The Alternative Host with the Most: Understanding the Ecology of Xanthomonas hortorum pv. carotae on Noncarrot Crops in Central Oregon.

Bacterial blight of carrot, caused by Xanthomonas hortorum pv. carotae (Xhc), is an economically important disease in carrot (Daucus carota subsp. sativus) seed production. The objectives of this study were to determine if Xhc was present on noncarrot crops grown in central Oregon and, if detected, evaluate its ability to colonize alternative hosts. Surveys of three carrot seed fields and adjacent fields of rye (Secale cereale), alfalfa (Medicago sativa), parsley root (Petroselinum crispum var. tuberosum), and Kentucky bluegrass (Poa pratensis) demonstrated that Xhc was present on noncarrot crops. Greenhouse experiments were conducted to determine the ability of Xhc to colonize crops cultivated in the region. Carrot, alfalfa, curly parsley (Petroselinum crispum), Kentucky bluegrass, mint (Mentha × piperita), parsley root, roughstalk bluegrass (Poa trivialis), and wheat (Triticum aestivum) plants were spray-inoculated with Xhc and destructively sampled at 1, 7, 14, and 28 or 25 days post-inoculation. Xhc populations were quantified using viability quantitative PCR and dilution plating. A significant (P ≤ 0.03) effect of crop was observed at 1, 14, and 28 or 25 days in both experiments. While carrot hosted the most Xhc at the final timepoint, other crops supported epiphytic Xhc populations including wheat and both bluegrasses. Mint, parsley root, and alfalfa hosted the least Xhc. Bacterial blight symptoms were observed on carrots but not on noncarrot crops. This suggests that crops grown in central Oregon have the potential to be asymptomatically colonized by Xhc and may serve as reservoirs of the pathogen in carrot seed production systems.

First report of Spiroplasma citri associated with disease symptoms in field-grown hemp (Cannabis sativa L.) in the Pacific Northwest.

Hemp (Cannabis sativa L.) is grown for cannabinoid oil production in Oregon. During the 2021 and 2022 growing seasons, plants with leaf curling, puckering, chlorotic mosaic, fasciation, and vein clearing were observed in disease surveys of Oregon hemp fields (Fig. 1). Symptoms were present on 1-10% of 2-4-month-old plants in fields located in Benton, Clackamas, Deschutes, Jackson, Josephine, Lane, Linn, Marion, Morrow, and Polk counties between July and September. Leaf and stem samples were collected from 38 symptomatic plants. Symptoms resembled those caused by beet leafhopper-vectored (BLH, Circulifer tenellus Baker) curtovirus or phytoplasma infection. Therefore, total nucleic acids were extracted (Dellaporta et al. 1983) from leaf material, and PCR conducted on all 38 samples to detect curtovirus coat proteins (BCTV-1/2 primers; Rondon et al. 2016) and 16S rRNA of phytoplasmas (nested primers P1/P7 followed by FU/RU; Lorenz et al. 1995). No curtoviruses or phytoplasmas were detected in any samples. Spiroplasma citri is also transmitted by BLH, so PCR was conducted with primers targeting the putative P89 adhesin gene and spiralin gene of S. citri (Yokomi et al. 2008). For all samples, PCR using P89F/R primers resulted in a 707 bp amplicon, and a 675 bp amplicon with Spiralin-f/r primers. PCR products were purified with ExoSapIT (Applied Biosystems, Waltham, MA), and two representative samples (ScH1; ScH2) were Sanger sequenced (EuroFins, Lancaster, PA) in the forward and reverse direction. Pairwise aligned P89 sequences were found to be 99 to 100% (ScH1: 633/637 bp; ScH2: 654/654 bp) identical to S. citri accession KT377386. Aligned/span>spiralin sequences were found to be 99 to 100% (ScH1: 661/664 bp; ScH2: 647/647 bp) identical to S. citri accession CP013197 in the NCBI GenBank Database. All sequences were deposited into GenBank (accession no. OQ969983, OQ992766, OQ969984, OQ969986). Frozen leaf material from one sample was used to culture S. citri according to Lee and Davis (1984). Leaf tissue was surface sterilized for 60 s in 1% NaOCl in 70% ethanol, cut into pieces in LD8 broth, and incubated for 25 min at 25˚C. The solution was passed through a 0.45um filter and incubated for 14 days at 30°C with constant shaking (150 rpm). Then, cultures were centrifuged for 2 min at 12,000 rpm to pelletize, resuspended in 100µL sterile distilled water and passed through a 0.2um filter. From this culture, 2 µL of broth was used as template for both the P89F/R and Spiralin-f/r primer sets. Amplicons were purified and sequenced as above (accession nos. OQ969982, OQ969985). Leaf tissue from both representative samples (ScH1 and ScH2) were positive for S. citri using double antibody sandwich ELISA (Agdia, Inc., Elkhart, IN) following manufacturer instructions. These results from sequencing, culturing, and ELISA testing indicate the hemp samples were infected with S. citri. To our knowledge this is the first report of disease symptoms in hemp associated with S. citri in Oregon and the Pacific Northwest. Infection by S. citri can limit yield by reducing photosynthetic capability of the plant and distortion of plant growth. Other pathogens like curtoviruses and phytoplasmas have also been detected in hemp in Oregon and the U.S. (Hu 2021; Rivedal et al. 2022), and the addition of S. citri associated with disease symptoms indicates a need for BLH management research. This discovery has implications for arid regions with other S. citri hosts including cruciferous plants, carrots, and tree fruit crops, all of which were grown near the sampled hemp fields in this study.

