scholarly journals Activity of the Novel Succinate Dehydrogenase Inhibitor Fungicide Pydiflumetofen Against SDHI-Sensitive and SDHI-Resistant Isolates of Botrytis cinerea and Efficacy Against Gray Mold

Plant Disease ◽  
2020 ◽  
Vol 104 (8) ◽  
pp. 2168-2173 ◽  
Author(s):  
Leiming He ◽  
Kaidi Cui ◽  
Yufei Song ◽  
Tongtong Li ◽  
Ning Liu ◽  
...  
Keyword(s):  
Succinate Dehydrogenase ◽  
Botrytis Cinerea ◽  
Gray Mold ◽  
Effective Control ◽  
The Novel ◽  
Baseline Sensitivity ◽  
Single Spore ◽  
Significant Difference ◽  
Low Levels ◽  
Succinate Dehydrogenase Inhibitor

Succinate dehydrogenase inhibitor (SDHI) fungicides are currently the most frequently used fungicides for controlling gray mold. However, isolates of Botrytis cinerea resistant to SDHI fungicides have emerged in the field. Pydiflumetofen is a new SDHI fungicide that can control a variety of fungal diseases, but its efficacy against gray mold and whether the activity of pydiflumetofen is affected by the current SDHI-resistant isolates is currently unknown. The sensitivity of 291 single-spore B. cinerea isolates collected from 2017 to 2019 to pydiflumetofen was determined by spore germination inhibition assays. The mean EC50 value (fungicide concentration resulting in a 50% inhibition compared with that of the control) of pydiflumetofen was 0.06 ± 0.01, 0.07 ± 0.02, and 0.05 ± 0.02 mg/liter in 2017, 2018, and 2019, respectively. There was no significant difference in the sensitivity of B. cinerea to pydiflumetofen among the 3 years. Furthermore, pydiflumetofen at 300 mg/liter effectively controlled gray mold on cucumber leaves (80.9%), and its efficacy was superior to that of boscalid at 400 mg/liter (42.7%). The isolates carrying P225F, N230I, H272Y, and H272R mutations in the SdhB subunit were associated with the less sensitivity of B. cinerea to SDHI fungicides. After establishing the baseline sensitivity of B. cinerea to pydiflumetofen (EC50 of 0.03 ± 0.003 mg/liter), we found that the P225F and H272Y mutant isolates showed low to moderate levels of resistance to pydiflumetofen, and the H272R and N230I mutant isolates showed low levels of resistance. The reduced sensitivity to pydiflumetofen resulted from the positive correlation of pydiflumetofen with the other four SDHI fungicides (i.e., boscalid, fluopyram, isopyrazam, and benzovindiflupyr). These results suggest that pydiflumetofen provides effective control for the management of gray mold but must be used with caution.

scholarly journals Resistance to Fluopyram, Fluxapyroxad, and Penthiopyrad in Botrytis cinerea from Strawberry

Plant Disease ◽  
2014 ◽  
Vol 98 (4) ◽  
pp. 532-539 ◽  
Author(s):  
Achour Amiri ◽  
Stacy M. Heath ◽  
Natalia A. Peres
Keyword(s):  
Succinate Dehydrogenase ◽  
Botrytis Cinerea ◽  
High Resistance ◽  
Gray Mold ◽  
Cross Resistance ◽  
The Novel ◽  
Resistance Development ◽  
Dehydrogenase Gene ◽  
Resistance Patterns ◽  
Subunit B

Succinate dehydrogenase inhibitors (SDHIs) constitute a mainstay in management of gray mold caused by Botrytis cinerea in strawberry and several other crops. In this study, we investigated the risks of resistance development to three newer SDHIs (i.e., fluopyram, fluxapyroxad, and penthiopyrad) and their cross-resistance with the previously registered boscalid. We investigated the mutations in the SdhB subunit and evaluated their impact on microbial fitness in field populations of B. cinerea. Amino acid substitutions associated with resistance to SDHIs were detected at three codons of the SdhB subunit (BH272R/Y/L, BP225F, and BN230I) in the succinate dehydrogenase gene of field isolates from Florida. The BH272R, BH272Y, BH272L, BP225F, and BN230I mutations were detected at frequencies of 51.5, 28.0, 0.5, 2.5, and 4%, respectively. Strong cross-resistance patterns were evident between boscalid and fluxapyroxad and penthiopyrad but not with fluopyram, except in BH272L, BP225F, and BN230I mutants. All five mutations conferred moderate to very high resistance to boscalid whereas the BH272Y conferred resistance to fluxapyroxad and penthiopyrad. The BH272L, BN230I, and BP225F mutations conferred high resistance to all four SDHIs tested. Resistance monitoring following the first use of penthiopyrad in strawberry fields in Florida in 2013 suggests potential for quick selection for highly resistant populations and warrants careful use of the newer SDHIs. No evidence of major fitness costs due to the mutations in the SdhB subunit was found, which indicates the potential ability of the mutants to survive and compete with wild-type isolates. Our study suggests high risks for rapid widespread occurrence of B. cinerea populations resistant to the novel SDHIs unless appropriate rotation strategies are implemented immediately upon registration.

