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1.
Pestic Biochem Physiol ; 158: 121-127, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31378347

RESUMO

Iminoctadine tris (albesilate) is a bis-guazatine fungicide, and its specific modes of action and efficacy against C. cassiicola are not yet clear. In this study, baseline sensitivity data for mycelial growth showed that the frequency distribution curve of iminoctadine tris (albesilate) EC50 values is unimodal. The EC50 values ranged from 0.1151 to 1.2101 µg/mL, with a mean of 0.5775 ±â€¯0.2677 µg/mL. Iminoctadine tris (albesilate) affected the morphological development of C. cassiicola, causing increased branching of the mycelium. The significant increase in membrane permeability and malondialdehyde content after iminoctadine tris (albesilate) treatment indicated that this fungicide caused severe damage to the membrane structure. Furthermore, 0.4 µg/mL iminoctadine tris (albesilate) could decrease the spore density of C. cassiicola from 2.6200 × 104 to 1.4967 × 104/cm2 on average in vitro, indicating that the fungicide had great potential to reduce secondary infection with C. cassiicola in the field. Additionally, 120 µg/mL iminoctadine tris (albesilate) provided over 95% curative efficacy and 81.17% preventative efficacy on detached cucumber leaves inoculated with C. cassiicola. In field trials, iminoctadine tris (albesilate) at a dose of 120 g a.i./ha exhibited 72.92% and 80.92% control efficacy in 2017 and 2018, respectively. However, the efficacy supplied by the reference fungicide azoxystrobin at 250 g a.i./ha was only approximately 50% due to the development of fungicide resistance in C. cassiicola. Taken together, the findings above provide a solid foundation for the exploration of the action mechanisms of iminoctadine tris (albesilate) against C. cassiicola and provide overwhelming evidence for the use of iminoctadine tris (albesilate) as an excellent potential alternative fungicide in the management of cucumber Corynespora leaf spot.


Assuntos
Ascomicetos/efeitos dos fármacos , Fungicidas Industriais/farmacologia , Guanidinas/farmacologia , Cucumis sativus/microbiologia , Folhas de Planta/microbiologia
2.
Pestic Biochem Physiol ; 158: 128-134, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31378348

RESUMO

Indoleacetic acid (IAA)-carbendazim was synthesized to assess whether this conjugate could retain the fungicidal activity of carbendazim and gain root-inducing properties upon the addition of an indoleacetic acid group. An indoor virulence test demonstrated that the conjugate retained the fungicidal activity of carbendazim towards Cylindrocladium parasiticum. The conjugate was detected in roots after soaking Ricinus communis L. leaves into a solution of the IAA-carbendazim, which confirmed its phloem mobility. The activities of the cellulase, polygalacturonase and xylanase produced by Cylindrocladium parasiticum treated with different concentrations of the conjugate were determined, and the peak activities appeared at 72 h or 96 h. More importantly, the conjugate showed the ability to promote root growth. These results revealed that indoleacetic acid-carbendazim may be useful in preventing Cylindrocladium parasiticum and other diseases.


Assuntos
Ascomicetos/efeitos dos fármacos , Benzimidazóis/farmacologia , Carbamatos/farmacologia , Fungicidas Industriais/farmacologia , Ácidos Indolacéticos/farmacologia , Celulase/metabolismo , Floema/efeitos dos fármacos , Raízes de Plantas/efeitos dos fármacos , Poligalacturonase/metabolismo , Ricinus/efeitos dos fármacos
3.
Pestic Biochem Physiol ; 158: 175-184, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31378354

RESUMO

Succinate dehydrogenase (SDH), an essential component of cellular respiratory chain and tricarboxylic acid (or Krebs) cycle, has been identified as one of the most significant targets for pharmaceutical and agrochemical. Herein, with the aim of discovery of new antifungal lead structures, a class of novel N-(4-fluoro-2-(phenylamino)phenyl)-pyrazole-4-carboxamides were designed, synthesized and evaluated for their biological activities. They were bioassayed against seven phytopathogenic fungi, Rhizoctonia solani, Phytophthora infestans, Fusarium oxysporum f. sp. vasinfectum, Botryosphaeria dothidea, Gibberella zeae, Alternaria alternate and Fusarium oxysporum f. sp. niveum. The results indicated that most of the compounds displayed good antifungal activities, especially against R. solani. Among them, compounds 7 and 12 exhibited higher antifungal activities against R. solani in vitro with EC50 value of 0.034 mg/L and 0.021 mg/L, being superior to the commercially available fungicide bixafen (EC50 = 0.043 mg/L). Pot tests against R. solani showed that in vivo EC50 values of compounds 7 (2.694 mg/L) and 12 (2.331 mg/L) were higher than that of bixafen (3.724 mg/L). In addition, inhibitory activity of compound 12 against SDH indicated compound 12 (IC50 = 1.836 mg/L) showed good inhibitory activity against SDH, being close to bixafen's inhibitory activity (IC50 = 1.222 mg/L). And, molecular modeling of the SDH-compound 12 complex suggested that compound 12 could strongly bind to and interact with the binding site of the SDH. The results of the present work showed that N-(4-fluoro-2-(phenylamino)phenyl)-pyrazole-4-carboxamides were a new fungicides for discovery of SDH inhibitors and worth further study.


