scholarly journals Downy Mildew from Lake Erie Vineyards is Diverse for the G143A SNP Conferring Resistance to Quinone Outside Inhibitor Fungicides

2013 ◽  
Vol 14 (1) ◽  
pp. 23
Author(s):  
Christopher T. Gee ◽  
Stephanie Chestnut ◽  
Eilene Duberow ◽  
Andrea Collins ◽  
Michael A. Shields

Downy mildew (Plasmopara viticola) is a significant problem in grape vineyards throughout the growing season. Control of downy mildew is carried out with a combination of host tolerance and chemical applications. Even in vineyards planted with very tolerant varieties (e.g., Concord), control is important in years with ideal pathogen conditions. Fungicides with a single mode of action possess a very high potential for the development of resistance. Resistance has been observed often in the Quinone outside inhibitor (QoI) fungicides, such as strobilurins. We ascertained the levels of QoI resistance in downy mildew colonies on diseased leaves using CAPS-PCR to detect the glycine to alanine mutation (G143A) known to confer a qualitative level of resistance in fungal pathogens. Our data uncovered a small percentage of samples that contain G143A, suggesting an overall low level of QoI resistance. The low prevalence of the resistant single nucleotide polymorphism (SNP) suggests that QoI fungicides should remain a viable control mechanism in Lake Erie vineyards. Additionally, it appears that a viticultural region where tolerant hosts predominant and QoI use is minimal, resistance buildup in the pathogen population will be minimal. Accepted for publication 15 January 2013. Published 22 April 2013.

2020 ◽  
Vol 21 (4) ◽  
pp. 256-261
Author(s):  
Sarah E. Campbell ◽  
Phillip M. Brannen ◽  
Harald Scherm ◽  
Marin T. Brewer

Grapevine downy mildew, caused by Plasmopara viticola, is among the most damaging diseases of grapes globally and in the viticultural regions of Georgia (U.S.A.). Although management of this disease typically involves fungicide applications, resistance development in P. viticola can render chemical management ineffective. The objective of this study was to survey fungicide sensitivities of P. viticola populations in vineyards across three regions in Georgia. Samples were collected from eight vineyards in 2017 and four in 2018 and tested phenotypically for sensitivity to quinone outside inhibitor (QoI), carboxylic acid amide (CAA), and phenylamide (PA) fungicides using leaf disc bioassays. In addition, DNA was extracted from P. viticola samples collected from 2015 through 2018 in these 12 vineyards and six others for a total of 18 vineyards sampled. All available DNA samples were tested for the presence of the known resistance-causing mutations G143A (QoI) and G1105S (CAA) using polymerase chain reaction (PCR). This study documented widespread occurrence of QoI resistance in P. viticola in Georgia, with 83.0% of isolates collected in 2017 and 2018 testing positive for QoI resistance based on bioassay and 82.9% of isolates collected from 2015 to 2018 testing positive for QoI resistance through PCR testing. In total, 94.4% (17/18) of surveyed vineyards had confirmed QoI resistance by the conclusion of the survey period. No reduced sensitivity to CAA or PA fungicides was identified. Due to widespread QoI resistance, these fungicides should not be relied upon for downy mildew management in Georgia vineyards with a long history of their use.


Plant Disease ◽  
2014 ◽  
Vol 98 (2) ◽  
pp. 276-276 ◽  
Author(s):  
N. A. Ward Gauthier ◽  
B. Amsden

