Identification, pathogenicity, and fungicide sensitivity of Eutiarosporella dactylidis associated with leaf blight on maize in China

Maize(Zea mays L.) is an economically vital grain crop that is cultivated worldwide. In 2011, a maize foliar disease was detected in Lingtai and Lintao counties in Gansu Province, China. The characteristic signs and symptoms of this disease include irregular chlorotic lesions on the tips and edges of infected leaves and black punctate fruiting bodies in dead leaf tissues. Given favourable environmental conditions, this disease spread to areas surrounding Gansu. In this study, infected leaves were collected from Gansu and Ningxia Hui Autonomous Region between 2018and 2020 to identify the disease-causing pathogen. Based on morphological features, pathogenicity tests, and multilocus phylogenetic analysis involving internal transcribed spacer(ITS), 18S small subunit rDNA(SSU), 28S large subunit rDNA(LSU), translation elongation factor 1-alpha(TEF), and β-tubulin(TUB) sequences, Eutiarosporella dactylidis was identified as the causative pathogen of this newly discovered leaf blight. Furthermore, an in vitro bioassay was conducted on representative strains using six fungicides, and both fludioxonil and carbendazim were found to significantly inhibit the mycelial growth of E. dactylidis. The results of this study provide a reference for the detection and management of Eutiarosporella leaf blight.

Research Progress on Detection Techniques of Fungicide Residues in Chinese Chives

Chinese chive is a kind of medicinal and edible plant,with many diseases,and chemical fungicides are usually used for control.In order to find out the risk of pesticide residues in Chinese chives,this paper summarized relevant literatures published in recent years,and sorted out and analyzed the types of pesticides used and detection techniques of common diseases in Chinese chives.

Toxicity of 26 Fungicides against and Synergistic Effects of Binary Mixtures on Oidium heveae

[Objective] This study was conducted to determine the toxicity of 26 fungicides against Oidium heveae and the synergistic effects of their binary mixtures.[Method] The sensitivity of O.heveae to 26 fungicides was determined in vitro and in vivo by calculating EC50 values.Horsfall＇s toxicity rate was preliminarily employed to predict the type of joint action of 180 two-way mixtures,based on which the interaction level of the potentially synergistic mixtures was validated by Sun and Johnson＇s co-toxicity coefficient.[Result] Triflumizole,pyraoxystrobin,flufenoxystrobin,epoxiconazole,pyrametostrobin,flusilazole,pyraclostrobin and tetraconazole were the most potent inhibitors of both spore germination and mycelial growth.An ‘in vitro-in vivo＇ integrated strategy is proposed for screening the inhibitors of O.heveae,according to the fact that data obtained in germination experiment are able to reflect to some extent the effectiveness of fungicide confirmed by pot experiment.Results achieved by Horsfall＇s were in line with that validated by Sun and Johnson＇s,the Horsfall＇s seems to be a convenient means to assess rapidly the potential interaction type and level of fungicides in a binary mixture.Nineteen mixtures interact synergistically：epoxiconazole ＋ pyraclostrobin at the mixing ratio of 6：4,8：2;tetraconazole ＋ flufenoxystrobin at 1：9,2：8,3：7 and 4：6;triadimefon ＋ flufenoxystrobin at 9：1;triflumizole ＋ flufenoxystrobin at 8：2;flusilazole ＋pyrametostrobin at 7：3,8：2 and 9：1;tetraconazole ＋ pyrametostrobin at 6：4;triadimefon ＋ pyrametostrobin at 5：5,8：2;triflumizole ＋ pyrametostrobin at 7：3,8：2;tetraconazole ＋ pyraoxystrobin at 9：1;triflumizole ＋ pyraoxystrobin at 8：2,9：1.[Conclusion] The single fungicides and their binary mixtures are expected to serve as the alternatives to triadimefon and the alternate use of them could delay the buildup of resistant O.heveae isolate.

