Proteomics of Aspergillus fumigatus Conidia-containing Phagolysosomes Identifies Processes Governing Immune Evasion.

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Schmidt, Hella; Vlaic, Sebastian; Krüger, Thomas; Schmidt, Franziska; Balkenhol, Johannes; Dandekar, Thomas; Guthke, Reinhard; Kniemeyer, Olaf; Heinekamp, Thorsten; Brakhage, Axel A

2018-06-01

Invasive infections by the human pathogenic fungus Aspergillus fumigatus start with the outgrowth of asexual, airborne spores (conidia) into the lung tissue of immunocompromised patients. The resident alveolar macrophages phagocytose conidia, which end up in phagolysosomes. However, A. fumigatus conidia resist phagocytic degradation to a certain degree. This is mainly attributable to the pigment 1,8-dihydroxynaphthalene (DHN) melanin located in the cell wall of conidia, which manipulates the phagolysosomal maturation and prevents their intracellular killing. To get insight in the underlying molecular mechanisms, we comparatively analyzed proteins of mouse macrophage phagolysosomes containing melanized wild-type (wt) or nonmelanized pksP mutant conidia. For this purpose, a protocol to isolate conidia-containing phagolysosomes was established and a reference protein map of phagolysosomes was generated. We identified 637 host and 22 A. fumigatus proteins that were differentially abundant in the phagolysosome. 472 of the host proteins were overrepresented in the pksP mutant and 165 in the wt conidia-containing phagolysosome. Eight of the fungal proteins were produced only in pksP mutant and 14 proteins in wt conidia-containing phagolysosomes. Bioinformatical analysis compiled a regulatory module, which indicates host processes affected by the fungus. These processes include vATPase-driven phagolysosomal acidification, Rab5 and Vamp8-dependent endocytic trafficking, signaling pathways, as well as recruitment of the Lamp1 phagolysosomal maturation marker and the lysosomal cysteine protease cathepsin Z. Western blotting and immunofluorescence analyses confirmed the proteome data and moreover showed differential abundance of the major metabolic regulator mTOR. Taken together, with the help of a protocol optimized to isolate A. fumigatus conidia-containing phagolysosomes and a potent bioinformatics algorithm, we were able to confirm A. fumigatus conidia

Proteomic analysis of early phase of conidia germination in Aspergillus nidulans.

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Oh, Young Taek; Ahn, Chun-Seob; Kim, Jeong Geun; Ro, Hyeon-Su; Lee, Chang-Won; Kim, Jae Won

2010-03-01

In order to investigate proteins involved in early phase of conidia germination, proteomic analysis was performed using two-dimensional gel electrophoresis (2D-GE) in conjunction with MALDI-TOF mass spectrometry (MS). The expression levels of 241 proteins varied quantitatively with statistical significance (Pproteomic analysis of early phase of conidia germination and will contribute to a better understanding of the molecular events involved in conidia germination process. Copyright (c) 2009 Elsevier Inc. All rights reserved.

Optimizing growth and conidia production of Cercospora medicaginis

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Naceur DJEBALI

2010-09-01

Full Text Available The fungus Cercospora medicaginis is pathogenic to annual and perennial Medicago species. It grows slowly and produces only few conidia. To test the pathogenicity or virulence of C. medicaginis and to breed resistant lines of Medicago truncatula we optimized in vitro the growth and the conidium production of four isolates of C. medicaginis derived from M. truncatula and M. polymorpha. Of the eight media tested, that with wheat bran juice (WBJ yielded optimal growth and conidium production with most strains. The optimum growth temperature on WBJ medium was 25–30°C. Growth and conidia production were better in conditions of alternating light and darkness than with constant darkness. The best growth in the liquid WBJ medium occurred at pH 6–7, but the greatest number of conidia in that medium was obtained at pH 8–9.

A long natural-antisense RNA is accumulated in the conidia of Aspergillus oryzae.

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Tsujii, Masaru; Okuda, Satoshi; Ishi, Kazutomo; Madokoro, Kana; Takeuchi, Michio; Yamagata, Youhei

2016-01-01

Analysis of expressed sequence tag libraries from various culture conditions revealed the existence of conidia-specific transcripts assembled to putative conidiation-specific reductase gene (csrA) in Aspergillus oryzae. However, the all transcripts were transcribed with opposite direction to the gene csrA. The sequence analysis of the transcript revealed that the RNA overlapped mRNA of csrA with 3'-end, and did not code protein longer than 60 amino acid residues. We designated the transcript Conidia Specific Long Natural-antisense RNA (CSLNR). The real-time PCR analysis demonstrated that the CSLNR is conidia-specific transcript, which cannot be transcribed in the absence of brlA, and the amount of CSLNR was much more than that of the transcript from csrA in conidia. Furthermore, the csrA deletion, also lacking coding region of CSLNR in A. oryzae reduced the number of conidia. Overexpression of CsrA demonstrated the inhibition of growth and conidiation, while CSLNR did not affect conidiation.

Chronological aging in conidia of pathogenic Aspergillus: Comparison between species.

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Oliveira, Manuela; Pereira, Clara; Bessa, Cláudia; Araujo, Ricardo; Saraiva, Lucília

2015-11-01

Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus and Aspergillus niger are common airborne fungi, and the most frequent causative agents of human fungal infections. However, the resistance and lifetime persistence of these fungi in the atmosphere, and the mechanism of aging of Aspergillus conidia are unknown.With this work, we intended to study the processes underlying conidial aging of these four relevant and pathogenic Aspergillus species. Chronological aging was therefore evaluated in A. fumigatus, A. flavus, A. terreus and A. niger conidia exposed to environmental and human body temperatures. The results showed that the aging process in Aspergillus conidia involves apoptosis,with metacaspase activation, DNA fragmentation, and reactive oxygen species production, associated with secondary necrosis. Distinct results were observed for the selected pathogenic species. At environmental conditions, A. niger was the species with the highest resistance to aging, indicating a higher adaption to environmental conditions, whereas A. flavus followed by A. terreus were the most sensitive species. At higher temperatures (37 °C), A. fumigatus presented the longest lifespan, in accordance with its good adaptation to the human body temperature. Altogether,with this work new insights regarding conidia aging are provided, which may be useful when designing treatments for aspergillosis.

Sensitivity of Beauveria bassiana conidia to abiotic factors after successive in vitro subcultures

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Janaina Zorzetti

2014-09-01

Full Text Available The aim of this study was to evaluate the effect of in vitro successive subcultures of Beauveria bassiana isolates in different nutritional conditions on conidia heat and UV radiation tolerance and on sensitivity to azoxystrobin fungicide. The fungus was initially inoculated in Alphitobius diaperinus (Coleoptera: Tenebrionidae adults and after its development, the conidia were cultivated in PDA (potato dextrose agar and MPE (medium to produce Beauveria spp. conidia originating the first subculture 1st (A conidia, which were successively multiplied in their media until the 20th cultivation. Conidia taken from the 20th subculture were again inoculated in the insect and after sporulation they were once more multiplied in the media, producing the 1st (B conidia. For the tests, 1st (A, 10th, 20th and 1st (B conidia from the CG 152 and Unioeste 40 isolates were used. Successive subcultures and the medium nutritional conditions interfered on the fungus physiology, and the isolates showed different behaviors. The subcultures on PDA showed higher thermotolerance to Unioeste 40 in all subcultures and to CG 152 just after inoculation in the host. B. bassiana was susceptible to UV radiation, and this characteristic was influenced by the subcultures in different media. Both isolates were more tolerant to UV radiation when subcultivated on MPE, while just for the 40 Unioeste this tolerance increased after host inoculation. The B. bassiana sensitivity to azoxystrobin fungicide was affected after successive subcultures in both medium. Conidia originated in MPE were more resistant to fungicide and the host inoculation increased this tolerance. These results show that it is possible to enhance the fungi efficiency to control pests, through the selection of a virulent isolate, and handling their nutritional conditions during cultivation and host inoculation.

Production of Conidia by the Fungus Metarhizium anisopliae Using Solid-State Fermentation.

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Loera-Corral, Octavio; Porcayo-Loza, Javier; Montesinos-Matias, Roberto; Favela-Torres, Ernesto

2016-01-01

This chapter describes the production of conidia by Metarhizium anisopliae using solid-state fermentation. Before production of conidia, procedures for strains conservation, reactivation, and propagation are essential in order to provide genetic stability of the strains. The strain is conserved in freeze-dried vials and then reactivated through insect inoculation. Rice is used as a substrate for the conidia production in two different bioreactors: plastic bags and tubular bioreactor. The CO2 production in the tubular bioreactors is measured with a respirometer; this system allows calculating indirect growth parameters as lag time (tlag) (25-35 h), maximum rate of CO2 production (rCO2 max) (0.5-0.7 mg/gdm h), specific rate of CO2 production (μ) (0.10-0.15 1/h), and final CO2 production (CO2) (100-120 mg/gdm). Conidial yield per gram of dry substrate (gdm) should be above 1 × 10(9) conidia/gdm after 10 days of incubation. Germination and viability of conidia obtained after 10 days of incubation should be above 80 % and 75 %, respectively. Bioassays using of Tenebrio molitor as a host insect should yield a final mortality above 80 %.

Microencapsuling aerial conidia of Trichoderma harzianum through spray drying at elevated temperatures

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Trichoderma conidia are mostly produced by solid fermentation systems. Inoculum is produced by liquid culturing, and then transferred to solid substrate for aerial conidial production. Aerial conidia of T. harzianum are hydrophilic in nature, and it is difficult to separate them from the solid subst...

Citric acid production using immobilized conidia of Aspergillus niger TMB 2022

Energy Technology Data Exchange (ETDEWEB)

Tsay, S.S.; To, K.Y.