Fungicides in the quinone outside inhibitor class (QoI, also referred to as strobilurins, FRAC group 11) are relied upon for management of a wide range of diseases, including anthracnose, black rot, downy mildew, and powdery mildew on grape. In June 2012, a grape grower from Anderson County in central Kentucky reported a planting of grapevines (Vitis vinifera cv. Vidal Blanc) with 90% downy mildew (Plasmopara viticola) incidence that would not respond to applications of the QoI fungicide Abound 2.08F (22.9% azoxystrobin, Syngenta Crop) or the QoI-containing fungicide Pristine (12.8% pyraclostrobin + 25.2% boscalid, BASF Corporation). Symptoms included yellow irregular lesions or brownish red angular lesions with necrosis on upper sides of leaves. Undersides of leaves contained dense white sporulation. The grower confirmed usage of 4 to 5 applications each of Abound and Pristine fungicides between 2011 and 2012, which exceeded the maximum number of applications allowed per season, as expressed on individual labels. Samples were collected from throughout the 0.4-ha vineyard, and the pathogen was identified as Plasmopara viticola based on morphology of sporangia and sporangiophores (2). Fungicide sensitivity was determined using methods published by Wong and Wilcox (3). Leaves were selected from the 3rd to 6th leaf position from disease-free plants. Nine-millimeter leaf discs were surface disinfested and treated with fungicide concentrations ranging from one-half of the lowest labeled rate to twice the highest label rate (850, 170, 270, and 540 mg/l azoxystrobin and 40, 80, 120, and 240 mg/l pyraclostrobin). Leaf discs were inoculated by placing 10-μl droplets of sporangial suspensions (1 × 104 sporangia per ml) and then incubated at room temperature (22 to 24°C) under fluorescent lights with a 12-h photoperiod (1,3). Dense white fungal growth developed within 10 days; discs treated with water did not develop signs of disease. Fungicide sensitivity was evaluated by determination of the effective concentration (EC50) (3). Leaf discs were examined under a dissecting microscope after 14 days to determine presence of fungal growth; those with visible sporangia and/or sporangiophores were considered diseased. Resulting EC50 concentrations were 420 and 390 mg a.i./l for Abound (azoxystrobin) and Pristine (pyraclostrobin), respectively. This was higher than EC50 ranges of resistant isolates reported by Baudoin et al. (100 and 25 mg/l for azoxystrobin and pyraclostrobin, respectively) (1). Additionally, ranges were higher than EC50 values of isolates not exposed to QoI fungicides reported by Baudoin et al. (1) and higher (14× and 39×, respectively) than those previously determined from P. viticola from Kentucky that were not exposed to QoI fungicides (Ward, unpublished). These EC50 ranges were also much higher than recommended label application rates, which ranged from 170 to 270 mg a.i./l for Abound and from 80 to 120 mg a.i./l for Pristine. Results indicated that P. viticola from this vineyard became insensitive to the fungicides Abound and Pristine. This will lead to future fungicide failures and increased incidences of downy mildew in vineyards. Although QoI-resistant P. viticola has been reported in Europe and elsewhere in the United States, this is the first documented report of QoI-resistant P. viticola in Kentucky. A complete survey is necessary to determine whether this phenomenon is widespread within the state. References: (1) A. Baudoin et al. Plant Health Progress doi:10.1094/PHP-2008-0211-02-RS, 2008. (2) R. C. Pearson and A. C. Goheen, eds. Compendium of Grape Diseases, 4th ed. The American Phytopathological Society, St. Paul, MN, 1998. (3) F. P. Wong and W. F. Wilcox. Plant Dis. 84: 275, 2000.


2021 ◽  
Vol 12 ◽  
Author(s):  
Demetrio Marcianò ◽  
Valentina Ricciardi ◽  
Elena Marone Fassolo ◽  
Alessandro Passera ◽  
Piero Attilio Bianco ◽  
...  

Downy mildew, caused by the oomycete Plasmopara viticola, is one of the diseases causing the most severe economic losses to grapevine (Vitis vinifera) production. To date, the application of fungicides is the most efficient method to control the pathogen and the implementation of novel and sustainable disease control methods is a major challenge. RNA interference (RNAi) represents a novel biotechnological tool with a great potential for controlling fungal pathogens. Recently, a candidate susceptibility gene (VviLBDIf7) to downy mildew has been identified in V. vinifera. In this work, the efficacy of RNAi triggered by exogenous double-stranded RNA (dsRNA) in controlling P. viticola infections has been assessed in a highly susceptible grapevine cultivar (Pinot noir) by knocking down VviLBDIf7 gene. The effects of dsRNA treatment on this target gene were assessed by evaluating gene expression, disease severity, and development of vegetative and reproductive structures of P. viticola in the leaf tissues. Furthermore, the effects of dsRNA treatment on off-target (EF1α, GAPDH, PEPC, and PEPCK) and jasmonic acid metabolism (COI1) genes have been evaluated. Exogenous application of dsRNA led to significant reductions both in VviLBDIf7 gene expression, 5 days after the treatment, and in the disease severity when artificial inoculation was carried out 7 days after dsRNA treatments. The pathogen showed clear alterations to both vegetative (hyphae and haustoria) and reproductive structures (sporangiophores) that resulted in stunted growth and reduced sporulation. Treatment with dsRNA showed signatures of systemic activity and no deleterious off-target effects. These results demonstrated the potential of RNAi for silencing susceptibility factors in grapevine as a sustainable strategy for pathogen control, underlying the possibility to adopt this promising biotechnological tool in disease management strategies.