Field Efficacy of Six Kinds of Fungicides for Control and Prevention of Apple Powdery Mildew(Podosphaera leucotricha)

[Objective] This study aimed to screen effective fungicide for prevention and control of apple powdery mildew. [Method] Field efficacy of six kinds of fungi- cides with 18 concentrations against apple powdery mildew was investigated. [Re- sult] 4 000-fold triflumizole WP exhibited the highest control efficacy of 82.96%, fol- lowed by 3 000-fold triflumizole WP with control efficacy of 77.75%, while 800-fold and 1 000-fold 70% thiophanate methyl WP exhibited relatively low control efficacy, which was both lower than 60%. [Conclusion] In actual production, 4 000-fold triflu- mizole WP and 3 000-fold triflumizole WP should be used alternately to control and prevent apple powdery mildew.

Inhibition of Chlamydospore Germination and Mycelial Growth of Trichoderma spp. by Chemical Fungicides

For exploring the influences of application and residue of chemical fungi- cides on chlamydospore preparations of Trichoderma spp., the effects of seven chemical fungicides on chlamydospore germination and mycelia growth of two bio- control fungi T. harzianum 610 and T. Iongibrachiatum 758 were studied. Carben- dazim, tebuconazole and difenoconazole showed strong toxicities, thiram and car- bexin showed moderate toxicities, and metalaxyl showed Mycelia growth of the two strains was more sensitive to most tested fungicides than those of chlamydospore germination. Chlamydospore germination of 610 was more sensitive to tested fungicides than those of 758, and mycelia growth of 758 was more sensitive to most tested fungi- cides than those of 610. Among the seven fungicides, 98% carbendazim had the strongest effects （ECru values were 1.64 and 0.05μpg/ml）, and 70% pentachloroni- trobenzene had the weakest effects （EC50 values were 1.64 and 0,05 μg/ml） for chlamydospore germination and mycelia growth of 610. As for 758, 98% carbendaz- im had the strongest inhibitory effects and 95% metalaxyl had the weakest inhibitory for chlamydospore germination of 756 （EC50 values were 0.62 and 1 108.61 μg/ml respectively）, whereas 96.2% tebuconazole showed the strongest inhibitory effects for mycelia growth of 758 （EC= value was 0.32μg/ml）, and 95% metalaxyl was the weakest （EC= value was 1 206.29 μg/ml）. According to the applied concentration of different fungicides in practice, we concluded that chlamydospore preparations of 610 and 758 could not be combined with carbendazim, tebuconazole, thiram and carboxin for controlling plant diseases, and the pesticide residues to the biocontrol effects should be kept in mind. Chlamydospore preparations of 610 and 758 can be and difenoconazole for controlling plant dis- eases, 758 chlamydospore preparations and germinated chlamydospore of 610 can be combined with metalaxyl for controlling plant diseases, and pesticide residue risk was not serious.

Antibiotic Activity of Seven Fungicides against Alternaria panax Whetz

[Objective] This study was conducted to screen suitable fungicides to con-trol ginseng leaf blight caused by Alternaria panax_Whetz. [Method] The antifungal activity of seven fungicides against A. panax_ was determined based on mycelial growth rate in vitro. [Result] The results of in vitro antibiotic activity assay showed that there were significant differences in inhibition rate among different concentration treatments of each of the seven fungicides. Toxicity test results showed that among the seven fungicides, difenoconazole had the smal est EC50 （0.61 mg/L）, fol owed by streptomycin and captan, with EC50 value lower than 100 mg/L. [Conclusion] A fungicide which had strong antifungal activity on A. panax was screened out, and the results wil provide a theoretical basis for further field trial.

