Development of a sequential sampling plan for classifying Thrips tabaci (Thysanoptera: Thripidae) populations in onion fields.

Onion thrips, Thrips tabaci Lindeman, is a global pest of onion crops, causing substantial economic damage by diminishing bulb yields and transmitting plant pathogens. Insecticides are used to manage T. tabaci infestations with control decisions traditionally based on action thresholds that require visually counting thrips on a fixed, predetermined number of onion plants per field. However, this approach for treatment decisions is inefficient when thrips populations are well above or below the action threshold. The aim of this research was to develop a sequential sampling plan that would provide a rapid and reliable classification of thrips populations in commercial onion fields above or below prespecified management thresholds. The study was conducted in a total of 24 commercial onion fields in New York in 2021 and 2022. Taylor's power law and Wald's Sequential Probability Ratio Test were used in concert to develop each sampling plan. Simulated and historical field data of thrips populations were used to further validate the efficacy of each sampling plan. Results demonstrated the sequential sampling plan required an average of 78% fewer samples to make a control decision compared with the traditional fixed-sampling approach. Treatment decisions were reached in 72% of cases after inspecting only 10 plants, while only 6% of the cases required examining more than 25 plants. Comparisons with fixed-sample sizes ranging from 23 to 68 plants revealed a 96% agreement in decision-making and a 78% reduction in sampling effort when using the sequential sampling plans.

Identifying Onion Fields at Risk of Iris Yellow Spot Virus in New York.

Iris yellow spot virus (IYSV) poses a significant threat to dry bulb onion, Allium cepa L., production and can lead to substantial yield reductions. IYSV is transmitted by onion thrips, Thrips tabaci (Lindeman), but not via seed. Transplanted onion fields have been major early season sources of IYSV epidemics. As onion thrips tend to disperse short distances, seeded onion fields bordering transplanted onion fields may be at greater risk of IYSV infection than seeded fields isolated from transplanted ones. Additionally, seeded onion fields planted early may be at greater risk of IYSV infection than those seeded later. In a 2-year study in New York, we compared IYSV incidence and onion thrips populations in seeded onion fields relative to their proximity to transplanted onion fields. In a second study, we compared IYSV incidence in onion fields with either small or large plants during midseason. Results showed similar IYSV incidence and onion thrips populations in seeded onion fields regardless of their proximity to transplanted onion fields, while IYSV incidence was over four times greater in large onion plants than in small ones during midseason. These findings suggest a greater risk of onion thrips-mediated IYSV infection in onion fields with large plants compared with small ones during midseason and that proximity of seeded fields to transplanted ones is a poor indicator of IYSV risk. Our findings on IYSV spread dynamics provided valuable insights for developing integrated pest and disease management strategies for New York onion growers.

Spatiotemporal Patterns of Iris Yellow Spot Virus and Its Onion Thrips Vector, Thrips tabaci, in Transplanted and Seeded Onion Fields in New York.

Onion thrips, Thrips tabaci (Lindeman), transmits iris yellow spot virus (IYSV) and is one of the most important pests of Allium crops. IYSV is a member of the species Tospovirus iridimaculaflavi in the genus Orthotospovirus of the family Tospoviridae. This virus typically reduces overall onion bulb quality and weight but can also prematurely kill onion plants. IYSV is neither seed nor mechanically transmitted. Onion fields are typically established via seeds and transplants. A decade ago, onion thrips tended to colonize transplanted fields before seeded fields because plants in transplanted fields were larger and more attractive to thrips than smaller onions in seeded fields. Therefore, we hypothesized that the incidence of IYSV in transplanted fields would be detected early in the season and be spatially aggregated, whereas IYSV would be absent from seeded fields early in the season and initial epidemic patterns would be spatially random. In 2021 and 2022, IYSV incidence and onion thrips populations were quantified in 12 onion fields (four transplanted fields and eight seeded fields) in New York. Fields were scouted four times throughout the growing season (n = 96 samples), and a geospatial and temporal analysis of aggregation and incidence was conducted to determine spatiotemporal patterns in each field type. Results indicated that spatial patterns of IYSV incidence and onion thrips populations were similar early in the season, indicating that transplanted onion fields are no longer the dominant early-season source of IYSV in New York. These findings suggest the need to identify other important early-season sources of IYSV that impact New York onion fields.