1987-02-20

Conidia of Aspergillus niger TMB 2022 were immobilized in calcium alginate for the production of citric acid. A 1-ml condidia suspension containing ca. 2.32 x 10/sup 8/ conidia were entrapped into sodium alginate solution in order to prepare 3% Ca-alginate (w/v) gel bead. Immobilized conidia were inoculated into productive medium containing 14% sucrose, 0.25% (NH/sub 4/)/sub 2/CO/sub 3/, 0.25% KH/sub 2/PO/sub 4/, and 0.025% MgSO/sub 4/.7H/sub 2/O with addition of 0.06 mg/l CuSO/sub 4/.5H/sub 2/O, 0.25 mg/l ZnCl/sub 2/, 1.3 mg/l FeCl/sub 3/.6H/sub 2/O, pH 3.8, and incubated at 35 degrees C for 13 days by surface culture to produce 61.53 g/l anhydrous citric acid. Under the same conditions with a batchwise culture, it was found that immobilized conidia could maintain a longer period for citric acid production (31 days): over 70 g/l anhydrous citric acid from runs No. 2-4, with the maximum yield for anhydrous citric acid reaching 77.02 g/l for run No. 2. In contrast, free conidia maintained a shorter acid-producing phase, circa 17 days; the maximum yield for anhydrous citric acid was 71.07 g/l for run No. 2 but dropped quickly as the run number increased. 14 references.

Metabolic activity in dormant conidia of Aspergillus niger and developmental changes during conidial outgrowth

OpenAIRE

Novodvorska, Michaela; Stratford, Malcolm; Blythe, Martin J.; Wilson, Raymond; Beniston, Richard G.; Archer, David B.

2016-01-01

The early stages of development of Aspergillus niger conidia during outgrowth were explored by combining genome-wide gene expression analysis (RNAseq), proteomics, Warburg manometry and uptake studies. Resting conidia suspended in water were demonstrated for the first time to be metabolically active as low levels of oxygen uptake and the generation of carbon dioxide were detected, suggesting that low-level respiratory metabolism occurs in conidia for maintenance. Upon triggering of spore germ...

Aspergillus fumigatus viability drives allergic responses to inhaled conidia.

Science.gov (United States)

Nayak, Ajay P; Croston, Tara L; Lemons, Angela R; Goldsmith, W T; Marshall, Nikki B; Kashon, Michael L; Germolec, Dori R; Beezhold, Donald H; Green, Brett J

2018-04-13

Aspergillus fumigatus induced allergic airway disease has been shown to involve conidial germination in vivo but the immunological mechanisms remain uncharacterized. A subchronic murine exposure model was used to examine the immunological mediators that are regulated in response to either culturable or non-culturable A. fumigatus conidia. Female B6C3F1/N mice were repeatedly dosed via inhalation with 1 x 105 viable or heat inactivated conidia (HIC), twice a week for 13 weeks (26 exposures). Control mice inhaled HEPA-filtered air. The influence of A. fumigatus conidial germination on the pulmonary immunopathological outcomes was evaluated by flow cytometry analysis of cellular infiltration in the airways, assessment of lung mRNA expression, and quantitative proteomics and histopathology of whole lung tissue. Repeated inhalation of viable conidia, but not HIC, resulted in allergic inflammation marked by vascular remodeling, extensive eosinophilia, and accumulation of alternatively activated macrophages (AAMs) in the murine airways. More specifically, mice that inhaled viable conidia resulted in a mixed TH1 and TH2 (IL-13) cytokine response. Recruitment of eosinophils corresponded with increased Ccl11 transcripts. Furthermore, genes associated with M2 or alternatively activated macrophage polarization (e.g. Arg1, Chil3 and Retnla) were significantly upregulated in viable A. fumigatus exposed mice. In mice inhaling HIC, CD4+ T cells expressing IFN-γ (TH1) dominated the lymphocytic infiltration. Quantitative proteomics of the lung revealed metabolic reprogramming accompanied by mitochondrial dysfunction and endoplasmic reticulum stress stimulated by oxidative stress from repetitive microbial insult. Our studies demonstrate that A. fumigatus conidial viability in vivo is critical to the immunopathological presentation of chronic fungal allergic disease. Copyright © 2018. Published by Elsevier Inc.

Development of a disease risk prediction model for downy mildew (Peronospora sparsa) in boysenberry.

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Kim, Kwang Soo; Beresford, Robert M; Walter, Monika

2014-01-01

Downy mildew caused by Peronospora sparsa has resulted in serious production losses in boysenberry (Rubus hybrid), blackberry (Rubus fruticosus), and rose (Rosa sp.) in New Zealand, Mexico, and the United States and the United Kingdom, respectively. Development of a model to predict downy mildew risk would facilitate development and implementation of a disease warning system for efficient fungicide spray application in the crops affected by this disease. Because detailed disease observation data were not available, a two-step approach was applied to develop an empirical risk prediction model for P. sparsa. To identify the weather patterns associated with a high incidence of downy mildew berry infections (dryberry disease) and derive parameters for the empirical model, classification and regression tree (CART) analysis was performed. Then, fuzzy sets were applied to develop a simple model to predict the disease risk based on the parameters derived from the CART analysis. High-risk seasons with a boysenberry downy mildew incidence >10% coincided with months when the number of hours per day with temperature of 15 to 20°C averaged >9.8 over the month and the number of days with rainfall in the month was >38.7%. The Fuzzy Peronospora Sparsa (FPS) model, developed using fuzzy sets, defined relationships among high-risk events, temperature, and rainfall conditions. In a validation study, the FPS model provided correct identification of both seasons with high downy mildew risk for boysenberry, blackberry, and rose and low risk in seasons when no disease was observed. As a result, the FPS model had a significant degree of agreement between predicted and observed risks of downy mildew for those crops (P = 0.002).

effect of dry conidia formulations of metarhizium anisopliae

African Journals Online (AJOL)

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In preliminary studies,. S. zeamais adults sprayed with M. anisopliae ... Initial cultures were stored at 4°C and sub- culturing was made for ... Significant differences between treatment means were compared at .... conidia attachment. Journal of ...

Growth under visible light increases conidia and mucilage production and tolerance to UV-B radiation in the plant pathogenic fungus Colletotrichum acutatum.

Science.gov (United States)

de Menezes, Henrique D; Massola, Nelson S; Flint, Stephan D; Silva, Geraldo J; Bachmann, Luciano; Rangel, Drauzio E N; Braga, Gilberto U L

2015-01-01

Light conditions can influence fungal development. Some spectral wavebands can induce conidial production, whereas others can kill the conidia, reducing the population size and limiting dispersal. The plant pathogenic fungus Colletotrichum acutatum causes anthracnose in several crops. During the asexual stage on the host plant, Colletototrichum produces acervuli with abundant mucilage-embedded conidia. These conidia are responsible for fungal dispersal and host infection. This study examined the effect of visible light during C. acutatum growth on the production of conidia and mucilage and also on the UV tolerance of these conidia. Conidial tolerance to an environmentally realistic UV irradiance was determined both in conidia surrounded by mucilage on sporulating colonies and in conidial suspension. Exposures to visible light during fungal growth increased production of conidia and mucilage as well as conidial tolerance to UV. Colonies exposed to light produced 1.7 times more conidia than colonies grown in continuous darkness. The UV tolerances of conidia produced under light were at least two times higher than conidia produced in the dark. Conidia embedded in the mucilage on sporulating colonies were more tolerant of UV than conidia in suspension that were washed free of mucilage. Conidial tolerance to UV radiation varied among five selected isolates. © 2014 The American Society of Photobiology.

Infectivity of the conidia of the rice blast fungus treated with the different fungicidal solutions

Energy Technology Data Exchange (ETDEWEB)

Hashioka, Y; Ikegami, H

1959-01-01

Infectivity of the conidia of Piricularia oryzae (rice blast fungus) which had been treated with different fungicides was examined. Germination of conidia treated with phenylmercuric acetate was severely repressed. Copper sulfate had a mild inhibitory effect. Organosulfur compounds also had a slight inhibitory effect.

Time-resolved study of absorbing film assisted laser induced forward transfer of Trichoderma longibrachiatum conidia

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Hopp, B [Hungarian Academy of Sciences and University of Szeged, Research Group on Laser Physics, H-6720 Szeged, Dom ter 9 (Hungary); Smausz, T [Hungarian Academy of Sciences and University of Szeged, Research Group on Laser Physics, H-6720 Szeged, Dom ter 9 (Hungary); Barna, N [Department of Optics and Quantum Electronics, University of Szeged, H-6720 Szeged, Dom ter 9 (Hungary); Vass, Cs [Department of Optics and Quantum Electronics, University of Szeged, H-6720 Szeged, Dom ter 9 (Hungary); Antal, Zs [Hungarian Academy of Sciences and University of Szeged, Microbiological Research Group, PO Box 533, H-6701 Szeged (Hungary); Kredics, L [Hungarian Academy of Sciences and University of Szeged, Microbiological Research Group, PO Box 533, H-6701 Szeged (Hungary); Chrisey, D [Naval Research Laboratory, Washington, DC 20375 (United States)

2005-03-21

We have characterized the absorbing film assisted transfer of Trichoderma longibrachiatum conidia using a synchronized laser for illumination. The transfer laser used was a KrF excimer laser ({lambda} = 248 nm, FWHM = 30 ns) and the ejected material was illuminated parallel to the quartz plate by a nitrogen laser pumped Coumarine 153 dye laser beam ({lambda} = 453 nm, FWHM 1 ns) electronically delayed relative to the transfer UV pulse. Our time-resolved investigations determined that the ejection velocity front of the conidia plume from the donor surface during the transfer procedure was 1150 m s{sup -1} at 355 mJ cm{sup -2} applied laser fluence. On the basis of the measured data, the acceleration of the emitted conidia at the plume front was approximately 10{sup 9} x g. The conidia survived the absorbing film assisted forward transfer and associated mechanical shear without significant damages suggesting that the technique might be applicable to other more fragile types of biological objects and applications.

