scholarly journals First Report on Resistance to Pyraclostrobin, Thiophanate-methyl, Fenhexamid and Boscalid in Botrytis cinerea from Eucalyptus Seedlings in Florida Greenhouses

Plant Disease ◽  
2014 ◽  
Vol 98 (6) ◽  
pp. 851-851 ◽  
Author(s):  
A. Amiri ◽  
A. I. Zuniga ◽  
J. Mertely ◽  
N. A. Peres
Keyword(s):  
Botrytis Cinerea ◽  
Germ Tube ◽  
Crop Protection ◽  
Malt Extract ◽  
Thiophanate Methyl ◽  
First Report ◽  
Research Triangle ◽  
Extract Agar ◽  
Bayer Cropscience ◽  
Syngenta Crop Protection

Botryotinia fuckeliana de Bary (anamorph Botrytis cinerea Pers.) is an ubiquitous plant pathogen causing gray mold disease on more than 200 crops grown in the field or in greenhouses. Eucalyptus seedlings originating from three different greenhouses showing stem lesions were submitted to the Gulf Coast Research and Education Center Disease Clinic in June 2012. Ten single spore isolates of B. cinerea were obtained and tested for sensitivity using spore germination and germ tube elongation assays described previously (4). Fungicides tested were pyraclostrobin at 100 μg/ml (Cabrio, BASF, Research Triangle Park, NC), thiophanate-methyl at 100 μg/ml (Topsin-M, UPI, King of Prussia, PA), fenhexamid at 1 and 50 μg/ml (Elevate, Arysta Life Sciences, Cary, NC), fludioxonil at 0.1 and 10 μg/ml (Medallion, Syngenta Crop Protection, Research Triangle Park, NC), and iprodione at 5 and 50 μg/ml (Rovral, Bayer CropScience, Greensboro, NC) on 1% malt extract agar (MEA, 10 g malt extract and 15 g agar), and to cyprodinil at 1 and 25 μg/ml (Vanguard, Syngenta Crop Protection) on 0.5% sucrose agar (4). Sensitivity to the succinate dehydrogenase inhibitors (SDHIs) boscalid at 5 μg/ml (Endura, BASF), penthiopyrad at 1 and 3 μg/ml (Fontelis, DuPont Crop Protection, Willington, DE), and fluopyram at 3 μg/ml (Luna Privilege, Bayer CropScience) was evaluated on yeast bacto acetate agar (YBA) (3). The discriminatory dose for boscalid was adapted from (2) whereas those used for penthiopyrad and fluopyram were developed in this study. Isolates were grown on malt yeast extract agar for 7 to 10 days and spore suspensions were prepared in sterile distilled water and diluted to 106 conidia/ml. Respective media in 9-cm petri dishes were seeded with 7-μl droplets from each isolate allowing testing for all isolates on one plate. Two plates were used for each fungicide and sensitivity tests were repeated twice. Germination and germ tube growth were assessed microscopically after 16 to 24 h incubation at 22°C. The frequency of isolates resistant to two, three, and four fungicides was 90, 60, and 10%, respectively. Nine isolates (90%) were resistant to thiophanate-methyl and pyraclostrobin, simultaneously, whereas six (60%) and two isolates (20%) were resistant to boscalid and fenhexamid, respectively. All boscalid-resistant isolates were also resistant to pyraclostrobin and thiophanate-methyl, but one fenhexamid-resistant isolate was sensitive to the other three fungicides. Eight isolates that germinated at 5 μg/ml iprodione but not at 50 μg/ml were considered sensitive. All isolates were sensitive to the SDHIs penthiopyrad and fluopyram as well as to cyprodinil and fludioxonil. To our knowledge, this is the first report of resistance to pyraclostrobin, thiophanate-methyl, fenhexamid, and boscalid in B. cinerea from eucalyptus seedlings in Florida. The absence of resistance to fludioxonil and iprodione is likely because these fungicides are not registered in nurseries as well as fluopyram and penthiopyrad which were developed only recently. Management practices should be developed to limit the selection and spread of additional resistant populations in eucalyptus nurseries as has occurred in Florida strawberries where multi-fungicide resistance is widespread (1). References: (1) A. Amiri et al. Plant Dis. 97:393, 2013. (2) M. Leroch et al. Appl. Environ. Microbiol. 79:159, 2013. (3) G. Stammler and J. Speakman. J. Phytopathol. 154:508, 2006. (4) R. W. S. Weber and M. Hahn. J. Plant Dis. Prot. 118:17, 2011.

