scholarly journals Baseline Sensitivity and Cross-Resistance to Succinate-Dehydrogenase-Inhibiting and Demethylation-Inhibiting Fungicides in Didymella bryoniae

Plant Disease ◽  
2012 ◽  
Vol 96 (7) ◽  
pp. 979-984 ◽  
Author(s):  
A. Thomas ◽  
D. B. Langston ◽  
K. L. Stevenson

Didymella bryoniae, which causes gummy stem blight (GSB) of watermelon, has a history of developing resistance to fungicides, most recently the succinate-dehydrogenase-inhibiting (SDHI) fungicide boscalid. To facilitate fungicide resistance monitoring, baseline sensitivity distributions were established for demethylation-inhibiting (DMI) fungicides tebuconazole and difenoconazole and the SDHI fungicide penthiopyrad, and reestablished for the SDHI fungicide boscalid. In all, 71 isolates with no known prior exposure to SDHIs or DMIs were used to determine the effective concentration at which mycelial growth was inhibited by 50% (EC50). EC50 values for boscalid, penthiopyrad, tebuconazole, and difenoconazole were 0.018 to 0.064, 0.015 to 0.057, 0.062 to 0.385, and 0.018 to 0.048 μg/ml, with median values of 0.032, 0.026, 0.118, and 0.031 μg/ml, respectively. Significant positive correlations between the sensitivity to penthiopyrad and boscalid (P < 0.0001, r = 0.75) and between tebuconazole and difenoconazole (P < 0.0001, r = 0.59) indicate a potential for cross-resistance between chemically related fungicides. In 2009, 103 isolates from fungicidetreated watermelon fields were tested for sensitivity to boscalid and penthiopyrad using a discriminatory concentration of 3.0 μg/ml. Of the isolates tested, 82 were insensitive and 14 were sensitive to both fungicides. Because of the significant potential for cross-resistance between closely related fungicides, growers will be advised not to use both SDHIs or both DMIs successively in the same fungicide spray program.

Plant Disease ◽  
1997 ◽  
Vol 81 (2) ◽  
pp. 163-166 ◽  
Author(s):  
Katherine L. Reynolds ◽  
Timothy B. Brenneman ◽  
Paul F. Bertrand

Monoconidial isolates of the pecan scab fungus, Cladosporium caryigenum, were obtained in 1993 and 1994 from one pecan orchard each in Jeff Davis and Troup counties in Georgia, counties with no previous history of exposure to demethylation-inhibiting (DMI) fungicides. Isolates were grown on potato dextrose agar (PDA) amended with propiconazole or fenbuconazole at 0,0.0001, 0.0005, 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1.0, or 5.0 µg ml-1. After 4 to 5 weeks at 25°C, the diameter of each colony was measured. Relative growth was expressed as the colony diameter for each fungicide concentration as a percentage of the diameter on unamended PDA. (ED)50 values for both fungicides in all years and locations were lognormally distributed. Mean (ED)50 values ranged from 0.19 to 0.30 µg ml-1 for fenbuconazole and 0.12 to 0.17 µg ml-1 for propiconazole. There were highly significant positive correlations between sensitivity to propiconazole and sensitivity to fenbuconazole at all locations and years, indicating significant potential for development of cross-resistance to these compounds. A discriminatory concentration of 0.2 µg ml-1 propiconazole was selected for further monitoring of DMI sensitivity. With this discriminatory concentration, a sample size of 50 isolates was estimated to be sufficient to detect a difference of 8.3% in mean relative growth between two populations.


Plant Disease ◽  
2010 ◽  
Vol 94 (5) ◽  
pp. 604-612 ◽  
Author(s):  
Y. K. Kim ◽  
C. L. Xiao