Molecular and Alkaloid Characterization of Claviceps purpurea Sensu Lato From Grass Seed Production Areas of the U.S. Pacific Northwest.

Ergot, caused by Claviceps purpurea sensu lato, is an economically important seed replacement disease of Kentucky bluegrass (Poa pratensis) and perennial ryegrass (Lolium perenne) seed crops. C. purpurea sensu stricto is considered the primary Claviceps species responsible, but genetic diversity and cryptic species within C. purpurea sensu lato have previously been reported. Fifty-six C. purpurea sensu lato isolates collected from P. pratensis (n = 21) and L. perenne (n = 35) in Oregon and Washington between 2010 and 2014 were characterized via random amplified polymorphic DNA (RAPD), partial internal transcribed spacer (ITS), ß-tubulin and elongation factor-1α (EF-1α) sequences, conidial size, and ergot alkaloid chemotype. Based on RAPD analysis, seven isolates from P. pratensis and 33 isolates from L. perenne collected in Oregon corresponded to C. purpurea sensu stricto, and 13 isolates collected from P. pratensis in Washington and Oregon were identified as C. humidiphila. Partial ITS, ß-tubulin, and EF-1α sequences identified 10 isolates from P. pratensis as C. humidiphila, and seven isolates from P. pratensis and 33 isolates from L. perenne were identified as C. purpurea sensu stricto. Several isolates generated ambiguous RAPD bands or sequences that prevented identification. Ergot alkaloid chemotype profiling found that ergocornine and its epimer were predominant in sclerotia from P. pratensis, whereas ergotamine and its epimer were most abundant in sclerotia from L. perenne. This study confirms the presence of the C. purpurea sensu lato species complex in the U.S. Pacific Northwest and suggests that more research is needed to characterize and mitigate Claviceps spp. infection of grass seed crops in North America.

Population Genetic Structure of Claviceps purpurea in Cool-Season Grass Seed Crops of Oregon.