Activity of the Fungicide JS399-19 Against Fusarium Head Blight of Wheat and the Risk of Resistance

This report reviews the characteristics of JS399-19, a novel cyanoacrylate fungicide. JS399-19 strongly inhibits the mycelial growth of the fungal plant pathogens of the genus Fusarium and exhibits great potential in controlling Fusarium head blight （FHB） on wheat and other cereals. The mode of action of JS399-19 is evidently different from that of benzimidazole （for example, carbendazim） and other sort of fungicides, making it a possible replacement for carbendazim in China to manage carbendazim-resistant subpopulations of Fusarium graminearum and F. asiaticum. JS399-t9 has excellent protective and curative activity against these pathogens. Incorrect use of this fungicide, however, is likely to select for resistance. Among JS399-19-resistant mutants of F. asiaticum induced in the laboratory, the resistant level of mutants was high and the phenotype of resistance against JS399-19 was conferred by a major gene by genetic analysis. The fitness of laboratory-induced JS399-19-resistant mutants of F. asiaticum was nearly equal to that of their parents. JS399-19 lacks cross resistance with other sort fungicides. To control FHB with JS399-19 and to delay the development of the fungicide-resistance, farmers should use tank mixtures containing JS399-19 and carbendazim, metconazole, tebuconazole, or prothioconazole.

The Effect of Three New Fungicides against Rice Sheath Blight in Field Experiment

[Objective] The aim was to study the effect of three new fungicides against rice sheath blight in field experiment. [Methods] The experiment set up 7 treatments with three times of repetition and designed by random grouping. By using 5 sampling points in each plot, and investigating continuous 4 holes of each point, total plants, diseased plants and disease degrees were recorded. Then disease index and control efficiency were calculated, and variance analysis was carried out. [Results] 300 or 450 ml/hm^2 azoxystrobin ＋ difenoconazole 325 g/L SC had better control efficiency to rice sheath blight and had no phytotoxicity effect, we should use it at the initial disease stage and continuously spray 2-3 times. [Conclusion] The experiment provided a theoretical basis for controlling rice sheath blight using fungicides.

Pathogen Identification of the Anthracnose of Ligusticum chuanxiong Hort. and Inhibitory Effect of Four Fungicides on the Pathogen

[Objective] The research aimed to isolate and identify the pathogen of anthracnose of Ligusticum chuanxiong Hort,and make the antifungal test on four kinds of fungicides for selecting the optimum fungicides.[Method] The pathogenic fungi was separated according to Koch＇s rule and the shape characteristics were identified. The antifungal test of fungicides was made by using plate mycelium growth inhibition method.[Result] The pathogenic fungi was separated from leaves of Ligusticum chuanxiong Hort. and morphological characters were consistent with that of Colletotrichum gloeosporioides. In addition to chloroisobromine cyanuric acid,the other three fungicides could inhibit the mycelium growth. The growth inhibition rates of 1 000 times Metalaxyl＋mancozeb,zineb,Thiophanate-methyl for the mycelium were 26.8%,22.1%,59.8% respectively after 7 days.[Conclusion] The anthracnose was caused by Colletotrichum gloeosporioides. Ligusticum chuanxiong Hort. was a new record host plant of Colletotrichum gloeosporioides and the best inhibiting fungicide was Thiophanate methyl.

Biological Activity of Several Fungicides against Ustilaginoidea virens

[Objective] This study aims to screen for the high effective fungicides which could significantly decrease the disease incidence and disease index of rice false smut. [Method] The inhibitory activities of the fungicide against mycelial growth of Ustilaginoidea virens were measured to in vitro evaluate the ECho values. And 17 fungicides were sprayed to evaluate the efficacy and effect of the fungicides tested in the field trials on the rice characters, [Result] The results showed that epoxicona- zole, difenoconazole, propiconazole and procloraz exhibited high inhibitory activity against mycelial growth of Ustilaginoidea virens with the ECso values 0.04, 0.07, 0.12 and 0.11 pg/ml, respectively. The results of field trials showed that the efficacy of Wen- quning, and fungicides such as difenoconazole, prochloraz, propiconazole, epoxi- conazole and their mixtures in controlling rice false smut were all 70% or more. [Conclusion] The 17 tested fungicides behaved efficacy in controlling rice false smut and did not cause drug injury on leaves and grains of rice plants, sprayed when flag leaves of rice fully expanded.