Gray mold caused by Botrytis cinerea is a major postharvest disease of apple. Pristine, a formulated mixture of pyraclostrobin and boscalid, was recently registered for use on apple. Pristine applied within 2 weeks before harvest is effective in controlling gray mold in stored apple fruit. To determine the baseline sensitivity of B. cinerea populations to these fungicides, 40 isolates from organic and 80 from conventional apple orchards where Pristine had not been used were tested for mycelial growth or conidial germination on fungicide-amended media. To monitor fungicide resistance, gray-mold-decayed apple fruit originating from orchards in which Pristine had been used were sampled from a fruit packinghouse. Isolates of B. cinerea recovered from the fruit were tested for resistance to the two fungicides. In the in vivo study in the orchards, Pristine was applied to fruit 1 day before harvest. Fruit were then harvested, wounded, and inoculated with isolates exhibiting different fungicide-resistance phenotypes. Fruit were stored at 0°C for 8 weeks for decay development. The effective concentration that inhibits mycelial growth by 50% relative to the control (EC50) values for sensitive isolates ranged from 0.008 to 0.132 μg/ml (mean = 0.043, n = 116) for pyraclostrobin and from 0.003 to 0.183 μg/ml (mean = 0.075, n = 117) for Pristine in a mycelial growth assay on potato dextrose agar. The EC50 values of boscalid for sensitive isolates ranged from 0.065 to 1.538 μg/ml (mean = 0.631, n = 29) in a conidial germination assay on water agar. Four isolates were resistant to pyraclostrobin, with resistance factors (RFs) ranging from 12 to 4,193. Of the four pyraclostrobin-resistant isolates, one also was resistant to boscalid (RF = 14) and Pristine (RF = 373), and two exhibited reduced sensitivity to Pristine (RF = 16 and 17). The minimum inhibitory concentration for conidial germination (for boscalid) or mycelial growth (for pyraclostrobin and Pristine) of sensitive isolates was 5 μg/ml, which is thus recommended as a discriminatory concentration for phenotyping isolates for resistance to these fungicides. Of the 56 isolates obtained from decayed apple fruit that had been exposed to Pristine, 11 (approximately 20%) were resistant to both pyraclostrobin and boscalid and 1 was resistant only to pyraclostrobin. Of the additional 43 isolates obtained from decayed apple fruit originating from an organic orchard, 3 were resistant only to pyraclostrobin, 2 were resistant only to boscalid, and 2 were resistant to both fungicides. It appeared that there was no cross resistance between pyraclostrobin and boscalid because of the existence of isolates resistant only to either pyraclostrobin or boscalid. Pristine applied at label rate in the orchard failed to control gray mold on apple fruit inoculated with the Pristine-resistant isolates. This is the first report of multiple resistance to pyraclostrobin, boscalid, and Pristine in field populations of B. cinerea. Our results suggest that the development of dual resistance to pyraclostrobin and boscalid in B. cinerea populations could result in the failure to control gray mold with Pristine.


Plant Disease ◽  
2020 ◽  
pp. PDIS-03-20-0508
Author(s):  
Can Zhang ◽  
Muhammad Imran ◽  
Lu Xiao ◽  
Zhihong Hu ◽  
Guixiang Li ◽  
...  

Gray mold caused by Botrytis cinerea is one of the most important diseases in tomato. It can be controlled effectively by demethylation inhibitor (DMI) fungicides, but their resistance status after long-term use in the field is unclear. The baseline sensitivity to difenoconazole of 142 B. cinerea isolates from China with no history of DMI usage was characterized, with a mean effective concentration for 50% mycelial growth inhibition (EC50) of 0.97 ± 0.50 μg/ml. EC50 values for difenoconazole sensitivity of another 248 isolates collected in 2011 and 2016 ranged from 0.04 to 11.99 μg/ml, and the frequency of difenoconazole sensitivity formed a nonnormal distribution curve. Detached fruit studies revealed that isolates with EC50 values of approximately 6.00 μg/ml were not controlled effectively. The mean EC50 of the resistant isolates changed from 6.74 to 8.65 μg/ml between 2011 and 2016. Positive cross-resistance was only observed between difenoconazole and two DMIs. One dual resistant isolate and one triple resistant isolate were found among the difenoconazole-resistant isolates collected in 2016, associated with point mutations in corresponding target proteins of the fungicides azoxystrobin and fludioxonil. This indicated that B. cinerea not only showed higher difenoconazole resistance levels but gradually changed from single to multiple fungicide resistance over time. No amino acid variation was found in the CYP51 protein. In the absence of difenoconazole, the relative expression of CYP51 was not significantly different in sensitive and resistant isolates. Induced expression of CYP51 is an important determinant of DMI resistance in B. cinerea from tomato. However, nucleotide variants found in the upstream region had no association with the fungicide resistance phenotype. These results will be helpful for the management of B. cinerea in the field.


Plant Disease ◽  
2021 ◽  
Author(s):  
Hafiz Muhammad Usman ◽  
Qin Tan ◽  
Mohammad Mazharul Karim ◽  
Muhammad Adnan ◽  
Weixiao Yin ◽  
...  