Ergot, caused by Claviceps purpurea, is a primary disease concern in irrigated cool-season grass seed production systems of Oregon. In order to better understand the genetic diversity, population structure, and the epidemiology of C. purpurea in grasses grown for seed, 226 isolates were obtained using a hierarchical sampling strategy from two fields each of Kentucky bluegrass (n = 102) and perennial ryegrass (n = 124) and characterized using 12 microsatellite markers. A total of 194 unique multilocus genotypes (MLGs) were identified in this study. There were moderate levels of genotypic diversity (H = 3.43 to 4.23) and gene diversity (Hexp = 0.45 to 0.57) within fields. After clone correction, analysis of molecular variance revealed that 66% of the genetic variation occurred between the two C. purpurea isolates collected from the same seed head of individual plants, indicating that many of the seed heads bearing multiple sclerotia were infected by ascospores rather than conidia. However, the majority of the clonal isolates obtained in this study were collected from the same seed head (i.e., the two isolates were identical MLGs), indicating a role of conidia (honeydew) in secondary infections within seed heads. Genetic differentiation was observed between populations from different hosts (22%) but was confounded by geography. The standardized index of association ranged from 0.007 to 0.122 among the four populations, suggesting potential outcrossing and differences in the relative contribution of ascospores and conidia to ergot among the fields. The results from this study provide insights into the epidemiology of ergot in cool-season grass seed crops of Oregon.

Distribution of Xanthomonas hortorum pv. carotae Populations in Naturally Infested Carrot Seed Lots.

Bacterial blight of carrot (Daucus carota subsp. sativus), caused by the plant-pathogenic bacterium Xanthomonas hortorum pv. carotae, is a common seedborne disease of carrot wherever the crop is grown. Carrot seed lots were evaluated to determine the variability and distribution of populations of X. hortorum pv. carotae among individual carrot seeds. Twenty-four carrot seed lots harvested between 2014 and 2016 were subjected to a bulk seed wash dilution-plate assay to obtain mean X. hortorum pv. carotae levels. Mean infestation levels resulting from the bulk seed wash assays among the 24 seed lots ranged from 1.2 × 107 and 9.6 × 108 CFU/g seed and averaged 3.6 × 108 CFU/g seed. Individual seeds from the same 24 lots were also tested with a scaled-down wash assay of individual seeds. Among the 1,380 seeds that were individually assayed, 475 X. hortorum pv. carotae-positive seeds were detected (34.4%). Rates of X. hortorum pv. carotae detection on individual seed in seed lots ranged from 0% (not detected) to 97.9%, and the mean and median X. hortorum pv. carotae population on an individual seed was 8.3 × 104 and 6.3 × 101 CFU/seed, respectively. Among individual seeds, X. hortorum pv. carotae populations ranged from 2 (the limit of detection of the assay) to 3.6 × 107 CFU/seed. CFU data for 23 of the 24 seed lots were nonnormal and the Log-Logistic (3P) distribution best described populations of X. hortorum pv. carotae recovered from individual carrot seeds. The influence and impact of nonnormal distributions of X. hortorum pv. carotae in commercial carrot seed lots on seed health tests, seedborne transmission, and bacterial blight epidemiology requires further study.

Genetic Diversity of Verticillium dahliae Isolates From Mint Detected with Genotyping by Sequencing.

Verticillium wilt is the most important disease threatening the commercial production of mint grown for essential oil. An important long-term goal for mint breeders is the production of cultivars with resistance to Verticillium wilt. Before that can be accomplished, a better understanding of the genetic variation within and among populations of Verticillium dahliae is needed. We characterized the extent of phenotypic and genetic diversity present in contemporary and archival populations of V. dahliae from mint fields in Oregon and other production regions of the United States using genotyping by sequencing, PCR assays for mating type and pathogenic race, vegetative compatibility group (VCG) tests, and aggressiveness assays. We report that the population in the Pacific Northwest can be described as one common genetic group and four relatively rare genetic groups. Eighty-three percent of the isolates belonged to VCG2B, and all isolates possessed the MAT1-2 idiomorph and were characterized as pathogenic race 2. These results indicate low levels of genetic diversity and a negligible risk of sexual recombination in populations of this host-adapted pathogen population. Knowledge of the genetic structure of V. dahliae in the Pacific Northwest will inform breeders about the diversity of pathogenicity factors that may need to be considered in their breeding programs.

Detection and Quantification of Airborne Claviceps purpurea sensu lato Ascospores from Hirst-Type Spore Traps using Real-Time Quantitative PCR.