Assuntos
Antifúngicos/química , Antifúngicos/farmacologia , Succinato Desidrogenase/metabolismo , Alternaria/efeitos dos fármacos , Alternaria/enzimologia , Ascomicetos/efeitos dos fármacos , Ascomicetos/enzimologia , Fusarium/efeitos dos fármacos , Fusarium/enzimologia , Phytophthora infestans/efeitos dos fármacos , Phytophthora infestans/enzimologia , Rhizoctonia/efeitos dos fármacos , Rhizoctonia/enzimologia , Relação Estrutura-Atividade
4.
J Agric Food Chem ; 67(37): 10448-10457, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31453693

RESUMO

Carabrone is isolated from Carpesium macrocephalum Franch. et Sav, which has good fungicidal activity, especially for Gaeumannomyces graminis (Get). According to previous studies, we speculated that carabrone targets the mitochondrial enzyme complex III of Get. To elucidate the mode of action, we used carabrone to induce oxidative stress and apoptosis in Get. Incubation with carabrone reduced the burst of reactive oxygen species (ROS) and mitochondrial membrane potential, as well as phosphatidylserine release. Carabrone caused ROS accumulation in mycelia by inhibiting the activity of antioxidase enzymes, among which inhibition of glutathione reductase (GR) activity was most obvious. The catalytic center of GR consists of l-cysteine residues that react with the α-methylene-γ-butyrolactone active site of carabrone. Additionally, a positive TUNEL reaction led to diffusion of the DNA electrophoresis band and upregulation of Ggmet1 and Ggmet2. We propose that carabrone inhibits antioxidant enzymes and promotes ROS overproduction, which causes membrane hyperpermeability, release of apoptotic factors, activation of the mitochondria-mediated apoptosis pathway, and fungal cell apoptosis.


Assuntos
Apoptose/efeitos dos fármacos , Ascomicetos/efeitos dos fármacos , Fungicidas Industriais/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Ascomicetos/citologia , Ascomicetos/metabolismo , Asteraceae/química , Proteínas Fúngicas/metabolismo , Glutationa Redutase/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Espécies Reativas de Oxigênio/metabolismo
5.
Plant Dis ; 103(9): 2271-2276, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31287371

RESUMO

Sensitivity monitoring of Venturia effusa, cause of pecan scab, has revealed insensitivity to fentin hydroxide and tebuconazole, but recent research indicates that the insensitivity to fentin hydroxide is not stable. A study was undertaken to determine if a fitness cost may be responsible for this instability. In this study, experiments were conducted to evaluate fitness components and phenotypic stability of insensitivity of V. effusa to fentin hydroxide and tebuconazole. Conidial production, conidial germination, microcolony growth, sensitivity to osmotic stress, and sensitivity to oxidative stress in the absence of fungicide were compared for isolates with differing sensitivities to both fungicides. Percent conidial germination decreased linearly with increasing fentin hydroxide insensitivity, and microcolony growth on 1.0 mM H2O2 decreased linearly with increasing tebuconazole insensitivity. Stability of resistance was assessed on concentrations of 1.0, 3.0, and 10 µg/ml of both fungicides prior to and after five transfers on non-fungicide-amended medium. Tebuconazole insensitivity was stable after transfers, but fentin hydroxide insensitivity on 1.0 and 3.0 µg/ml decreased significantly after transfers, indicating instability. Here we provide evidence that in V. effusa tebuconazole insensitivity is stable and fentin hydroxide insensitivity is not. These results suggest that fentin-hydroxide-resistant V. effusa isolates have reduced conidial viability compared with sensitive isolates, which may allow the population to regain sensitivity in the absence of this frequently used fungicide.


Assuntos
Ascomicetos , Farmacorresistência Fúngica , Compostos Orgânicos de Estanho , Triazóis , Ascomicetos/efeitos dos fármacos , Compostos Orgânicos de Estanho/farmacologia , Triazóis/farmacologia
6.
Plant Dis ; 103(9): 2212-2220, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31306094