Time-resolved study of absorbing film assisted laser induced forward transfer of Trichoderma longibrachiatum conidia

International Nuclear Information System (INIS)

Hopp, B; Smausz, T; Barna, N; Vass, Cs; Antal, Zs; Kredics, L; Chrisey, D

2005-01-01

We have characterized the absorbing film assisted transfer of Trichoderma longibrachiatum conidia using a synchronized laser for illumination. The transfer laser used was a KrF excimer laser (λ = 248 nm, FWHM = 30 ns) and the ejected material was illuminated parallel to the quartz plate by a nitrogen laser pumped Coumarine 153 dye laser beam (λ = 453 nm, FWHM 1 ns) electronically delayed relative to the transfer UV pulse. Our time-resolved investigations determined that the ejection velocity front of the conidia plume from the donor surface during the transfer procedure was 1150 m s -1 at 355 mJ cm -2 applied laser fluence. On the basis of the measured data, the acceleration of the emitted conidia at the plume front was approximately 10 9 x g. The conidia survived the absorbing film assisted forward transfer and associated mechanical shear without significant damages suggesting that the technique might be applicable to other more fragile types of biological objects and applications

Metabolic activity in dormant conidia of Aspergillus niger and developmental changes during conidial outgrowth.

Science.gov (United States)

Novodvorska, Michaela; Stratford, Malcolm; Blythe, Martin J; Wilson, Raymond; Beniston, Richard G; Archer, David B

2016-09-01

The early stages of development of Aspergillus niger conidia during outgrowth were explored by combining genome-wide gene expression analysis (RNAseq), proteomics, Warburg manometry and uptake studies. Resting conidia suspended in water were demonstrated for the first time to be metabolically active as low levels of oxygen uptake and the generation of carbon dioxide were detected, suggesting that low-level respiratory metabolism occurs in conidia for maintenance. Upon triggering of spore germination, generation of CO2 increased dramatically. For a short period, which coincided with mobilisation of the intracellular polyol, trehalose, there was no increase in uptake of O2 indicating that trehalose was metabolised by fermentation. Data from genome-wide mRNA profiling showed the presence of transcripts associated with fermentative and respiratory metabolism in resting conidia. Following triggering of conidial outgrowth, there was a clear switch to respiration after 25min, confirmed by cyanide inhibition. No effect of SHAM, salicylhydroxamic acid, on respiration suggests electron flow via cytochrome c oxidase. Glucose entry into spores was not detectable before 1h after triggering germination. The impact of sorbic acid on germination was examined and we showed that it inhibits glucose uptake. O2 uptake was also inhibited, delaying the onset of respiration and extending the period of fermentation. In conclusion, we show that conidia suspended in water are not completely dormant and that conidial outgrowth involves fermentative metabolism that precedes respiration. Copyright © 2016. Published by Elsevier Inc.

Density, Viability Conidia And Symptoms of Metarhizium anisopliae infection on Oryctes rhinoceros larvae

Science.gov (United States)

Indriyanti, D. R.; Putri, R. I. P.; Widiyaningrum, P.; Herlina, L.

2017-04-01

M. anisopliae is parasitic fungus on insect pests; it is used as a biocontrol agent. M. anisopliae can be propagated on maize or rice substrate. M. anisopliae is currently sold in the form of kaolin powder formulations. Before it is used to check the density, viability and pathogenicity of M. anisopliae. However the problem is the kaolin powder very soft, so it difficult to distinguish between kaolin and conidia. This article gives information on how to calculate conidia density, viability and symptoms of M. anisopliae infection on Oryctes rhinoceros larvae. The study was conducted in the laboratory to determine the density and viability. The pathogenicity testing was done using pots. The Pot is containing soil substrate mixed with M. Anispoliae and ten tails O. Rhinoceros larvae per pot. The results showed that the density of M. anisopliae conidia was 1.81 x 108 conidia mL-1 and the viability was 94% within 24 hours. The larval mortality began to emerge in the 1st week, and all larvae died at the sixth week. The symptom of M. anisopliae infection on Oryctes rhinoceros larvae, there was a black spot on the larval integument. The larvae movements become slow and poor appetite; it will die within 3-7 days. The larvae die hard, and the white hyphae grow on the body surface that turns green.

Comparative study of airborne Alternaria conidia levels in two cities in Castilla-La Mancha (central Spain), and correlations with weather-related variables.

Science.gov (United States)

Sabariego, Silvia; Bouso, Veronica; Pérez-Badia, Rosa

2012-01-01

Alternaria conidia are among the airborne biological particles known to trigger allergic respiratory diseases. The presented paper reports on a study of seasonal variations in airborne Alternaria conidia concentrations in 2 cities in the central Spanish region of Castilla-La Mancha, Albacete and Toledo. The influence of weather-related variables on airborne conidia levels and distribution was also analysed. Sampling was carried out from 2008-2010 using a Hirst sampler, following the methodology established by the Spanish Aerobiology Network. Annual airborne Alternaria conidia counts were higher in Toledo (annual mean 3,936 conidia) than in Albacete (annual mean 2,268 conidia). Conidia were detected in the air throughout the year, but levels peaked between May-September. Considerable year-on-year variations were recorded both in total annual counts and in seasonal distribution. A significant positive correlation was generally found between mean daily Alternaria counts and both temperature and hours of sunlight, while a significant negative correlation was recorded for relative humidity, daily and cumulative rainfall, and wind speed. Regression models indicated that between 31%-52% of the variation in airborne Alternaria conidia concentrations could be explained by weather-related variables.

A novel computerised image analysis method for the measurement of production of conidia from the aphid pathogenic fungus Erynia neoaphidis.

Science.gov (United States)

Bonner, Tony J; Pell, Judith K; Gray, Simon N

2003-03-14

A semi-automated method has been developed for the quantification and measurement of conidia discharged by the aphid pathogen Erynia neoaphidis. This was used to compare conidiation by E. neoaphidis-mycosed pea aphid cadavers, mycelial plugs cut from agar plates, mycelial pellets from shake flasks and by mycelial pellets from different phases of liquid batch fermenter culture. Aphid cadavers discharged significantly more and significantly smaller conidia than plugs or pellets. The volume of conidia discharged was stable over the period of discharge (80 h), but more detailed analysis of the size frequency distribution showed that more very small and very large conidia were discharged after 5 h incubation than after 75 h incubation. Biomass harvested at the end of the exponential growth phase in batch fermenter culture produced significantly more conidia than biomass from any other growth phase. The implications of these findings for the development of production and formulation processes for E. neoaphidis as a biological control agent are discussed.

Evaluación molecular y fenotípica de la sensibilidad al fungicida fenamidone en aislamientos de peronospora sparsa

OpenAIRE

Argel, Luz Edith; Marín, Mauricio; Cotes, José Miguel; Jaramillo, Sonia; Guzmán, Edgar

2010-01-01

El Mildeo velloso es uno de los mayores limitantes del cultivo de la rosa en Colombia. Esta enfermedad es causada por el Oomycete holobiótrofo Peronospora sparsa que fue detectado en la década de 1970 en la Sabana de Bogotá. En los últimos años su efecto negativo sobre la floricultura aumentó conduciendo a los cultivadores al empleo excesivo de fungicidas con acción sistémica, muchos de los cuales no cuentan con estudios de líneas base de sensibilidad ni con el diseño de estrategias anti-resi...

A bioencapsulation and drying method increases shelf life and efficacy of Metarhizium brunneum conidia.

Science.gov (United States)

Przyklenk, Michael; Vemmer, Marina; Hanitzsch, Miriam; Patel, Anant

2017-08-01

This study reports the development of encapsulated and dried entomopathogenic fungus Metarhiuzm brunneum with reduced conidia content, increased conidiation, a high drying survival and enhanced shelf life. Dried beads prepared with the fillers corn starch, potato starch, carboxymethylcellulose or autoclaved baker's yeast, showed enhanced survival with increasing filler content. The maximum survival of 82% was found for beads with 20% corn starch at shelf life compared to non-formulated conidia. This "microfermenter" will pave the way for encapsulated fungi to be used as cost-effective biocontrol agents.

Daytime Solar Heating Controls Downy Mildew Peronospora belbahrii in Sweet Basil.

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Yigal Cohen

Full Text Available The biotrophic oomycete Peronospora belbahrii causes a devastating downy mildew disease in sweet basil. Due to the lack of resistant cultivars current control measures rely heavily on fungicides. However, resistance to fungicides and strict regulation on their deployment greatly restrict their use. Here we report on a 'green' method to control this disease. Growth chamber studies showed that P. belbahrii could hardly withstand exposure to high temperatures; exposure of spores, infected leaves, or infected plants to 35-45 °C for 6-9 hours suppressed its survival. Therefore, daytime solar heating was employed in the field to control the downy mildew disease it causes in basil. Covering growth houses of sweet basil already infected with downy mildew with transparent infra-red-impermeable, transparent polyethylene sheets raised the daily maximal temperature during sunny hours by 11-22 °C reaching 40-58 °C (greenhouse effect. Such coverage, applied for a few hours during 1-3 consecutive days, had a detrimental effect on the survival of P. belbahrii: killing the pathogen and/or suppressing disease progress while enhancing growth of the host basil plants.

A proteomic and ultrastructural characterization of Aspergillus fumigatus' conidia adaptation at different culture ages.

Science.gov (United States)

Anjo, Sandra I; Figueiredo, Francisco; Fernandes, Rui; Manadas, Bruno; Oliveira, Manuela

2017-05-24

The airborne fungus Aspergillus fumigatus is one of the most common agents of human fungal infections with a remarkable impact on public health. However, A. fumigatus conidia atmospheric resistance and longevity mechanisms are still unknown. Therefore, in this work, the processes underlying conidial adaptation were studied by a time course evaluation of the proteomics and ultrastructural changes of A. fumigatus' conidia at three time-points selected according to relevant changes previously established in conidial survival rates. The proteomics characterization revealed that conidia change from a highly active metabolic to a dormant state, culminating in cell autolysis as revealed by the increased levels of hydrolytic enzymes. Structural characterization corroborates the proteomics data, with noticeable changes observed in mitochondria, nucleus and plasma membrane ultrastructure, accompanied by the formation of autophagic vacuoles. These changes are consistent with both apoptotic and autophagic processes, and indicate that the changes in protein levels may anticipate those in cell morphology. The findings presented in this work not only clarify the processes underlying conidial adaptation to nutrient limiting conditions but can also be exploited for improving infection control strategies and in the development of new therapeutical drugs. Additionally, the present study was deposited in a public database and thus, it may also be a valuable dataset to be used by the scientific community as a tool to understand and identified other potential targets associated with conidia resistance. Copyright © 2017. Published by Elsevier B.V.