Phylogenetic relatedness can influence cover crop-based weed suppression.

Cover crops are plants grown to provide regulating, supporting, and cultural ecosystem services in managed environments. In agricultural systems, weed suppression services from cover crops can be an important tool to promote sustainability as reliance on herbicides and tillage for weed management has caused pollution, biodiversity loss, and human health issues. However, to effectively use weed suppression services from cover crops, farmers must carefully select species that fit within their rotations and suppress their problematic weeds. Understanding how the relatedness between cover crops and weeds affects their interactions will help farmers select cover crops for targeted weed management. The phylogenetic distance between species reflects their relatedness and was studied through a series of field experiments that compared weed suppression in winter and summer cover crops with tilled controls. This study demonstrates that cover crops can reduce up to 99% of weed biomass and alter weed community structure by suppressing phylogenetically related weed species. Results also suggest that cover crop planting season can influence weed community structure since only overwintering treatments affected the phylogenetic distance of weed communities. In an applied context, these results help develop cover crop-based weed management systems, demonstrating that problematic weeds can be managed by selecting phylogenetically related cover crop species. More broadly, this study provides a framework for evaluating weed communities through a phylogenetic perspective, which provides new insight into plant interactions in agriculture.

Enabling Population Biology Studies of Stemphylium vesicarium from Onion with Microsatellites.

Stemphylium leaf blight (SLB), caused by the fungus Stemphylium vesicarium, is dominant within the foliar disease complex affecting onion production in New York (NY). The disease causes premature defoliation and significant reductions in bulb weight and quality. Foliar diseases of onion are usually managed by an intensive fungicide program, but SLB management is complicated by resistance to multiple single-site modes of action. The design of integrated disease management strategies is limited by incomplete knowledge surrounding the dominant sources of S. vesicarium inoculum. To facilitate genomic-based studies of S. vesicarium populations, nine microsatellite markers were developed. The markers were multiplexed into two PCR assays containing four and five fluorescently labeled microsatellite markers. Initial testing of the S. vesicarium isolates found the markers were highly polymorphic and reproducible with an average of 8.2 alleles per locus. The markers were used to characterize 54 S. vesicarium isolates from major NY onion production regions in 2016 (n = 27) and 2018 (n = 27). Fifty-two multilocus genotypes (MLGs) were identified between these populations. Genotypic and allelic diversities were high in both the 2016 and 2018 populations. A greater degree of genetic variation was observed within populations than between years. No distinct pattern of MLGs according to population was identified and some MLGs were closely related between 2016 and 2018. The lack of evidence for linkage among loci also was strongly suggestive of clonal populations with only minor differences between the two populations. These microsatellite markers will be a foundational resource for the testing of hypotheses surrounding the population biology of S. vesicarium and therefore informing disease management.

Distinct Red Blotch Disease Epidemiological Dynamics in Two Nearby Vineyards.

Grapevine red blotch virus (GRBV) causes red blotch disease and is transmitted by the three-cornered alfalfa hopper, Spissistilus festinus. GRBV isolates belong to a minor phylogenetic clade 1 and a predominant clade 2. Spatiotemporal disease dynamics were monitored in a 1-hectare 'Merlot' vineyard planted in California in 2015. Annual surveys first revealed disease onset in 2018 and a 1.6% disease incidence in 2022. Ordinary runs and phylogenetic analyses documented significant aggregation of vines infected with GRBV clade 1 isolates in one corner of the vineyard (Z = -4.99), despite being surrounded by clade 2 isolates. This aggregation of vines harboring isolates from a non-prevalent clade is likely due to infected rootstock material at planting. GRBV clade 1 isolates were predominant in 2018-2019 but displaced by clade 2 isolates in 2021-2022, suggesting an influx of the latter isolates from outside sources. This study is the first report of red blotch disease progress immediately after vineyard establishment. A nearby 1.5-hectare 'Cabernet Sauvignon' vineyard planted in 2008 with clone 4 (CS4) and 169 (CS169) vines was also surveyed. Most CS4 vines that exhibited disease symptoms one-year post-planting, likely due to infected scion material, were aggregated (Z = -1.73). GRBV isolates of both clades were found in the CS4 vines. Disease incidence was only 1.4% in non-infected CS169 vines in 2022 with sporadic infections of isolates from both clades occurring via secondary spread. Through disentangling GRBV infections due to the planting material and S. festinus-mediated transmission, this study illustrated how the primary virus source influences epidemiological dynamics of red blotch disease.