scholarly journals Baseline Sensitivity and Toxic Action of the Sterol Demethylation Inhibitor Flusilazole Against Botrytis cinerea

Plant Disease ◽  
2020 ◽  
Vol 104 (11) ◽  
pp. 2986-2993
Author(s):  
Yong Wang ◽  
Miaomaio Wang ◽  
Letian Xu ◽  
Yang Sun ◽  
Juntao Feng
Keyword(s):  
Botrytis Cinerea ◽  
Plant Pathogens ◽  
Resistance Mechanism ◽  
Point Mutations ◽  
Gray Mold ◽  
Shaanxi Province ◽  
Cross Resistance ◽  
Baseline Sensitivity ◽  
Single Spore ◽  
Demethylation Inhibitor

In the present study, a total of 95 Botrytis cinerea single-spore strains collected from different hosts in Shaanxi Province of China were characterized for their sensitivity to the sterol demethylation inhibitor fungicide flusilazole. The effective concentration for 50% inhibition of mycelial growth (EC50) of flusilazole ranged from 0.021 to 0.372 µg/ml, with an average value of 0.093 µg/ml. Cross-resistance between flusilazole and commonly used fungicides was not detected, and no flusilazole-resistant mutants were induced. Both on detached strawberry leaves and in greenhouse experiments, flusilazole was more effective than the commonly used fungicide carbendazim at reducing gray mold. After culture on PDA plates or detached strawberry leaves, no difference in sclerotia production or pathogenicity was detected between two strains, WG12 (most sensitive to flusilazole) and MX18 (least sensitive to flusilazole). After treatment with flusilazole, however, the two strains lost the ability to produce sclerotia, and oxalic acid and ergosterol contents in mycelium decreased. Interestingly, the inhibition rate of ergosterol content in MX18 was significantly lower than that in WG12. Expression of Cyp51, BcatrD, and Bcmfs1 genes all increased after treatment with flusilazole, especially the Cyp51 and BcatrD genes. However, the expression of Cyp51 gene or BcatrD gene in WG12 and MX18 were significantly different from each other after treatment with flusilazole. In addition, no point mutations in Cyp51 gene were found in MX18. These data suggest flusilazole is a promising fungicide for resistance management of gray mold and also provided novel insights into understanding the resistance mechanism of flusilazole against plant pathogens.

scholarly journals Baseline Sensitivity of Botrytis cinerea to the Succinate Dehydrogenase Inhibitor Isopyrazam and Efficacy of this Fungicide

Plant Disease ◽  
2016 ◽  
Vol 100 (7) ◽  
pp. 1314-1320 ◽  
Author(s):  
Yingying Song ◽  
Zhengqun Zhang ◽  
Lele Chen ◽  
Leiming He ◽  
Hongbao Lu ◽  
...  
Keyword(s):  
Succinate Dehydrogenase ◽  
Botrytis Cinerea ◽  
Spore Germination ◽  
Mycelial Growth ◽  
Field Trials ◽  
Baseline Sensitivity ◽  
Control Efficacy ◽  
Frequency Distributions ◽  
Geographical Regions ◽  
Succinate Dehydrogenase Inhibitor

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.

scholarly journals First Report of Leaf Spot Caused by Botrytis cinerea on Cardamine hupingshanensis in China

Plant Disease ◽  
2021 ◽  
Author(s):  
Jun Guo ◽  
Jin Chen ◽  
Zhao Hu ◽  
Jie Zhong ◽  
Jun Zi Zhu
Keyword(s):  
Botrytis Cinerea ◽  
Morphological Characteristics ◽  
Gray Mold ◽  
Hunan Province ◽  
Conidial Suspension ◽  
Heat Shock Protein 60 ◽  
Basal Cells ◽  
Single Spore ◽  
Internal Transcribed Spacer Region ◽  
First Report

