scholarly journals Response to Fungicides of Colletotrichum spp. Isolated from Red Peppers in Sunchang, Korea

2021 ◽  
Vol 27 (4) ◽  
pp. 164-171
Author(s):  
Hyeon-Yeong Ju ◽  
Mi-Ra Son ◽  
Jin-Won Kim
Keyword(s):  
Fungicide Resistance ◽  
Mycelial Growth ◽  
Susceptibility Test ◽  
Continuous Treatment ◽  
Inhibition Rate ◽  
Strain Group ◽  
Resistant Strains ◽  
Colletotrichum Spp ◽  
Red Peppers

In August 2020, 377 anthracnose strains were isolated from anthracnose-infected peppers collected from 25 farms in Sunchang-gun, Jeollabuk-do. Inhibition rate of mycelial growth of 11 pepper anthracnose fungicides registered in Korea was investigated for 62 strains selected by region and the degree of susceptibility to each fungicide was investigated. As a result of the fungicide susceptibility test of anthracnose to the fungicide, no resistant strains were observed in fluazinam, prochloraz manganese, and benomyl, but resistant strains appeared in at least three areas for other fungicides, and all strains in all regions were resistant to dithianon. Through this, it was confirmed that the fungicide resistance was expressed in the strain group due to the continuous treatment of the fungicide in some areas. By region, resistant strains to seven pesticides appeared in Sunchang-eup and Paldeok-myeon, and resistant strains to six pesticides appeared in Geumgwa-myeon, Bokheung-myeon, Ssangchi-myeon, Yudeung-myeon, and Pungsan-myeon. There was no significant correlation between fungicide registration time and emergence of resistant strains.

scholarly journals Combination of Suspension Array and Mycelial Growth Assay for Detecting Multiple-Fungicide Resistance in Botrytis cinerea in Hebei Province in China

Plant Disease ◽  
2019 ◽  
Vol 103 (6) ◽  
pp. 1213-1219
Author(s):  
Zehua Su ◽  
Xin Zhang ◽  
Jianjiang Zhao ◽  
Wenqiao Wang ◽  
Lei Shang ◽  
...  
Keyword(s):  
Botrytis Cinerea ◽  
High Throughput ◽  
Fungicide Resistance ◽  
Mycelial Growth ◽  
Accurate Method ◽  
Hebei Province ◽  
Combination Method ◽  
Major Type ◽  
Suspension Array ◽  
Growth Assay

To provide a high-throughput, efficient, and accurate method to monitor multiple-fungicide resistance of Botrytis cinerea in the field, we used the suspension array, sequencing, and mycelial growth assay in our research. Discriminating-dose bioassays for detecting carbendazim, diethofencarb, boscalid, and iprodione resistance (CarR, DieR, BosR, and IprR, respectively) were used to analyze 257 isolates collected from Hebei Province in China during 2016 and 2017. High resistance frequencies to carbendazim (100%), diethofencarb (92.08%), and iprodione (86.59%) were detected. BosR isolates accounted for 11.67% of the total. In addition, 103 isolates were randomly selected for phenotype and genotype detection. The high-throughput suspension array was utilized to detect eight genotypes simultaneously, including BenA-E198, BenA-198A, SdhB-H272, SdhB-272Y, BcOS1-I365, BcOS1-365S, erg27-F412, and erg27-412S, which were associated with resistance toward carbendazim or diethofencarb, boscalid, iprodione, and fenhexamid (FenR), respectively. Most of the benzimidazole-resistant isolates (81.55%) possessed the E198V mutation in the BenA gene. Ninety-three isolates with dual resistance to carbendazim and diethofencarb showed the E198V/K mutation. All BosR isolates carried the H272R mutation in the SdhB gene. The I365S and Q369P+N373S (66.99%) mutations in the BcOS1 gene were more frequently observed. No mutation was detected in the erg27 gene in Hebei isolates. There were 13 resistance profile phenotypes. Phenotypes with triple resistance were the most common (83.50%), and CarRDieRBosSIprRFenS was the major type. CarR isolates that carried E198V/K/A were all highly resistant (HR) and only one F200Y mutant was moderately resistant (MR) to carbendazim. Isolates that possessed E198V/K were MR or HR to diethofencarb. BosR isolates that possessed H272R mutation were lowly resistant (LR). IprR isolates were all LR or MR. The distribution of half maximal effective concentrations of CarR isolates with E198V/K mutations and IprR isolates with Q369P+N373S mutations significantly increased from 2016 to 2017. Combined with our observations, a combination method of the high-throughput suspension array and the mycelial growth assay was suggested to accurately monitor multiple resistance of B. cinerea in the field.