The U.S. Pacific Northwest states of Oregon and Washington are major producers of cool-season grass seed. Ergot, caused by fungi in the Claviceps purpurea sensu lato group, is an important seed replacement disease of grass worldwide. Microscopic methods that are currently used to quantify airborne Claviceps ascospores captured by spore traps are not currently rapid enough to allow for detecting and reporting of spore numbers in a timely manner, hindering growers from using this information to help manage ergot. We developed a SYBR Green real-time quantitative polymerase chain reaction (qPCR)-based assay for fast and efficient detection and quantification of C. purpurea sensu lato ascospores from Hirst-type spore traps. Species-specificity of the qPCR assay was confirmed against 41 C. purpurea sensu lato isolates collected from six hosts and six other Claviceps spp. Significant relationships were observed between cycle threshold (Ct) values and standard curves of serial dilutions of DNA ranging from 1 pg to 10 ng (R2 = -0.99; P = 0.0002) and DNA extracted from a conidial suspension representing 8 to 80,000 conidia (R2 = -0.99; P = 0.0004). Ct values from qPCR were significantly correlated with results from microscopic examination of spore trap samples from the field (r = -0.68; P < 0.0001) and the procedure was able to detect a single ascospore from spore trap tape samples. The qPCR procedure developed in this study provided a means for quantifying airborne Claviceps ascospores that was highly specific and useful over a wide range of spore densities, and could be performed in a matter of hours instead of days. The qPCR assay developed in this study could be part of an integrated pest management approach to help grass seed growers make risk-based fungicide application decisions for ergot management in grass grown for seed.

Late-summer Disease Symptoms in Western Washington Red Raspberry Fields Associated with Co-Occurrence of Phytophthora rubi, Verticillium dahliae, and Pratylenchus penetrans, but not Raspberry bushy dwarf virus.

Sixty percent of the $109 million processed red raspberry industry of the United States occurs in northern Washington State. In 2012, late-summer symptoms of vascular wilt and root disease were observed in many raspberry plantings. These symptoms were initially attributed to Verticillium dahliae. However, diagnostic tests for the pathogen were often contradictory and other soilborne pathogens (Phytophthora rubi and Pratylenchus penetrans) or Raspberry bushy dwarf virus (RBDV) might also have been involved. Therefore, a survey was conducted in 2013 and 2014 to (i) establish the incidence and soil population levels of V. dahliae in red raspberry production fields, (ii) compare among diagnostic methods and laboratories for detecting and quantifying V. dahliae from raspberry field soil, and (iii) assess which pathogens are associated with late-summer disease symptoms of raspberry. Plant and soil samples were collected from 51 disease sites and 20 healthy sites located in 24 production fields. Samples were analyzed for the presence and quantity of each pathogen using traditional plating and extraction methods (V. dahliae, P. rubi, and P. penetrans), quantitative polymerase chain reaction (qPCR) (V. dahliae and P. rubi), and enzyme-linked immunosorbent assay (RBDV). Results showed that V. dahliae was present in 88% of the production fields and that detection of the pathogen differed by method and by laboratory: qPCR detected V. dahliae in the soil from approximately three times as many sites (51 of 71 total sites) as by plating on NP10 semi-selective medium (15 of 71 total sites). Soil populations of V. dahliae were slightly greater at disease sites, but the pathogen was detected with similar frequency from healthy sites and it was rarely isolated from diseased plants (4%). P. rubi, P. penetrans, and RBDV were also common in production fields (79, 91, and 53% of fields, respectively). Both P. rubi (soil and root samples) and P. penetrans (root populations only), but not RBDV, were more frequently found at disease sites than healthy sites, and the amount of P. rubi detected by qPCR was greater from disease sites than healthy sites. In addition, P. rubi was isolated from 27% of the symptomatic plants located at disease sites. Regardless of detection method, V. dahliae, P. rubi, and P. penetrans, either with or without RBDV, were more likely to co-occur at disease sites (73%) than healthy sites (35%), suggesting that a soilborne disease complex is present in raspberry production fields. Results indicate that P. rubi is the primary pathogen most strongly associated with late-summer symptoms of disease, but root populations of P. penetrans and higher soil populations of V. dahliae may also be of concern. Therefore, disease control methods should focus on all three soilborne pathogens.