Inhibitory Effect of 16 Fungicides on Persimmon Anthracnose Pathogen Colletotrichum horii

In this research,the inhibitory effect of 16 fungicides on Colletotrichum horri causing persimmon anthracnose was investigated using mycelial growth method and spore germination method. The results showed that among the 16 tested fungicides,10% of Difenoconazole WG,33. 5% of Copper quinolate SC,25%of Bromothalonil EC,70% of Mancozeb WP,430 g/L of Tebuconazole SC,50% of Prochloraz-manganese chloride and 400 g/L of Flusilazole EC achieved the best inhibitory effect on mycelial growth of C. horri,with the inhibition rate of 100%; 70% of Polyram WG,33. 5% of Copper quinolate SC,25% of Bromothalonil EC,70% of Mancozeb WP,50% of Chlorobromoisocyanuric acid AF,50% of Triram WP and 400 g/L of Flusilazole achieved the best inhibitory effect on spore germination of C. horri,with the germination rate of 0. In conclusion,33. 5% of Copper quinolate SC,25% of Bromothalonil EC,70% of Mancozeb WP and 400 g/L of Flusilazole EC achieved the best inhibitory effect both in mycelial growth and spore germination,which could be used as the preference fungicides for the control of persimmon anthracnose,and 70% of Polyram WG and 50% of Triram WP achieved the secondly best inhibitory effect,which could be used as alternative fungicides. The results of this research could provide scientific evidence for the effective control of persimmon anthracnose,and more optional pesticides for utilization in the production practice of persimmon industry.

A Preliminary Study on the Resistance to Different Fungicides in Phytophthora parasitica var. nicotianae

[Objective] This study aimed to investigate the resistance to different fungicides in Phytophthora parasitica var. nicotianae. [Method] Under indoor incubation conditions, the resistance to dimethomorph, metalaxyl-mancozeb, propamocarb and ovraclostrobin.dimethomorph in P. parasitica strain isolated from Zhenyuan County in Qiandongnan State was analyzed with colony growth measurement method. [Result] P. parasitica exhibited different levels of sensitivity to four fungicides. To be specific, P. parasitica exhibited the highest resistance to dimethomorph, and ECho reached 1.19 μg/ml. [Conclusion] In Zhenyuan tobacco-growing area, long-term single use of dimethomorph possesses certain resistance risk in prevention and control of black shank disease.

Assessing the impact of fungicide enostroburin application on bacterial community in wheat phyllosphere

Fungicides have been used extensively for controlling fungal pathogens of plants. However, little is known regarding the effects that fungicides upon the indigenous bacterial communities within the plant phyllosphere. The aims of this study were to assess the impact of fungicide enostroburin upon bacterial communities in wheat phyllosphere. Culture-independent methodologies of 16S rDNA clone library and 16S rDNA directed polymerase chain reaction with denaturing gradient gel electrophoresis （PCR-DGGE） were used for monitoring the change of bacterial community. The 16S rDNA clone library and PCR-DGGE analysis both confirmed the microbial community of wheat plant phyllosphere were predominantly of the γ-Proteobacteria phyla. Results from PCR-DGGE analysis indicated a significant change in bacterial community structure within the phyllosphere following fungicide enostroburin application. Bands sequenced within control cultures were predominantly of Pseudomonas genus, but those bands sequenced in the treated samples were predominantly strains of Pantoea genus and Pseudomonas genus. Of interest was the appearance of two DGGE bands following fungicide treatment, one of which had sequence similarities （98%） to Pantoea sp. which might be a competitor of plant pathogens. This study revealed the wheat phyllosphere bacterial community composition and a shift in the bacterial community following fungicide enostroburin application.

Effects of Priming in Combination with Fungicides on Germination and Infestation of Lettuce (Lactuca sativa L.)