RESUMO

White mold, caused by Sclerotinia sclerotiorum, is a yield-limiting disease of soybean in Brazil. Uniform fungicide trials have been conducted annually since 2009. Data from 74 cooperative field trials conducted over a 10-year period were assembled. We selected five fungicides applied two times around flowering: dimoxystrobin plus boscalid (DIMO+BOSC), carbendazim plus procymidone (CARB+PROC), fluazinam (FLUZ), fluopyram (FLUO), and procymidone (PROC). For comparison, thiophanate-methyl (TMET) applied four times was also included as a low-cost treatment. Network models were fitted to the log of white mold incidence (percentages) and log of sclerotia mass data (grams/hectare) and to the nontransformed yield data (kilograms/hectare) for each treatment, including the untreated check. Back-transformation of the meta-analytic estimates indicated that the lowest and highest mean (95% confidence interval [CI]) percent reductions in incidence and sclerotia mass were 54.2 (49.3 to 58.7) and 51.6% (43.7 to 58.3) for TMET and 83.8 (79.1 to 87.5) and 87% (81.9 to 91.6) for CARB+PROC, respectively. The overall mean (95% CI) yield responses ranged from 323 kg/ha (247.4 to 400.3) for TMET to 626 kg/ha (521.7 to 731.7) for DIMO+BOSC, but the variance was significantly reduced by a binary variable (30% threshold) describing disease incidence in the untreated check. On average, an increment of 352 kg/ha was estimated for trials where the incidence was >30% compared with the low-disease scenario. Hence, the probability of breaking even on fungicide costs for the high-disease scenario was >65% for the more effective, but more expensive fungicide (FLUZ) than TMET. For the low-disease scenario, profitability was less likely and depended more on variations in fungicide cost and soybean price.


Assuntos
Ascomicetos , Fungicidas Industriais , Doenças das Plantas , Soja , Agricultura , Ascomicetos/efeitos dos fármacos , Brasil , Fungicidas Industriais/economia , Fungicidas Industriais/farmacologia , Fungicidas Industriais/normas , Doenças das Plantas/prevenção & controle , Soja/microbiologia
7.
World J Microbiol Biotechnol ; 35(7): 106, 2019 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-31267229

RESUMO

Xenorhabdus nematophila HB310 secreted the insecticidal protein toxin complex. Two chitinase genes, chi60 and chi70, were found in X. nematophila toxin complex locus. In order to clarify the function of two chitinases, chi60 and chi70 genes were cloned and expressed in Escherichia coli Transetta (DE3). As a result, we found that the Chi60 and Chi70 belonged to glycoside hydrolases (GH) family 18 with a molecular mass of 65 kDa and 78 kDa, respectively. When colloidal chitin was treated as the substrate, Chi60 and Chi70 were proved to have the highest enzymatic activity at pH 6.0 and 50 °C. Chi60 and Chi70 had obvious growth inhibition effect against the second larvae of Helicoverpa armigera with growth inhibiting rate of 81.99% and 90.51%. Chi70 had synergistic effect with the insecticidal toxicity of Bt Cry 1Ac, but the Chi60 had no synergistic effect with Bt Cry 1Ac. Chi60 and Chi70 showed antifungal activity against Alternaria brassicicola, Verticillium dahliae and Coniothyrium diplodiella. The results increased our understanding of the chitinases produced by X. nematophila and laid a foundation for further studies on the mechanism of the chitinases.


Assuntos
Antifúngicos/farmacologia , Quitinases/antagonistas & inibidores , Quitinases/genética , Quitinases/metabolismo , Xenorhabdus/metabolismo , Alternaria/efeitos dos fármacos , Animais , Ascomicetos/efeitos dos fármacos , Quitina/metabolismo , Quitinases/classificação , Clonagem Molecular , Sinergismo Farmacológico , Ensaios Enzimáticos , Estabilidade Enzimática , Escherichia coli/genética , Expressão Gênica , Glicosídeo Hidrolases/genética , Concentração de Íons de Hidrogênio , Inseticidas/metabolismo , Inseticidas/farmacologia , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Peso Molecular , Mariposas/efeitos dos fármacos , Mariposas/crescimento & desenvolvimento , Micotoxinas/genética , Micotoxinas/metabolismo , Filogenia , Domínios Proteicos , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Temperatura Ambiente , Verticillium/efeitos dos fármacos , Xenorhabdus/genética
8.
Microbiol Res ; 226: 55-64, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31284945

RESUMO

Functional association between genomic loci and specific biological traits remains lacking in many fungi, including the African tree pathogen Ceratocystis albifundus. This is mainly because of the absence of suitable transformation systems for allowing genetic manipulation of this and other fungi. Here, we present an optimized protocol for Agrobacterium tumefaciens-mediated transformation of C. albifundus. Strain AGL-1 of A. tumefaciens and four binary T-DNA vectors (conferring hygromycin B or geneticin resistance and/or expressing the green fluorescent protein [GFP]) were used for transforming germinated conidia of three isolates of C. albifundus. Stable expression of these T-DNA-encoded traits was confirmed through sequential sub-culturing of fungal transformants on selective and non-selective media and by using PCR and sequence analysis. Single-copy integration of the respective T-DNAs into the genomes of these fungi was confirmed using Southern hybridization analysis. The range of experimental parameters determined and optimised included: (i) concentrations of hygromycin B and geneticin required for inhibiting growth of the wild type fungus and (ii) the dependence of transformation on acetosyringone for inducing the bacterium's virulence genes, as well as (iii) the duration of fungus-bacterium co-cultivation periods and (iv) the concentrations of fungal conidia and bacterial cells used for the latter. The system developed in this study is stable with a high-efficiency, yielding up to 400 transformants per 106 conidia. This is the first report of a transformation protocol for C. albifundus and its availability will be invaluable for functional studies in this important fungus.