Cardamine hupingshanensis is a selenium (Se) and cadmium (Cd) hyperaccumulator plant distributed in wetlands along the Wuling Mountains of China (Zhou et al. 2018). In March of 2020, a disease with symptoms similar to gray mold was observed on leaves of C. hupingshanensis in a nursery located in Changsha, Hunan Province, China. Almost 40% of the C. hupingshanensis (200 plants) were infected. Initially, small spots were scattered across the leaf surface or margin. As disease progressed, small spots enlarged to dark brown lesions, with green-gray, conidia containing mold layer under humid conditions. Small leaf pieces were cut from the lesion margins and were sterilized with 70% ethanol for 10 s, 2% NaOCl for 2 min, rinsed with sterilized distilled water for three times, and then placed on potato dextrose agar (PDA) medium at 22°C in the dark. Seven similar colonies were consistently isolated from seven samples and further purified by single-spore isolation. Strains cultured on PDA were initially white, forming gray-white aerial mycelia, then turned gray and produced sclerotia after incubation for 2 weeks, which were brown to blackish, irregular, 0.8 to 3.0 × 1.2 to 3.5 mm (n=50). Conidia were unicellular, globose or oval, colourless, 7.5 to 12.0 × 5.5 to 8.3 μm (n=50). Conidiophores arose singly or in group, straight or flexuous, septate, brownish to light brown, with enlarged basal cells, 12.5 to 22.1 × 120.7 to 310.3 μm. Based on their morphological characteristics in culture, the isolates were putatively identified as Botrytis cinerea (Ellis 1971). Genomic DNA of four representative isolates, HNSMJ-1 to HNSMJ-4, were extracted by CTAB method. The internal transcribed spacer region (ITS), glyceraldehyde-3-phosphate dehydrogenase gene (G3PDH), heat-shock protein 60 gene (HSP60), ATP-dependent RNA helicaseDBP7 gene (MS547) and DNA-dependent RNA polymerase subunit II gene (RPB2) were amplified and sequenced using the primers described previously (Aktaruzzaman et al. 2018) (MW820311, MW831620, MW831628, MW831623 and MW831629 for HNSMJ-1; MW314722, MW316616, MW316617, MW316618 and MW316619 for HNSMJ-2; MW820519, MW831621, MW831627, MW831624 and MW831631 for HNSMJ-3; MW820601, MW831622, MW831626, MW831625 and MW831630 for HNSMJ-4). BLAST searches showed 99.43 to 99.90% identity to the corresponding sequences of B. cinerea strains, such as HJ-5 (MF426032.1, MN448500.1, MK791187.1, MH727700.1 and KX867998.1). A combined phylogenetic tree using the ITS, G3PDH, HSP60 and RPB2 sequences was constructed by neighbor-joining method in MEGA 6. It revealed that HNSMJ-1 to HNSMJ-4 clustered in the B. cinerea clade. Pathogenicity tests were performed on healthy pot-grown C. hupingshanensis plants. Leaves were surface-sterilized and sprayed with conidial suspension (106 conidia/ mL), with sterile water served as controls. All plants were kept in growth chamber with 85% humidity at 25℃ following a 16 h day-8 h night cycle. The experiment was repeated twice, with each three replications. After 4 to 7 days, symptoms similar to those observed in the field developed on the inoculated leaves, whereas controls remained healthy. The pathogen was reisolated from symptomatic tissues and identified using molecular methods, confirming Koch’s postulates. B. cinerea has already been reported from China on C. lyrate (Zhang 2006), a different species of C. hupingshanensis. To the best of our knowledge, this is the first report of B. cinerea causing gray mold on C. hupingshanensis in China and worldwide. Based on the widespread damage in the nursery, appropriate control strategies should be adopted. This study provides a basis for studying the epidemic and management of the disease.

scholarly journals Timing of Preharvest Infection of Pear Fruit by Botrytis cinerea and the Relationship to Postharvest Decay

Plant Disease ◽  
2004 ◽  
Vol 88 (5) ◽  
pp. 468-473 ◽  
Author(s):  
C. L. Lennox ◽  
R. A. Spotts
Keyword(s):  
Botrytis Cinerea ◽  
Gray Mold ◽  
Economic Losses ◽  
Pear Fruit ◽  
Full Bloom ◽  
Postharvest Decay ◽  
Poor Predictor ◽  
The Mean ◽  
Low Levels ◽  
The Relationship

