Phylogeny and taxonomy of Podosphaera cerasi, sp. nov., and Podosphaera prunicola sensu lato.

Powdery mildew of Prunus spp. is a significant disease in most cherry growing regions of Washington, USA. Powdery mildews on Prunus virginiana and Pr. avium were previously assigned to Podosphaera clandestina s. lat. (= Po. oxyacanthae) or Po. prunicola. In this report, we confirm the presence of two distinct Podosphaera species on these hosts. Phylogenetic analyses of nuc rDNA sequences from the internal transcribed spacer region (ITS1-5.8S-ITS2 = ITS) and 28S subunit confirmed the presence of two distinct species. A morphological comparison with type material of Po. prunicola and additional collections demonstrated that the powdery mildew on Pr. virginiana (including var. demissa and var. melanocarpa) is in fact Po. prunicola. The powdery mildew on Pr. avium is genetically, morphologically, and biologically distinct from Po. prunicola and is described here as the new species Po. cerasi. Cross-inoculation experiments confirmed that these two species are host specific. Podosphaera prunicola was unable to colonize Pr. avium, whereas Po. cerasi was unable to colonize Pr. virginiana. Morphological reexamination of numerous specimens identified as Po. prunicola on a broad range of Prunus species suggests that Po. prunicola is probably confined to species in Prunus subgen.Padus (= Prunus subgen. Cerasus sect. Laurocerasus, including sect. Padus), with Pr. virginiana as the principal host. Podosphaera cerasi occurs on hosts in Prunus subgen. Cerasus, and our work confirms a newly described species of powdery mildew on Pr. avium. This work also includes the first documented and genetically proven European record of Po. prunicola on Pr. serotina and its widespread occurrence in the United States.

Distribution and Characterization of Podosphaera macularis Virulent on Hop Cultivars Possessing R6-Based Resistance to Powdery Mildew.

Host resistance, both quantitative and qualitative, is the preferred long-term approach for disease management in many pathosystems, including powdery mildew of hop (Podosphaera macularis). In 2012, an epidemic of powdery mildew occurred in Washington and Idaho on previously resistant cultivars whose resistance was putatively based on the gene designated R6. In 2013, isolates capable of causing severe disease on cultivars with R6-based resistance were confirmed in Oregon and became widespread during 2014. Surveys of commercial hop yards during 2012 to 2014 documented that powdery mildew is now widespread on cultivars possessing R6 resistance in Washington and Oregon, and the incidence of disease is progressively increasing. Pathogenic fitness, race, and mating type of R6-virulent isolates were compared with isolates of P. macularis lacking R6 virulence. All isolates were positive for the mating type idiomorph MAT1-1 and were able to overcome resistance genes Rb, R3, and R5 but not R1 or R2. In addition, R6-virulent isolates were shown to infect differential cultivars reported to possess the R6 gene and also the R4 gene, although R4 has not yet been broadly deployed in the United States. R6-virulent isolates were not detected from the eastern United States during 2012 to 2015. In growth chamber studies, R6-virulent isolates of P. macularis had a significantly longer latent period and produced fewer lesions on plants with R6 as compared with plants lacking R6, indicating a fitness cost to the fungus. R6-virulent isolates also produced fewer conidia when compared with isolates lacking R6 virulence, independent of whether the isolates were grown on a plant with or without R6. Thus, it is possible that the fitness cost of R6 virulence occurs regardless of host genotype. In field studies, powdery mildew was suppressed by at least 50% on plants possessing R6 as compared with those without R6 when coinoculated with R6-virulent and avirulent isolates. R6 virulence in P. macularis appears to be race specific and, at this time, imposes a measurable fitness penalty on the fungus. Resistance genes R1 and R2 appear to remain effective against R6-virulent isolates of P. macularis in the U.S. Pacific Northwest.

Interaction of Basal Foliage Removal and Late-Season Fungicide Applications in Management of Hop Powdery Mildew.