Identification of Environmental Factors Related to Claviceps purpurea Ascospore Production in Perennial Ryegrass Seed Fields and Development of Predictive Models.

Claviceps purpurea, the causal agent of ergot of perennial ryegrass seed crops, overwinters as sclerotia in the soil and releases airborne ascospores in the spring that infect flower ovaries and replace seed with sclerotia. Burkard spore traps were used to quantify the dispersal phenology and concentration of ascospores in perennial ryegrass seed fields in the Columbia Basin of Oregon. Weather factors were measured concurrently with spore trapping. Nonparametric regression, box-and-whisker plots, and univariate analysis were used to visualize and identify trends between ascospore concentrations and weather variables. Most ascospores (75.4%) were trapped when minimum soil temperatures were between 16.2 and 20.4°C. Over 67% of the total ascospores trapped were observed when minimum air temperatures were between 6.8 and 12.4°C and 64% of ascospores were trapped when daily mean dew point was between 3.7 and 8.2°C. Environmental favorability index (EFI) models were developed and validated based on their ability to predict ascospore occurrence. The EFI models were able to predict ascospore occurrence with an accuracy of 71.7 to 87.5% depending on the year. The models were up to 79.8% accurate when validated using three years of historical spore trap data not used in the EFI model development. Ninety-four percent of ascospores were trapped when cumulative air degree days, using lower and upper thresholds of 10 and 25°C, respectively, were between 230 and 403. These results suggest that weather parameters can be used to model C. purpurea ascospore occurrence and potentially improve the timing and efficacy of fungicide applications by identifying when plant protection is most needed.

Spatial Patterns of Ergot and Quantification of Sclerotia in Perennial Ryegrass Seed Fields in Eastern Oregon.

Ergot, caused by Claviceps purpurea, is a major disease of perennial ryegrass grown for seed in eastern Oregon. The objective of this research was to quantify and describe the spatial patterns of ergot severity in each of three 50-ha commercial fields of perennial ryegrass grown for seed in 2012 and 2013. In total, 1,433 and 1,405 quadrats were sampled among the three fields in 2012 and 2013, respectively, and the percentage of quadrats with ergot ranged from 59 to 90%. The mean incidence of infected seed heads in each quadrat ranged between 13 and 29%, while mean severity in each quadrat ranged from 0.2 to 0.5 sclerotia per seed head. Significant autocorrelation and clustering were observed in all three fields in both years, as indicated by Moran's I and spatial analysis by distance indices of aggregation. The mean number of ergot sclerotia collected from each field after harvest ranged between 4 and 15 sclerotia m-2 in 2012 and 18 and 119 sclerotia m-2 in 2013. Sclerotia left in perennial fields after harvest are a significant source of inoculum that should be targeted for control. This is the first study to quantify spatial patterns of ergot in perennial ryegrass and provides insights into possible mechanisms that contribute to ergot etiology and epidemiology.

Verticillium dahliae populations from mint and potato are genetically divergent with predominant haplotypes.

In total, 286 Verticillium dahliae isolates from mint, potato, and other hosts and substrates were characterized for mating type, vegetative compatibility group (VCG), and multilocus microsatellite haplotype to determine population genetic structure among populations infecting mint and potato. Populations from mint and potato fit a clonal reproductive model, with all isolates a single mating type (MAT1-2) and multiple occurrences of the same haplotypes. Haplotype H02 represented 88% of mint isolates and was primarily VCG2B, while haplotype H04 represented 70% of potato isolates and was primarily VCG4A. Haplotypes H02 and H04 typically caused severe disease on mint and potato, respectively, in greenhouse assays regardless of host origin. Principal coordinate analysis and analysis of molecular variance indicated that mint and potato populations were significantly genetically diverged (P = 0.02), and identification of private alleles and estimation of migration rates suggested restricted gene flow. Migration was detected between infected potato plants and seed tubers, infested tare soil, and field soils. Genetic differentiation of V. dahliae from mint and potato may be due to the occurrence of a single mating type and differences in VCG. Populations of V. dahliae in potato and mint were characterized by the presence of aggressive, clonally reproducing haplotypes which are widely distributed in commercial mint and potato production.