Botrytis cinerea causes significant levels of postharvest decay in the winter pear cultivar d'Anjou. The objectives of this study were to determine the timing of B. cinerea infection of pear stems and calyxes in the orchard during the growing season, to investigate the development of gray mold in storage, and to determine whether preharvest levels of B. cinerea in pear stems and calyxes can be used as predictors of gray mold levels observed in storage. Very low levels of B. cinerea were isolated from stem tissue prior to harvest. In a single year repeat experiment, stems sampled at harvest had higher levels of infection than those sampled earlier in the season. Little or no stem end gray mold was detected in fruit after 3 months in air-storage; however, incidence increased between 6 and 8 months. Calyx end gray mold was detected at low levels in fruit stored for up to 8 months. The mean incidence of stem end gray mold was 3.6 and 2.0%, and incidence of calyx end gray mold was 1.2 and 0.2%, in 1996 and 1997, respectively. Calyxes were susceptible to infection soon after full bloom; however, inoculation of calyxes in April or May did not result in higher levels of calyx end gray mold in storage. Therefore, preharvest level of calyx infection is a poor predictor of calyx end gray mold in storage. In addition, application of benomyl in the orchard reduced the level of B. cinerea in blossoms but had no effect on levels of calyx end gray mold of fruit in storage. Packing and shipping fruit within 3 to 6 months of harvest may mitigate economic losses due to gray mold.

scholarly journals Amino Acid Polymorphism in Succinate Dehydrogenase Subunit C Involved in Biological Fitness of Botrytis cinerea

2020 ◽  
Vol 33 (4) ◽  
pp. 580-589
Author(s):  
Wenyong Shao ◽  
Jingtao Sun ◽  
Xiaoke Zhang ◽  
Changjun Chen
Keyword(s):  
Amino Acid ◽  
Succinate Dehydrogenase ◽  
Botrytis Cinerea ◽  
Tricarboxylic Acid Cycle ◽  
Small Population ◽  
Oxidative Stress Response ◽  
Carbon Source Utilization ◽  
Amino Acid Polymorphism ◽  
Baseline Sensitivity ◽  
Subunit C

Succinate dehydrogenase (SDH) is an important respiratory enzyme which participates in the tricarboxylic acid cycle and oxidative phosphorylation. A previous study of the baseline sensitivity of Botrytis cinerea against SDH inhibitors (SDHIs) showed that intrinsic sensitivity of the small population against the SDHIs exhibited significant differences. In the sequencing assay, we found five kinds of amino acid polymorphism in SDH subunit C (SdhC) of B. cinerea isolates which were never exposed to the SDHIs. To validate that amino acid polymorphism in the SdhC of B. cinerea confers intrinsic sensitivity against the SDHIs, the replacement mutants containing each kind of amino acid polymorphism of SdhC exhibited phenotype differences in intrinsic sensitivity to SDHIs, mycelial growth, sporulation, virulence, oxidative stress response, and carbon source utilization. These results indicated that SdhC of B. cinerea experienced positive selection during evolution and resulted in amino acid polymorphism which is involved in intrinsic sensitivity to SDHIs and biological fitness.

scholarly journals Expression of Putative Defense Responses in Cannabis Primed by Pseudomonas and/or Bacillus Strains and Infected by Botrytis cinerea

2020 ◽  
Vol 11 ◽  
Author(s):  
Carole Balthazar ◽  
Gabrielle Cantin ◽  
Amy Novinscak ◽  
David L. Joly ◽  
Martin Filion
Keyword(s):  
Botrytis Cinerea ◽  
Cannabis Sativa ◽  
Molecular Level ◽  
Quantitative Polymerase Chain Reaction ◽  
Defense Responses ◽  
Gray Mold ◽  
Control Measures ◽  
Effective Control ◽  
Defense Genes ◽  
Polymerase Chain

Cannabis (Cannabis sativa L.) offers many industrial, agricultural, and medicinal applications, but is commonly threatened by the gray mold disease caused by the fungus Botrytis cinerea. With few effective control measures currently available, the use of beneficial rhizobacteria represents a promising biocontrol avenue for cannabis. To counter disease development, plants rely on a complex network of inducible defense pathways, allowing them to respond locally and systemically to pathogens attacks. In this study, we present the first attempt to control gray mold in cannabis using beneficial rhizobacteria, and the first investigation of cannabis defense responses at the molecular level. Four promising Pseudomonas (LBUM223 and WCS417r) and Bacillus strains (LBUM279 and LBUM979) were applied as single or combined root treatments to cannabis seedlings, which were subsequently infected by B. cinerea. Symptoms were recorded and the expression of eight putative defense genes was monitored in leaves by reverse transcription quantitative polymerase chain reaction. The rhizobacteria did not significantly control gray mold and all infected leaves were necrotic after a week, regardless of the treatment. Similarly, no systemic activation of putative cannabis defense genes was reported, neither triggered by the pathogen nor by the rhizobacteria. However, this work identified five putative defense genes (ERF1, HEL, PAL, PR1, and PR2) that were strongly and sustainably induced locally at B. cinerea’s infection sites, as well as two stably expressed reference genes (TIP41 and APT1) in cannabis. These markers will be useful in future researches exploring cannabis defense pathways.

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