Canopy management is an important aspect of control of powdery mildew diseases and may influence the intensity of fungicide applications required to suppress disease. In hop, powdery mildew (caused by Podosphaera macularis) is most damaging to cones when infection occurs during bloom and the juvenile stages of cone development. Experiments were conducted over 3 years to evaluate whether fungicide applications could be ceased after the most susceptible stages of cone development (late July) without unduly affecting crop yield and quality when disease pressure was moderated with varying levels of basal foliage removal. In experimental plots of 'Galena' hop, the incidence of leaves with powdery mildew was similar whether fungicides were ceased in late July or made in late August. Disease levels on leaves were unaffected by the intensity of basal foliage removal, whereas the intensity of basal foliage removal interacted with the duration of fungicide applications to affect disease levels on cones. Similar experiments conducted in large plots of 'Tomahawk' hop in a commercial hop yard similarly found no significant impact on disease levels on leaves from either the duration of fungicide applications or intensity of basal foliage removal. In contrast, on cones, application of fungicides into August had a modest, suppressive effect on powdery mildew. There was also some evidence that the level of powdery mildew on cones associated with fungicide treatment was influenced by the intensity of basal foliage removal. When fungicide applications ceased in late July, there was a progressive decrease in the incidence of cones with powdery mildew with increasing intensity of basal foliage removal. Removing basal foliage two to three times allowed fungicide applications to be terminated in late July rather than late August without diminishing disease control on cones, yield, or cone quality factors. Thus, this study further establishes that fungicide applications made during the early stages of hop cone development have the strongest effect on suppression of powdery mildew on cones. The additive effect of fungicide applications targeted to the periods of greatest cone susceptibility and canopy management to reduce disease favorability may obviate the need for fungicide applications later in the season. This appears to be a viable strategy in mature hop yards of certain cultivars when disease pressure is not excessively high.

Hop Powdery Mildew Control Through Alteration of Spring Pruning Practices.

Podosphaera macularis, the causal agent of hop powdery mildew, is a recurrent threat to hops in the Pacific Northwest because of the potential to reduce cone yield and quality. Early-season pruning is a common practice in hop production for horticultural reasons. Studies were conducted over a 3-year period in a commercial hop yard to quantify the effect of pruning method and timing on disease development, yield, and cone quality factors. A 4-week delay in pruning reduced the incidence of leaves with powdery mildew from 46 to 10% and cones from 9 to 1%, with the specific effect being season dependent. Pruning using chemical desiccants rather than by mechanical means had similar effects on disease levels on leaves. On cones, though, chemical pruning had a small but significant reduction in the incidence of powdery mildew compared with mechanical pruning. Cone yield, levels of bittering-acids, and color were not negatively affected in any individual year or cumulatively over three seasons when pruning treatments were applied repeatedly to the same plots during the study period. Delayed pruning may offer a low-cost means of reducing both the incidence of powdery mildew and early-season fungicide inputs in certain cultivars.

Meta-Analysis Reveals a Critical Period for Management of Powdery Mildew on Hop Cones.

Results of 28 field trials conducted over a 12-year period investigating management of hop powdery mildew caused by Podosphaera macularis were quantitatively summarized by meta-analysis to compare product efficacy and use patterns by mode of action as defined by Fungicide Resistance Action Committee (FRAC) groups. Availability of original observations enabled individual participant data meta-analysis. Differences in control of powdery mildew on leaves and cones were apparent among fungicide FRAC groups when individual products were evaluated over the course of a growing season. FRAC groups 13, 3, and U13 provided the most efficacious control of powdery mildew on leaves. Percent disease control on cones was influenced by midseason foliar disease and fungicide mode-of-action. FRAC 13 provided significantly better disease control on cones than all other groups except U13, 3, and premixes of 7 with 11. Disease control on leaves was similar when a rotational program of fungicides was used, independent of the modes of action, but improved on cones if FRAC groups 13 and 3 were both included compared with programs consisting of FRAC groups 11 and 3, 11 and 5, or 3 and 5. Disease control on cones was improved from 32 to 52%, on average, when the fungicide quinoxyfen (FRAC 13) was applied at least once during the early stages of cone development, defined in this analysis as 20 July to 10 August, as compared with all other treatments. Efficacy of disease control on cones by quinoxyfen was moderated by and interacted with the incidence of leaves with powdery mildew. Disease control on cones was further improved if two applications of quinoxyfen were made during this period. Collectively, these findings suggest that disease control during juvenile stages of cone development largely influences the success of fungicide programs and point to the critical importance of focusing management efforts during this stage of development, independent of what actual management strategy is employed.