Incidence and impact of Verticillium dahliae in soil associated with certified potato seed lots.

Verticillium dahliae causes Verticillium wilt of potato and can be found in soil associated with potato seed tubers. The purpose of this research was to quantify V. dahliae in soil associated with certified seed tubers and determine if this potential inoculum source is related to disease development in the field. Approximately 68% of seed lots assayed contained V. dahliae-infested soil on seed tuber surfaces (seed tuber soil). Over 82% of seed lots contained V. dahliae in loose seed lot soil obtained from bags and trucks used to transport seed tubers. Most samples contained ≤50 CFU/g but some contained >500 CFU/g. Most isolates (93%) were vegetative compatibility group 4A. Populations of V. dahliae in stem sap increased with increasing inoculum densities in field soils only when V. dahliae concentrations in seed tuber soil were low. High concentrations of V. dahliae in seed tuber soil resulted in greater stem sap colonization when V. dahliae inoculum densities in field soil were low (P < 0.01) and resulted in greater pathogen inoculum densities in postharvest field soils (P = 0.04). Seed tubers contaminated with V. dahliae-infested soils may introduce the pathogen into fields not previously cropped to potato or recontaminate those which have received preplant management practices. Long-term management of V. dahliae requires reducing propagules in soil associated with seed lots.

Roles of Infected Seed Tubers and Soilborne Inoculum on Verticillium Wilt of 'Russet Burbank' Potato.

Verticillium dahliae, causal agent of Verticillium wilt of potato, persists in soil as microsclerotia and can be found in infected tubers used for seed. The effects of naturally infected tubers and soilborne inoculum on Verticillium wilt symptoms were compared in the greenhouse. Infected and noninfected tubers were grown in infested and noninfested potting soil. Chlorosis and necrosis were measured and converted to area under senescence progress curves (AUSPC). Aboveground stems and progeny tubers were assayed for V. dahliae. Plants from infested soils exhibited significantly greater AUSPC than plants from noninfested soil. Plants grown from infected and noninfected tubers had similar AUSPC and interactions between infected tubers and infested soil were not observed. The pathogen was isolated from the vascular system of 94% of plants grown in infested soils and 8% of plants grown from infected tubers in noninfested soil. Plants grown in infested soil contained microsclerotia on 46% of stems while plants grown from infected tubers in noninfested soils exhibited microsclerotia on <1% of stems. Infected progeny tubers were only recovered from plants grown in infested soil. Seed tuber infection did not contribute to premature senescence or potential inoculum production, indicating that management efforts should focus on reducing soilborne inoculum.

Impact of Seed Lot Infection on the Development of Black Dot and Verticillium Wilt of Potato in Washington.

The fungi Colletotrichum coccodes and Verticillium dahliae are two pathogens of potato which are prevalent in potato field soils and seed tubers. The objective of this study was to investigate the influence of seed lot infection by C. coccodes and V. dahliae on disease severity and yield using infested seed lots and fields with and without potato in their recent cropping histories. Greater incidence of C. coccodes in seed led to more root colonization in 2008 (both field types) but not in 2007 and did not influence tuber quantity or yield. Incidence of infection in seed lots by V. dahliae was not related to canopy necrosis, infection of progeny tubers, or yield. However, seed lots exhibited more V. dahliae aboveground vascular colonization, canopy necrosis, infested plant debris, and infected progeny tubers in 5-year potato rotation fields compared with long potato rotation fields. In general, foliar disease symptoms and yield reductions were not related to levels of either pathogen in seed lots. Symptoms of both diseases were more severe and yields were lower in fields with potato in their recent cropping histories, emphasizing the importance of soilborne inoculum in the development of black dot and Verticillium wilt of potato.