Sampling, a New iOS Application for Assessment of Damage by Diseases and Insect Pests Using Sequential Sampling Plans.

Regular scouting for plant diseases and insect pests by growers, crop consultants, extension educators, and researchers (herein defined as stakeholders) is the cornerstone of integrated pest management practices. Sequential sampling plans have the potential to save time and labor in field scouting and reduce the frequency of errors surrounding decision-making. The incorporation of the algorithms behind sequential sampling plans into mobile devices can make scouting for diseases and insect pests more straightforward, practical, and enjoyable. Here, we introduce an iOS application called Sampling. The application was designed for stakeholders to use on a mobile device for assessing disease and insect pest incidence in the field using sequential sampling plans. The application allows users to select a disease or insect pest from a prepopulated list and specify the objective of sampling: Estimation or classification. Conducting sequential sampling depends upon different precision levels and action thresholds within each objective. Detailed instructions for each sequential sampling plan are available as a guide. When sampling begins, users enter the number of diseased individuals at each sampling unit. The specific algorithm developed for the disease or insect pest will inform the user when to stop sampling for the desired goal and return the final incidence and precision or threshold achieved. Results are automatically saved in the application, and the user can inspect and share results by exporting them to a range of compatible programs. The initial version of Sampling (1.1) was released with the sequential sampling plans for Cercospora leaf spot of table beet. Sequential sampling plans for additional diseases or pests will be added to Sampling in subsequent versions. Sampling is available as a free download from the Apple Store (https://apple.co/3pUiYKy) and is compatible with iOS 14.0 or greater on the iPhone or iPad.

Control of Phoma Leaf Spot and Root Decay of Table Beet in New York.

Disease caused by Neocamarosporium betae (syn. Phoma betae, Pleospora betae) results in reductions in plant populations, foliar disease (Phoma leaf spot [PLS]), and root disease and decay in table beet. Disease caused by N. betae has reemerged as prevalent in organic table beet production in New York. The disease can also cause substantial issues in conventional table beet production. To evaluate in-field control options for conventional and organic table beet production, small-plot, replicated trials were conducted in each of two years (2019 and 2021). The fungicides, propiconazole and difenoconazole, and premixtures, pydiflumetofen + fludioxonil or pydiflumetofen + difenoconazole, provided excellent PLS and root decay control. Azoxystrobin provided excellent (69.9%) control of PLS in 2019 and lesser (40%) control in 2021. Field trial results complemented in vitro sensitivity testing of 30 New York N. betae isolates that were all highly sensitive to azoxystrobin (mean effective concentration to reduce mycelial growth by 50%, EC50 = 0.0205 µg/ml) and propiconazole (mean EC50 = 0.0638 µg/ml). Copper octanoate and microbial biopesticides containing either Bacillus amyloliquefaciens D747 or B. mycoides strain J provided moderate (68.5 to 74.6%) PLS control as reflected in epidemic progress. The Gompertz model provided the best fit to PLS epidemics reflecting a polycyclic epidemic. Reductions in PLS severity were associated with significant decreases in Phoma root decay and increases in canopy health and the time-to-death of leaves compared with nontreated control plots. Prolonging leaf survival is critical for mechanical harvest of roots. These findings underpin the design of programs for foliar disease control in conventional and organic table beet production. Assessment of PLS severity in the field will better inform postharvest management decisions.