scholarly journals First Report of Fludioxonil Resistance in Botrytis cinerea from a Blackberry Field in Georgia

Plant Disease ◽  
2014 ◽  
Vol 98 (6) ◽  
pp. 848-848 ◽  
Author(s):  
D. Fernández-Ortuño ◽  
A. Grabke ◽  
P. K. Bryson ◽  
E. D. Beasley ◽  
L. A. Fall ◽  
...  
Keyword(s):  
South Carolina ◽  
Botrytis Cinerea ◽  
Crop Protection ◽  
The United States ◽  
Gray Mold ◽  
Resistant Isolate ◽  
Malt Extract ◽  
First Report ◽  
Control Fruit ◽  
Lesion Diameter

Botrytis cinerea Pers. is an important plant-pathogenic fungi responsible for gray mold on more than 230 plant species worldwide, including blackberry (Rubus). One of the main strategies to control the disease involves the application of different classes of fungicides. The phenylpyrrole fludioxonil is currently marketed in combination with the anilinopyrimidine cyprodinil as Switch 62.5WG (Syngenta Crop Protection Inc., Greensboro, NC) for gray mold control. In August 2013, blackberries affected with symptoms resembling gray mold were collected from a field located in Berrien County (Georgia), where Switch 62.5WG had been used extensively over the last 5 years. Three single-spore isolates, each from a different fruit, were obtained and identified as B. cinerea on the basis of morphology and confirmed by a 238-bp PCR amplification product obtained with primer set G3PDH-F1 (5′-GGACCCGAGCTAATTTATGTCACGT-3′), G3PDH-F2 (5′-GGGTGTCAACAACGAGACCTACACT-3′), and G3PDH-R (5′-ACCGGTGCTCGATGGGATGAT-3′). In vitro sensitivity to fludioxonil (Scholar SC, Syngenta) was determined on 1% malt extract agar (MEA) using a conidial germination assay as previously described (4). One isolate was moderately resistant due to growth on medium amended with the discriminatory dose of 0.1 μg/ml fludioxonil and residual growth at 10 μg/ml (4). To assess performance of fludioxonil in detached fruit assays, commercially grown strawberries (24 in total for each isolate and treatment) were rinsed with water, dried, and sprayed 4 h prior to inoculation with either water (control fruit) or 2.5 ml/liter of Scholar SC to runoff using a hand mister. Scholar SC was used because fludioxonil was the sole active ingredient in this product and strawberries were used because latent infections in fresh blackberry fruit interfered with inoculation experiments. This dose reflects the rate recommended for postharvest gray mold control according to the Scholar label. Fruit was stab-wounded with a sterile syringe and inoculated with a 30-μl droplet of conidia suspension (106 spores/ml) of the two sensitive or the resistant isolate. After inoculation, the fruit were kept at 22°C for 4 days. The sensitive isolates developed gray mold on non-treated (2.7 cm lesion diameter) but not on Scholar SC-treated fruit (0.0 cm lesion diameter). The resistant isolate developed gray mold disease on the water-treated control fruit (2.5 cm lesion diameter) and the fungicide-treated fruit (1.8 cm lesion diameter). EC50 values were determined in microtiter assays as described previously (3) using the concentrations of 0.01, 0.04, 0.12, 0.37, 1.1, 3.3, and 10 μg/ml fludioxonil. Values were 0.02 and 0.05 μg/ml for the two sensitive isolates and 3.15 μg/ml for the resistant isolate. All experiments were performed twice. This is the first report of fludioxonil resistance in B. cinerea from blackberry in Georgia. Prior to this study, resistance to fludioxonil in B. cinerea was reported in France, Germany, and only a few states in the United States including Maryland, South Carolina, Virginia, and Washington (1,2). The emergence of resistance to fludioxonil emphasizes the importance of resistance management strategies. References: (1) D. Fernández-Ortuño et al. Plant Dis. 97:848, 2013. (2) D. Fernández-Ortuño et al. Plant Dis. 98:692, 2013. (3) M. Kretschmer et al. PLOS Pathog. 5:e1000696, 2009. (4) R. W. S. Weber and M. Hahn. J. Plant Dis. Prot. 118:17, 2011.

scholarly journals First Report of Streptomyces galilaeus Associated with Common Scab in China

