Management of grape powdery mildew with an intelligent sprayer and sulfur

Wine grapes are an important agricultural commodity in the Pacific Northwest where grape powdery mildew (GPM) is one of the main disease problems. The efficacy of different sulfur concentrations and different output volumes from an air blast sprayer retrofitted with the Intelligent Spray System (ISS) were evaluated for the management of GPM. The ISS consists of a LiDAR sensor, Doppler speed sensor, embedded computer, flow controller, and individual pulse-width-modulation solenoid valves at each nozzle. GPM cluster severity ranged from 55% to 75% across all trials in the study when using the ISS at its default spray rate of 62.5 ml m-3 and micronized sulfur at 6 g L-1, which was significantly higher than all other fungicide treatments, but lower than non-treated controls. Similarly, leaf incidence values were highest on non-treated vines, followed by micronized sulfur at 6 g L-1 applied at 62.5 ml m-3 , with all other fungicide treatments being significantly lower in all trials. Using the ISS at the 62.5 ml m-3 rate and a rotation of locally systemic fungicides resulted in the lowest observed GPM leaf incidence, and average cluster severity of 11% in both 2019 and 2020, the lowest cluster severity of all fungicide treatments tested. GPM control using the ISS and micronized sulfur was equivalent to a constant-rate air blast treatment at 6 g L-1 when the spray rate of the ISS was increased to 125ml m-3, or if the concentration of sulfur was increased to 24 g L-1. In those cases, the amount of sulfur applied to vines was at or above the minimum label rate from bloom until the end of the season, or the entire season, respectively. This study has shown that sufficient disease control cannot always be expected when mixing pesticides at the same rate as would be used for a constant-rate sprayer in a variable rate sprayer, especially when using contact fungicides like sulfur . With appropriate adjustments, the variable-rate ISS can be a useful tool to reduce pesticide quantities, water required for mixing, and as a result labor, as fewer trips to refill for a given spray event are required.

Characterization of the grape powdery mildew genetic resistance loci in Muscadinia rotundifolia Trayshed

Muscadinia rotundifolia cv. Trayshed is a valuable source of resistance to grape powdery mildew (PM). It carries two PM resistance (R) loci, Run1.2 on chromosome 12 and Run2.2 on chromosome 18. This study identified the nucleotide-binding leucine-rich repeat (NLR) genes composing each R locus and their associated defense mechanisms. Evaluation of PM disease development showed that introgression of both loci confers resistance to PM in a V. vinifera background, but with varying speed and intensity of the response. To better understand the effect of NLR composition on PM resistance, both haplotypes of each R locus were reconstructed and the gene models within each haplotype were manually refined. We found that the number and classes of NLR genes differed between Run1.2 and Run2.2 loci and between the haplotypes of each R locus. In addition, NLR genes composing Run1.2b or Run2.2 loci exhibited different levels of gene expression, pointing to candidate NLR genes responsible for PM resistance in Trayshed. Finally, a transcriptomic analysis that included six additional R loci showed differences in the defense mechanisms associated with Run1.2b and Run2.2 in response to PM and at constitutive level. Altogether, our results reveal that Trayshed's R loci are composed of distinct NLRs that trigger different plant defense mechanisms in response to PM and at constitutive level, which would explain the variation of pathogen restriction between the two loci.

The mitochondrial genome of the grape powdery mildew pathogen Erysiphe necator is intron rich and exhibits a distinct gene organization

Powdery mildews are notorious fungal plant pathogens but only limited information exists on their genomes. Here we present the mitochondrial genome of the grape powdery mildew fungus Erysiphe necator and a high-quality mitochondrial gene annotation generated through cloning and Sanger sequencing of full-length cDNA clones. The E. necator mitochondrial genome consists of a circular DNA sequence of 188,577 bp that harbors a core set of 14 protein-coding genes that are typically present in fungal mitochondrial genomes, along with genes encoding the small and large ribosomal subunits, a ribosomal protein S3, and 25 mitochondrial-encoded transfer RNAs (mt-tRNAs). Interestingly, it also exhibits a distinct gene organization with atypical bicistronic-like expression of the nad4L/nad5 and atp6/nad3 gene pairs, and contains a large number of 70 introns, making it one of the richest in introns mitochondrial genomes among fungi. Sixty-four intronic ORFs were also found, most of which encoded homing endonucleases of the LAGLIDADG or GIY-YIG families. Further comparative analysis of five E. necator isolates revealed 203 polymorphic sites, but only five were located within exons of the core mitochondrial genes. These results provide insights into the organization of mitochondrial genomes of powdery mildews and represent valuable resources for population genetic and evolutionary studies.