Plant Disease ◽  
2020 ◽  
Vol 104 (3) ◽  
pp. 893-903 ◽  
Author(s):  
Keevan J. MacKenzie ◽  
Katia V. Xavier ◽  
Aimin Wen ◽  
Sujan Timilsina ◽  
Heather M. Adkison ◽  
...  

Target spot of tomato caused by Corynespora cassiicola is one of the most economically destructive diseases of tomato in Florida. A collection of 123 isolates from eight counties in Florida were evaluated for sensitivity to azoxystrobin and fenamidone based on mycelial growth inhibition (MGI), spore germination (SG), detached leaflet assays (DLAs), and sequence-based analysis of the cytochrome b gene (cytb). Cleavage of cytb by restriction enzyme (Fnu4HI) revealed the presence of a mutation conferring a glycine (G) to alanine (A) mutation at amino acid position 143 (G143A) in approximately 90% of the population, correlating with quinone outside inhibitor (QoI) resistance based on MGI (<40% at 5 μg/ml), SG (<50% at 1 and 10 μg/ml), and DLA (<10% severity reduction). The mutation conferring a phenylalanine (F) to leucine (L) substitution at position 129 (F129L) was confirmed in moderately resistant isolates (#9, #19, and #74) based on MGI (40 to 50% at 5 μg/ml), SG (<50% at 1 μg/ml and >50% at 10 μg/ml), and DLA (>10% and <43% severity reduction) for both QoI fungicides, whereas sensitive isolates (#1, #4, #7, #28, #29, #46, #61, #74, #75, #76, #91, #95, and #118) based on MGI (>50% at 5 μg/ml), SG (>50% at 1 μg/ml and 10 μg/ml), and DLA (>50% severity reduction) correlated to non-mutation-containing isolates or those with a silent mutation. This study indicates that QoI resistance among C. cassiicola isolates from tomato is widespread in Florida and validates rapid screening methods using MGI or molecular assays to identify resistant isolates in future studies.


2021 ◽  
Vol 34 ◽  
pp. 02001
Author(s):  
Elena Ilnitskaya ◽  
Marina Makarkina ◽  
Valeriy Petrov

Downy mildew (Plasmopara viticola) and powdery mildew (Erysiphe necator) are the most common and economically significant fungal diseases in vineyards. The task of this work is to study the genotypes of new promising hybrid forms of table grapes for the presence of resistance genes to downy mildew (Rpv10 and Rpv3) and powdery mildew (Ren9) using DNA-markers. The study was carried out on table grape hybrids under the working names Agat dubovskiy, Akelo, Arabella, Artek, Dubovskiy rozovyi, Gamlet, Ispolin, Kishmish dubovskiy, Kurazh, Pestryi, Valensiya and registered variety Liviya. The studied genes were analyzed using markers UDV305 and UDV737 (Rpv3), GF09-46 (Rpv10), CenGen6 (Ren9). The following cultivars were used as reference genotypes: Saperavi severnyi (carries Rpv10 gene) and Regent (Rpv3 and Ren9). It was established that Rpv3 gene is carried by hybrids Kishmish dubovskiy, Agat dubovskiy, Kurazh, Valensiya, Akelo, Gamlet, Dubovskiy rozovyi, Pestryi. Ren9 gene was found in Artek, Agat dubovskiy, Kurazh, Ispolin, Valensiya, Arabella, Gamlet, Dubovskiy rozovyi, Pestryi. The Rpv10 gene was not detected in any of the analyzed grapevine samples. genotypes Agat dubovskiy, Kurazh, Gamlet, Dubovskiy rozovyi, Pestryi, Valensiya carry Rpv3 and Ren9 genes simultaneously. These grapevines have an elegant bunch and large berries that are attractive to consumers.