Evaluation of Sulfur-Based Biostimulants for the Germination of Sclerotium cepivorum Sclerotia and Their Interaction with Soil.

White rot is an economically significant disease of Allium crops. The pathogen Sclerotium cepivorum produces long-lived sclerotia that germinate in response to sulfur-containing compounds released from Allium roots. Diallyl disulfide (DADS) was the primary organic sulfur compound detected in the rhizosphere soil of two garlic cultivars, "California Early and Late", growing in greenhouse conditions. DADS, dimethyl trisulfide (DMTS), dimethyl disulfide (DMDS), isopropyl disulfide (IPDS), dipropyl disulfide (DPDS), diethyl disulfide (DEDS), together with garlic oil, garlic juice, garlic powder, raw onion pieces, cabbage pieces, and Chinese cabbage pieces were investigated for their activities toward germinating dormant sclerotia. Results showed that DADS and other volatile sulfur compounds could stimulate sclerotial germination, and a dose-response was observed. In addition, garlic juice, powder, raw onion, and the two cabbages could stimulate sclerotial germination. Furthermore, the laboratory soil incubation experiments demonstrated the strong interaction of organic sulfur compounds with soil.

Verticillium Wilt of Skullcap and Potential for Pathogen Dissemination via Seeds and Stems.

A commercial skullcap (Scutellaria lateriflora, family Lamiaceae) crop with wilted and necrotic plants was examined in Washington State in 2008. Three fungal isolates were obtained and identified as Verticillium dahliae based on morphology and sequencing of the internal transcribed spacer DNA region. All three skullcap isolates caused typical Verticillium wilt symptoms on skullcap and two peppermint cultivars. Inoculations of skullcap with the V. dahliae isolates from skullcap and an isolate from peppermint resulted in severe symptoms and a 21 to 78% reduction in aboveground biomass. Isolates from skullcap caused severe symptoms on the susceptible peppermint 'Black Mitcham' and reduced yield by up to 82%. One skullcap isolate caused severe symptoms on the moderately resistant 'Redefined Murray' in three of four trials and reduced biomass up to 71% compared with noninoculated control plants. The pathogen was recovered from 43 to 69% of skullcap stems from plants inoculated with skullcap or peppermint isolates, and was isolated from 2.5% of seed harvested from skullcap plants inoculated with the peppermint isolate of V. dahliae. This is the first report of V. dahliae infecting skullcap, and the first demonstration of V. dahliae isolates from skullcap and peppermint causing symptoms on both hosts, as well as the seedborne nature of V. dahliae in skullcap.

Development and Suppression of Aerial Stem Rot in Commercial Potato Fields.

The development of aerial stem rot of potato was quantified in relation to soil type, previous crop, and atmospheric temperature in the Columbia Basin. Incidence of bacterial stem rot was assessed for several weeks beginning before row closure in 18 commercial potato fields and once at the estimated peak of disease development in 38 fields over 4 years. Aerial stem rot was first observed between 25 to 36 days following row closure for 78% of the fields assessed multiple times per season. Slopes of disease progress curves initially increased very rapidly. The partial slope estimate for mean daily maximum temperature indicated an increase of disease with an increase of mean daily maximum temperature (P < 0.0001). Estimates of partial slopes for soil type (sandy loam versus silt loam) and previous crop (sweet or field corn versus other) were significant at P < 0.001 and P = 0.046, respectively, suggesting that cultivation of potato in sandy loam-type soils or following sweet or field corn as a previous crop increases the probability of aerial stem rot. Incidence of bacterial stem rot was significantly reduced in the sections of fields treated with famoxadone plus mancozeb and famoxadone plus mancozeb plus copper hydroxide mixtures.

Evaluation of Verticillium Wilt Resistance in Mentha arvensis and M. longifolia Genotypes.