Plant Disease ◽  
2014 ◽  
Vol 98 (5) ◽  
pp. 683-683 ◽  
Author(s):  
F. L. Guo ◽  
H. Y. Zhang ◽  
X. M. Yu ◽  
W. Q. Zhao ◽  
D. Q. Liu ◽  
...  
Keyword(s):  
16S Rdna ◽  
Common Scab ◽  
Potato Scab ◽  
Rdna Sequence ◽  
Malt Extract ◽  
16S Rdna Sequence ◽  
First Report ◽  
Carbon And Nitrogen ◽  
Physiological Characterization ◽  
Extract Agar

During a survey of potato scab pathogens in China from 2003 to 2012, a new pathogen was found in Shanxi and Neimenggu provinces. The incidence was approximately 20% of all recovered strains. The lesions caused by the pathogen were slightly raised and similar to those caused by Streptomyces scabies (3). Lesions were excised (approximately 10 mm3) from 40 infected tubers, surface-disinfested with 0.3% NaOCl for 30 s, rinsed in sterile water three times, cut into 5 mm3, then sliced into 1-mm pieces, and plated on water agar amended with ampicillin (50 μg/ml). Plates were incubated at 28°C in the dark for 4 days. The spores of Streptomyces sp. strains growing from the tuber pieces were collected from single bacterial colonies and cultured on oatmeal agar. To fulfill Koch's postulates, one strain, CPS-2, was grown at 28°C for 10 days and the spores were washed from the plates as inoculum. One hundred milliliters of inoculum (1 × 105 CFU/ml) was mixed with autoclaved soil and vermiculite (1:1) in each pot (15 cm in diameter). Cut tubers were planted in the pots (potato cv. Favorita, one plant per pot, five replicates) and grown under greenhouse conditions (22 ± 5°C). Typical common scab symptoms consisting of small, brown, raised lesions developed on potato tubers 12 weeks after planting. The same strain was re-isolated from the lesions of the new scabby tubers. Non-inoculated plants, treated as described above, but without strain CPS-2, remained healthy. The CPS-2 strain was identified based on morphological and physiological characterization and 16S rDNA sequence. On yeast-malt extract agar, the test strain produced grayish-white aerial hypha, reddish brown substrate mycelium and pigments, and loose spiral spore chains. Spores were smooth and were 0.8 to 0.9 × 1.1 to 1.2 μm in size (diameter and length). The ability of the strain to use single sources of carbon and nitrogen was verified according to the International Streptomyces project (4). The strain grew in media supplemented with L-arabinose, D-fructose, D-glucose, rhamnose, raffinose, meso-inositol, sucrose, and D-xylose, but not D-mannitol. It used L-hydroxyproline, L-methionine, and L-histidine, and produced melanin on tyrosine and peptone yeast extract agar. The strain did not grow at a pH less than 5.0 and was sensitive to streptomycin (20 μg/ml), phenol (0.1%), and crystal violet (0.5 μg/mL), but not to penicillin (10 IU/ml). The strain also produced hydrogen sulfide. The biological characteristics of strain CPS-2 were in accord with Streptomyces galilaeus. CPS-2 produced thaxtomin A in oatmeal liquid medium and the txt AB gene fragment was successfully amplified using specific primers (2). The 16S rDNA sequence of CPS-2 was amplified by PCR with primers 16S1-F: 5′-CATTCACGGAGAGTTTGATCC-3′ and 16S1-R: 5′-AGAAAGGAGGTGATCCAGCC-3′ (1) and sequenced. A BLAST search of the 16S rDNA sequence for CPS-2 was conducted using the NCBI GenBank database, resulting in 99.8% similarity to S. galilaeus (NR_040857). The 16S rDNA sequence for CPS-2 (1,388 bp) was deposited in GenBank (AY621378). To our knowledge, this is the first report of S. galilaeus causing common scab of potato in China. References: (1) R. A. Bukhalid et al. Appl. Environ. Microbiol. 68:738, 2002. (2) R. Flores-González et al. Plant Pathol. 57:162, 2008. (3) D. H. Lambert and R. Loria. Int. J. Syst. Bacteriol. 39:387, 1989. (4) E. B. Shirling and D. Gottlieb. Int. J. Syst. Bacteriol. 16:313, 1966.

scholarly journals First Report of Fludioxonil Resistance in Botrytis cinerea, the Causal Agent of Gray Mold, from Strawberry Fields in Maryland and South Carolina