Anthracnose, mainly caused by Colletotrichum gloeosporioides species complex including C. fructicola and C. siamense, is a devastating disease of peach. The chemical control has been widely used for years and management failures have increased towards commonly used fungicides. Therefore, screening of sensitivity of Colletotrichum spp. to fungicides with different modes of action is needed to make proper management strategies for peach anthracnose. In this study, sensitivity of 80 isolates of C. fructicola and C. siamense was screened for pyraclostrobin, procymidone, prochloraz and fludioxonil based on mycelial growth inhibition at discriminatory doses. Results showed that C. fructicola and C. siamense isolates were highly resistant to procymidone and fludioxonil with 100% resistance frequencies to both fungicides, but sensitive to prochloraz, i.e., no resistant isolates were found. For pyraclostrobin, 74% of C. fructicola isolates showed high resistance and 26 % were low resistant, all of the C. siamense isolates were low resistant. No positive cross-resistance was observed between pyraclostrobin and azoxystrobin, even they are members of the same quinone outside inhibitor (QoI) fungicide group, and between pyraclostrobin and non-QoIs. Resistant isolates to QoI fungicides were evaluated for the fitness penalty. Results showed that no significant differences except for mycelial growth rates were detected between highly resistant and low-resistant isolates of C. fructicola. Molecular characterization of Cyt b gene revealed that the G143A point mutation was the determinant of the high resistance in C. fructicola. This study demonstrated the current resistance status of C. fructicola and C. siamense to different fungicides and their future perspectives. Demethylation inhibitor (DMI) fungicides are the best option among different chemicals to control peach anthracnose in China.


Plant Disease ◽  
2021 ◽  
Author(s):  
Gangming Zhan ◽  
Fan Ji ◽  
Jun Zhao ◽  
Yue Liu ◽  
Aihong Zhou ◽  
...  

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is a destructive disease of wheat that seriously threatens production safety in wheat-producing areas worldwide. In China, the disease has been largely controlled with fungicide triadimefon. Although high levels of fungicide resistance in other fungal pathogens have been reported, failure to control Pst with any fungicides has seldomly been reported and fungicide sensitivity of Pst has not been evaluated in China. The distribution of triadimefon-resistant Pst isolates was investigated in the present study. The baseline sensitivity of 446 Pst isolates across the country to triadimefon was determined, and the concentration for 50% of maximal effect (EC50) showed a unimodal distribution curve, with a mean value of 0.19 μg mL-1. The results indicated a wide range of sensitivity to triadimefon, with more insensitive isolates collected from Pst winter-increasing areas and northwest over-summering areas, whereas more sensitive isolates were collected from southwest over-summering areas and epidemic areas of Xinjiang and Tibet. The majority of the tested Pst isolates were sensitive to triadimefon; only 6.79% had developed varying degrees of resistance. Characterization of parasitic fitness revealed that the triadimefon-resistant isolates exhibited strong adaptive traits in urediniospore germination rate, latent period, sporulation intensity, and lesion expansion rate. Positive cross-resistance was observed between triadimefon and tebuconazole or hexaconazole, but not between pyraclostrobin or flubeneteram. The point mutation Y134F in the 14α-demethylase enzyme (CYP51) was detected in triadimefon-resistant isolates. A molecular method (Kompetitive Allele Specific PCR) was established for the rapid detection of Y134F mutants in the Pst population. Two genotypes with one point mutation Y134F conferred resistance to triadimefon in Pst. The risk of resistance to triadimefon in Pst may be low to moderate. This study provided important data for establishment of high throughput molecular detection methods, fungicide resistance risk management, and the development of new target fungicides.


2019 ◽  
Vol 109 (12) ◽  
pp. 2096-2106 ◽  
Author(s):  
Qin Peng ◽  
Zhiwen Wang ◽  
Yuan Fang ◽  
Weizhen Wang ◽  
Xingkai Cheng ◽  
...  