Verticillium wilt, caused by Verticillium dahliae, is a major constraint to mint (Mentha spp.) production in the United States, and the use of resistant cultivars is an important component of Verticillium wilt management. Two Mentha arvensis and four M. longifolia genotypes were evaluated for resistance to Verticillium wilt in the greenhouse using V. dahliae isolates obtained from different hosts and belonging to different vegetative compatibility groups. Isolates of V. dahliae obtained from peppermint (M. × piperita) caused significantly higher disease severity, plant mortality, and yield reduction than isolates obtained from other hosts. Disease severity, plant mortality, and pathogen incidence in aboveground stems were higher and yields lower in peppermint, the susceptible standard, compared with the resistant standard, native spearmint (M. spicata). Root-dip inoculations of M. arvensis and M. longifolia with isolates of V. dahliae obtained from peppermint produced severe symptoms; however, both species displayed the ability to recover from infection and produce asymptomatic growth from rhizomes. Both M. arvensis cultivars exhibited lower mean disease severity ratings following cutback and regrowth and were not significantly different than native spearmint. The restriction of pathogen movement in aboveground tissue and ability to recover from infection may be important components of V. dahliae resistance in perennial mint cropping systems.

Verticillium Wilt of Mint in the United States of America.

Verticillium wilt, caused by the fungus Verticillium dahliae, is the most important and destructive disease of mint (Mentha spp.) in the United States (U.S.). The disease was first observed in commercial mint fields in the Midwestern U.S. in the 1920s and, by the 1950s, was present in mint producing regions of the U.S. Pacific Northwest. Verticillium wilt continues to be a major limiting factor in commercial peppermint (Mentha x piperita) and Scotch spearmint (Mentha x gracilis) production, two of the most important sources of mint oil in the U.S. The perennial aspect of U.S. mint production, coupled with the soilborne, polyetic nature of V. dahliae, makes controlling Verticillium wilt in mint a challenge. Studies investigating the biology and genetics of the fungus, the molecular mechanisms of virulence and resistance, and the role of soil microbiota in modulating host-pathogen interactions are needed to improve our understanding of Verticillium wilt epidemiology and inform novel disease management strategies. This review will discuss the history and importance of Verticillium wilt in commercial U.S. mint production, as well as provide a format to highlight past and recent research advances in an effort to better understand and manage the disease.

Simple sequence repeat markers that identify Claviceps species and strains.

BACKGROUND: Claviceps purpurea is a pathogen that infects most members of Pooideae, a subfamily of Poaceae, and causes ergot, a floral disease in which the ovary is replaced with a sclerotium. When the ergot body is accidently consumed by either man or animal in high enough quantities, there is extreme pain, limb loss and sometimes death. RESULTS: This study was initiated to develop simple sequence repeat (SSRs) markers for rapid identification of C. purpurea. SSRs were designed from sequence data stored at the National Center for Biotechnology Information database. The study consisted of 74 ergot isolates, from four different host species, Lolium perenne, Poa pratensis, Bromus inermis, and Secale cereale plus three additional Claviceps species, C. pusilla, C. paspali and C.fusiformis. Samples were collected from six different counties in Oregon and Washington over a 5-year period. Thirty-four SSR markers were selected, which enabled the differentiation of each isolate from one another based solely on their molecular fingerprints. Discriminant analysis of principle components was used to identify four isolate groups, CA Group 1, 2, 3, and 4, for subsequent cluster and molecular variance analyses. CA Group 1 consisting of eight isolates from the host species P. pratensis, was separated on the cluster analysis plot from the remaining three groups and this group was later identified as C. humidiphila. The other three groups were distinct from one another, but closely related. These three groups contained samples from all four of the host species. These SSRs are simple to use, reliable and allowed clear differentiation of C. humidiphila from C. purpurea. Isolates from the three separate species, C. pusilla, C. paspali and C.fusiformis, also amplified with these markers. CONCLUSIONS: The SSR markers developed in this study will be helpful in defining the population structure and genetics of Claviceps strains. They will also provide valuable tools for plant breeders needing to identify resistance in crops or for researchers examining fungal movements across environments.