An Overview of Pathogens Associated with Biotic Stresses in Hemp Crops in Oregon, 2019 to 2020.

Hemp (Cannabis sativa) acreage in Oregon has increased by approximately 240 times in the last 5 years, and a greater number of hemp diseases have been observed. This special report documents pathogens, particularly those causing virus and virus-like diseases, that have been detected from field and greenhouse-grown hemp crops in Oregon, based on plant samples submitted to the Hermiston Agricultural Research and Extension Center Plant Clinic of Oregon State University in 2019 and 2020. Symptoms and signs were used to evaluate disease types and determine diagnostic assays used on each submission. Plants with signs or symptoms of fungal or oomycete infection were cultured to isolate pathogenic organisms and plants with symptoms suspected to be caused by virus infection were assayed for the presence of Beet curly top virus (BCTV), viroids, and phytoplasmas using PCR, or reverse transcription (RT)-PCR. Diseases with fungal or oomycete, and virus causes accounted for 26.5 and 42.9% of submissions, respectively; coinfection of viral and fungal or oomycete pathogens were detected from 6.1% of submissions between 2019 and 2020. BCTV, a curtovirus, and hop latent viroid (HLVd) were the predominant pathogens detected from field and indoor grown hemp. Worland-like strains of BCTV represented 93% of all curtovirus detections. Eighty percent of HLVd detections occurred from plants that originated from indoor growing facilities. Based on BCTV vector, beet leafhopper, prevalence, field-grown hemp in western production regions may be affected by curly top and increasing hemp acreage in the landscape may have potential implications on other crops affected by curtoviruses. Virus and virus-like diseases could be a limiting factor for hemp production in some regions of the United States.

Factors influencing aster leafhopper (Hemiptera: Cicadellidae) abundance and aster yellows phytoplasma infectivity in Wisconsin carrot fields.

In Wisconsin, vegetable crops are threatened annually by infection of the aster yellows phytoplasma (AYp), the causal agent of aster yellows (AY) disease, vectored by the aster leafhopper, Macrosteles quadrilineatus Forbes. Aster leafhopper abundance and infectivity are influenced by processes operating across different temporal and spatial scales. We applied a multilevel modeling approach to partition variance in multifield, multiyear, pest scouting data sets containing temporal and spatial covariates associated with aster leafhopper abundance and infectivity. Our intent was to evaluate the relative importance of temporal and spatial covariates to infer the relevant scale at which ecological processes are driving AY epidemics and identify periods of elevated risk for AYp spread. The relative amount of aster leafhopper variability among and within years (39%) exceeded estimates of variation among farm locations and fields (7%). Similarly, time covariates explained the largest amount of variation of aster leafhopper infectivity (50%). Leafhopper abundance has been decreasing since 2001 and reached its minimum in 2010. The average seasonal pattern indicated that periods of above average abundance occurred between 11 June and 1 August. Annual infectivity appears to oscillate around an average value of 2% and seasonal periods of above average infectivity occur between 19 May and 15 July. The coincidence of the expected periods of high leafhopper abundance and infectivity increases our knowledge of when the insect moves into susceptible crop fields and when it spreads the pathogen to susceptible crops, representing a seasonal interval during which management of the insect can be focused.

Seasonal patterns of aster leafhopper (Hemiptera: Cicadellidae) abundance and aster yellows phytoplasma infectivity in Wisconsin carrot fields.

In Wisconsin, vegetable crops are threatened annually by the aster yellows phytoplasma (AYp), which is obligately transmitted by the aster leafhopper. Using a multiyear, multilocation data set, seasonal patterns of leafhopper abundance and infectivity were modeled. A seasonal aster yellows index (AYI) was deduced from the model abundance and infectivity predictions to represent the expected seasonal risk of pathogen transmission by infectious aster leafhoppers. The primary goal of this study was to identify periods of time during the growing season when crop protection practices could be targeted to reduce the risk of AYp spread. Based on abundance and infectivity, the annual exposure of the carrot crop to infectious leafhoppers varied by 16- and 70-fold, respectively. Together, this corresponded to an estimated 1,000-fold difference in exposure to infectious leafhoppers. Within a season, exposure of the crop to infectious aster leafhoppers (Macrosteles quadrilineatus Forbes), varied threefold because of abundance and ninefold because of infectivity. Periods of above average aster leafhopper abundance occurred between 11 June and 2 August and above average infectivity occurred between 27 May and 13 July. A more comprehensive description of the temporal trends of aster leafhopper abundance and infectivity provides new information defining when the aster leafhopper moves into susceptible crop fields and when they transmit the pathogen to susceptible crops.

Effects of landscape composition and rotation distance on Leptinotarsa decemlineata (Coleoptera: Chrysomelidae) abundance in cultivated potato.