Assuntos
Agrobacterium tumefaciens/genética , Ascomicetos/genética , Transformação Genética , Ascomicetos/citologia , Ascomicetos/efeitos dos fármacos , Ascomicetos/crescimento & desenvolvimento , Southern Blotting , Carbenicilina/farmacologia , Técnicas de Cocultura , DNA Bacteriano , Regulação Fúngica da Expressão Gênica , Gentamicinas/farmacologia , Proteínas de Fluorescência Verde/genética , Higromicina B/farmacologia , Canamicina/farmacologia , Reação em Cadeia da Polimerase , Análise de Sequência , Virulência/genética
9.
J Sci Food Agric ; 99(13): 6060-6065, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31226223

RESUMO

BACKGROUND: Chestnuts are gluten-free, low-fat, cholesterol-free products. Postharvest decay reduces chestnut shelf life and can cause severe economic losses. In this study we investigated the effect of ozone (O3 ) gaseous treatment on chestnut rot caused by Gnomoniopsis castanea and the quality parameters of chestnuts. RESULTS: The results showed that ozone treatment (150 ppb during the day, and 300 ppb during the night) reduced the decay of chestnuts and had a fungistatic effect on isolates of G. castanea. The exposure of chestnuts to ozone did not alter weight losses, sugar content and titratable acidity. The concentration of total phenolics decreased during the storage period, both for treated and untreated nuts. However, after 150 days of treatment the polyphenol content of the chestnuts exposed to ozone was significantly higher than in control nuts. CONCLUSIONS: Our results suggested that ozone is an appropriate and economical tool to maximize the quality of chestnut shelf life, enabling it to be stored for long periods. © 2019 Society of Chemical Industry.


Assuntos
Fagaceae/química , Conservação de Alimentos/métodos , Conservantes de Alimentos/farmacologia , Nozes/química , Ozônio/farmacologia , Ascomicetos/efeitos dos fármacos , Ascomicetos/fisiologia , Carboidratos/química , Fagaceae/microbiologia , Conservação de Alimentos/instrumentação , Conservantes de Alimentos/química , Armazenamento de Alimentos , Nozes/microbiologia , Ozônio/química , Fenóis/química , Controle de Qualidade
10.
Rev Inst Med Trop Sao Paulo ; 61: e31, 2019 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-31241660

RESUMO

Chromoblastomycosis is a chronic fungal infection. Itraconazole and terbinafine are the most recommended antifungal drugs for chromoblastomycosis, while amphotericin B is not usually recommended. A patient with chromoblastomycosis in our hospital showed poor clinical responses to itraconazole and terbinafine. The fungus isolated from the lesions of this patient was identified as Fonsecaea nubica and numbered zssy0803. In vitro antifungal susceptibilities of F. nubica zssy0803 to terbinafine, amphotericin B, itraconazole, voriconazole and caspofungin were evaluated, as well as the combinations of terbinafine with the other four antifungals. The combined effect of terbinafine and amphotericin B on other 20 clinical F. nubica strains was also evaluated. The minimal inhibitory concentrations of terbinafine, amphotericin B, itraconazole, voriconazole and caspofungin on F. nubica zssy0803 were 0.25 µg/mL, 2 µg/mL, 1 µg/mL, 4 µg/mL and 8 µg/mL, respectively. The combination of terbinafine and amphotericin B showed the lowest fractional inhibitory concentration index of 0.28 to F. nubica zssy0803 in comparison with combinations of terbinafine and the other four antifungal drugs. The combination of terbinafine and amphotericin B was also synergistic for all the other 20 F. nubica strains. Then, the combination of oral terbinafine (500 mg/day) and intralesional injections of amphotericin B (1 mg/mL) was used to treat this patient. After this combined therapy for 25 weeks and terbinafine monotherapy for additional 12 weeks, the patient was cured. These findings indicate for the first time that terbinafine and amphotericin B are synergistic in killing F. nubica both in vitro and in vivo.