Comparing the Fungicide Sensitivity of Sclerotinia sclerotiorum Using Mycelial Growth and Ascospore Germination Assays.

The infection of the floral tissues of snap bean and other crops by Sclerotinia sclerotiorum, the causative agent of white mold, is by ascospores. Irrespective of the fungicide mode of action being evaluated, in vitro fungicide sensitivity tests are conducted almost exclusively using mycelial growth assays. This is likely because of difficulties and time involved in sclerotial conditioning required to produce apothecia and ascospores. The objective of this research was to compare estimates of fungicide sensitivity between mycelial growth and ascospore germination assays for S. sclerotiorum. Sensitivity assays were conducted using serial doses of three fungicides commonly used to control white mold: boscalid, fluazinam, and thiophanate-methyl. A total of 27 isolates were evaluated in replicated trials conducted for each fungicide and assay type. The effective concentration to reduce mycelial growth or ascospore germination by 50% (EC50) was estimated for each isolate, fungicide, and assay type. The median EC50 values obtained from ascospore germination assays were 52.7, 10.0, and 2.7 times higher than those estimated from the mycelial growth for boscalid, fluazinam, and thiophanate-methyl, respectively. No significant correlation was found between EC50 values estimated by the two methods. These findings highlight differences that may be important in evaluating the sensitivity of S. sclerotiorum given the fungicide mode of action and how they will be used in the field.

Spatiotemporal Dynamics of Stemphylium Leaf Blight and Potential Inoculum Sources in New York Onion Fields.

Stemphylium leaf blight (SLB) caused by Stemphylium vesicarium is the dominant foliar disease affecting large-scale onion production in New York. The disease is managed by fungicides, but control failures are prevalent and are attributed to fungicide resistance. Little is known of the relative role of inoculum sources in initiation and spread of SLB epidemics. Plate testing of 28 commercially available organic onion seedlots from 2016 and 2017 did not detect S. vesicarium. This finding suggests that although S. vesicarium has been reported as seed-transmitted, this is unlikely to be a significant inoculum source in commercially available organic seed lots and even less so in fungicide-treated seed used to establish conventional fields. The spatial and spatiotemporal dynamics of SLB epidemics in six onion fields were evaluated along linear transects in 2017 and 2018. Average SLB incidence increased from 0 to 100% throughout the cropping seasons with an average final lesion length of 28.3 cm. Disease progress was typical of a polycyclic epidemic and the logistic model provided the best fit to 83.3% of the datasets. Spatial patterns were better described by the beta-binomial than binomial distribution in half of the datasets (50%) and random patterns were more frequently observed by the index of dispersion (59%). Geostatistical analyses also found a low frequency of datasets with aggregation (60%). Spatiotemporal analysis of epidemics detected that the aggregation was influenced by disease incidence. However, diseased units were not frequently associated with the previous time period according to the spatiotemporal association function of spatial analyses by distance indices. Variable spatial patterns suggested mixed inoculum sources dependent upon location, and likely an external inoculum source at the sampling scale used in this study. A small-plot replicated trial was also conducted in each of 2 years to quantify the effect of S. vesicarium-infested onion residue on SLB epidemics in a field isolated from other onion fields. SLB incidence was significantly reduced in plots without residue compared with those in which residue remained on the soil surface. Burial of infested residue also significantly reduced epidemic progress in 1 year. The effect of infested onion residue on SLB epidemics in the subsequent onion crop suggests rotation or residue management may have a substantial effect on epidemics. However, the presence of an inoculum source external to fields in onion production regions, as indicated by a lack of spatial aggregation, may reduce the efficacy of in-field management techniques.

Development of a Sequential Sampling Plan using Spatial Attributes of Cercospora Leaf Spot Epidemics of Table Beet in New York.