scholarly journals Development and Significance of Dicarboximide Resistance in Sclerotinia minor Isolates from Commercial Lettuce Fields in California

Plant Disease ◽  
1997 ◽  
Vol 81 (2) ◽  
pp. 148-153 ◽  
Author(s):  
J. C. Hubbard ◽  
K. V. Subbarao ◽  
S. T. Koike
Keyword(s):  
Fungicide Resistance ◽  
Disease Incidence ◽  
Vegetative Mycelium ◽  
Wild Type ◽  
In Vitro Production ◽  
Field Isolates ◽  
Resistant Strains ◽  
Measurable Growth ◽  
Vitro Production

Three growth stages of each of 20 Sclerotinia minor isolates were tested for resistance to iprodione. Sclerotia and both vegetative and mature mycelium of each isolate were plated on potato-dextrose agar (PDA) amended with 0, 1, 5, 10, 25, and 100 µg of the fungicide per ml, and radial growth was measured. All wild-type field isolates were initially sensitive and did not grow in the presence of iprodione. However, fungicide resistance arose readily in vitro. All 20 isolates produced measurable growth (≥2 mm) on iprodione at 5 µg/ml after 2 weeks when started from mature mycelium, and 18 of 20 isolates produced measurable growth after 4 weeks when started from vegetative mycelium. In general, fungicide-resistant growth arose more frequently and mean colony diameters were significantly greater (P ≤ 0.05) with mature mycelium than with vegetative mycelium at all times and concentrations. In sclerotial germination tests, at least 1% of sclerotia germinated in 18 of 20 isolates after 2 weeks on iprodione at 5 µg/ml, and in 19 of 20 isolates after 5 weeks on 100 µg/ml. Of growth produced on 79 plates containing iprodione, 73 remained viable on PDA after 5 months, and 71 retained resistance to 5 µg/ml. Seventy of these also exhibited cross-resistance to vinclozolin, another dicarboximide fungicide. Pathogenicity of five fungicide-resistant strains was tested in greenhouse, microplot, and field experiments, with and without iprodione. Two months after in vitro production, one of the five resistant strains was avirulent, but disease incidence for the other four ranged from 40 to 75%, compared with 40% for the wild-type isolates. However, the virulence of the fungicide-resistant strains declined over time. Ten months after their production, two of the isolates were avirulent and disease incidence for the others ranged from 3 to 17%, compared with 40 to 90% for the wild-type isolates. The strains that remained virulent 5 and 7 months after in vitro production were not significantly controlled by iprodione sprayed according to labeled rates, although disease was significantly controlled by the fungicide in treatments inoculated with wild-type field isolates (P > 0.05). In experiments in commercial fields to determine the efficacy of fungicide sprays on the wild-type S. minor population, there was no evidence that fungicide resistance was the cause of lack of lettuce drop control observed in many coastal California fields. Application of fungicides at a less than optimal time may account for some incidents of control failure.

scholarly journals Identification of Resistance to Multiple Fungicides in Field Populations of Venturia inaequalis

Plant Disease ◽  
2011 ◽  
Vol 95 (8) ◽  
pp. 921-926 ◽  
Author(s):  
Kimberly S. Chapman ◽  
George W. Sundin ◽  
Janna L. Beckerman
Keyword(s):  
Fungicide Resistance ◽  
Mycelial Growth ◽  
Venturia Inaequalis ◽  
Apple Scab ◽  
Field Resistance ◽  
Single Spore ◽  
Mycelium Growth ◽  
Thiophanate Methyl ◽  
Development Of Resistance

Venturia inaequalis, the causal agent of apple scab, is controlled primarily by fungicides. Long-term, extensive fungicide use has led to the development of resistance to multiple fungicides. To assess fungicide resistance, isolates of V. inaequalis were collected from Indiana and Michigan orchards. Single-spore derived isolates were evaluated by mycelium growth assays with previously determined discriminatory doses on media containing dodine, kresoxim-methyl, myclobutanil, or thiophanate-methyl. Of 195 isolates tested, 5.2, 0.7, 57.0, and 92.6% of isolates were found to be resistant to dodine, kresoxim-methyl, myclobutanil, and thiophanate-methyl, respectively. This is the first report of kresoxim-methyl field resistance in these states. Isolates resistant or shifted to a single fungicide were often found to have multiple fungicide resistance. Of all isolates tested, 38% were identified as resistant or shifted to two fungicides, and 12% were resistant or shifted to all four fungicides tested. No fitness penalty was found for isolates resistant to multiple fungicides based on a statistical analysis of mycelial growth and conidial production.