Assuntos
Anfotericina B/administração & dosagem , Antifúngicos/administração & dosagem , Ascomicetos/efeitos dos fármacos , Cromoblastomicose/tratamento farmacológico , Terbinafina/administração & dosagem , Ascomicetos/isolamento & purificação , Sinergismo Farmacológico , Quimioterapia Combinada , Feminino , Humanos , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade
11.
Plant Dis ; 103(8): 1884-1888, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31161931

RESUMO

It is a common practice to add salicylhydroxamic acid (SHAM) into artificial medium in the in vitro sensitivity assay of fungal phytopathogens to the quinone outside inhibitor (QoI) fungicides. The rationale for adding SHAM is to inhibit fungal alternative oxidase, which is presumed to be inhibited by secondary metabolites of plants. Therefore, the ideal characteristics of SHAM should be almost nontoxic to phytopathogens and have no significant effect on control efficacy of fungicides. However, this study showed that the average effective concentration for 50% inhibition (EC50) of mycelial growth values of SHAM were 97.5 and 401.4 µg/ml for Sclerotinia sclerotiorum and Botrytis cinerea, respectively. EC50 values of the three QoI fungicides azoxystrobin, kresoxim-methyl, and trifloxystrobin in the presence of SHAM at 20 and 80 µg/ml for S. sclerotiorum and B. cinerea, respectively, declined by 52.7 to 78.1% compared with those without SHAM. For the dicarboximide fungicide dimethachlone, the average EC50 values in the presence of SHAM declined by 18.2% (P = 0.008) for S. sclerotiorum and 35.9% (P = 0.012) for B. cinerea. Pot experiments showed that SHAM increased control efficacy of the three QoI fungicides against the two pathogens by 43 to 83%. For dimethachlone, SHAM increased control efficacy by 134% for S. sclerotiorum and 86% for B. cinerea. Biochemical studies showed that SHAM significantly inhibited peroxidase activity (P = 0.024) of B. cinerea and esterase activity (P = 0.015) of S. sclerotiorum. The strong inhibitions of SHAM per se on mycelial growth of B. cinerea and S. sclerotiorum and significant influences on the sensitivity of the two pathogens to both the QoI fungicides and dimethachlone as well as inhibitions on peroxidase and esterase indicate that SHAM should not be added in the in vitro assay of sensitivity to the QoI fungicides.


Assuntos
Ascomicetos , Botrytis , Farmacorresistência Fúngica , Fungicidas Industriais , Salicilamidas , Ascomicetos/efeitos dos fármacos , Botrytis/efeitos dos fármacos , Farmacorresistência Fúngica/efeitos dos fármacos , Fungicidas Industriais/farmacologia , Salicilamidas/farmacologia
12.
Plant Dis ; 103(8): 2076-2082, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31194616

RESUMO

Stem-end rot caused by Botryodiplodia theobromae is a destructive disease of mango. B. theobromae field isolates resistant to carbendazim (MBC) were collected in Hainan Province, China. In this study, the characteristics of these field isolates with resistance to MBC were investigated. The resistance of B. theobromae isolates to MBC was stably inherited. Both the MBC-resistant and MBC-sensitive isolates had similar mycelial growth rates, pathogenicity, sensitivity to high glucose, glycerol content, and peroxidase activity. Compared with MBC-sensitive isolates, MBC-resistant isolates were more sensitive to low temperature and had a significant decrease in sensitivity to high NaCl and a significant increase in catalase (CAT) and glutathione S-transferase (GST) activities. After MBC treatment, the cell membrane permeability of the sensitive isolates was markedly increased compared with that of the resistant isolates. Analysis of the ß-tubulin gene sequence revealed point mutations resulting in substitutions at codon 198 from glutamic acid (GAG) to alanine (GCG) in moderately resistant isolates, and at codon 200 from phenylalanine (TTC) to tyrosine (TAC) in highly resistant isolates. These ß-tubulin gene mutations were consistently associated with MBC resistance. Overall, we infer that the altered cell membrane permeability and the increase in CAT and GST activities of the resistant isolates are linked to MBC resistance.


Assuntos
Ascomicetos , Benzimidazóis , Carbamatos , Farmacorresistência Fúngica , Fungicidas Industriais , Ascomicetos/efeitos dos fármacos , Ascomicetos/enzimologia , Ascomicetos/genética , Ascomicetos/metabolismo , Benzimidazóis/farmacologia , Carbamatos/farmacologia , China , Farmacorresistência Fúngica/genética , Fungicidas Industriais/farmacologia , Genes Fúngicos/genética
13.
Plant Dis ; 103(7): 1515-1524, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31059385