Plant Disease ◽  
2016 ◽  
Vol 100 (2) ◽  
pp. 337-344 ◽  
Author(s):  
Jeneylyne F. Colcol ◽  
Anton B. Baudoin

The sensitivity of downy mildew (DM, Plasmopara viticola) and powdery mildew (PM, Erysiphe necator) of grape (Vitis sp.) to commonly used nondemethylation inhibitor, single-site fungicides in and near Virginia was determined from 2005 to 2007, with more limited additional sampling in subsequent years. In grape leaf disc bioassays, 92% of the P. viticola isolates were quinone outside inhibitor (QoI, azoxystrobin) resistant but none were resistant to mefenoxam. In all, 82% of the E. necator isolates were QoI resistant. Most of the QoI-resistant P. viticola and E. necator isolates contained >95% of the G143A point mutation, which confers high levels of QoI resistance. In contrast, QoI-sensitive P. viticola isolates contained less than 1% of G143A. In total, 1 of 145 and 14 of 154 QoI-resistant P. viticola and E. necator isolates (able to grow on azoxystrobin concentration ≥1 μg/ml), respectively, contained <1% G143A. In total, 61 E. necator isolates from 23 locations were tested against thiophanate methyl, and the majority grew well on leaf tissue treated with 50 and 250 μg/ml. Through 2012, none of the E. necator isolates were resistant to boscalid and quinoxyfen. However, in 2013, quinoxyfen-resistant E. necator was detected in one vineyard experiencing difficulties with powdery mildew control. No 50% effective concentration value could be calculated but these isolates tolerated labeled rates with only limited inhibition. QoI (E. necator and P. viticola) and benzimidazole (E. necator) resistance were widespread in Virginia, rendering these materials inadvisable for control of these diseases. The practical importance and current distribution of quinoxyfen resistance needs further investigation.


Plant Disease ◽  
2014 ◽  
Vol 98 (1) ◽  
pp. 67-76 ◽  
Author(s):  
Byron Vega ◽  
Megan M. Dewdney

Chemical control, based on copper and quinone outside inhibitor (QoI) fungicides, has been essential for the management of brown spot of citrus, caused by Alternaria alternata. However, QoI control failures were detected recently in Florida. From 2008 to 2012, 817 monoconidial isolates of A. alternata from 46 citrus orchards were examined for sensitivity to azoxystrobin (AZ) and pyraclostrobin (PYR). Of the isolates, 57.6% were resistant to both fungicides, with effective concentration to inhibit 50% growth (EC50) values greater than 5 μg/ml for AZ and 1 μg/ml for PYR. The mean EC50 values for sensitive isolates were 0.139 and 0.020 μg/ml for AZ and PYR, respectively. The EC50 values of both fungicides were highly correlated (P < 0.0001), indicating cross resistance. The proportion of resistant isolates differed significantly (P < 0.0001) among cultivars and with QoI application frequency (P < 0.0001). However, resistance was not significantly related (P = 0.364) to disease severity in the field (low, moderate, and high) or isolate virulence (P = 0.397). The molecular basis for QoI resistance was determined for a subset of 235 isolates using polymerase chain reaction restriction fragment length polymorphism of the cytochrome b gene. All resistant isolates showed the point mutation G143A. Based on the presence of one or two introns, isolates were classified as profile I and profile II, respectively. The resistance frequency was significantly higher (P < 0.0001) in isolate profile II, suggesting a higher selection pressure for resistant population profile II.


Plant Disease ◽  
2014 ◽  
Vol 98 (8) ◽  
pp. 1122-1130 ◽  
Author(s):  
Zachary A. Frederick ◽  
Sara M. Villani ◽  
Daniel R. Cooley ◽  
Alan R. Biggs ◽  
Jessica J. Raes ◽  
...  