Evaluación molecular y fenotípica de la sensibilidad al fungicida fenamidone en aislamientos de Peronospora sparsa

OpenAIRE

Argel Luz Edith; Marín Mauricio; Cotes José Miguel; Jaramillo Sonia; Guzmán Edgar

2007-01-01

El Mildeo velloso es uno de los mayores limitantes del cultivo de la rosa en Colombia. Esta enfermedad es causada por el Oomycete holobiótrofo Peronospora sparsa que fue detectado en la década de 1970 en la Sabana de Bogotá. En los últimos años su efecto negativo sobre la floricultura aumentó conduciendo a los cultivadores al empleo excesivo de fungicidas con acción sistémica, muchos de los cuales no cuentan con estudios de líneas base de sensibilid...

Efecto del fotoperiodo y la intensidad lumínica sobre la esporulación de peronospora sparsa berkeley, bajo condiciones controladas

OpenAIRE

Soto S., Johana Carolina; Filgueira D., Juan José

2009-01-01

El mildeo velloso de la rosa, producido por Peronospora sparsa, es una de las enfermedades más limitantes y costosas en los cultivos comerciales de rosa en Colombia. La estrecha relación de la enfermedad con las condiciones climáticas ha llevado a realizar estudios del parásito en condiciones microclimáticas controladas. Esta investigación buscó ahondar en el conocimiento de la biología del mildeo velloso de la rosa, evaluando para ello el efecto del fotoperiodo y la intensidad lumínica sobre...

Preincubation of Penicillium commune conidia under modified atmosphere conditions: Influence on growth potential as determined by an impedimetric method

DEFF Research Database (Denmark)

Haasum, Iben; Nielsen, Per Væggemose

1996-01-01

The combined effect of preincubation time, relative humidity (r.h.), headspace carbon dioxide (CO2) and oxygen (O2) on subsequent growth potential of conidia from Penicillium commune was studied using Response Surface Modelling (RSM). Native conidia were preincubated under modified atmosphere...... conditions in sealed vials for 14, 35 and 56 d. Lag time and growth rates were determined using impedance microbiology on a Bactometer. Conidia survived and some swelling was observed during all experimental preincubation conditions. Regression analysis of the subsequent growth responses showed that relative...... humidity in the vials was the most significant factor affecting lag time of the conidia after preincubation for 14 and 35 d. Storage for 35 d extended lag times by 15 h when the level of r.h. was increased from 41% to 80%. After prolonged storage (56 d) r.h and CO2 levels elicited a significant effect...

Germination of fungal conidia after exposure to low concentration ozone atmospheres.

Science.gov (United States)

The germinability of conidia of Alternaria alternata, Aspergillus flavus, Aspergillus niger, Penicillium digitatum, Penicillium expansum, or Penicillium italicum was determined periodically during exposure for approximately 100 days to a humid atmosphere of air alone or air containing 150 ppb ozone ...

Efecto del fotoperiodo y la intensidad lumínica sobre la esporulación de Peronospora sparsa Berkeley, bajo condiciones controladas

OpenAIRE

Soto S. Johana Carolina; Filgueira D. Juan José

2009-01-01

El mildeo velloso de la rosa, producido por Peronospora sparsa, es una de las enfermedades más limitantes y costosas en los cultivos comerciales de rosa en Colombia. La estrecha relación de la enfermedad con las condiciones climáticas ha llevado a realizar estudios del parásito en condiciones microclimáticas controladas. Esta investigación buscó ahondar en el conocimiento de la biología del mildeo velloso de la rosa, evalu...

Human Neutrophils Use Different Mechanisms To Kill Aspergillus fumigatus Conidia and Hyphae: Evidence from Phagocyte Defects.

Science.gov (United States)

Gazendam, Roel P; van Hamme, John L; Tool, Anton T J; Hoogenboezem, Mark; van den Berg, J Merlijn; Prins, Jan M; Vitkov, Ljubomir; van de Veerdonk, Frank L; van den Berg, Timo K; Roos, Dirk; Kuijpers, Taco W

2016-02-01

Neutrophils are known to play a pivotal role in the host defense against Aspergillus infections. This is illustrated by the prevalence of Aspergillus infections in patients with neutropenia or phagocyte functional defects, such as chronic granulomatous disease. However, the mechanisms by which human neutrophils recognize and kill Aspergillus are poorly understood. In this work, we have studied in detail which neutrophil functions, including neutrophil extracellular trap (NET) formation, are involved in the killing of Aspergillus fumigatus conidia and hyphae, using neutrophils from patients with well-defined genetic immunodeficiencies. Recognition of conidia involves integrin CD11b/CD18 (and not dectin-1), which triggers a PI3K-dependent nonoxidative intracellular mechanism of killing. When the conidia escape from early killing and germinate, the extracellular destruction of the Aspergillus hyphae needs opsonization by Abs and involves predominantly recognition via Fcγ receptors, signaling via Syk, PI3K, and protein kinase C to trigger the production of toxic reactive oxygen metabolites by the NADPH oxidase and myeloperoxidase. A. fumigatus induces NET formation; however, NETs did not contribute to A. fumigatus killing. Thus, our findings reveal distinct killing mechanisms of Aspergillus conidia and hyphae by human neutrophils, leading to a comprehensive insight in the innate antifungal response. Copyright © 2016 by The American Association of Immunologists, Inc.

Proteome profiling and functional classification of intracellular proteins from conidia of the human-pathogenic mold Aspergillus fumigatus.

Science.gov (United States)

Teutschbein, Janka; Albrecht, Daniela; Pötsch, Maria; Guthke, Reinhard; Aimanianda, Vishukumar; Clavaud, Cécile; Latgé, Jean-Paul; Brakhage, Axel A; Kniemeyer, Olaf

2010-07-02

Aspergillus fumigatus is a ubiquitously distributed filamentous fungus that has emerged as one of the most serious life-threatening pathogens in immunocompromised patients. The mechanisms for its pathogenicity are poorly understood. Here, we analyzed the proteome of dormant A. fumigatus conidia as the fungal entity having the initial contact with the host. Applying two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), we established a 2-D reference map of conidial proteins. By MALDI-TOF mass spectrometry, we identified a total number of 449 different proteins. We show that 57 proteins of our map are over-represented in resting conidia compared to mycelium. Enzymes involved in reactive oxygen intermediates (ROI) detoxification, pigment biosynthesis, and conidial rodlet layer formation were highly abundant in A. fumigatus spores and most probably account for their enormous stress resistance. Interestingly, pyruvate decarboxylase and alcohol dehydrogenase were detectable in dormant conidia, suggesting that alcoholic fermentation plays a role during dormancy or early germination. Moreover, we show that enzymes for rapid reactivation of protein biosynthesis and metabolic processes are preserved in resting conidia, which therefore feature the potential to immediately respond to an environmental stimulus by germination. The generated data lay the foundations for further proteomic analyses and a better understanding of fungal pathogenesis.

Gene co-expression analysis identifies gene clusters associated with isotropic and polarized growth in Aspergillus fumigatus conidia.

Science.gov (United States)

Baltussen, Tim J H; Coolen, Jordy P M; Zoll, Jan; Verweij, Paul E; Melchers, Willem J G

2018-04-26

Aspergillus fumigatus is a saprophytic fungus that extensively produces conidia. These microscopic asexually reproductive structures are small enough to reach the lungs. Germination of conidia followed by hyphal growth inside human lungs is a key step in the establishment of infection in immunocompromised patients. RNA-Seq was used to analyze the transcriptome of dormant and germinating A. fumigatus conidia. Construction of a gene co-expression network revealed four gene clusters (modules) correlated with a growth phase (dormant, isotropic growth, polarized growth). Transcripts levels of genes encoding for secondary metabolites were high in dormant conidia. During isotropic growth, transcript levels of genes involved in cell wall modifications increased. Two modules encoding for growth and cell cycle/DNA processing were associated with polarized growth. In addition, the co-expression network was used to identify highly connected intermodular hub genes. These genes may have a pivotal role in the respective module and could therefore be compelling therapeutic targets. Generally, cell wall remodeling is an important process during isotropic and polarized growth, characterized by an increase of transcripts coding for hyphal growth and cell cycle/DNA processing when polarized growth is initiated. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Contributions towards a monograph of Phoma (Coelomycetes) — I. 2. Section Phoma: Additional taxa with very small conidia and taxa with conidia up to 7 µm long

NARCIS (Netherlands)

Gruyter, de J.; Noordeloos, M.E.; Boerema, G.H.

1993-01-01

Twenty-seven taxa in section Phoma with conidia not exceeding 7 µm in length are keyed out and described on account of their characteristics in vitro. Four new species are introduced: Phoma aurea de Gruyter, Noordel. & Boerema, Phoma crystallifer de Gruyter, Noordel. & Boerema, Phoma flavescens de

Consecutive monitoring of lifelong production of conidia by individual conidiophores of Blumeria graminis f. sp. hordei on barley leaves by digital microscopic techniques with electrostatic micromanipulation.