RESUMO

Powdery mildew, caused by the fungus Podosphaera xanthii, is one of the most economically important diseases affecting cucurbit crops in Spain. Currently, chemical control offers the most efficient management of the disease; however, P. xanthii isolates resistant to multiple classes of site-specific fungicides have been reported in the Spanish cucurbit powdery mildew population. In previous studies, resistance to the fungicides known as methyl benzimidazole carbamates (MBCs) was found to be caused by the amino acid substitution E198A on ß-tubulin. To detect MBC-resistant isolates in a faster, more efficient, and more specific way than the traditional methods used to date, a loop-mediated isothermal amplification (LAMP) system was developed. In this study, three sets of LAMP primers were designed. One set was designed for the detection of the wild-type allele and two sets were designed for the E198A amino acid change. Positive results were only obtained with both mutant sets; however, LAMP reaction conditions were only optimized with primer set 2, which was selected for optimal detection of the E198A amino acid change in P. xanthii-resistant isolates, along with the optimal temperature and duration parameters of 65°C for 75 min, respectively. The hydroxynaphthol blue (HNB) metal indicator was used for quick visualization of results through the color change from violet to sky blue when the amplification was positive. HNB was added before the amplification to avoid opening the lids, thus decreasing the probability of contamination. To confirm that the amplified product corresponded to the ß-tubulin gene, the LAMP product was digested with the enzyme LweI and sequenced. Our results show that the LAMP technique is a specific and reproducible method that could be used for monitoring MBC resistance of P. xanthii directly in the field.


Assuntos
Ascomicetos , Farmacorresistência Fúngica , Doenças das Plantas , Ascomicetos/efeitos dos fármacos , Ascomicetos/genética , Ascomicetos/fisiologia , Benzimidazóis/farmacologia , Carbamatos/farmacologia , Técnicas de Amplificação de Ácido Nucleico , Espanha
14.
Plant Dis ; 103(7): 1657-1664, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31082320

RESUMO

Adding adjuvants improved the affinity of fungicide droplets to cucumber leaves infected with powdery mildew (Podosphaera xanthii) and subsequent efficacy of fungicide treatments in reducing the disease. The affinity of oil adjuvants was quantified by adhesional tension and "work of adhesion". Oil adjuvant-fungicide mixtures were applied to plants in field experiments to evaluate their effectiveness in disease prevention. Both the adhesional tension and work of adhesion of the adjuvants at selected concentrations increased on powdery-mildew-infected cucumber leaves more than on healthy cucumber leaves. The adjuvant GY-Tmax (GYT) displayed the best surface activity or "surfactivity" in enhancing the affinity and adherence of droplets to powdery-mildew-infected cucumber leaves, while epoxidized soybean oil (ESO), methyl oleate, and biodiesel exhibited much lower effects in terms of the surface tension, contact angle, adhesional tension, and work of adhesion. Field experiments determined that the combination of GYT at 1,000 mg liter-1 and pyraclostrobin (150 g a.i. ha-1) was most effective (91.52%) in controlling cucumber powdery mildew. Pyraclostrobin with ESO was also highly effective (ranging from 77.54 to 89.65%). The addition of oil adjuvants, especially GYT and ESO, to fungicide applications can be an effective strategy to enhance the efficacy of pesticides in controlling plant diseases by modifying the affinity of fungicide droplets to symptomatic leaves.


Assuntos
Ascomicetos , Cucumis sativus , Óleos , Estrobilurinas , Ascomicetos/efeitos dos fármacos , Ascomicetos/fisiologia , Fungicidas Industriais/farmacologia , Óleos/farmacologia , Folhas de Planta/microbiologia , Estrobilurinas/farmacologia
15.
Sci Total Environ ; 679: 260-269, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31082599

RESUMO

Heavy metals are known for their negative impact on the physiological processes of lichen photobiont. In spite of this, certain lichens are known to be effective pioneers of polluted sites. Cladonia cariosa, C. rei, and Diploschistes muscorum are prominent examples of lichens that spontaneously colonise post-industrial wastes. We examined the effect of total and intracellular Zn, Pb, Cd, As, Cu, and Ni accumulation in the thalli of these species on the physiological parameters of photobiont. Increased accumulation of Zn, Cd, Cu, and Ni in D. muscorum and of Zn and Ni in C. rei negatively affected contents of photosynthetic pigments, whereas concentrations of Pb had a positive effect in all lichen species. Moreover, pigment contents were positively associated with the concentrations of most examined elements in C. cariosa. The results indicate that even if chlorophyll contents reduced, its degradation does not progress. This suggests that metal stress may exert a negative effect on the synthesis rather than on the integrity of chlorophyll. Most importantly, lichen samples of each of the species from polluted sites proved to possess significantly higher FV/FM ratios than those from a reference site; moreover, the contents of elements of lichen thalli positively influenced this parameter. The efficient functioning of the algal component under heavy-metal stress conditions indicates that the examined lichens are well adapted to extremely contaminated substrates.