scholarly journals Tandem Repeat of a Transcriptional Enhancer Upstream of the Sterol 14α-Demethylase Gene (CYP51) inPenicillium digitatum

2000 ◽  
Vol 66 (8) ◽  
pp. 3421-3426 ◽  
Author(s):  
Hiroshi Hamamoto ◽  
Koji Hasegawa ◽  
Ryoji Nakaune ◽  
Young Jin Lee ◽  
Yoshiyuki Makizumi ◽  
...  
Keyword(s):  
Filamentous Fungi ◽  
Tandem Repeat ◽  
Resistant Strain ◽  
Fungicide Resistance ◽  
Promoter Region ◽  
Resistance Mechanism ◽  
Constitutive Expression ◽  
Structural Genes ◽  
Transcriptional Enhancer ◽  
Resistant Strains

ABSTRACT We investigated the mechanism of resistance to demethylation inhibitors (DMI) in Penicillium digitatum by isolating theCYP51 gene, which encodes the target enzyme (P45014DM) of DMI, from three DMI-resistant and three DMI-sensitive strains. The structural genes of all six strains were identical, but in the promoter region, a unique 126-bp sequence was tandemly repeated five times in the DMI-resistant strains and was present only once in the DMI-sensitive strains. Constitutive expression of CYP51 in the resistant strains was about 100-fold higher than that in the sensitive strains. We introduced CYP51, including the promoter region, from a DMI-resistant strain into a DMI-sensitive strain, which rendered the transformants DMI resistant and increased CYP51 expression. We also found that if the number of copies of the repeat was reduced to two, resistance andCYP51 expression also decreased. These results indicate that the 126-bp unit acts as a transcriptional enhancer and that a tandem repeat of the unit enhances CYP51 expression, resulting in DMI resistance. This is a new fungicide resistance mechanism for filamentous fungi.

scholarly journals Molecular Characterization, Pathogenicity and Biological Characterization of Colletotrichum Species Associated with Anthracnose of Camellia yuhsienensis Hu in China

Forests ◽  
2021 ◽  
Vol 12 (12) ◽  
pp. 1712
Author(s):  
Xinggang Chen ◽  
Lingyu Jiang ◽  
Anhua Bao ◽  
Changlin Liu ◽  
Junang Liu ◽  
...  
Keyword(s):  
Mycelial Growth ◽  
Morphological Characteristics ◽  
Biological Characteristics ◽  
Camellia Oleifera ◽  
Biological Characterization ◽  
Pathogenicity Tests ◽  
Colletotrichum Spp ◽  
And Control ◽  
First Time

Camellia yuhsienensis Hu, a species of tea oil tree with resistance to anthracnose, is widely used to breed disease-resistant Camellia varieties. In 2019, anthracnose symptoms were observed on Ca. yuhsienensis for the first time. However, the species and biological characteristics of Colletotrichum spp. isolated from Ca. yuhsienensis (YX-Colletotrichum spp.) have not been elucidated. In this study, five isolates (YX2-5-2, 2YX-3-1, 2YX-5-1, 2YX-8-1-1 and 2YX-8-1-2), which were consistent with the morphological characteristics of Colletotrichum spp., were obtained from Ca. yuhsienensis. A phylogenetic analysis demonstrated that YX2-5-2, 2YX-3-1 and 2YX-8-1-2 belonged to first clade along with Colletotrichum fructicola. 2YX-8-1-1 belonged to the second clade along with Colletotrichum siamense. 2YX-5-1 belonged to the third clade with Colletotrichum camelliae. Pathogenicity tests revealed that the pathogenicity of YX-Colletotrichum spp. was stronger than that of Colletotrichum spp. isolated from Camellia oleifera (GD-Colletotrichum spp.). Biological characteristics illustrated that the mycelial growth of YX-Co. camelliae (2YX-5-1) was slower than that of GD-Co. camelliae when the temperature exceeded 20 °C. In addition, in the presence of ions, the mycelial growth of YX-Co. fructicola (YX2-5-2) and YX-Co. siamense (2YX-8-1-1) was also slower than that of GD-Co. fructicola and GD-Co. siamense. Furthermore, the ability of YX-Colletotrichum spp. to utilize lactose and mannitol was weaker than that of GD-Colletotrichum spp., while the ability to utilize NH4+ was generally stronger than that of GD-Colletotrichum spp. This is the first report of anthracnose of Ca. yuhsienensis induced by Co. fructicola, Co. siamense and Co. camelliae in China. These results will provide theoretical guidance for the study of the pathogenesis and control of anthracnose on Ca. yuhsienensis.