Science.gov (United States)

Moriura, Nobuyuki; Matsuda, Yoshinori; Oichi, Wataru; Nakashima, Shinya; Hirai, Tatsuo; Sameshima, Takeshi; Nonomura, Teruo; Kakutani, Koji; Kusakari, Shin-Ichi; Higashi, Katsuhide; Toyoda, Hideyoshi

2006-01-01

Conidial formation and secession by living conidiophores of Blumeria graminis f. sp. hordei on barley leaves were consecutively monitored using a high-fidelity digital microscopic technique combined with electrostatic micromanipulation to trap the released conidia. Conidial chains formed on conidiophores through a series of septum-mediated division and growth of generative cells. Apical conidial cells on the conidiophores were abstricted after the conidial chains developed ten conidial cells. The conidia were electrically conductive, and a positive charge was induced in the cells by a negatively polarized insulator probe (ebonite). The electrostatic force between the conidia and the insulator was used to attract the abstricted conidia from the conidiophores on leaves. This conidium movement from the targeted conidiophore to the rod was directly viewed under the digital microscope, and the length of the interval between conidial septation and secession, the total number of the conidia produced by a single conidiophore, and the modes of conidiogenesis were clarified. During the stage of conidial secession, the generative cells pushed new conidial cells upwards by repeated division and growth. The successive release of two apical conidia was synchronized with the successive septation and growth of a generative cell. The release ceased after 4-5 conidia were released without division and growth of the generative cell. Thus, the life of an individual conidiophore (from the erection of the conidiophore to the release of the final conidium) was shown to be 107 h and to produce an average of 33 conidia. To our knowledge, this is the first report on the direct estimation of life-long conidial production by a powdery mildew on host leaves.

Phytopharmacological studies on the rice diseases. III. Fungitoxicity of the different dust fungicides evaluated by protoplasm coagulation in conidia of Cochliobolus miyabeanus

Energy Technology Data Exchange (ETDEWEB)

Hashioka, Y; Ikegami, H

1956-01-01

The modes of reaction of fungal conidia to fungicides (mercury and copper compounds) varied with the concentration of fungicide. At lower fugicide concentrations, restriction of elongation of germ-tubes was noted. At high concentrations, there was no germination accompanied by vacuolation. At the highest concentrations, conidia protoplasm was coagulated.

The pbrB gene encodes a laccase required for DHN-melanin synthesis in conidia of Talaromyces (Penicillium) marneffei.

Science.gov (United States)

Sapmak, Ariya; Boyce, Kylie J; Andrianopoulos, Alex; Vanittanakom, Nongnuch

2015-01-01

Talaromyces marneffei (Basionym: Penicillium marneffei) is a significant opportunistic fungal pathogen in patients infected with human immunodeficiency virus in Southeast Asia. T. marneffei cells have been shown to become melanized in vivo. Melanins are pigment biopolymers which act as a non-specific protectant against various stressors and which play an important role during virulence in fungi. The synthesis of the two most commonly found melanins in fungi, the eumelanin DOPA-melanin and the allomelanin DHN-melanin, requires the action of laccase enzymes. The T. marneffei genome encodes a number of laccases and this study describes the characterization of one of these, pbrB, during growth and development. A strain carrying a PbrB-GFP fusion shows that pbrB is expressed at high levels during asexual development (conidiation) but not in cells growing vegetatively. The pbrB gene is required for the synthesis of DHN-melanin in conidia and when deleted results in brown pigmented conidia, in contrast to the green conidia of the wild type.

Influence of temperature and humidity on the viability of Ophiognomonia clavigignenti-juglandacearum Conidia

Science.gov (United States)

M.J. Moore; M.E. Ostry

2015-01-01

Butternut canker, caused by the fungus Ophiognomonia clavigignenti-juglandacearum, primarily kills butternut (Juglans cinerea). Rain splash and local air currents are the primary means of conidia dispersal but that does not explain its long-distance spread and infection of isolated trees. Dispersal by insect or animal vectors...

Nutritional improvement of the rose handling Peronospora Sparsa Berkeley, causal agent of downy mildew

International Nuclear Information System (INIS)

Castillo Carlos Fernando; Alvarez Elizabeth; Gomez Eduardo; Llano, German A; Castano Zapata, Jairo

2010-01-01

Nutritional solutions, silica, resistance, downy mildew. The downy mildew (Peronospora sparsa), is one of the most important diseases of rose in Colombia, causing losses up to 8%. The objective of this research was to determine a preventive control of the disease, through the improvement of the nutritional balance of the plant. The first phase, involved the effect of different concentrations of N, K, Ca, B and Mn, on the incidence and severity of the disease, and in the second one, the evaluation of the best five treatments of the first phase. The treatments were applied to the varieties Charlotte, Classy and Malibu during 4 weeks, using a splitting plot design with six replications. After one month the plants were inoculated with the fungus at a concentration of 3x104 sporangia mL.The results of the area under the disease progress curve (AUDPC), showed that the plants of Charlotte with 200 ppm of Si had the lowest expression of the disease. In Classy, the best treatment was the standard solution, demonstrating that the effects of the treatments depend of the variety. Charlotte and Malibu showed susceptibility, while Classy, partial resistance to the disease.

Nutritional improvement of the rose handling Peronospora Sparsa Berkeley, causal agent of downy mildew

International Nuclear Information System (INIS)

Castillo, Carlos Fernando; Alvarez, Elizabeth; Gomez, Eduardo; Llano, German; Castano Zapata, Jairo

2010-01-01

The downy mildew (Peronospora sparsa), is one of the most important diseases of rose in Colombia, causing losses up to 8%. The objective of this research was to determine a preventive control of the disease, through the improvement of the nutritional balance of the plant. The first phase, involved the effect of different concentrations of N, K, Ca, B and Mn, on the incidence and severity of the disease, and in the second one, the evaluation of the best five treatments of the first phase. The treatments were applied to the varieties Charlotte, Classy and Malibu during 4 weeks, using a splitting plot design with six replications. After one month the plants were inoculated with the fungus at a concentration of 3x104 sporangia mL. The results of the area under the disease progress curve (AUDPC), showed that the plants of Charlotte with 200 ppm of Si had the lowest expression of the disease. In Classy, the best treatment was the standard solution, demonstrating that the effects of the treatments depend of the variety. Charlotte and Malibu showed susceptibility, while Classy, partial resistance to the disease.

Modification of a Pollen Trap Design To Capture Airborne Conidia of Entomophaga maimaiga and Detection of Conidia by Quantitative PCR.

Science.gov (United States)

Bittner, Tonya D; Hajek, Ann E; Liebhold, Andrew M; Thistle, Harold

2017-09-01

The goal of this study was to develop effective and practical field sampling methods for quantification of aerial deposition of airborne conidia of Entomophaga maimaiga over space and time. This important fungal pathogen is a major cause of larval death in invasive gypsy moth ( Lymantria dispar ) populations in the United States. Airborne conidia of this pathogen are relatively large (similar in size to pollen), with unusual characteristics, and require specialized methods for collection and quantification. Initially, dry sampling (settling of spores from the air onto a dry surface) was used to confirm the detectability of E. maimaiga at field sites with L. dispar deaths caused by E. maimaiga , using quantitative PCR (qPCR) methods. We then measured the signal degradation of conidial DNA on dry surfaces under field conditions, ultimately rejecting dry sampling as a reliable method due to rapid DNA degradation. We modified a chamber-style trap commonly used in palynology to capture settling spores in buffer. We tested this wet-trapping method in a large-scale (137-km) spore-trapping survey across gypsy moth outbreak regions in Pennsylvania undergoing epizootics, in the summer of 2016. Using 4-day collection periods during the period of late instar and pupal development, we detected variable amounts of target DNA settling from the air. The amounts declined over the season and with distance from the nearest defoliated area, indicating airborne spore dispersal from outbreak areas. IMPORTANCE We report on a method for trapping and quantifying airborne spores of Entomophaga maimaiga , an important fungal pathogen affecting gypsy moth ( Lymantria dispar ) populations. This method can be used to track dispersal of E. maimaiga from epizootic areas and ultimately to provide critical understanding of the spatial dynamics of gypsy moth-pathogen interactions. Copyright © 2017 American Society for Microbiology.

Metacridamides A and B, bioactive macrocycles from conidia of the entomopathogenic fungus Metarhizium acridum

Science.gov (United States)

Metarhizium acridum, an entomopathogenic fungus, has been commercialized and used successfully for biocontrol of grasshopper pests in Africa and Australia. Its conidia produce two novel 17-membered macrocycles, metacridamides A (1) and B (2), which consist of a Phe unit condensed with a nonaketide....

Partitioning of ochratoxin A in mycelium and conidia of Aspergillus carbonarius and the impact on toxin contamination of grapes and wine

International Nuclear Information System (INIS)

Atoui, A.; Mathieu, F.; Lebrihi, A.; Mitchell, D.; Magan, N.

2007-01-01

Aspergillus carbonarius is an important ochratoxin A (OTA)-producing fungus which is responsible for toxin contamination of grapes and wine. The objectives of this study were to examine the partitioning of OTA in mycelium and conidia of a range of A. carbonarius strains on artificial grape juice and defined media, to determine the excretion patterns of OTA from these spores, and the effect of organic acids used in wine production on OTA excretion from conidia. The results showed that 60-70% of the OTA was accumulated in the conidia of a number of different isolates of A. carbonarius. Calculations showed that on different defined media, an amount of 0011- to 01-pg OTA was present per conidium. The OTA in spores was found to be rapidly excreted into the medium during the initial few hours after conidial germination leading to an increase of OTA in must during maceration for wine production. The presence of tartaric acid inhibited OTA production, but malic acid enhanced this production during mycelial growth. These acids were also shown to affect the time course of germination and the rate of OTA excretion from conidia during germination. Conclusions: This study is the first to examine and show the partitioning of OTA into spores of strains of A. carbonarius and that rapid excretion of OTA from spores could be a reason for OTA accumulation in musts during wine production.Significance and Impact of the Study: Conidia of A. carbonarius could be a major source of OTA contamination of grapes used in wine production. This information could help in the development of effective prevention strategies to minimize wine contamination with this important mycotoxin. (author)

Contributions towards a monograph of Phoma (Coelomycetes) — I. 2. Section Phoma: Additional taxa with very small conidia and taxa with conidia up to 7 µm long

OpenAIRE

Gruyter, de, J.; Noordeloos, M.E.; Boerema, G.H.