Quinone-outside-inhibitor (QoI) fungicides are a safe and effective means of managing apple scab caused by Venturia inaequalis. To determine the prevalence of both quantitative (partial) and qualitative (complete) QoI resistance in V. inaequalis in the northeastern United States, we sampled single-lesion conidial isolates (n = 4,481) from 120 commercial and research orchards from 2004 to 2011 with a range of exposure to QoI fungicides from none to several applications a year. In all, 67% of these orchard populations of V. inaequalis were sensitive to QoI fungicides, 28% exhibited QoI practical resistance, and 5% were not sensitive QoI fungicides but had not become practically resistant. Isolates with qualitative QoI resistance, conferred by the G143A cytochrome b gene mutation, were found in 13 of the 34 QoI-resistant orchard populations. To evaluate the stability of the G143A mutation, 27 isolates were selected from different orchard populations to represent the scope of regional populations. These isolates were subcultured continuously in the presence or absence of the QoI fungicide trifloxystrobin. All isolates that initially possessed qualitative resistance maintained the resistant genotype (G143A) for six transfers over 6 months in both the absence and presence of trifloxystrobin. Given the observed QoI resistance in orchard populations of V. inaequalis and the stability of the G143A mutation in individual isolates, apple scab management paradigms must encompass strategies to limit selection of QoI resistance in the sensitive orchard populations remaining in the region.


Plant Disease ◽  
2012 ◽  
Vol 96 (9) ◽  
pp. 1262-1270 ◽  
Author(s):  
Byron Vega ◽  
Daniele Liberti ◽  
Philip F. Harmon ◽  
Megan M. Dewdney

Chemical management of Alternaria brown spot of citrus is based upon the timely application of site-specific fungicides, many of which are vulnerable to the development of fungicide resistance. A rapid microtiter bioassay based on the colorimetric changes of resazurin (RZ) dye was developed to evaluate the sensitivity of Alternaria alternata to quinone outside inhibitor (QoI) fungicides. Four liquid media (complete medium, minimal medium, potato dextrose broth, and yeast peptone dextrose broth), five conidia concentrations (from 101 to 105 conidia/ ml), and five RZ concentrations (10, 20, 30, 40, and 50 μM) were evaluated. Complete medium at 105 conidia/ml and 40 μM RZ were identified as optimal for measuring RZ reduction. The effective concentration of two QoI fungicides (azoxystrobin and pyraclostrobin) needed to reduce RZ by 50% (EC50) was calculated and compared with those obtained from conidia germination tests on fungicide-amended media. Concordant EC50 values were observed (R2 = 0.923; P < 0.0001) from both methods. Resistant phenotypes were further characterized by the partial sequencing of the cytochrome b gene. Genetic variability associated with the presence or absence of two introns was observed among isolates. The identified resistant isolates had the amino acid substitution G143A, typical of QoI resistance in other fungi.


2008 ◽  
Vol 9 (1) ◽  
pp. 25 ◽  
Author(s):  
Anton Baudoin ◽  
Gilberto Olaya ◽  
François Delmotte ◽  
Jeneylyne F. Colcol ◽  
Helge Sierotzki

Of 20 Plasmopara viticola isolates collected in four locations in Virginia and northwest North Carolina in 2005, 16 were resistant to QoI fungicides. The resistance factor was over 100, and label rates of formulated azoxystrobin and pyraclostrobin provided little or no control of these isolates. Additional sampling in 2006 revealed at least 15 additional vineyards with QoI-resistant P. viticola in Virginia, Maryland, and Pennsylvania. Of 22 isolates of Erysiphe (Uncinula) necator collected in 2005 from five Virginia locations, 20 isolates from 4 locations showed resistance to QoI fungicides. The G143A mutation for resistance was detected in several isolates of both pathogens. This is the first detection of this type of resistance in P. viticola in North America, and the second North American report of QoI resistance in E. necator. Accepted for publication 26 November 2007. Published 11 February 2008.


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