Plant Disease ◽  
2014 ◽  
Vol 98 (5) ◽  
pp. 692-692 ◽  
Author(s):  
D. Fernández-Ortuño ◽  
A. Grabke ◽  
P. K. Bryson ◽  
R. J. Rouse ◽  
P. Rollins ◽  
...  
Keyword(s):  
United States ◽  
South Carolina ◽  
Botrytis Cinerea ◽  
Crop Protection ◽  
The United States ◽  
Gray Mold ◽  
Causal Agent ◽  
First Report ◽  
Lesion Diameter

Botrytis cinerea Pers. is the causal agent of gray mold and one of the most economically important plant-pathogenic fungi affecting strawberry (Fragaria × ananassa). Control of gray mold mainly depends on the use of site-specific fungicides, including the phenylpyrrole fludioxonil. This fungicide is currently registered in combination with cyprodinil in form of Switch 62.5WG (Syngenta Crop Protection, Greensboro, NC) for gray mold control of small fruits in the United States. In June 2013, strawberries affected with symptoms resembling gray mold were observed despite the application of Switch in one field located in Federalsburg, MD, and one located near Chesnee, SC. Ten single-spore isolates, each from a different fruit, were obtained from each location and confirmed to be B. cinerea using cultural and molecular tools as described previously (3). In vitro sensitivity to fludioxonil (Scholar SC, 20.4% [v/v] active ingredient, Syngenta Crop Protection, Greensboro, NC) was determined using a conidial germination assay as previously described (4). Eight of the 20 isolates (six from Maryland and two from South Carolina) were moderately resistant to fludioxonil, i.e., they grew on medium amended with 0.1 μg/ml fludixonil and showed residual growth at 10 μg/ml (4). The in vitro assay was repeated obtaining the same results. To assess in vivo sensitivity on fungicide-treated fruit, commercially grown strawberries were rinsed with water, dried, and sprayed 4 h prior to inoculation with either water or 2.5 ml/liter of Scholar SC to runoff using a hand mister. Fruit was stab-wounded with a sterile syringe and inoculated with a 30-μl droplet of conidia suspension (106 spores/ml) of either two sensitive or four resistant isolates (two isolates from Maryland and two isolates from South Carolina). Each isolate/treatment combination consisted of 24 mature but still firm strawberry fruit with three 8-fruit replicates. The fruit were kept at 22°C and lesion diameters were measured after 4 days of inoculation. The sensitive isolates developed gray mold symptoms on nontreated (2.5 cm lesion diameter) but not on Scholar SC-treated fruit. The resistant isolates developed gray mold on both, the water-treated control (2.3 cm lesion diameter), and the fungicide-treated fruit (1.8 cm lesion diameter). The experiment was performed twice. To our knowledge this is the first report of fludioxonil resistance in B. cinerea from strawberry fields in Maryland and South Carolina. Resistance to fludioxonil is still rare in the United States and has only been reported in B. cinerea isolates from a Virginia strawberry field (1). The increase in occurrence of resistance to fludioxonil may be a result of increased use of Switch following reports of resistance to other chemical classes in this pathogen in southern strawberry fields (2). References: (1) D. Fernández-Ortuño et al. Plant Dis. 97:848, 2013. (2) D. Fernández-Ortuño et al. Plant Dis. 96:1198, 2012. (3) D. Fernández-Ortuño et al. Plant Dis. 95:1482, 2011. (4) R. W. S. Weber and M. Hahn. J. Plant Dis. Prot. 118:17, 2011.

scholarly journals First Report of Gray Mold in Tomato Caused by Botrytis cinerea in Baja California, Mexico

Plant Disease ◽  
2005 ◽  
Vol 89 (5) ◽  
pp. 528-528 ◽  
Author(s):  
R. J. Holguín-Peña ◽  
F. G. Arcos
Keyword(s):  
Botrytis Cinerea ◽  
Baja California ◽  
Polymerase Chain Reaction Amplification ◽  
Random Amplified Polymorphic Dna ◽  
Gray Mold ◽  
Malt Extract ◽  
Conidial Suspension ◽  
Sterile Water ◽  
Tomato Production ◽  
First Report