1993-01-01

Twenty-seven taxa in section Phoma with conidia not exceeding 7 µm in length are keyed out and described on account of their characteristics in vitro. Four new species are introduced: Phoma aurea de Gruyter, Noordel. & Boerema, Phoma crystallifer de Gruyter, Noordel. & Boerema, Phoma flavescens de Gruyter, Noordel. & Boerema and Phoma subherbarum de Gruyter, Noordel. & Boerema. As new names are proposed Phoma chenopodiicola de Gruyter, Noordel. & Boerema, Phoma pereupyrena de Gruyter, Noordel...

A Non-canonical Melanin Biosynthesis Pathway Protects Aspergillus terreus Conidia from Environmental Stress.

Science.gov (United States)

Geib, Elena; Gressler, Markus; Viediernikova, Iuliia; Hillmann, Falk; Jacobsen, Ilse D; Nietzsche, Sandor; Hertweck, Christian; Brock, Matthias

2016-05-19

Melanins are ubiquitous pigments found in all kingdoms of life. Most organisms use them for protection from environmental stress, although some fungi employ melanins as virulence determinants. The human pathogenic fungus Aspergillus fumigatus and related Ascomycetes produce dihydroxynaphthalene- (DHN) melanin in their spores, the conidia, and use it to inhibit phagolysosome acidification. However, biosynthetic origin of melanin in a related fungus, Aspergillus terreus, has remained a mystery because A. terreus lacks genes for synthesis of DHN-melanin. Here we identify genes coding for an unusual NRPS-like enzyme (MelA) and a tyrosinase (TyrP) that A. terreus expressed under conidiation conditions. We demonstrate that MelA produces aspulvinone E, which is activated for polymerization by TyrP. Functional studies reveal that this new pigment, Asp-melanin, confers resistance against UV light and hampers phagocytosis by soil amoeba. Unexpectedly, Asp-melanin does not inhibit acidification of phagolysosomes, thus likely contributing specifically to survival of A. terreus conidia in acidic environments. Copyright © 2016 Elsevier Ltd. All rights reserved.

Identification of Aspergillus fumigatus Surface Components That Mediate Interaction of Conidia and Hyphae With Human Platelets.

Science.gov (United States)

Rambach, Günter; Blum, Gerhard; Latgé, Jean-Paul; Fontaine, Thierry; Heinekamp, Thorsten; Hagleitner, Magdalena; Jeckström, Hanna; Weigel, Günter; Würtinger, Philipp; Pfaller, Kristian; Krappmann, Sven; Löffler, Jürgen; Lass-Flörl, Cornelia; Speth, Cornelia

2015-10-01

Platelets were recently identified as a part of innate immunity. They are activated by contact with Aspergillus fumigatus; putative consequences include antifungal defense but also thrombosis, excessive inflammation, and thrombocytopenia. We aimed to identify those fungal surface structures that mediate interaction with platelets. Human platelets were incubated with Aspergillus conidia and hyphae, isolated wall components, or fungal surface mutants. Interaction was visualized microscopically; activation was quantified by flow cytometry of specific markers. The capacity of A. fumigatus conidia to activate platelets is at least partly due to melanin, because this effect can be mimicked with "melanin ghosts"; a mutant lacking melanin showed reduced platelet stimulating potency. In contrast, conidial hydrophobin masks relevant structures, because an A. fumigatus mutant lacking the hydrophobin protein induced stronger platelet activation than wild-type conidia. A. fumigatus hyphae also contain surface structures that interact with platelets. Wall proteins, galactomannan, chitin, and β-glucan are not the relevant hyphal components; instead, the recently identified fungal polysaccharide galactosaminogalactan potently triggered platelet activation. Conidial melanin and hydrophobin as well as hyphal galactosaminogalactan represent important pathogenicity factors that modulate platelet activity and thus might influence immune responses, inflammation, and thrombosis in infected patients. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Susceptibility testing of terbinafine alone and in combination with amphotericin B, itraconazole, or voriconazole against conidia and hyphae of dematiaceous molds.

Science.gov (United States)

Biancalana, Fernanda Simas Corrêa; Lyra, Luzia; Moretti, Maria Luiza; Schreiber, Angélica Zaninelli

2011-12-01

Studies have demonstrated excellent in vivo efficacy of terbinafine combined with other antifungal agents against dematiaceous molds; however, there is a lack of in vitro studies. Most studies evaluated conidia inocula, but susceptibility testing of hyphae could mimic the fungal status in infected tissues and might reflect the therapeutic potential of the agent. We investigated the in vitro susceptibility of terbinafine alone and in combination with amphotericin B, itraconazole, or voriconazole against conidia by microdilution and dynamic measurement of hyphae growth of dematiaceous molds. The MIC values for hyphae were, until 3 dilutions, below the MIC obtained for conidia. The results indicated 100% synergistic interactions between terbinafine and azoles or amphotericin B in all tests, but lower MICs for hyphae. In conclusion, our findings allow us to say that the hyphal form of tested dematiaceous molds showed high susceptibility to all antifungal agents evaluated, alone and in combination with terbinafine. Copyright © 2011 Elsevier Inc. All rights reserved.

Impact of two treatments of a formulation of Beauveria bassiana (Deuteromycota: Hyphomycetes) conidia on Varroa mites (Acari: Varroidae) and on honeybee (Hymenoptera: Apidae) colony health.

Science.gov (United States)

Meikle, William G; Mercadier, Guy; Holst, Niels; Girod, Vincent

2008-12-01

Bee colonies in southern France were treated with conidia (asexual spores) from two strains of Beauveria bassiana, an entomopathogenic fungus. One strain was commercial (GHA) and the other had been isolated from Varroa mites in the region (Bb05002). Objectives were to evaluate treatment effect on colony weight, adult bee mass, capped brood, and on Varroa fall onto sticky boards. Treatments included conidia formulated with either carnauba or candelilla wax powder, candelilla wax powder alone, or control; in two treatment groups formulation was applied a second time after one week. Treatment did not affect colony health. Colonies treated twice with Bb05002 conidia and carnauba wax powder had significantly higher mite fall compared to colonies treated with blank candelilla wax powder. The proportion of fallen mites that were infected in both conidia treatments was higher than controls for 18 days after the second treatment. The number of fungal propagules on the bees themselves remained elevated for about 14 days after the second treatment. These results were compared to published results from previous experiments with regard to infection duration.

Surface structure characterization of Aspergillus fumigatus conidia mutated in the melanin synthesis pathway and their human cellular immune response.

Science.gov (United States)

Bayry, Jagadeesh; Beaussart, Audrey; Dufrêne, Yves F; Sharma, Meenu; Bansal, Kushagra; Kniemeyer, Olaf; Aimanianda, Vishukumar; Brakhage, Axel A; Kaveri, Srini V; Kwon-Chung, Kyung J; Latgé, Jean-Paul; Beauvais, Anne

2014-08-01

In Aspergillus fumigatus, the conidial surface contains dihydroxynaphthalene (DHN)-melanin. Six-clustered gene products have been identified that mediate sequential catalysis of DHN-melanin biosynthesis. Melanin thus produced is known to be a virulence factor, protecting the fungus from the host defense mechanisms. In the present study, individual deletion of the genes involved in the initial three steps of melanin biosynthesis resulted in an altered conidial surface with masked surface rodlet layer, leaky cell wall allowing the deposition of proteins on the cell surface and exposing the otherwise-masked cell wall polysaccharides at the surface. Melanin as such was immunologically inert; however, deletion mutant conidia with modified surfaces could activate human dendritic cells and the subsequent cytokine production in contrast to the wild-type conidia. Cell surface defects were rectified in the conidia mutated in downstream melanin biosynthetic pathway, and maximum immune inertness was observed upon synthesis of vermelone onward. These observations suggest that although melanin as such is an immunologically inert material, it confers virulence by facilitating proper formation of the A. fumigatus conidial surface. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Germination of Aspergillus niger conidia is triggered by nitrogen compounds related to L-amino acids.

Science.gov (United States)

Hayer, Kimran; Stratford, Malcolm; Archer, David B

2014-10-01

Conidial germination is fundamentally important to the growth and dissemination of most fungi. It has been previously shown (K. Hayer, M. Stratford, and D. B. Archer, Appl. Environ. Microbiol. 79:6924-6931, 2013, http://dx.doi.org/10.1128/AEM.02061-13), using sugar analogs, that germination is a 2-stage process involving triggering of germination and then nutrient uptake for hyphal outgrowth. In the present study, we tested this 2-stage germination process using a series of nitrogen-containing compounds for the ability to trigger the breaking of dormancy of Aspergillus niger conidia and then to support the formation of hyphae by acting as nitrogen sources. Triggering and germination were also compared between A. niger and Aspergillus nidulans using 2-deoxy-D-glucose (trigger), D-galactose (nontrigger in A. niger but trigger in A. nidulans), and an N source (required in A. niger but not in A. nidulans). Although most of the nitrogen compounds studied served as nitrogen sources for growth, only some nitrogen compounds could trigger germination of A. niger conidia, and all were related to L-amino acids. Using L-amino acid analogs without either the amine or the carboxylic acid group revealed that both the amine and carboxylic acid groups were essential for an L-amino acid to serve as a trigger molecule. Generally, conidia were able to sense and recognize nitrogen compounds that fitted into a specific size range. There was no evidence of uptake of either triggering or nontriggering compounds over the first 90 min of A. niger conidial germination, suggesting that the germination trigger sensors are not located within the spore. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Eficiencia del ácido ascórbico, ácido salicílico y extracto de Dulcamara (Bryophyllum gastonis B.) en la prevención de Mildeo velloso (Peronospora pulverulenta) en Gypsophila (Gypsophila paniculata) variedad Party Time

OpenAIRE

Puma Quinchuango, Lupe Marlene

2010-01-01

This research was conducted at Finca Santa Martha to the application of salicylic acid, ascorbic acid, dulcamara extract and mix them to control downy mildew ( Peronospora pulverulenta) in the cultivation of (Gypsophila paniculata) variety Party Time. 8 treatments were evaluated during the crop phenological stages such as induction, elongation and flowering. We utilized an experimental design with 3 replications DBCA was evaluated in incidence and severity. Results and discu...