Ethaboxam is a β-tubulin inhibitor registered for the control of oomycete pathogens. The current study was established to determine the ethaboxam sensitivity of the plant pathogen Phytophthora sojae and investigate the potential for the emergence of fungicide resistance. The effective concentration for 50% inhibition (EC50) of 112 Phytophthora sojae isolates exhibited a unimodal distribution with a mean EC50 for ethaboxam of 0.033 µg/ml. Establishing this baseline sensitivity provided critical data for monitoring changes in ethaboxam-sensitivity in field populations. The potential for fungicide resistance was investigated using adaptation on ethaboxam-amended V8 agar, which resulted in the isolation of 20 resistant mutants. An assessment of the biological characteristics of the mutants including mycelial growth, sporulation, germination rate and pathogenicity indicated that the resistance risk in Phytophthora sojae was low to medium with no cross-resistance between ethaboxam and cymoxanil, metalaxyl, flumorph, and oxathiapiprolin being detected. However, positive cross-resistance was found between ethaboxam and zoxamide for Q8L and I258V but negative cross-resistance for C165Y. Further investigation revealed that the ethaboxam-resistant mutants had point mutations at amino acids Q8L, C165Y, or I258V of their β-tubulin protein sequences. CRISPR/Cas9-mediated transformation experiments confirmed that the Q8L, C165Y, or I258V mutations could confer ethaboxam resistance in Phytophthora sojae and that the C165Y mutation induces high levels of resistance. Taken together, the results of the study provide essential data for monitoring the emergence of resistance and resistance management strategies for ethaboxam, as well as for improving the design of novel β-tubulin inhibitors for future development.


Plant Disease ◽  
2020 ◽  
Author(s):  
Shengming Liu ◽  
Liuyuan Fu ◽  
Huanhuan Tan ◽  
Jia Jiang ◽  
Zhiping Che ◽  
...  

Grey mold, caused by the fungus Botrytis cinerea Pers ex Fr., is one of the most destructive spoilage diseases, severely affecting tomato production in Henan Province, China. Spraying fungicides from the flowering to the harvest stage is a necessary measure to reduce losses associated with B. cinerea infection. However, B. cinerea has developed resistance to fungicides in many countries. Boscalid is a succinate dehydrogenase inhibitor (SDHI) fungicide, and was registered for the control of grey mold. In this study, a total of 269 B. cinerea isolates were collected from tomato in commercial greenhouses in different locations of Henan Province, in 2014 and 2015. The sensitivity and resistance of B. cinerea field isolates were determined based on mycelial growth. The effective concentration 50 (EC50) ranged from 0.11 to 15.92 μg ml−1 and 0.16 to 8.54 μg ml−1, in 2014 and 2015, respectively. The frequency of low resistance to boscalid was 12.6% and 7.6%, and moderate resistance were 2.7% and 1.3%, in 2014 and 2015, respectively. No high-resistant isolates were found in Henan Province, China. Mycelial growth, mycelial dry weight, spore production, and pathogenicity were not significantly different between resistant and sensitive phenotypes of the B. cinerea isolates. The results of cross-resistance test showed no correlation between boscalid and carbendazim, procymidone, pyrimethanil, fluazinam or fluopyram. In this study, the succinate dehydrogenase gene B (sdhB), C (sdhC), and D (sdhD) were analyzed and compared in sensitive, low and moderately resistant B. cinerea isolates to boscalid. Results showed point mutations occurred simultaneously at sdhC amino acid positions 85 (G85A), 93 (I93V), 158 (M158V), and 168 (V168I) in 4 out of 10 sensitive isolates, 23 out of 26 low and 5 out of 5 moderately resistant B. cinerea isolates to boscalid. No point mutations were found in the sdhB and sdhD genes of all isolates. Furthermore, no point mutations were found in sdhB, sdhC and sdhD genes in 3 out of 26 low resistant B. cinerea isolates to boscalid. Therefore, we speculate the simultaneous point mutations in the sdhC gene may not be related to the resistance of B. cinerea to boscalid. These results suggested that there might be a substitution mechanism for the resistance of B. cinerea to the SDHI fungicide boscalid.


2011 ◽  
Vol 101 (9) ◽  
pp. 1104-1111 ◽  
Author(s):  
Yang Bi ◽  
Xiaolan Cui ◽  
Xiaohong Lu ◽  
Meng Cai ◽  
Xili Liu ◽  
...  