Association of Spring Pruning Practices with Severity of Powdery Mildew and Downy Mildew on Hop.

Downy mildew (caused by Pseudoperonospora humuli) and powdery mildew (caused by Podosphaera macularis) are important diseases of hop in the Pacific Northwest United States, and cultural practices may affect the severity of both diseases. The association of spring pruning quality and timing with severity of downy mildew and powdery mildew was assessed through analysis of survey data collected from commercial hop yards in Oregon and Washington. Among 149 hop yards surveyed, the most common pruning method was chemical desiccation (48% of yards), mechanical pruning (23%), or a combination of these practices (15%). The quality of pruning was assessed using a three-category ordinal scale ("excellent", "moderate", or "poor") based on the amount of foliage remaining on plants following pruning. Excellent pruning quality was attained more often in yards pruned twice (74.6 to 82.1% of yards) versus once (33.8% of yards), independent of pruning method. Seasonal severity of downy mildew in Oregon increased approximately twofold with reduction in pruning quality from excellent to moderate to poor. Pruning quality was not significantly related to levels of powdery mildew on leaves or cones in Oregon. Under more severe disease pressure in Washington, however, seasonal severity of powdery mildew on leaves and the incidence of cones with powdery mildew were significantly greater in yards that had poor pruning compared with excellent pruning. Moreover, yards that had excellent pruning quality received, on average, 1.1 to 1.5 fewer fungicide applications per season for downy mildew or powdery mildew compared with yards that had moderate or poor pruning quality. This savings was associated with delayed initiation of the first application by 7.5 to 14.2 days in yards with excellent pruning quality. Replicated experiments in commercial yards in Oregon quantified the effect of delaying pruning timing 5 to 21 days compared with growers' standard practices on the diseases and yield. Downy mildew suppression by delayed pruning was dependent on cultivar and year of sampling, being significantly reduced fivefold only in 'Willamette' in 2007. Severity of powdery mildew and cone yield was similar between plots that received the delayed or standard pruning timing treatments. Collectively, these studies emphasize that early spring sanitation measures are associated with reduced primary inoculum and are critically important for managing both downy mildew and powdery mildew. A savings of at least one fungicide application per year appears achievable when spring pruning is conducted thoroughly and slightly delayed compared with growers' current practices.

Dynamic thermal time model of cold hardiness for dormant grapevine buds.

BACKGROUND AND AIMS: Grapevine (Vitis spp.) cold hardiness varies dynamically throughout the dormant season, primarily in response to changes in temperature. The development and possible uses of a discrete-dynamic model of bud cold hardiness for three Vitis genotypes are described. METHODS: Iterative methods were used to optimize and evaluate model parameters by minimizing the root mean square error between observed and predicted bud hardiness, using up to 22 years of low-temperature exotherm data. Three grape cultivars were studied: Cabernet Sauvignon, Chardonnay (both V. vinifera) and Concord (V. labruscana). The model uses time steps of 1 d along with the measured daily mean air temperature to calculate the change in bud hardiness, which is then added to the hardiness from the previous day. Cultivar-dependent thermal time thresholds determine whether buds acclimate (gain hardiness) or deacclimate (lose hardiness). KEY RESULTS: The parameterized model predicted bud hardiness for Cabernet Sauvignon and Chardonnay with an r(2) = 0·89 and for Concord with an r(2) = 0·82. Thermal time thresholds and (de-)acclimation rates changed between the early and late dormant season and were cultivar dependent but independent of each other. The timing of these changes was also unique for each cultivar. Concord achieved the greatest mid-winter hardiness but had the highest deacclimation rate, which resulted in rapid loss of hardiness in spring. Cabernet Sauvignon was least hardy, yet maintained its hardiness latest as a result of late transition to eco-dormancy, a high threshold temperature required to induce deacclimation and a low deacclimation rate. CONCLUSIONS: A robust model of grapevine bud cold hardiness was developed that will aid in the anticipation of and response to potential injury from fluctuations in winter temperature and from extreme cold events. The model parameters that produce the best fit also permit insight into dynamic differences in hardiness among genotypes.

