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Verticillium dahliae is a pathogen of spinach (Spinacia oleracea) during spinach seed crop production but not in vegetative leafy spinach crops, because plants remain asymptomatic until bolting has been initiated (conversion from vegetative to reproductive growth). The objectives of this research were to evaluate a set of USDA spinach germplasm accessions for resistance to Verticillium wilt and to determine the range in incidence of natural seed infection/infestation in a subset of the USDA spinach accessions screened for resistance. A total of 120 Spinacia spp. accessions from the USDA North Central Regional Plant Introduction Station spinach germplasm collection and 10 commercial S. oleracea hybrids were screened for resistance to V. dahliae in Trials 1, 2, and 3 in 2006, 2007, and 2008, respectively, in greenhouse evaluations. Each accession was grown in either V. dahliae-infested or non-infested potting mix and rated weekly on a seven-step scale from 1 week before bolting to 4 weeks after bolting. A wide range of disease severity ratings was observed among the accessions evaluated. Most of the accessions were highly susceptible. There was no evidence of qualitative resistance but some showed greater levels of quantitative resistance than others. Plants in soil infested with V. dahliae senesced faster and had significantly reduced biomass compared with plants in non-infested soil of the same accession. In addition, in Trial 2 (2007), 34% (20 of 59) of the seed samples assayed of the accessions were infested or infected with V. dahliae; and in Trial 3, (2008) 16% (21 of 130) of the seed samples of the USDA accessions evaluated were infested or infected with V. dahliae, V. tricorpus, or Gibellulopsis nigrescens (formerly known as V. nigrescens). These results are valuable for characterizing potential genetic variability within spinach germplasm for resistance to V. dahliae.

Iglesias-Garcia, A. M., Villarroel-Zeballos, M. I., Feng, C, du Toit, L. J., and Correll, J. C. 2013. Pathogenicity, virulence, and vegetative compatibility grouping of Verticillium isolates from spinach seed. Plant Dis. 97:1457-1469.In 2005, Verticillium dahliae was first reported to be pathogenic to spinach seed crops in the Pacific Northwest, with symptoms only developing after initiation of the reproductive stage of plant growth, and to be prevalent on commercial spinach seed lots produced in Denmark, The Netherlands, and the United States. In this study, the genetic diversity, pathogenicity, and virulence were examined for a collection of isolates of Verticillium spp. from spinach as well as other hosts (alfalfa, cotton, lettuce, mint, peppermint, potato, radish, and tomato) from various countries and from different vegetative compatibility groups (VCGs). Of a total of 210 isolates of V. dahliae obtained from spinach seed produced in Denmark, the Netherlands, New Zealand, or the United States, 128 were assigned to VCG 4B (89% of 91 U.S. isolates, 86% of 42 isolates from the Netherlands, 19% of 43 Denmark isolates, and 8% of 13 New Zealand isolates), 65 to VCG 2B (92% of the New Zealand isolates, 79% ofthe Denmark isolates, 14% ofthe Netherlands isolates, and 9% of the U.S. isolates), and 3 to VCG 2A (2% of each of the Denmark and U.S. isolates, and 0% of the Netherlands and New Zealand isolates); 14 isolates could not be assigned to a VCG. Although little variation in the sequence of the internal transcribed spacer (ITS) region of ribosomal DNA was observed among isolates within each Verticillium sp., the ITS region readily differentiated isolates of the species V. dahliae, V. tricorpus, and Gibellulopsis nigrescens (formerly V. nigrescens) obtained from spinach seed. Greenhouse pathogenicity assays on spinach, cotton, lettuce, and tomato plants using isolates of V. dahliae (n = 29 to 34 isolates), V. tricorpus {n = 3), G. nigrescens (n = 2), and V albo-atrum (n = 1) originally obtained from these hosts as well as from alfalfa, mint, peppermint, potato, and radish, revealed a wide range in virulence among the isolates. Isolates of V. tricorpus and G. nigrescens recovered from spinach seed and an isolate of V. albo-atrum from alfalfa were not pathogenic on spinach. In addition, isolates of V. dahliae from mint and peppermint were not pathogenic or only weakly virulent on the hosts evaluated. Although there was a wide range in virulence among the isolates of V. dahliae tested, all of the V. dahliae isolates caused Verticillium wilt symptoms on spinach, lettuce, tomato, and cotton. None of the isolates of V. dahliae showed host specificity. These results indicate that Verticillium and related species associated with spinach seed display substantial variability in virulence and pathogenicity to spinach and other plants but the V. dahliae isolates were restricted to three VCGs.

Downy mildew of spinach, caused by the obligate pathogen Peronospora effusa, remains the most important constraint in the major spinach production areas in the United States. This disease can potentially be initiated by asexual sporangiospores via “green bridges”, sexually derived oospores from seed or soil, or dormant mycelium. However, the relative importance of the various types of primary inoculum is not well known. The ability of P. effusa sporangiospores to withstand abiotic stress, such as desiccation, and remain viable during short- and long-distance dispersal and the ability of oospores to germinate and infect seedlings remain unclear. Thus, the primary objectives of this research were to evaluate the impact of desiccation on sporangiospore survival and infection efficiency and examine occurrence, production, and germination of oospores. Results indicate that desiccation significantly reduces sporangiospore viability as well as infection potential. Leaf wetness duration of 4 h was needed for disease establishment by spinach downy mildew sporangiospores. Oospores were observed in leaves of numerous commercial spinach cultivars grown in California in 2018 and Arizona in 2019. Frequency of occurrence varied between the two states-years. The presence of opposite mating types in spinach production areas in the United States was demonstrated by pairing isolates in controlled crosses and producing oospores on detached leaves as well as intact plants. Information from the study of variables that affect sporangiospore viability and oospore production will help in improving our understanding of the epidemiology of this important pathogen, which has implications for management of spinach downy mildew.

Downy mildew, caused by the obligate oomycete pathogen Peronospora effusa, is the most economically important disease of spinach. In the past 30 years, 14 new races and 13 strains with novel virulence have been identified. However, the mechanism(s) driving the rapid evolution of virulence remains unknown. To understand reproductive strategies potentially driving the emergence of new races in P. effusa, 67 composite isolates (a collection of symptomatic leaves from a single cultivar grown in a defined area) of P. effusa obtained from 13 states between 2010 and 2018 were used to analyze the population genetic diversity hierarchically. Genotypes at 33 SNP loci of 719 lesions from these 67 isolates were determined by targeted sequencing. Diversity was then evaluated among individual lesions within the composite isolates, between isolates, host cultivars, geographic locations, and years of isolates collected. A total of 380 genotypes were identified from 719 individual lesions. Of the 380 genotypes, 350 (92%) were unique while the most common genotype was identified in 110 lesions of 16 isolates collected from 13 cultivars from CA and AZ in 2016. Variation within composite isolates ranged from none (a single genotype among lesions from a composite isolate) to 38 unique genotypes recovered from 39 lesions of a composite isolate. An index of association analysis suggested asexual (clonal) and sexual reproduction play important roles in population structure. Based on discriminant analysis of principal components, four distinct subpopulations were identified. Host cultivar, origin, and time of collection had an effect on population differentiation, and genotypes specific to a certain location or collection period were identified. Some subpopulations were unique to certain areas, and were only detected after 2014-2016. The co-existence of sexual and asexual reproduction strategies may partially explain the rapid emergence and spread of new races and novel strains of P. effusa.