Sampling strategies that effectively assess disease intensity in the field are important to underpin management decisions. To develop a sequential sampling plan for the incidence of Cercospora leaf spot (CLS), caused by Cercospora beticola, 31 table beet fields were assessed in the state of New York. Assessments of CLS incidence were performed in six leaves arbitrarily selected in 51 sampling locations along each of three to six linear transects per field. Spatial pattern analyses were performed, and results were used to develop sequential sampling estimation and classification models. CLS incidence (p) ranged from 0.13 to 0.92 with a median of 0.31, and beta-binomial distribution, which is reflective of aggregation, best described the spatial patterns observed. Aggregation was commonly detected (>95%) by methods using the point-process approach, runs analyses, and autocorrelation up to the fourth spatial lag. For Spatial Analysis by Distance Indices, or SADIE, 45% of the datasets were classified as a random pattern. In the sequential sampling estimation and classification models, disease units are sampled until a prespecified target is achieved. For estimation, the goal was sampling CLS incidence with a preselected coefficient of variation (C). Achieving the C = 0.1 was challenging with <51 sampling units, and only observed on datasets with incidence >0.3. Reducing the level of precision, i.e., increasing C to 0.2, allowed the preselected C to be achieved with a lower number of sampling units and with an estimated incidence ([Formula: see text]) close to the true value of p. For classification, the goal was to classify the datasets above or below prespecified thresholds (pt) used for CLS management. The average sample number, or ASN, was determined by Monte Carlo simulations, and was between 20 and 45 at disease incidence values close to pt, and approximately 11 when far from pt. Correct decisions occurred in >76% of the validation datasets. Results indicated these sequential sampling plans can be used to effectively assess CLS incidence in table beet fields.

Cercospora beticola: The intoxicating lifestyle of the leaf spot pathogen of sugar beet.

Cercospora leaf spot, caused by the fungal pathogen Cercospora beticola, is the most destructive foliar disease of sugar beet worldwide. This review discusses C. beticola genetics, genomics, and biology and summarizes our current understanding of the molecular interactions that occur between C. beticola and its sugar beet host. We highlight the known virulence arsenal of C. beticola as well as its ability to overcome currently used disease management strategies. Finally, we discuss future prospects for the study and management of C. beticola infections in the context of newly employed molecular tools to uncover additional information regarding the biology of this pathogen. TAXONOMY: Cercospora beticola Sacc.; Kingdom Fungi, Phylum Ascomycota, Class Dothideomycetes, Order Capnodiales, Family Mycosphaerellaceae, Genus Cercospora. HOST RANGE: Well-known pathogen of sugar beet (Beta vulgaris subsp. vulgaris) and most species of the Beta genus. Reported as pathogenic on other members of the Chenopodiaceae (e.g., lamb's quarters, spinach) as well as members of the Acanthaceae (e.g., bear's breeches), Apiaceae (e.g., Apium), Asteraceae (e.g., chrysanthemum, lettuce, safflower), Brassicaceae (e.g., wild mustard), Malvaceae (e.g., Malva), Plumbaginaceae (e.g., Limonium), and Polygonaceae (e.g., broad-leaved dock) families. DISEASE SYMPTOMS: Leaves infected with C. beticola exhibit circular lesions that are coloured tan to grey in the centre and are often delimited by tan-brown to reddish-purple rings. As disease progresses, spots can coalesce to form larger necrotic areas, causing severely infected leaves to wither and die. At the centre of these spots are black spore-bearing structures (pseudostromata). Older leaves often show symptoms first and younger leaves become infected as the disease progresses. MANAGEMENT: Application of a mixture of fungicides with different modes of action is currently performed although elevated resistance has been documented in most employed fungicide classes. Breeding for high-yielding cultivars with improved host resistance is an ongoing effort and prudent cultural practices, such as crop rotation, weed host management, and cultivation to reduce infested residue levels, are widely used to manage disease. USEFUL WEBSITE: https://www.ncbi.nlm.nih.gov/genome/11237?genome_assembly_id=352037.

Optimizing Cercospora Leaf Spot Control in Table Beet Using Action Thresholds and Disease Forecasting.