Two seed lots of lettuce (Lactuca sativa L.) cv. Krlowa Majowych were primed in polyethylene glycol (PEG) 8000 at -1.25MPa for 7 days in dark at 20C. Two fungicide treatments combined with PEG were used to control fungi: 1) Funaben T(0.1%) and Apron 35 SD (0.02%) were added to -1.25 MPa PEG solution (PEG+F); 2) After seed priming and drying,Funaben T was mixed at 5 g kg-1 and Apron 35 SD at 1 g kg-1 of seeds (PEG then F dust). PEG, PEG+F and PEG then F dusttreatments reduced significantly thermodormancy and increased the seed germination at 35C in both lots. PEG then Fdust treatment had the best effect on reducing seed thermodormancy at 35C, and increased seed germination to 64.5% inlot 1 and 72.0% in lot 2 compared to 0 in untreated. PEG and PEG then F dust treatments have accelerated germinationspeed at 20 and 15C in both lots. There was a significant reduction in seed infestation in which the seeds were treatedwith PEG+F and PEG then F dust compared to untreated seeds and PEG primed seeds. PEG priming enhanced seedinfestation considerably by Alternaria alternata, Penicillium spp., Rhizopus spp. and Ulocladium spp. Mycologicalanalysis showed higher frequency of Alternaria alternata and Ulocladium spp. in untreated seeds at 20C. Priming incombination with fungicides significantly alleviated the infestation of fungi, especially Alternaria alternata infestation.

Effect of <i>Trichoderma harzianum</i>in Combination with Fungicides in Controlling Gray Mould Disease (<i>Botrytis cinerea</i>) of Strawberry

The effects of biofungicide formula containing the biocontrol agent Trichoderma harzianum (Jn14) as wettable powder in combination with the fungicides pyrimethanil and a mixture of cyprodinil and flydioxonil on Botrytis cinerea of strawberry in vitro, in vivo and in greenhouse were studied. The pathogen B. cinerea was more affected in vitro by low concentrations of the fungicides tested above 0.4 μg&middot;ml-1 than the bioagent T. harzianum (Jn14). The later was almost insensitive to pyrimethanil. In the same direction, gray mould disease severity on strawberry detached leaves and whole plants was reduced up to 89% by the tested fungicides, compared to the control, while the application of T. harzianum (Jn14) alone reduced disease severity up to 45% on strawberry detached leaves, compared to the control. In the integrated control approach, the combination of T. harzianum (Jn14) with higher concentrations of the tested fungicides (ED50) completely inhibited strawberry gray mould disease severity in pots and the greenhouse.

Sensitivity assessment of Phytophthora nicotianae to fungicides of fluazinam and its mixture with carboxylic acid amide

Tobacco black shank caused by Phytophthora parasitica var.nicotianae(Breda de Hean)is one of the most serious diseases in tobacco growing,and few effective fungicides are available currently.The purpose of this study was to assess the feasibility of using fluazinam to treat tobacco black shank disease.In vitro toxicities of fluazinam to 62 isolates of P.nicotianae were determined by mycelial growth rate method.The EC50 values ranged from 0.294 6 to 2.389 9 μg/mL with a mean of 1.084 0 μg/mL.The sensitivity distribution was unimodal,and there was no resistant isolate in P.nicotianae population.The carboxylic acid amide(CAA)fungicides were single-site in action and activity to oomycetes.The synergistic interactions were tested by Wadley method between fluazinam and dimethomorph,flumorph,mandipropamid,respectively.The mixture containing dimethomoph:Huazinam at 1:2 ratio showed additive actions,with the synergy ratio of 1.2.The mixture containing flumorph and fluazinam at 1:5 ratio had the highest synergy ratio(SR 2.07).Mandipropamid and fluazinam had an additive interaction and the proportion of 2:1 had the lowest EC50 value(0.020 7 μg/mL).This study demonstrated the sensitivity of P.nicotianae to fluazinam,and provided the experimental foundation to use fluazinam for tobacco black shank management.

Control Effects of Different Fungicides on Pear Scab

[Objective] The paper was to select different fungicides for field efficacy test against pear scab in summer.[Method] Five treatments were designed in the test,including 10% difenoconazole WDG 6 000 times dilution,50% captan WP 600 times dilution,30% pyraclostrobin SC 2 000 times dilution,430 g/L tebuconazole 4 000 times dilution,and water(control).[Result] 10% Difenoconazole WDG 6 000 times dilution,50% captan WP 600 times dilution,30% pyraclostrobin SC 2 000 times dilution,430 g/L tebuconazole SC 4 000 times dilution had good control effects against pear scab,and the efficacies were 89.13%-93.51%.[Conclusion] The results will provide the theoretical basis for field control of pear scab in summer.