Laboratory experiments were conducted to determine the baseline sensitivity of Phytophthora capsici and its risk for developing resistance to zoxamide. In total, 158 P. capsici isolates were collected from China. All 158 isolates were sensitive to zoxamide, with effective concentrations for 50% inhibition of mycelial growth of 0.023 to 0.383 μg/ml and a mean of 0.114 μg/ml, which showed a skewed unimodal distribution. Zoxamide-resistant mutants of P. capsici were obtained by either treating mycelial culture and zoospores with ultraviolet irradiation or adapting a culture on zoxamide-amended plates. The frequency of resistance selection averaged 1.8 × 10–7. Resistant isolates were also derived by selfing or crossing two sexually compatible isolates, resulting in a mean selection frequency of 0.47. The resistance factor (RF) for zoxamide was 25 to 100 in P. capsici mutants. Through 10 culture transfers, the mutants maintained high levels of RF (between 14 and 134) and had almost equal fitness as their wild-type parents in mycelial growth, sporulation, and virulence. There was no cross resistance between zoxamide and either flumorph, metalaxyl, azoxystrobin, or etridiazole. Based on the results above, P. capsici can develop resistance to zoxamide, and the risk is predicted to be moderate in nature.


Plant Disease ◽  
2012 ◽  
Vol 96 (3) ◽  
pp. 416-422 ◽  
Author(s):  
F. P. Chen ◽  
J. R. Fan ◽  
T. Zhou ◽  
X. L. Liu ◽  
J. L. Liu ◽  
...  

Sterol 14α-demethylase inhibitors (DMIs) continue to be important in the management of brown rot of Monilinia spp. worldwide. In this study, the sensitivity of 100 Monilinia fructicola isolates from four unsprayed orchards and two packinghouses in Beijing, China, to the new DMI fungicide SYP-Z048 was evaluated and ranged from 0.003 to 0.039 and 0.016 to 0.047 μg/ml, respectively. Laboratory mutants resistant to SYP-Z048 were generated using UV irradiation but no mutants occurred spontaneously. Resistance was stable after 10 weekly consecutive transfers on fungicide-free medium. Three parameters, including growth rate, sporulation in vitro, and lesion area, were significantly different when sensitive isolates and resistant mutants were analyzed as groups. Mutants grew more slowly and developed significantly smaller lesions on detached fruit, and their sporulation ability in vitro was reduced. Cross resistance was found between SYP-Z048 and propiconazole (ρ = 0.82, P < 0.0001) but not between SYPZ048 and tridemorph, carbendazim, procymidone, azoxystrobin, or pyrimethanil. SYP-Z048 resistance in mutants exhibiting 50% mycelial growth inhibition values greater than 0.3 μg/ml was correlated with the presence of a mutation in the CYP51 gene that encodes the target protein for DMI fungicides. The mutation caused an amino acid change from tyrosine to phenylalanine at position 136 (Y136F). To our knowledge, this is the first baseline sensitivity of M. fructicola collected from China to a DMI fungicide. The inability of M. fructicola to generate spontaneous DMI-resistant mutants coupled with reduced fitness of Y136F mutants can explain why this target site mutation has not yet emerged as a DMI fungicide resistance determinant in M. fructicola field populations worldwide.


Plant Disease ◽  
2016 ◽  
Vol 100 (7) ◽  
pp. 1314-1320 ◽  
Author(s):  
Yingying Song ◽  
Zhengqun Zhang ◽  
Lele Chen ◽  
Leiming He ◽  
Hongbao Lu ◽  
...  

Isopyrazam is a new broad-spectrum, foliar-absorbed and -translocated succinate dehydrogenase inhibitor fungicide. In this study, 159 Botrytis cinerea isolates collected from different geographical regions of Shandong Province of China were characterized for baseline sensitivity to isopyrazam. Furthermore, the protective and curative activity of isopyrazam on strawberry fruit and the control efficacy in the field were also determined. In contrast to its mycelial growth, the spore germination of B. cinerea was inhibited completely by lower concentrations of isopyrazam, about 1 μg ml−1 on yeast-peptone-acetate medium. Frequency distributions of isopyrazam 50% effective concentration (EC50) values were unimodal curves, with mean EC50 values of 0.07 ± 0.04 (standard deviation) and 0.68 ± 0.36 μg ml−1 for the inhibition of spore germination and mycelial growth, respectively. In addition, there was no positive multiple resistance between isopyrazam and other classes of botryticides such as diethofencarb, iprodione, pyrimethanil, or SYP-Z048. In field trials conducted during 2014 and 2015, isopyrazam used at a concentration of active ingredient at 150 and 200 g ha−1 provided a control efficacy ranging from 76.7 to 87.8% on leaves and from 81.5 to 90.7% on fruit. These results suggest that isopyrazam has the potential to play an important role in the management of gray mold.


Sign in / Sign up

Export Citation Format

Share Document