Cercospora leaf spot (CLS), caused by the fungus Cercospora beticola, is the dominant foliar disease affecting table-beet production in New York. CLS epidemics occur annually and, if uncontrolled, will rapidly lead to defoliation. In broad-acre production, season-long maintenance of healthy leaves is important to facilitate harvest by top-pulling. Fungicides are the dominant means of CLS control and applications are initiated at an action threshold of 1 CLS lesion/leaf. Regular fungicide application occurs thereafter without regard for scheduling based on weather-based risk. The current action threshold was evaluated with selected fungicides in two replicated field trials. Copper oxychloride + copper hydroxide and propiconazole significantly improved CLS control if initiated prior to infection. Pydiflumetofen + difenoconazole significantly reduced area under the disease progress stairs compared with other fungicides tested and was most efficacious when applications began at 1 CLS lesion/leaf. Six replicated field trials also evaluated the utility of scheduling fungicides on weather-based risk rather than a calendar approach. Two risk thresholds (moderate and high) integrating the accumulation of daily infection values based on temperature and relative humidity from a forecaster for CLS in sugar beet were evaluated. Applications of pydiflumetofen + difenoconazole were reduced from three to two by using the forecaster at either risk threshold compared with calendar applications without affecting CLS control. For propiconazole, the moderate risk threshold provided CLS control equivalent to calendar applications and saved one spray per season. Thus, there was substantial scope to reduce spray frequency by scheduling based on weather-based risk rather than calendar applications. The optimal risk thresholds for pydiflumetofen + difenoconazole and propiconazole were high and moderate, respectively. In these trials, periods of high risk occurred less frequently than moderate risk, increasing the reapplication intervals and, hence, represented a less conservative approach to disease management.

Detection of Cercospora beticola and Phoma betae on Table Beet Seed using Quantitative PCR.

Cercospora beticola and Phoma betae are important pathogens of table beet, sugar beet, and Swiss chard (Beta vulgaris subsp. vulgaris), causing Cercospora leaf spot (CLS) and Phoma leaf spot, root rot, and damping-off, respectively. Both pathogens may be seedborne; however, limited evidence is available for seed infestation by C. beticola. Due to the limitations of culture-based seed assessment methods, detection of these pathogens was investigated using PCR. A P. betae-specific quantitative PCR assay was developed and used in conjunction with a C. beticola-specific assay to assess the presence of pathogen DNA in 12 table beet seed lots. DNA of C. beticola and P. betae was detected in four and eight seed lots, respectively. Plate tests and BIO-PCR confirmed the viability of each pathogen; however, competitive growth of other microbes and low incidence limited the frequency and sensitivity of detection in some seed lots. The results for P. betae support previously described infestation of seed. Further investigation of C. beticola-infested seed lots indicated the ability of seedborne C. beticola to cause CLS on plants grown from infested seed. Detection of viable C. beticola on table beet seed demonstrates the potential for pathogen dispersal and disease initiation via infested seed, and provides valuable insight into the epidemiology of CLS. Surveys of commercial table beet seed are required to determine the frequency and source of C. beticola seed infestation and its role as primary inoculum for epidemics, and to evaluate the effectiveness of seed treatments.

Genome Resource for Two Stemphylium vesicarium Isolates Causing Stemphylium Leaf Blight of Onion in New York.

Stemphylium leaf blight caused by Stemphylium vesicarium was recently identified as an emerging disease and dominant in the foliar disease complex affecting onion in New York. Here, we report the genomes of two isolates of S. vesicarium, On16-63 and On16-391. The availability of the genomes will accelerate genomic studies of S. vesicarium, including population biology, sexual reproduction, and fungicide resistance. Additionally, comparative genomics with the other published genome of S. vesicarium causing brown spot of pear will help understand pathogen biology and underpin the development of management strategies for this disease.

Emergence of Stemphylium Leaf Blight of Onion in New York Associated With Fungicide Resistance.