scholarly journals A Rapid Method to Quantify Fungicide Sensitivity in the Brown Rot Pathogen Monilinia fructicola

Plant Disease ◽  
2009 ◽  
Vol 93 (4) ◽  
pp. 328-331 ◽  
Author(s):  
Kerik D. Cox ◽  
Kacie Quello ◽  
Ryan J. Deford ◽  
Janna L. Beckerman
Keyword(s):  
Rapid Method ◽  
Fungicide Resistance ◽  
Mycelial Growth ◽  
Brown Rot ◽  
Spore Density ◽  
Monilinia Fructicola ◽  
Chromogenic Substrate ◽  
Alamar Blue ◽  
Resistance Screening ◽  
Fungicide Sensitivity

Management of brown rot of stone fruit relies upon the application of effective fungicides that may be compromised by the development of fungicide resistance. We evaluated fungicide resistance in the brown rot pathogen, Monilinia fructicola, using Alamar blue (AB) dye, or resazurin, a chromogenic substrate that can be used as an indicator of respiration, in a 96-well microtiter format. We compared the AB method to traditional mycelial growth assays for resistance screening using 10 isolates of M. fructicola that represented a range of sensitivities to fenbuconazole. Using traditional mycelial growth assays, isolate sensitivity ranged from 17.7 to 115.3% growth on medium amended with fenbuconazole at 0.03 μg/ml relative to that on nonamended medium. Concordant results between both assays were obtained (R2 = 0.9943, P < 0.0001), but the AB method provided results within 24 h, as opposed to the 3- to 5-day period required for mycelial growth assays. We found that sensitive isolates reduced AB less than resistant isolates in the presence of fungicide. Spore density influenced the reduction of AB by M. fructicola; spectrophotometric discrimination of fungicide sensitivity was best achieved at a density of 105 spores/ml.

Assessment of quinone outside inhibitor sensitivity and frogeye leaf spot race of Cercospora sojina in Georgia soybean

Plant Disease ◽  
2021 ◽  
Author(s):  
Bennett Harrelson ◽  
Bikash Ghimire ◽  
Robert Kemerait ◽  
Albert Culbreath ◽  
Zenglu Li ◽  
...  
Keyword(s):  
Leaf Spot ◽  
Fungicide Resistance ◽  
Mycelial Growth ◽  
The United States ◽  
In Vitro Assays ◽  
Cercospora Sojina ◽  
Quinone Outside Inhibitor ◽  
Frogeye Leaf Spot ◽  
Differential Cultivars

Frogeye leaf spot (FLS), caused by the fungal pathogen Cercospora sojina K. Hara, is a foliar disease of soybean (Glycine max L. (Merr.)) responsible for yield reductions throughout the major soybean producing regions in the world. In the United States, management of FLS relies heavily on the use of resistant cultivars and in-season fungicide applications, specifically within the class of quinone outside inhibitors (QoIs), which has resulted in the development of fungicide resistance in many states. In 2018 and 2019, 80 isolates of C. sojina were collected from six counties in Georgia and screened for QoI fungicide resistance using molecular and in vitro assays, with resistant isolates being confirmed from three counties. Additionally, 50 isolates, including a “baseline isolate” with no prior fungicide exposure, were used to determine the percent reduction of mycelial growth to two fungicides, azoxystrobin and pyraclostrobin, at six concentrations: 0.0001, 0.001, 0.01, 0.1, 1, and 10 g ml-1. Mycelial growth observed for resistant isolates varied significantly from both the sensitive isolates and the baseline isolate for azoxystrobin concentrations of 10, 1, 0.1, and 0.01 g ml-1 and for pyraclostrobin concentrations of 10, 1, 0.1, 0.01 and 0.001 g ml-1. Moreover, 40 isolates were used to evaluate pathogen race on six soybean differential cultivars by assessing susceptible or resistant reactions. Isolate reactions suggested 12 races of C. sojina present in Georgia, four of which have not been previously described. Species richness indicators (rarefaction and abundance-based coverage estimator - ACE) indicated that within-county C. sojina race numbers were undersampled in the present study, suggesting the potential for the presence of either additional undescribed races or known but unaccounted for races in Georgia. However, no isolates were pathogenic on differential cultivar ‘Davis’, carrying the Rcs3 resistance allele, suggesting the gene is still an effective source of resistance in Georgia.

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