Mitochondrial dynamics caused by QoIs and SDHIs fungicides depended on FgDnm1 in Fusarium graminearum

Fusarium head blight(FHB) caused by Fusarium graminearum is a devastating fungal disease on small grain cereal crops,because it reduces yield and quality and causes the mycotoxin contamination to the grain.Dynamins and dynamin-related proteins(DRPs) are large GTPase superfamily members,which are typically involved in the budding and division of vesicles in eukaryotic cells,but their roles in Fusarium spp.remain unexplored.Here,we found that FgDnm1,a DRP and homolog to Dnm1 in Saccharomyces cerevisiae,contributes to the normal fungal growth,sexual reproduction and sensitivity to fungicides.In addition,we found FgDnm1 co-localizes with mitochondria and is involved in toxisome formation and deoxynivalenol(DON) production.Several quinone outside inhibitors(QoIs) and succinate dehydrogenase inhibitors(SDHIs) cause fragmentated morphology of mitochondria.Importantly,the deletion of FgDnm1displays filamentous mitochondria and blocks the mitochondrial fragmentation induced by QoIs and SDHIs.Taken together,our studies uncover the effect of mitochondrial dynamics in fungal normal growth and how such events link to fungicides sensitivity and toxisome formation.Thus,we concluded that altered mitochondrial morphology induced by QoIs and SDHIs depends on FgDnm1.

Sensitivity of Four Yeasts to Fungicides and CO2 Concentrations and Their Antagonistic Ability in Combination with Fungicide to Pathogenic Fungi in vitro

The yeasts Cryptococcus laurentii, Trichosporon pullulans, Rhodotorula glutinis andTrichosporon sp.were used to investigate their sensitivity to four fungicides anddifferent concentrations of CO2, as well as their antagonistic ability to Penicilliumexpansumand Alternariaalternata in vitro when applied with fungicide. There were significantdifferences in sensitivity to the fungicides among the different yeasts(P=0.05). R.glutinis was more sensitive to Deccocil, Iprodione and Stroby as compared to otheryeasts. Combination antagonistic yeasts with fungicide could more significantly enhancebiocontrol ability of the yeasts against the pathogenic fungi in vitro(P = 0.05). C.laurentii was the most effective antagonist among the four yeasts and could completelycontrol spore germination of P.expansum and A.alternata when combined with Stroby at theconcentration of 100 L L-1. The yeasts, except R.glutinis, could grow well on nutrientyeast dextrose agar (NYDA) after 8 d incubation even at 20% CO2 concentration at 25℃.Particularly Trichosporon sp. showeda better adaptability to low temperature as comparedto other antagonists.

Phytochemical Analysis and Antifungal Activity of Extracts from Leaves and Fruit Residues of Brazilian Savanna Plants Aiming Its Use as Safe Fungicides

The increasing demand for safe food without preservatives or pesticides residues has encouraged several studies on natural products with antifungal activity and low toxicity.In this study,ethanolic extracts from leaves and fruit residues(peel and seeds)of three Brazilian savanna species(Acrocomia aculeata,Campomanesia adamantium and Caryocar brasiliense)were evaluated against phytopathogenic fungi.Additionally,the most active extract was chemically characterized by ESI-MS and its oral acute toxicity was evaluated.Extracts from C.brasiliense(pequi)peel and leaves were active against Alternaria alternata,Alternaria solani and Venturia pirina with minimal inhibitory concentrations between 350 and 1000 lg/mL.When incorporated in solid media,these extracts extended the lag phase of A.alternata and A.solani and reduced the growth rate of A.solani.Pequi peel extract showed better antifungal activity and their ESI-MS analysis revealed the presence of substances widely reported as antifungal such as gallic acid,quinic acid,ellagic acid,glucogalin and corilagin.The oral acute toxicity was relatively low,being considered safe for use as a potential natural fungicide.