A complex of foliar diseases affects onion production in New York, including Botrytis leaf blight (Botrytis squamosa), purple blotch (Alternaria porri), Stemphylium leaf blight (SLB; Stemphylium vesicarium), and downy mildew (Peronospora destructor). Surveys were conducted in 2015 and 2016 to evaluate the cause of severe premature foliar dieback in New York onion fields. SLB was the most prevalent disease among fields with the greatest incidence, surpassing downy mildew, purple blotch, and Botrytis leaf blight. Sequencing of the internal transcribed spacer region of ribosomal DNA and the glyceraldedyhe-3-phosphate dehydrogenase and calmodulin genes identified S. vesicarium as the species most commonly associated with SLB. S. vesicarium was typically associated with a broad range of necrotic symptoms but, most commonly, dieback of leaf tips and asymmetric lesions that often extended over the entire leaf. Because of the intensive use of fungicides for foliar disease control in onion crops in New York, the sensitivity of S. vesicarium populations to various fungicides with site-specific modes of action was evaluated. Sensitivity of S. vesicarium isolates collected in 2016 to the quinone outside inhibitor (QoI) fungicide, azoxystrobin, was tested using a conidial germination assay. Isolates representing a broad range of QoI sensitivities were selected for sequencing of the cytochrome b gene to evaluate the presence of point mutations associated with insensitivity to azoxystrobin. The G143A mutation was detected in all 74 S. vesicarium isolates with an azoxystrobin-insensitive phenotype (effective concentrations reducing conidial germination by 50%, EC50 = 0.2 to 46.7 µg of active ingredient [a.i.]/ml) and was not detected in all 31 isolates with an azoxystrobin-sensitive phenotype (EC50 = 0.01 to 0.16 µg a.i./ml). The G143A mutation was also associated with insensitivity to another QoI fungicide, pyraclostrobin. Sensitivity to other selected fungicides commonly used in onion production in New York was evaluated using a mycelial growth assay and identified isolates with insensitivity to boscalid, cyprodinil, and pyrimethanil, but not difenoconazole. The frequency of isolates sensitive to iprodione, fluxapyroxad, and fluopyram was high (93.5 to 93.6%). This article discusses the emergence of SLB as dominant in the foliar disease complex affecting onion in New York and the complexities of management posed by resistance to fungicides with different modes of action.

Development of a Species-Specific PCR for Detection and Quantification of Meloidogyne hapla in Soil Using the 16D10 Root-Knot Nematode Effector Gene.

The Northern root-knot nematode (Meloidogyne hapla) is an important soilborne pathogen of numerous agricultural crops in temperate regions. Accurate detection and quantification is vital to supporting informed pest management decisions. However, traditional methods of manual nematode extraction and morphology-based identification are time-consuming and require highly specialized training. Molecular methods may expand the diagnostician's toolkit beyond those methods that rely on this disappearing specialized skillset. However, molecular assays targeting the internal transcribed spacer region may lead to inaccurate results because of intraspecific variability. The Meloidogyne spp. effector gene 16D10 was assessed as a target for a SYBR Green I quantitative PCR (qPCR) assay for detection and quantification of M. hapla. M. hapla-specific qPCR primers were developed and evaluated for specificity against five M. hapla isolates and 14 other plant-parasitic nematodes. A standard curve was generated by relating the quantification cycle (Cq) to the log of M. hapla population densities artificially introduced into soil. The influence of soil inhibitors on quantitative amplification was assessed by generating a dilution series from DNA extracted from pure nematode cultures and inoculated soil. Extracts from soil produced significantly higher Cq values than those produced from pure culture extracts. The utility of the qPCR was evaluated using soil samples collected from three naturally infested potato fields, resulting in a significant positive relationship between populations estimated using qPCR and populations derived from manual counting. The qPCR developed in this study provides a useful method for detecting and quantifying M. hapla in soil and demonstrates the utility of effector genes in plant-parasitic nematode diagnostics. The ability to use effector genes as targets for qPCR and other molecular detection and quantification methods may open additional avenues of novel research and support development of improved species-level diagnostics.

Alternative Hosts of Cercospora beticola in Field Surveys and Inoculation Trials.