Effect of Fungicide Mobility and Application Timing on the Management of Grape Powdery Mildew

Grape powdery mildew (GPM) fungicide programs consist of 5 to 15 applications, depending on region or market, in an attempt to achieve the high fruit quality standards demanded by the market. Understanding how fungicides redistribute and targeting redistributing fungicide to critical crop phenological stages could improve fungicide protection of grape clusters. This study evaluated fungicide redistribution in grapevines from major fungicide groups labeled for GPM control. Translaminar and xylem redistribution was examined by placing fungicide-impregnated filter disks on the adaxial or abaxial leaf surface of detached leaves for 10 min and then incubating for 48 h before inoculating the abaxial surface with conidia. Vapor redistribution used Teflon disks sprayed with fungicides and placed on the abaxial leaf surface of detached leaves 48 h before inoculation. Disease development was rated 10 days later. Translaminar movement through calyptra was tested using flowering potted vines. All fungicides tested redistributed through at least one mechanism. Fungicide timing at critical phenological stages (early, mid, and late bloom) was assessed in small plots of cultivar Pinot noir vines. The application of trifloxystrobin, quinoxyfen, or fluopyram at different bloom stages showed that applications initiated at end of bloom resulted in the lowest berry infection probabilities of 0.073, 0.097, and 0.020, respectively. The results of this study suggest that integrating two carefully timed applications of redistributing fungicides initiated at end of bloom into a fungicide program may be an effective strategy for wine grape growers in western Oregon to produce fruit with low GPM infection.

Evaluation of Quinoxyfen Resistance of Erysiphe necator (Grape Powdery Mildew) in a Single Virginia Vineyard

The protectant fungicide quinoxyfen has been used against grape powdery mildew (Erysiphe necator) in the United States since 2003. In 2013, isolates of grape powdery mildew with reduced quinoxyfen sensitivity (here designated as quinoxyfen lab resistance or QLR) were detected in a single vineyard in western Virginia, USA. Field trials were conducted in 2014, 2015, and 2016 at the affected vineyard to determine to what extent quinoxyfen might still contribute to disease control. Powdery mildew control by quinoxyfen was similar to, or only slightly less than, that provided by myclobutanil and boscalid in all three years. In 2016, early- versus late-season applications of quinoxyfen were compared to test the hypothesis that early-season applications were more effective, but differences were small. A treatment with two early quinoxyfen applications, at bloom and 2 weeks later, followed by a myclobutanil-boscalid plus a low dose of sulfur rotation provided slightly better control of foliar disease incidence than treatments containing four quinoxyfen applications or two midseason or two late quinoxyfen applications supplemented by myclobutanil and boscalid applications; severity differences were small and nonsignificant. Metrafenone and benzovindiflupyr generally provided excellent powdery mildew control. The frequency of QLR in vines not treated with quinoxyfen slowly declined from 65% in 2014 to 46% in 2016. Further research is needed to explain how, despite this QLR frequency, quinoxyfen applied to grapes in the field was still able to effectively control powdery mildew.

Development of a quantitative loop-mediated isothermal amplification assay for the field detection ofErysiphe necator

Plant pathogen detection systems have been useful tools to monitor inoculum presence and initiate management schedules. More recently, a loop-mediated isothermal amplification (LAMP) assay was successfully designed for field use in the grape powdery mildew pathosystem; however, false negatives or false positives were prevalent in grower-conducted assays due to the difficulty in perceiving the magnesium pyrophosphate precipitate at low DNA concentrations. A quantitative LAMP (qLAMP) assay using a fluorescence resonance energy transfer-based probe was assessed by grape growers in the Willamette Valley of Oregon. Custom impaction spore samplers were placed at a research vineyard and six commercial vineyard locations, and were tested bi-weekly by the lab and by growers. Grower-conducted qLAMP assays used a beta-version of the Smart-DART handheld LAMP reaction devices (Diagenetix, Inc., Honolulu, HI, USA), connected to Android 4.4 enabled, Bluetooth-capable Nexus 7 tablets for output. Quantification by a quantitative PCR assay was assumed correct to compare the lab and grower qLAMP assay quantification. Growers were able to conduct and interpret qLAMP results; however, theErysiphe necatorinoculum quantification was unreliable using the beta-Smart-DART devices. The qLAMP assay developed was sensitive to one spore in early testing of the assay, but decreased to >20 spores by the end of the trial. The qLAMP assay is not likely a suitable management tool for grape powdery mildew due to losses in sensitivity and decreasing costs and portability for other, more reliable molecular tools.