Assuntos
Ascomicetos/efeitos dos fármacos , Poluição Ambiental/efeitos adversos , Líquens/efeitos dos fármacos , Metais Pesados/efeitos adversos , Ascomicetos/crescimento & desenvolvimento , Líquens/crescimento & desenvolvimento , Polônia , Estresse Fisiológico
16.
Plant Dis ; 103(8): 1910-1917, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31140926

RESUMO

The occurrence of fungal brown spot, caused by Bipolaris oryzae, has increased in cultivated wild rice (Zizania palustris) paddies in spite of the use of azoxystrobin-based fungicides. The active ingredient blocks electron transfer at the quinone outside inhibitor (QoI) site in the mitochondrial cytochrome b within the bc1 complex, thus obstructing respiration. The in vitro averaged EC50 of baseline isolates collected in 2007 before widespread fungicide use was estimated to be 0.394 µg/ml with PROBIT and 0.427 µg/ml with linear regression analyses. Isolates collected during 2008, 2015, and 2016 had a range of sensitivity as measured by relative spore germination (RG) at a discriminatory dose of 0.4 µg/ml azoxystrobin. Isolates with a higher (≥80%) and lower RG (≤40%) had the wild type nucleotides at amino acid positions F129, G137, and G143 of cytochrome b, sites known to be associated with QoI fungicide resistance. Two Group I introns were found in the QoI target area. The splicing site for the second intron was found immediately after the codon for G143. A mutation for fungicide resistance at this location would hinder splicing and severely reduce fitness. B. oryzae expresses an alternative oxidase in vitro, which allows the fungus to survive inhibition of respiration by azoxystrobin. This research indicates that B. oryzae has not developed resistance to QoI fungicides, although monitoring for changes in sensitivity should be continued. Judicious use of QoI fungicides within an integrated disease management system will promote an effective and environmentally sound control of the pathogen in wild rice paddies.


Assuntos
Ascomicetos , Farmacorresistência Fúngica , Oryza , Pirimidinas , Estrobilurinas , Ascomicetos/efeitos dos fármacos , Oryza/microbiologia , Pirimidinas/farmacologia , Estrobilurinas/farmacologia
17.
Int J Food Microbiol ; 301: 9-18, 2019 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-31055161

RESUMO

A large amount of GRAS (generally recognized as safe) salts and concentrations were evaluated in in vitro tests (inhibition of mycelial growth on PDA dishes) against Lasiodiplodia theobromae, the causal agent of citrus Diplodia stem-end rot. Ammonium carbonate (AC, 0.2%), potassium sorbate (PS, 2.0%), potassium carbonate (PC, 0.2%), sodium methylparaben (SMP, 0.1%), sodium ethylparaben (SEP, 0.1%), sodium benzoate (SB, 2.0%), and potassium silicate (PSi, 2.0%) were selected as the most effective. Disease control ability of edible composite coatings formulated with hydroxypropyl methylcellulose (HPMC), beeswax (BW), and these selected antifungal GRAS salts was assessed in in vivo experiments with 'Ortanique' mandarins and 'Barnfield' oranges artificially inoculated with L. theobromae. Coatings containing 2% PS, 0.1% SEP, or 2% SB were the most effective reducing disease severity (up to 50% reduction) and were also applied to non-inoculated and cold-stored 'Barnfield' oranges to determine their effect on postharvest fruit quality. After periods of 21 and 42 d at 5 °C followed by 7 d of shelf life at 20 °C, coatings containing SEP and SB significantly reduced weight loss and did not adversely affect the physicochemical quality attributes (firmness, soluble solid content, titratable acidity, and ethanol and acetaldehyde content) and sensory flavor with respect to uncoated control fruit. Although the internal gas concentration (CO2 level) of coated fruit increased, the coatings did not induce off-flavors.


Assuntos
Ascomicetos/efeitos dos fármacos , Citrus/microbiologia , Microbiologia de Alimentos/métodos , Conservação de Alimentos/métodos , Derivados da Hipromelose/farmacologia , Sais/farmacologia , Antifúngicos/farmacologia , Frutas/microbiologia , Lipídeos/química , Lipídeos/farmacologia
18.
Plant Dis ; 103(7): 1613-1620, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30998449

RESUMO

Sclerotinia sclerotiorum, which can cause Sclerotinia stem rot, is a prevalent plant pathogen. This study aims to evaluate the application potential of benzovindiflupyr, a new generation of succinate dehydrogenase inhibitor (SDHI), against S. sclerotiorum. In our study, 181 isolates collected from different crops (including eggplant [n = 34], cucumber [n = 27], tomato [n = 29], pepper [n = 35], pumpkin [n = 32], and kidney bean [n = 25]) in China were used to establish baseline sensitivity to benzovindiflupyr. The frequency distribution of the 50% effective concentration (EC50) values of benzovindiflupyr was a unimodal curve, with mean EC50 values of 0.0260 ± 0.011 µg/ml, and no significant differences in mean EC50 existed among the various crops (P > 0.99). Benzovindiflupyr can effectively inhibit mycelial growth, sclerotial production, sclerotial shape, and myceliogenic and carpogenic germination of the sclerotia of S. sclerotiorum. In addition, benzovindiflupyr showed good systemic translocation in eggplant. Using benzovindiflupyr at 100 µg/ml yielded efficacies of 71.3 and 80.5% for transverse activity and cross-layer activity, respectively, which were higher than those of acropetal and basipetal treatments (43.6 and 44.7%, respectively). Greenhouse experiments were then carried out at two experimental sites for verification. Applying benzovindiflupyr at 200 g a.i. ha-1 significantly reduced the disease incidence and severity of Sclerotinia stem rot. Overall, the results demonstrated that benzovindiflupyr is a potential alternative product to control Sclerotinia stem rot.