San Quintin Valley, a 60-mile-long coastal plain (30°30′N, 116°W) in the Baja California Peninsula, is one of the major fresh tomato (Lycopersicon esculentum Mill.) production areas in Mexico with more than 8,000 ha. During the last 10 years, the valley's tomato production has declined because of gray mold and stem canker diseases. Flower rot, reddish brown margins on the leaves and stems, and fruit with a gray mold were observed on field-grown tomato plants (Roma type cv. Tequila) in the autumn of 2003. Severity ranging from 55 to 60% was observed at harvest. Infected tissues were sampled and disinfested by immersion in 1% NaOCl for 1 min, rinsed in sterile water, and placed on malt extract agar at 22°C. Fungal conidia were then transferred to 2% potato dextrose agar (PDA). The resulting fungal colonies were definitively identified as Botrytis cinerea Pers.:Fr. The colonies of B. cinerea were first hyaline and white and became dark gray after 96 h. Mycelia were septate with dark branched conidiophores. Conidia were unicellular, ellipsoid, and ranged from 5 to 8 × 8 to 14 μm. Profuse black sclerotia developed in 7-day-old cultures. Infection site analyses in diseased flowers at different stages during the bloom were done with scanning electron microscopy. Fungal hyphae were located predominantly on the receptacle areas, whereas conidia were located in the ovaries as described previously (3). The identity of B. cinerea was confirmed by a restriction digest with ApoI of the 413-kb polymerase chain reaction amplification product obtained with BA2f/BA1r primers (1) and random amplified polymorphic DNA banding patterns (2). Pathogenicity tests were done by spray inoculation of 1-ml aqueous conidial suspension (106 CFU/ml) on 20 healthy plants during the blossom stage. An equal number of plants sprayed with sterile water was used as the control. Plants were incubated at 20 ± 2°C for 5 days. The fungus was reisolated from diseased flowers and peduncles after surface disinfestation (2.5% NaOCl) and plating on PDA. No symptoms were observed in the noninoculated controls. To our knowledge, this is the first report of B. cinerea causing gray mold disease on tomato in Baja California. References: (1) K. Nielsen et al. Plant Dis. 86:682, 2002. (2) S. Rigotti et al. FEMS Microbiol. Lett. 209:169, 2002. (3) O. Viret et al. Phytopathology 94:850, 2004.

scholarly journals First Report of Botrytis cinerea on Kenaf in South Africa

Plant Disease ◽  
2001 ◽  
Vol 85 (9) ◽  
pp. 1032-1032
Author(s):  
W. J. Swart ◽  
M. T. Tesfaendrias ◽  
J. Terblanche
Keyword(s):  
South Africa ◽  
El Salvador ◽  
Botrytis Cinerea ◽  
Fiber Quality ◽  
Gray Mold ◽  
Hibiscus Cannabinus ◽  
Malt Extract ◽  
First Report ◽  
Significant Difference ◽  
Stem Lesions

Kenaf (Hibiscus cannabinus L.) (Malvaceae) is a source of high-quality cellulose fibers and is being investigated in South Africa with a view to commercial production. In April 2001, 20 to 30% of 5-month-old kenaf plants grown from seed in experimental plots near Rustenburg, Northwest Province, South Africa, were affected by gray mold caused by Botrytis cinerea Pers.:Fr. Infected plants displayed brown necrotic areas that girdled the stem, resulting in wilting and lodging in at least 50% of observed cases. Symptoms included extensive growth of mycelia and gray conidia on stem lesions. Microscopic examination revealed hyaline, one-celled conidia and conidiophores conforming to the description of B. cinerea. Plating of diseased stem tissue on malt extract agar (MEA) consistently yielded B. cinerea. Koch's postulates were satisfied by applying toothpick tips (5 mm) colonized by B. cinerea on MEA to the stems of 10 120-day-old greenhouse-grown plants of each of five kenaf cultivars. A colonized toothpick tip was placed on the stem of each of five plants per cultivar at a point ≍15 cm above soil level. Another five plants of each cultivar were wounded once using a sharp dissecting needle, and a colonized toothpick tip was placed on top of each wound. Corresponding control treatments consisted of five additional plants per cultivar, each wounded and mock-inoculated with sterile toothpick tips. Inoculation points were wrapped in Parafilm. The experiment was conducted twice. Developing lesions were measured after 7 days. Mean lesion lengths for the two treatments, nonwounded and wounded, on the five cultivars were, respectively: 32.4 and 35.2 mm for Everglades 41; 14.9 and 53.8 mm for Cuba 108; 39.5 and 55.8 mm for El Salvador; 19.0 and 44.3 mm for SF459; and 12.4 and 43.9 mm for Tainung 2. The Newman-Keuls multiple comparison test revealed no significant difference (P < 0.05) in means among cultivars for the wounded treatment. For the nonwounded treatment, Everglades 41 and El Salvador were significantly more susceptible (P < 0.05) than the three remaining cultivars. No lesions developed on control treatments. The fungus was reisolated on MEA from all artificially inoculated plants. The pathogen is reported to cause serious losses in yield and fiber quality of kenaf in Spain (1). This is the first report of B. cinerea on kenaf in South Africa, and its potential impact on kenaf production in this country should be taken seriously. Reference: (1) A. De Cal and P. Melgarejo. Plant Dis. 76:539, 1992.