Germination of Penicillium paneum Conidia Is Regulated by 1-Octen-3-ol, a Volatile Self-Inhibitor

NARCIS (Netherlands)

Chitarra, G.S.; Abee, T.; Rombouts, F.M.; Posthumus, M.A.; Dijksterhuis, J.

2004-01-01

Penicillium paneum is an important contaminant of cereal grains which is able to grow at low temperature, low pH, high levels of carbon dioxide, and under acid conditions. P. paneum produces mycotoxins, which may be harmful to animals and humans. We found that conidia in dense suspensions showed

Germination of penicillium paneum Conidia is regulated by 1-octen-3-ol, a volatile self-inhibitor

NARCIS (Netherlands)

Chitarra, Gilma S; Abee, Tjakko; Rombouts, Frank M; Posthumus, Maarten A; Dijksterhuis, Jan

Penicillium paneum is an important contaminant of cereal grains which is able to grow at low temperature, low pH, high levels of carbon dioxide, and under acid conditions. P. paneum produces mycotoxins, which may be harmful to animals and humans. We found that conidia in dense suspensions showed

Effect of zinc compounds on Fusarium verticillioides growth, hyphae alterations, conidia, and fumonisin production.

Science.gov (United States)

Savi, Geovana D; Vitorino, Vinícius; Bortoluzzi, Adailton J; Scussel, Vildes M

2013-10-01

Several strategies are used to eliminate toxigenic fungi that produce fumonisins in grains. Fusarium verticillioides can be controlled by the application of synthetic fungicides in the field or during storage. However, there may also be residuals, which may remain in the foods. Inorganic compounds such as zinc are cheap, stable and could present strong antifungal activity. Some Zn compounds can be utilized as dietary supplements and are authorized for the fortification of foods. Knowing the advantages and that low concentrations of Zn can have antimicrobial activity, our objective was to evaluate the effects of Zn compounds on the growth of F. verticillioides and the production of fumonisin and conidia. In addition, we aimed to verify that Zn compounds cause morphological alterations of the hyphae, mortality and production of reactive oxygen species. Zn compounds efficiently reduced fungal growth and fumonisin production. Treatment using zinc perchlorate gave the best results. All treatments inhibited conidia production and caused morphological alterations of the hyphae. It was possible to observe cell death and production of reactive oxygen species. Zn compounds have advantages compared to other antifungal compounds. In particular, they are non-toxic for the organism in appropriate amounts. They could be studied further as potential fungicides in agriculture. © 2013 Society of Chemical Industry.

Hypocrea rufa/Trichoderma viride: a reassessment, and description of five closely related species with and without warted conidia.

Science.gov (United States)

Jaklitsch, Walter M; Samuels, Gary J; Dodd, Sarah L; Lu, Bing-Sheng; Druzhinina, Irina S

2006-01-01

The type species of the genus Hypocrea (Hypocreaceae, Hypocreales, Ascomycota, Fungi), H. rufa, is re-defined and epitypified using a combination of phenotype (morphology of teleomorphs and anamorphs, and characteristics in culture) and phylogenetic analyses of the translation-elongation factor 1alpha gene. Its anamorph, T. viride, the type species of Trichoderma, is re-described and epitypified. Eidamia viridescens is combined as Trichoderma viridescens and is recognised as one of the most morphologically and phylogenetically similar relatives of T. viride. Its teleomorph is newly described as Hypocrea viridescens. Contrary to frequent citations of H. rufa and T. viride in the literature, this species is relatively rare. Although both T. viride and T. viridescens have a wide geographic distribution, their greatest genetic diversity appears to be in Europe and North America. Hypocrea vinosa is characterised and its anamorph, T. vinosum sp. nov., is described. Conidia of T. vinosum are subglobose and warted. The new species T. gamsii is proposed. It shares eidamia-like morphology of conidiophores with T. viridescens, but it has smooth, ellipsoidal conidia that have the longest L/W ratio that we have seen in Trichoderma. Trichoderma scalesiae, an endophyte of trunks of Scalesia pedunculata in the Galapagos Islands, is described as new. It only produces conidia on a low-nutrient agar to which filter paper has been added. Additional phylogenetically distinct clades are recognised and provisionally delimited from the species here described. Trichoderma neokoningii, a T. koningii-like species, is described from a collection made in Peru on a fruit of Theobroma cacao infected with Moniliophthora roreri.

Studies on the effect of Osmia rufa L. (Apoidea, Megachilidae on the effectiveness of pod and seed development in the subgenus Glycine

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Halina Skorupska

2014-01-01

Full Text Available Three abundantly blooming forms of Glycogen tabacina and one G. tomentella form were studied. The experiment was conducted under isolated conditions. The effect of Osmia rufa L. on the fertility of raceme flowers was studied. It was found that the G. tabacina and G. tomentella flowers were intensively penetrated by the insects. A very clear increase (3-4 fold in pod development was observed. The results ol the experiment indicate that geitonogamic pollination has a favorable influence on the effectiveness of the blooming of the chasmogamic flowers of the studied species.

Screening of Metarhizium and Beauveria spp. conidia with exposure to simulated sunlight and a range of temperatures

International Nuclear Information System (INIS)

Morley-Davies, J.; Moore, D.; Prior, C.

1996-01-01

Conidia of 14 isolates of the entomopathogenic fungi Beauveria bassiana, Metarhizium flavoviride and M. anisopliae were formulated by suspending in paraffinic oil or as dry powder. Non-indicating silica gel was added to both formulations which were stored at 13 °C for at least 2 wk before exposure to a range of temperatures: −10°, 10°, 20°, 30°, 40° and 50° to determine the effects of temperature of storage on viability. At 50° the isolate studied in most detail (M. flavoviride, IMI 330189) initially showed a gradual decline in viability with 73% germination for oil samples after 60 d from an initial level of 93%, whereas the dry samples typically showed higher germination rates (initially 96% germination, dropping to 80% after 60 d). Subsequently there was a rapid decline and both oil formulated and dry conidia had lost almost all viability by 90 d. Samples of IMI 330189 stored dry or in oil, at 40° and below showed > 79% germination after 90 d. M. flavoviride 191–660 exhibited the highest temperature tolerance with >40% germination of the dry stored conidia after 90 d at 50°. Some isolates of both B. bassiana and Metarhizium spp. showed markedly lower tolerance of high temperatures. Samples stored dry usually showed greater percentage germination than samples in oil for all isolates, at all temperatures. The isolates were also exposed to 4, 8, 16 and 24 h uv light from a sunlight simulator at 40°. Conidial viability decreased markedly in all isolates with increasing uv exposure. Germination ranged between 10 and 50% after 24 h exposure to uv, 191–660 retaining highest viability. (author)

Sterile Males of Ceratitis Capitata (Diptera: Tephritidae) as Disseminators of Beauveria Bassiana Conidia for IPM Strategies

Energy Technology Data Exchange (ETDEWEB)

Flores, Salvador; Campos, Sergio; Montoya, Pablo [Programa Moscafrut SAGARPA-IICA, Camino a los Cacahotales S/N, Metapa de Dominguez, Chiapas 30860 (Mexico); Villasenor, Antonio; Valle, Alvaro; Enkerlin, Walther [Codireccion Mexico, Programa Regional Moscamed Mexico- Guatemala-Estados Unidos (Guatemala); Toledo, Jorge; Liedo, Pablo [Departamento Agricultura Sociedad y Ambiente, El Colegio de la Frontera Sur. Carretera Antiguo Aeropuerto, Tapachula, Chiapas 30700 (Mexico)

2014-01-15

Full text: Sterile Mediterranean fruit fly, Ceratitis capitata (Wied.), males were evaluated as vectors to spread Beauveria bassiana (Bals) conidia to wild C. capitata populations under field conditions. The inoculated sterile males were released by air, using the chilled adult technique over 7000 ha of coffee growing in Chimaltenango, Guatemala, Central America. The impact of releases was determined using dry traps baited with a food attractant. The effects of these releases on Apis mellifera, Linnaeus (honey bee), Hypothenemus hampei, Ferrari (coffee berry borer) and the parasitic mite Varroa destructor (Oudeman) were also evaluated. Inoculated sterile males were able to transmit fungal spores to 44% of the wild C. capitata flies captured in traps, which likely were infected through intra- and intersexual interactions during leks, mating or mating attempts. There was no transmission of the fungal spores to non- target insect species such as coffee berry borer, honey bees or varroa. We conclude that sterile males of Mediterranean fruit fly inoculated with B. bassiana can act as effective vectors of conidia to wild populations, constituting a safe, environmentally friendly and selective alternative for suppressing the medfly under a Sterile Insect Technique-based IPM approach. (author)

Evaluation of Strains of Metarhizium anisopliae and Beauveria bassiana against Spodoptera litura on the Basis of Their Virulence, Germination Rate, Conidia Production, Radial Growth and Enzyme Activity.

