pv. carotae in and on carrot seed was killed by soaking the seed in hot water (52°C for 25 min), and a subsequent PMA treatment of these hot-water-treated seed washes suppressed detection of the pathogen with both the real-time PCR and LAMP assays. For 36 commercial seed lots treated with PMA but not hot water, regression of colony counts of X. hortorum pv. carotae measured by dilution plating on a semiselective agar medium versus estimates of pathogen CFU determined by the molecular assays resulted in significant (P ≤ 0.05) linear relationships (R2 = 0.68 for the real-time PCR assay and 0.79 for the LAMP assay). The molecular assays provided quantitative estimates of X. hortorum pv. carotae infestations in carrot seed lots in <24 h, which is a significant improvement over the 7 to 14 days required to obtain results from the traditional dilution-plating assay.

The first detection in the United States of isolates of Phytophthora infestans having metalaxyl insensitivity and complex pathotypes occurred in western Washington during the early 1990s. To determine the genetic structure of the current population in western Washington, a total of 115 isolates of P. infestans were obtained during 1996 from infected tubers or foliage of potato, tomato, nightshade, and bittersweet throughout the region. An additional 45 isolates were collected from a single field. Based on mating type, metalaxyl-insensitivity, and molecular markers (allozymes of glucose-6-phosphate isomerase, peptidase, and RG57 DNA fingerprint), all of the isolates were A1 mating type and had the US-11 multilocus genotype. Analyses of an additional 120 isolates collected during 1997 from potato, tomato, and nightshade were performed. As in 1996, US-11 was the predominant genotype detected on all three hosts. However, three additional A2 mating type genotypes were also detected: US-7, US-8, and US-14. These three genotypes represent the first A2 mating type isolates detected in western Washington. Most of a subset of 60 isolates infected 4 to 7 of the 10 potato differentials tested. This included 90% of the isolates collected in 1996 (all US-11), plus 72% of the US-11 and 100% of the US-8 and US-14 isolates collected during 1997. Virulence phenotypes in this region are complex even without the selection pressure of R-genes in the local commercial cultivars. The apparent increase in genetic variation observed in populations of P. infestans in western Washington from 1996 to 1997 most likely occurred by migration rather than by sexual recombination.