scholarly journals First Report of Occurrence of Benomyl Resistance in Botrytis cinerea Isolates on Raspberry in Serbia

Plant Disease ◽  
2010 ◽  
Vol 94 (4) ◽  
pp. 486-486 ◽  
Author(s):  
B. Tanović ◽  
M. Ivanović
Keyword(s):  
Botrytis Cinerea ◽  
Pcr Amplification ◽  
Rubus Idaeus ◽  
Fruit Rot ◽  
Single Spore ◽  
Thiophanate Methyl ◽  
First Report ◽  
Benomyl Resistance ◽  
Development Of Resistance ◽  
Resistant Strains

Botrytis fruit rot, caused by Botrytis cinerea, is one of the major diseases limiting production of raspberries (Rubus idaeus) in Serbia. Yield losses in commercial fields can exceed 50%, especially during periods of rainy, wet weather before harvest. Development of resistance to fungicides with site-specific modes of action is a serious problem in the control of B. cinerea worldwide. To insure the longest possible useful life of a fungicide, an early detection of shifts of sensitivity in pathogen population is crucial (1). The goal of this study was to evaluate sensitivity of B. cinerea isolates from commercial raspberry fields in Serbia to several fungicides that are frequently used: vinclozolin, benomyl, pyrimethanil, and fenhexamid. Initial isolation was done from sporulating berries during harvest. Single-spore isolates were identified based on colony and conidial morphology and by PCR amplification of an expected 0.7-kbp DNA fragment using B. cinerea-specific primer pair C729+/729- (3). Sensitivity of 130 isolates from six localities (20 to 30 isolates per locality) was determined on potato dextrose agar (PDA) amended with fungicides at discriminatory concentrations (1 and 10 mg/liter). Fungicides were suspended in sterile distilled water and added to autoclaved media that had cooled to 50°C. Inverted mycelial plugs (10-mm diameter), which had been cut from the edge of 4-day-old colonies on PDA, were placed on fungicide amended media and incubated for 48 h at 20°C. Treatments were replicated four times and the experiment repeated once. Strain SAS 56, which is sensitive to benzimidazoles and dicarboximides, and strain SAS 405, which is resistant to these fungicide classes, originating from German Collection of Microorganisms and Cell Cultures, were used as standards in the experiment. Isolates that did not grow at 1 mg/liter were designated as sensitive, those that grew at 10 mg/liter were considered highly resistant, and those that grew at 1 mg/liter but not at 10 mg/liter were classified as weakly resistant to all fungicides tested. Values of EC50 for all highly resistant strains were determined in radial growth experiments on PDA supplemented with a range of concentrations (5,000, 2,500, 1,000, and 500 mg/liter) of benomyl or thiophanate-methyl, according to the method described by Leroux and Gredt (2). All tested isolates were sensitive to vinclozolin, pyrimethanil, and fenhexamid. Nine of 130 isolates were highly resistant to benomyl with EC50 values between 1,056 and 1,523 mg/liter. The reference strain SAS 56 had an EC50 value of 0.17 mg/liter, compared to an EC50 value for SAS 405 strain of 1,548 mg/liter. All benomyl resistant isolates were also resistant to thiophanate-methyl and EC50 values ranged from 2,328 to 7,699 mg/liter. To our knowledge, this is the first report of benomyl resistance in isolates of B. cinerea on raspberry in Serbia. References: (1) H. Ishii. Jarq 40:205, 2006. (2) P. Leroux and M. Gredt. Page 1 in: Laboratoire de Phytopharmacie, INRA, Versailles, 1972. (3) S. Rigotti et al. FEMS Microbiol. Lett. 209:169, 2002.

scholarly journals First Report of Fludioxonil Resistance in Botrytis cinerea from a Strawberry Field in Virginia