Assuntos
Ascomicetos , Norbornanos , Pirazóis , Ascomicetos/efeitos dos fármacos , China , Produtos Agrícolas/microbiologia , Fungicidas Industriais/farmacologia , Norbornanos/farmacologia , Pirazóis/farmacologia , Succinato Desidrogenase/antagonistas & inibidores
19.
Plant Dis ; 103(7): 1458-1463, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31025906

RESUMO

Botryosphaeria dothidea is an important fungal pathogen that causes apple ring rot, which can significantly reduce apple yield. Fungicide applications are the main control measure of apple ring rot worldwide. Pyraclostrobin is a quinone outside inhibitor (QoI) fungicide that has yet to be registered for control of B. dothidea in China. Baseline sensitivity of B. dothidea to pyraclostrobin (EC50 of mycelial growth inhibition) was assessed for 97 isolates collected in Shandong Province. The EC50 values ranged from 0.7010 to 7.1378 µg/ml with the mean value of 3.0870 µg/ml and displayed a unimodal frequency distribution. After cultured on fungicide-free PDA medium or on apples for multiple generations, the B. dothidea-resistant isolates (RST) remained resistant to pyraclostrobin, but exhibited similar virulence as the susceptible isolates (ST). Cross-resistance investigation revealed that pyraclostrobin was not cross-resistant to tebuconazole, flusilazole, carbendazim, and iprodione. Field evolution showed that pyraclostrobin at 200 and 250 g a.i./ha provided greater than 80% control efficacy against apple ring rot disease when applied as a therapeutic or preventive fungicide. The efficacy was similar to fungicides that have been registered for apple.


Assuntos
Ascomicetos , Malus , Estrobilurinas , Ascomicetos/efeitos dos fármacos , China , Fungicidas Industriais/farmacologia , Concentração Inibidora 50 , Malus/microbiologia , Estrobilurinas/farmacologia
20.
Plant Dis ; 103(6): 1112-1118, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30995420

RESUMO

Cherry leaf spot (CLS), caused by the fungus Blumeriella jaapii, is a major disease of tart cherry (Prunus cerasus L.) trees, leading to early defoliation that results in uneven ripening and poor fruit quality in the current season, reduced fruit set in the following season, and increased potential for winter injury and tree death. Pristine (BASF Corporation, Research Triangle Park, NC), a commonly used fungicide for CLS management in Michigan, is a premix of boscalid, a succinate dehydrogenase inhibitor, and pyraclostrobin, a quinone outside inhibitor. Reduced efficacy of Pristine for CLS control was observed in field trials and commercial orchards and highlights the importance of fungicide resistance monitoring. A total of 1,189 isolates from 31 commercial orchards in Michigan, 111 isolates from nontreated trees (four locations in Michigan and two locations in Ohio), and 133 isolates from a research orchard were collected during 2010, 2011, and 2012 and assayed on boscalid-amended media at concentrations ranging from 0 to 25 µg ml-1. Because of the very slow growth rate of B. jaapii in culture, we determined the minimum inhibitory concentration (MIC) of boscalid as opposed to the effective concentration that inhibits mycelial growth to 50% of the control. Isolates from nontreated trees had MIC values ranging from 0.1 to 0.5 µg ml-1; the MIC of isolates from commercial orchards ranged from 0.1 to >25 µg ml-1, and isolates from the research orchard ranged from 2.5 to >25 µg ml-1. Isolates with MIC values ≥25 µg ml-1 were considered boscalid resistant and comprised 0% of the nontreated isolates, 30.4% of the commercial isolates, and 42.1% of the research orchard isolates. Sequencing of the sdhB gene of resistant isolates led to the detection of the amino acid mutation H260R, which is known to confer boscalid resistance in other phytopathogenic fungi. Our results indicate that the occurrence of the H260R mutation in Michigan populations of B. jaapii is correlated with the reduction in sensitivity to boscalid observed in commercial orchards.


Assuntos
Ascomicetos , Compostos de Bifenilo , Farmacorresistência Fúngica , Niacinamida/análogos & derivados , Ascomicetos/efeitos dos fármacos , Ascomicetos/genética , Compostos de Bifenilo/farmacologia , Farmacorresistência Fúngica/genética , Genes Fúngicos/genética , Michigan , Testes de Sensibilidade Microbiana , Niacinamida/farmacologia , Ohio , Prunus/microbiologia
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