Cercospora beticola, the cause of Cercospora leaf spot (CLS) of sugar beet and table beet, has a broad range of potential alternative hosts. The role of these hosts as inoculum sources in the field is unclear and has had limited investigation since the advent of DNA-based pathogen identification. The presence of C. beticola on alternative hosts associated with table beet fields of New York was assessed in field surveys during 2016. Lesions were collected, and 71 cercosporoid conidia were isolated for phylogenetic comparison. C. beticola was identified from Solanum ptycanthum (n = 4), Chenopodium album (n = 2), and Spinacia oleracea (n = 1), whereas C. chenopodii was identified on Chenopodium album (n = 51). Artificial inoculation of 21 plants species demonstrated that C. beticola was pathogenic to Brassica kaber, Chenopodium album, Carthamus tinctorius, Rumex obtusifolius, and Spinacia oleracea. These results indicate that although C. beticola may be pathogenic to a range of plant species, the role of symptomatic tissue for inoculum production on alternative hosts in the field appears limited. Observations of C. beticola on necrotic and naturally senescent tissue suggest saprophytic survival on plant debris of a range of species, which has implications for CLS epidemics and disease management.

Genetic Diversity and Differentiation in Phoma betae Populations on Table Beet in New York and Washington States.

Phoma betae is an important seedborne pathogen of table beet worldwide that is capable of causing foliar, root, and damping-off diseases. Ten microsatellite and mating type markers were developed to investigate the genetics of P. betae populations in table beet root crops in New York and in table beet seed crops in Washington, from where table beet seed is predominantly sourced. The markers were used to characterize 175 isolates comprising five P. betae populations (two from New York and three from Washington), and they were highly polymorphic with an allelic range of 4 to 33 and an average of 11.7 alleles per locus. All populations had high genotypic diversity (Simpson's complement index = 0.857 to 0.924) and moderate allelic diversity (Nei's unbiased gene diversity = 0.582 to 0.653). Greater differentiation observed between populations from the two states compared with populations within the same state suggested that an external inoculum source, such as windblown ascospores, may be homogenizing the populations. However, most genetic diversity (87%) was among individual isolates within populations (pairwise index of population differentiation = 0.127; P = 0.001), suggesting that local within-field inoculum source(s), such as infested field debris or infected weeds, may also be important in initiating disease outbreaks. Standardized index of association, proportion of compatible pairs of loci, and mating type ratio calculations showed evidence for a mixed reproduction mode in all populations. These findings could be useful in designing more effective management strategies for diseases caused by P. betae in table beet production.

Evaluation of App-Embedded Disease Scales for Aiding Visual Severity Estimation of Cercospora Leaf Spot of Table Beet.

Two diagrammatic ordinal scales are available in the Estimate app (2017 version) for Cercospora leaf spot (CLS) severity on table beet: 10% linear (linear-based diagrammatic scale [LIN]) and logarithmic based (Horsfall-Barratt [HB]). These allow for estimating severity data of four types depending on the system used. A group of 30 raters assigned percentage severity on 30 photographs of diseased table beet leaves during five rounds first without an aid and then using each of the four rating systems in Estimate. In two, the perceived ordinal score of the HB or LIN scale was assigned where severity of the subject fit best. HB2 and LIN2 involved a second choice of unitary severity within the perceived score interval. There was large variation in unaided ability of raters to estimate severity: 13% were accurate (Lin's concordance correlation [LCC] > 0.9), 23% were inaccurate (LCC < 0.7), and the remaining had moderate accuracy. Larger disparities between assigned and actual ordinal scores (mostly overestimates) occurred using the LIN compared with the HB. The LIN2 produced the most accurate estimates (Lin's concordance correlation coefficient, ρc = 0.96; generalized bias parameter, Cb = 0.99; Pearson's correlation coefficient r = 0.95) and the greatest interrater reliability (overall concordance correlation coefficient and intraclass correlation coefficient > 0.93). The two-step process using the 10% linear scale is recommended for severity estimates of CLS in table beet.