Plant Disease ◽  
2013 ◽  
Vol 97 (6) ◽  
pp. 848-848 ◽  
Author(s):  
D. Fernández-Ortuño ◽  
P. K. Bryson ◽  
A. Grabke ◽  
G. Schnabel
Keyword(s):  
South Carolina ◽  
Botrytis Cinerea ◽  
Disease Incidence ◽  
Crop Protection ◽  
Gray Mold ◽  
Resistant Isolate ◽  
Single Spore ◽  
First Report ◽  
Single Spore Isolate

Gray mold caused by Botrytis cinerea Pers.:Fr. is one of the most economically important diseases of cultivated strawberry (Fragaria × ananassa) worldwide. Control of gray mold mainly depends on fungicides, including the phenylpyrrole fludioxonil, which is currently marketed in combination with cyprodinil as Switch 62.5WG (Syngenta Crop Protection, Research Triangle Park, Raleigh, NC). In 2012, 790 strains of B. cinerea were collected from 76 strawberry fields in eight states, including Arkansas, Florida, Georgia, Kansas, Maryland, North Carolina, South Carolina, and Virginia. Strains were collected from sporulating flowers and fruit and sensitivity to fludioxonil was determined using a conidial germination assay as previously described (2). Only one isolate from a farm located in Westmoreland County, Virginia, grew on medium amended with the discriminatory dose of 0.1 μg/ml fludioxonil and was therefore considered low resistant. The isolate did not grow on 10 μg/ml. All other 789 isolates did not grow at either of the two doses. This assay was repeated twice with a single-spore culture of the same strain. In both cases, residual growth was observed on the fludioxonil-amended medium of 0.1 μg/ml. The single spore isolate was confirmed to be B. cinerea Pers. using cultural and molecular tools as described previously (1). To assess resistance in vivo, commercially grown ripe strawberry fruit were rinsed with sterile water, dried, placed into plastic boxes (eight strawberries per box for each of the three replicates per treatment), and sprayed 4 h prior to inoculation with either water or 2.5 ml/liter of fludioxonil (Scholar SC, Syngenta) to runoff using a hand mister. This dose reflects the rate recommended for gray mold control according to the Scholar label. Each fruit was stabbed at three equidistant points, each about 1 cm apart and 1 cm deep using a syringe tip. Wounds were injected with a 30-μl droplet of conidia suspension (106 spores/ml) of either 5 sensitive or the resistant isolate. Control fruit were inoculated with water. After inoculation, the fruit were kept at 22°C for 4 days. In two independent experiments, sensitive and low resistant isolates were indistinguishable in pathogenicity on detached, unsprayed fruit. The low resistant isolate developed gray mold disease on all treated and untreated fruit (100% disease incidence) as determined by the absence or presence of gray mold symptoms. The sensitive isolates only developed disease on untreated fruit. The EC50 values, determined in microtiter assays with concentrations of 0.01, 0.03, 0.1, 0.3, 1, 3, and 10 μg/ml fludioxonil, were 0.01 μg/ml for the sensitive isolates and 0.26 μg/ml for the resistant isolate. To our knowledge, this is the first report of fludioxonil resistance in B. cinerea from strawberry in North America. Our monitoring results indicate that resistance is emerging 10 years after the introduction of fludioxonil and stress the importance of chemical rotation for gray mold control. References: (1) X. P. Li et al. Plant Dis. 96:1634, 2012. (2) R. W. S. Weber and M. Hahn. J. Plant Dis. Prot. 118:17, 2011.

Isolation of endophytes from two species of palm, from Bermuda

1997 ◽  
Vol 43 (8) ◽  
pp. 789-792 ◽  
Author(s):  
K. A. Southcott ◽  
J. A. Johnson
Keyword(s):  
Malt Extract Agar ◽  
Malt Extract ◽  
Fungal Isolation ◽  
First Report ◽  
Extract Agar ◽  
Fungal Isolates ◽  
Significant Difference ◽  
Livistona Chinensis

This is the first report of endophytes from the indigenous Bermudian palmetto (Sabal bermudana Bailey) and the introduced Chinese palmetto (Livistona chinensis Jacquin), from Bermuda. Fronds were surface sterilized and 8-mm-diameter disks were removed and placed on 2% malt extract agar. Fungal isolates were obtained from 76 of the 375 disks from both species of palm. Idriella (two species) was the most common taxon isolated from both species of palm, making up 22 of the total 76 isolates, while Aspergillus accounted for 17 of the total 76 isolates. An unidentified isolate (BCP95-A), found in the Bermudian and the Chinese palmettos, accounted for 8 of the total 76 isolates. No statistically significant difference was found between fungal isolation frequencies of the two species of palm.Key words: endophyte, palm, leaves, Bermuda.