Knowledge of the Colorado potato beetle's, Leptinotarsa decemlineata (Say), relationship to previous potato crops has contributed to the development of a pest management strategy focused upon crop rotation. Previous investigations revealed that potato rotations exceeding 0.4 km were effective in reducing colonization in current season potato. The current study examines the relationship between beetle abundance in potato (Solanum tuberosum L.) and distance from multiple, previous year potato fields in Wisconsin, and integrates information about the influence of natural habitats adjacent to previous season potato. Colorado potato beetle count data were collected in 1998 and 2008 and distance to previous potato, field areas, and landscape classes were estimated using maps from 1997 and 2007. Poisson regression was used to relate counts to combinations of distance and local landscape characteristics calculated for all fields within 1,500 m of sampled potato. In 1998, beetle counts measured in current season potato declined significantly with increasing distance from previous potato fields and field size did not influence these counts. However, there was no relationship between beetle abundance and distance to prior year potatoes in 2008. In both years, increased proportions of surrounding habitats, previously described as preferred for diapause sites (e.g., wooded field boundaries), did not relate significantly to counts. However, grassland habitat was negatively correlated with counts. Results indicate that distance from previous potato remains an important factor to reduce the magnitude of colonization. This analysis further suggests that certain landscape components (e.g., grassland) may influence infestation, which may be useful for refining future integrated pest management programs.

Detection and variability of aster yellows phytoplasma titer in its insect vector, Macrosteles quadrilineatus (Hemiptera: Cicadellidae).

The aster yellows phytoplasma (AYp) is transmitted by the aster leafhopper, Macrosteles quadrilineatus Forbes, in a persistent and propagative manner. To study AYp replication and examine the variability of AYp titer in individual aster leafhoppers, we developed a quantitative real-time polymerase chain reaction assay to measure AYp concentration in insect DNA extracts. Absolute quantification of AYp DNA was achieved by comparing the amplification of unknown amounts of an AYp target gene sequence, elongation factor TU (tuf), from whole insect DNA extractions, to the amplification of a dilution series containing known quantities of the tuf gene sequence cloned into a plasmid. The capabilities and limitations of this method were assessed by conducting time course experiments that varied the incubation time of AYp in the aster leafhopper from 0 to 9 d after a 48 h acquisition access period on an AYp-infected plant. Average AYp titer was measured in 107 aster leafhoppers and, expressed as Log10 (copies/insect), ranged from 3.53 (+/- 0.07) to 6.26 (+/- 0.11) occurring at one and 7 d after the acquisition access period. AYp titers per insect and relative to an aster leafhopper chromosomal reference gene, cp6 wingless (cp6), increased approximately 100-fold in insects that acquired the AYp. High quantification cycle values obtained for aster leafhoppers not exposed to an AYp-infected plant were interpreted as background and used to define a limit of detection for the quantitative real-time polymerase chain reaction assay. This method will improve our ability to study biological factors governing AYp replication in the aster leafhopper and determine if AYp titer is associated with frequency of transmission.

Seasonal phenology of Aphis glycines (Hemiptera: Aphididae) and other aphid species in cultivated bean and noncrop habitats in Wisconsin.

The occurrence of aphid-transmitted viruses in agricultural crops of the Midwest and northeastern United States has become more frequent since the arrival and establishment of the soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae). A. glycines is a competent vector of plant viruses and may be responsible for recent virus epidemics in Wisconsin snap bean, Phaseolus vulgaris L., fields. To determine whether vegetation surrounding crop fields could serve as sources of virus inocula, we examined the settling activity ofA. glycines and other aphid species in agricultural crops and noncrop field margins adjacent to snap bean fields. Noncrop field margins were made up of numerous virus-susceptible plant species within 10 m from snap bean field edges. During summers 2006 and 2007, horizontal pan traps were placed in commercial soybean [Glycine max (L.) Merr.], snap bean, and surrounding field margins to characterize aphid flight activity patterns in the different habitat types. Alate abundance and peak occurrence across years varied between crop and noncrop field margins and differed among patches of plants in field margins. Overall aphid activity peaked late in the season (21 August in 2006 and 28 July in 2007); with the majority (52%) of total aphids trapped in all habitats being A. glycines. Susceptibility to viral infection and confirmed visitation of A. glycines to these forage plants suggests the importance ofnoncrop habitats as potential sources of primary virus inoculum. Viral disease onset followed peak aphid flights and further implicates A. glycines as a likely vector of viruses in commercial bean and other crops in Wisconsin.

Considerations for Verticillium Wilt Resistance Evaluation in Potato.

Verticillium wilt (Vw), caused by the soilborne fungi Verticillium dahliae and V. albo-atrum, is an important disease of potato (Solanum tuberosum). Host plant resistance is a promising method of Vw control. Culture-based methods that quantify the pathogen in host tissue often are used for Vw resistance screening. To evaluate the processing time, accuracy, and precision of these methods, 46 clones were planted in a field naturally infested with V. dahliae to collect data on visual disease symptoms, pathogen colonization, and yield. In 2002, disease severity explained 4.34% of the variability of yield loss, but the linear relationship between stem colonization and yield loss was not significant. In 2003, stem colonization explained 57.5% of the variability of yield loss, whereas disease severity explained 1.7% of the variability of yield loss. Correlations comparing clone ranks from repeated pathogen measurements indicated that culturing sap from individual stems or bulked stems generated more repeatable clone rankings than culturing dried stems. Clone rankings were more repeatable between years if pathogen measurements were made earlier in the growing season. The results indicate a need to characterize the effect of the environment on the relationship among pathogen population sizes in planta, disease symptoms, and yield loss.