A web-based decision support system to enhance IPM programs in Washington tree fruit.

BACKGROUND: Integrated pest management (IPM) decision-making has become more information intensive in Washington State tree crops in response to changes in pesticide availability, the development of new control tactics (such as mating disruption) and the development of new information on pest and natural enemy biology. The time-sensitive nature of the information means that growers must have constant access to a single source of verified information to guide management decisions. RESULTS: The authors developed a decision support system for Washington tree fruit growers that integrates environmental data [140 Washington State University (WSU) stations plus weather forecasts from NOAA], model predictions (ten insects, four diseases and a horticultural model), management recommendations triggered by model status and a pesticide database that provides information on non-target impacts on other pests and natural enemies. A user survey in 2008 found that the user base was providing recommendations for most of the orchards and acreage in the state, and that users estimated the value at $ 16 million per year. CONCLUSIONS: The design of the system facilitates education on a range of time-sensitive topics and will make it possible easily to incorporate other models, new management recommendations or information from new sensors as they are developed.

Persistence of Phenylamide Insensitivity in Pseudoperonospora humuli.

Downy mildew, caused by Pseudoperonospora humuli, is an important disease of hop in most production regions in the northern hemisphere. Insensitivity to phenylamide fungicides was detected in isolates of P. humuli in production regions in Oregon and Idaho in 1992, and these fungicides since have been used on a limited basis. In this study, the prevalence of phenylamide insensitivity among isolates of P. humuli collected from 2005 to 2007 in the northwestern United States was quantified using a leaf disk assay with a discriminating dose (25 µg/ml) of either metalaxyl or mefenoxam with inoculum derived from 201 systemically infected diseased shoots (basal spikes) collected from 6, 10, and 11 hop yards in Idaho, Washington, and Oregon, respectively. A subset of 47 basal spike isolates and 42 monosporic isolates collected from two yards in Idaho and nine yards in Oregon during 2006 and 2007 were assayed using a dilution series of metalaxyl to determine the effective dose that inhibited 50% of the incidence of sporulation (ED50). Insensitivity to mefenoxam was detected in 31 of 74 (41.9%) basal spikes collected from 13 hop yards. Insensitivity to the related compound metalaxyl was detected in 52 of 80 (65%) spikes collected from nine hop yards, including four hop yards in Washington. Log ED50 values ranged from -2.25 to 2.67 for basal spike isolates and -2.27 to 2.98 for monosporic isolates and had a similar distribution. Log ED50 values for monosporic isolates and entire basal spike isolates were significantly associated. However, the slope of the regression line was less than 1, indicating that the log ED50 values obtained from entire basal spike isolates were greater than the corresponding log ED50 values obtained from a monosporic isolate obtained from that spike. This research suggests that insensitivity to phenylamide fungicides is a stable phenotype in P. humuli. Management of downy mildew should rely on measures other than these fungicides in yards or regions where phenylamide insensitivity is prevalent, which is now known to include certain hop yards in Idaho, Oregon, and Washington.

Detection of Erysiphe necator in Air Samples Using the Polymerase Chain Reaction and Species-Specific Primers.