scholarly journals First Report of Cylindrocladium buxicola on Buxus sempervirens in Spain

Plant Disease ◽  
2009 ◽  
Vol 93 (6) ◽  
pp. 670-670 ◽  
Author(s):  
C. Pintos Varela ◽  
B. González Penalta ◽  
J. P. Mansilla Vázquez ◽  
O. Aguín Casal
Keyword(s):  
Uv Light ◽  
Malt Extract Agar ◽  
Terminal Branch ◽  
Malt Extract ◽  
Tubulin Gene ◽  
First Report ◽  
Potted Plants ◽  
Extract Agar ◽  
Buxus Sempervirens ◽  
Β Tubulin

Cylindrocladium buxicola Henricot, included in the EPPO alert list until November 2008, causes a dangerous foliar disease on Buxus spp. that has been recorded in several European countries and New Zealand (3,4). Buxus sempervirens L. (common boxwood) is one of the oldest ornamental garden plants in Europe. In September 2008, we received 10 2- to 3-year-old potted plants of B. sempervirens cv. Suffruticosa from a nursery in Galicia (northwest Spain) where ≈60% of the plants were affected and had finally defoliated. Diseased plants showed dark brown-to-black spots on the leaves and black streaks on the stems (3,4). To induce sporulation, diseased leaves and stem pieces were incubated in damp chambers at 22°C. A Cylindrocladium sp. was obtained. Four single conidial isolates were plated onto carnation leaf agar under near-UV light at 25°C for 7 days (2,3). Only conidiophores of the isolates growing on the surface of the carnation leaves were examined microscopically (1,3). Macroconidiophores were comprised of a stipe, a stipe extension, a terminal vesicle, and a penicillate arrangement of fertile branches (2). The stipe extension was septate, hyaline, and 90 to 165 × 2 to 4.5 μm (from the highest primary branch to the vesicle tip) (1) terminating in an ellipsoidal vesicle (6 to 11 μm in diameter) with a papillate apex. The widest part of the vesicle was above the middle. Primary branches were mainly aseptate or one septate (12 to 35 × 3 to 6 μm), secondary branches were aseptate (11 to 21 × 3 to 6 μm), and tertiary branches were rare. Each terminal branch produced two to five phialides (9 to 20 × 2.5 to 5 μm) that were reniform and aseptate. Conidia were cylindrical, straight, and one septate (56 to 75 × 4 to 6 μm). Chlamydospores were dark brown and aggregated to form microsclerotia. Cardinal temperatures of Cylindrocladium isolates on 2% malt extract agar ranged from 7 to 28°C (optimum 25°C). The 5′ end of the β-tubulin gene was amplified using primers T1 and Bt2b (3), and PCR products were sequenced directly and deposited in GenBank (Accession No. FJ696535). Comparison of the sequence with others available in GenBank showed 100% homology with those previously identified as C. buxicola (Accession Nos. AY078123 and AY078118). Pathogenicity of one representative isolate was confirmed by inoculating stems and leaves of four 3- to 4-year–old plants of B. sempervirens cv. Suffruticosa. Leaves were inoculated by spraying a spore suspension of the fungus (1 × 106 conidia per ml). For the stems, agar pieces of 1-week-old cultures grown on malt extract agar were placed and sealed with Parafilm. As a control, four plants were inoculated with agar malt plugs and sterile distilled water. Plants were incubated at 22°C and 95% humidity. Symptoms identical to ones previously described appeared 4 days after inoculation. C. buxicola was reisolated from inoculated plants but not from the controls. On the basis of morphological and physiological characteristics, pathogenicity, and the DNA sequencing of the β-tubulin gene, the isolates obtained from B. sempervirens were identified as C. buxicola (3). To our knowledge, this is the first report of C. buxicola on B. sempervirens in Spain. References: (1) P. W. Crous. Taxonomy and Pathology of Cylindrocladium (Calonectria) and Allied Genera. The American Phytopathological Society, St. Paul, MN, 2002. (2) P. W. Crous and M. J. Wingfield. Mycotaxon 51:341, 1994. (3) B. Henricot and A. Culham. Mycologia 94:980, 2002. (4) B. Henricot et al. Plant Pathol. 49:805, 2000.

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