ABSTRACT A polymerase chain reaction (PCR) assay employing species-specific primers was developed to differentiate Erysiphe necator from other powdery mildews common in the northwest United States. DNA was extracted from mycelia, conidia, and/or chasmothecia that were collected from grape leaves with a Burkard cyclonic surface sampler. To differentiate E. necator from other erysiphaeceous fungi, primer pairs Uncin144 and Uncin511 were developed to select unique sequences of the internal transcribed spacer regions of E. necator. Using these primers in PCR amplifications, a 367-bp amplicon specific to E. necator was generated, but no amplicons were generated from other erysiphaceous species collected from 48 disparate hosts representing 26 vascular plant families. The PCR limit of detection was one to five conidia of E. necator placed directly into reaction mixtures or 100 to 250 conidia placed on glass rods coated with silicon grease. During field studies, this PCR assay facilitated the detection of E. necator inoculum in air samples within hours of sample rod collection and prior to disease onset. Amplification of E. necator DNA did not occur when the PCR assay was conducted on vineyard air samples collected while grapes were dormant or during periods when vine growth occurred but E. necator remained dormant. The initial PCR detection of E. necator of the season occurred during seasonal ascospore releases caused by precipitation events between bud burst and the prebloom period during the 3 years of the study. Detection ceased for 7 to 11 days following ascospore release and then resumed several days prior to the observance of microscopic symptoms and signs of powdery mildew in the field. Results of this study represent the initial step toward the goal of incorporating an inoculum availability component into current and future grapevine powdery mildew risk assessment models.

Population Biology of Pseudoperonospora humuli in Oregon and Washington.

Pseudoperonospora humuli populations from Oregon and Washington were analyzed for genetic variation using random amplified polymorphic DNA (RAPD) and DNA amplification fingerprinting (DAF) markers. The genetic structure of the Oregon and Washington populations differed considerably. There was little genetic diversity in Washington, with only five RAPD and six DAF groups detected among 40 isolates tested. One genotype was predominant in Washing-ton. In contrast, 18 RAPD and 34 DAF groups were found among the 40 isolates tested from Oregon. No unique band profile associated with host cultivar was observed. It is suggested that the distinct difference in population structure between the two geographic regions might be due to climatic differences resulting in a higher frequency of sexual reproduction of P. humuli in Oregon than in Washington.

Production and Dispersal of Conidia of Leucostoma cinctum in Peach and Cherry Orchards Under Irrigation in Eastern Washington.

The objectives of this study were to determine the seasonal production patterns of Leucostoma cinctum conidia in peach and cherry orchards and the effects of irrigation on dispersal of conidia. Conidia were trapped throughout the year and were present in highest numbers during spring and summer on both cherry and peach. The pathogen was dispersed 1.0 m by water drops that simulated irrigation drops that would impact cankers during over-the-canopy irrigation. Spore production and dispersal also resulted from the use of over-the-canopy and under-tree impact sprinklers. Although the two irrigation methods were not directly compared, the latter mode dispersed propagules of L. cinctum over horizontal distances of about 6 m, a distance sufficient to account for the movement of the pathogen between trees within and across orchard rows. It is concluded that disease management could be improved by alteration of horticultural and irrigation practices.

Meteorological Factors Affecting Airborne Conidia Concentrations and the Latent Period of Podosphaera clandestina on Sweet Cherry.

Conidia of Podosphaera clandestina were trapped beginning about 1 month after bud burst and continuing through the majority of the fruit development period and beyond. The presence of airborne conidia generally followed a diurnal pattern, with peak concentrations occurring in late morning to early afternoon. The three general spore dispersal patterns described by Sutton and Jones for P. leucotricha were observed: diurnal dispersal with concentrations peaking about midday (type I dispersal); random dispersal in which the spore concentrations were not correlated with any measured meteorological parameter (type II dispersal); and dispersal associated with the onset of rain (type III dispersal). Positive correlations with wind speed and temperature, and negative correlations with relative humidity, were characteristic of days when spore dispersal was diurnal. Consistent correlations with meteorological parameters were not detected on days with random or rain-onset dispersal. The latent period of P. clandestina on cherry foliage was found to be shortest at 20 to 25°C and was influenced by temperature and incubation time.