Science.gov (United States)

Petlamul, Wanida; Prasertsan, Poonsuk

2012-06-01

Ten strains of the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana were evaluated to find the most effective strain for optimization studies. The first criterion tested for strain selection was the mortality (> 50%) of Spodoptera litura larvae after inoculation of the fungus for 4 days. Results on several bioassays revealed that B. bassiana BNBCRC showed the most virulence on mortality S. litura larvae (80% mortality). B. bassiana BNBCRC also showed the highest germination rate (72.22%). However, its conidia yield (7.2 × 10(8) conidia/mL) was lower than those of B. bassiana B 14841 (8.3 × 10(8) conidia/mL) and M. anisopliae M6 (8.2 × 10(8) conidia/mL). The highest accumulative radial growth was obtained from the strain B14841 (37.10 mm/day) while the strain BNBCRC showed moderate radial growth (24.40 mm/day). M. anisopliae M6 possessed the highest protease activity (145.00 mU/mL) while M. anisopliae M8 possessed the highest chitinase activity (20.00 mU/mL) during 96~144 hr cultivation. Amongst these criteria, selection based on virulence and germination rate lead to the selection of B. bassiana BNBCRC. B. bassiana B14841 would be selected if based on growth rate while M. anisopliae M6 and M8 possessed the highest enzyme activities.

DIVERSIDAD GENÉTICA DE Peronospora sparsa (PERONOSPORACEAE EN CULTIVOS DE ROSA DE COLOMBIA

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MARILUZ AYALA-VÁSQUEZ

2008-01-01

Full Text Available El mildeo velloso de la rosa es la enfermedad más limitante de este cultivo en Colombia. Aunque algunos trabajos de investigación se han adelantado recientemente sobre la epidemiología de esta enfermedad, uno de los campos menos estudiados es el nivel de variabilidad de su agente causal. El conocimiento de este aspecto es fundamental para definir las estrategias de manejo, especialmente aquellas relacionadas con la resistencia genética y el control químico. Los objetivos de esta investigación fueron confirmar la identidad taxonómica del agente causal del mildeo velloso de la rosa y profundizar en el conocimiento de su estructura poblacional en Colombia. Para esto se colectaron 34 aislamientos del pseudohongo en cultivos de rosas ubicados en Antioquia y la sabana de Bogotá y se extrajo su ADN, para proceder a la amplificación mediante PCR de la región ITS del ADNr utilizando los primers especieespecíficos PS3 y PS1. Dichos productos se emplearon tanto para su secuenciación directa como para la evaluación de variabilidad genética mediante la técnica PCRRFLP, resultados que se complementaron con la utilización de los marcadores RAPD y RAMS. La presencia de un amplicón de aproximadamente 700 pb que compartió un porcentaje de identidad del 100% con secuencias depositadas en el GenBank, permitió confirmar a los aislamientos como pertenecientes a Peronospora sparsa, mientras que el análisis de RFLP, RAPD y RAMS indicó un muy bajo nivel de variabilidad entre todos los aislamientos, concluyéndose que la población de P. sparsa en Colombia es predominantemente clonal

The polyene antimycotics nystatin and filipin disrupt the plasma membrane, whereas natamycin inhibits endocytosis in germinating conidia of Penicillium discolor

NARCIS (Netherlands)

Leeuwen, van M.R.; Golovina, E.A.; Dijksterhuis, J.

2009-01-01

To investigate the differences in membrane permeability and the effect on endocytosis of the polyene antimycotics nystatin, filipin and natamycin on germinating fungal conidia. Methods and Results: The model system was Penicillium discolor, a food spoilage fungus. Filipin resulted in

Liquid culture production of microsclerotia and submerged conidia by Trichoderma harzianum active against damping-off disease caused by Rhizoctonia solani.

Science.gov (United States)

Kobori, Nilce N; Mascarin, Gabriel M; Jackson, Mark A; Schisler, David A

2015-04-01

Media and culturing protocols were identified that supported the formation of submerged conidia and microsclerotia (MS) by Trichoderma harzianum Rifai strain T-22 using liquid culture fermentation. Liquid media with a higher carbon concentration (36 g L(-1)) promoted MS formation at all C:N ratios tested. Hyphae aggregated to form MS after 2 d growth and after 7 d MS were fully melanized. This is the first report of MS formation by T. harzianum or any species of Trichoderma. Furthermore, submerged conidia formation was induced by liquid culture media, but yields, desiccation tolerance, and storage stability varied with C:N ratio and carbon rate. Air-dried MS granules (culture production, stabilization process, and bioefficacy of the hitherto unreported MS of T. harzianum as a potential biofungicide for use in integrated management programs against soilborne diseases. Copyright © 2014 The British Mycological Society. All rights reserved.

Photodynamic inactivation of conidia of the fungus Colletotrichum abscissum on Citrus sinensis plants with methylene blue under solar radiation.

Science.gov (United States)

Gonzales, Júlia C; Brancini, Guilherme T P; Rodrigues, Gabriela B; Silva-Junior, Geraldo José; Bachmann, Luciano; Wainwright, Mark; Braga, Gilberto Ú L

2017-11-01

Antimicrobial photodynamic treatment (APDT) is a promising light based approach to control diseases caused by plant-pathogenic fungi. In the present study, we evaluated the effects of APDT with the phenothiazinium photosensitizer methylene blue (MB) under solar radiation on the germination and viability of conidia of the pathogenic fungus Colletotricum abscissum (former Colletotrichum acutatum sensu lato). Experiments were performed both on petals and leaves of sweet orange (Citrus sinensis) in different seasons and weather conditions. Conidial suspensions were deposited on the leaves and petals surface, treated with the PS (25 or 50μM) and exposed to solar radiation for only 30min. The effects of APDT on conidia were evaluated by counting the colony forming units recovered from leaves and petals and by direct evaluating conidial germination on the surface of these plant organs after the treatment. To better understand the mechanistic of conidial photodynamic inactivation, the effect of APDT on the permeability of the conidial plasma membrane was assessed using the fluorescent probe propidium iodide (PI) together with flow cytometry and fluorescence microscopy. APDT with MB and solar exposure killed C. abscissum conidia and prevented their germination on both leaves and petals of citrus. Reduction of conidial viability was up to three orders of magnitude and a complete photodynamic inactivation was achieved in some of the treatments. APDT damaged the conidial plasma membrane and increased its permeability to PI. No damage to sweet orange flowers or leaves was observed after APDT. The demonstration of the efficacy of APDT on the plant host represents a further step towards the use of the method for control phytopathogens in the field. Copyright © 2017 Elsevier B.V. All rights reserved.

A novel approach of utilization of the fungal conidia biomass to remove heavy metals from the aqueous solution through immobilization

Science.gov (United States)

Cai, Chun-Xiang; Xu, Jian; Deng, Nian-Fang; Dong, Xue-Wei; Tang, Hao; Liang, Yu; Fan, Xian-Wei; Li, You-Zhi

2016-11-01

The biomass of filamentous fungi is an important cost-effective biomass for heavy metal biosorption. However, use of free fungal cells can cause difficulties in the separation of biomass from the effluent. In this study, we immobilized the living conidia of the heavy metal-resistant Penicillium janthinillum strain GXCR by polyvinyl alcohol (PVA)-sodium alginate (SA) beads to remove heavy metals from an aqueous solution containing a low concentration (70 mg/L) of Cu, Pb, and Cd. The PVA-SA-conidia beads showed perfect characters of appropriate mechanical strength suitable for metal removal from the dynamic wastewater environment, an ideal settleability, easy separation from the solution, and a high metal biosorption and removal rate even after four cycles of successive sorption-desorption of the beads, overcoming disadvantages when fungal biomasses alone are used for heavy metal removal from wastewater. We also discuss the major biosorption-affecting factors, biosorption models, and biosorption mechanisms.

Quinoa from Valley (Chenopodium quinoa Willd.: Valuable source of genetic resistance to powdery mildew(Peronospora farinosa Willd.

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Gabriel Julio

2013-02-01

Full Text Available In order to identify quinoa resistant cultivars to powdery mildew (Peronospora farinosa of high yield and large grain size, 36 cultivars were evaluated under two different fungicide applications and a control in the Valle Bajo from Cochabamba. We determined the realtive Area under Develop Progress Curve of Peronosporafarinosa (AUDPCPF relative, the yield and another eleven quantitative variables. The results showed that cultivars 01Tardía, 08Tardía, 12Tardía, 04Tardía, 11Tardía 10Tardía, 19Tardía y 18Tardía were susceptible and the cultivars H172, A26, A03, A16, A22, A14 and H171 were resistant. The cultivars A40, H177, A26, H172, A25, A1 y H176 showed yields from 3.4 to 6.34 t ha-1. The cultivars 15 Tardía, 03 Tardía, 14 Tardía, H173, H171, A25, H176 and H172 with chemical control strategy and the tricobalreacted favorably against powdery mildew, which was associated with levels of resistance in each cultivar. Finally, there was a high significant negative correlation between the variables AUDPCPF relative and physiological maturity, plant length, panicle length, stem diameter, panicle diameter and weight of 100 seeds. This showed that when the attack of mildewis severe, also affects the grains yield.

Inhibition of Pseudogymnoascus destructans growth from conidia and mycelial extension by bacterially produced volatile organic compounds.

Science.gov (United States)

Cornelison, Christopher T; Gabriel, Kyle T; Barlament, Courtney; Crow, Sidney A

2014-02-01

The recently identified causative agent of white-nose syndrome (WNS), Pseudogymnoascus destructans, has been implicated in the mortality of an estimated 5.5 million North American bats since its initial documentation in 2006 (Frick et al. in Science 329:679-682, 2010). In an effort to identify potential biological and chemical control options for WNS, 6 previously described bacterially produced volatile organic compounds (VOCs) were screened for anti-P. destructans activity. The compounds include decanal; 2-ethyl-1-hexanol; nonanal; benzothiazole; benzaldehyde; andN,N-dimethyloctylamine. P. destructans conidia and mycelial plugs were exposed to the VOCs in a closed air space at 15 and 4 °C and then evaluated for growth inhibition. All VOCs inhibited growth from conidia as well as inhibiting radial mycelial extension, with the greatest effect at 4 °C. Studies of the ecology of fungistatic soils and the natural abundance of the fungistatic VOCs present in these environments suggest a synergistic activity of select VOCs may occur. The evaluation of formulations of two or three VOCs at equivalent concentrations was supportive of synergistic activity in several cases. The identification of bacterially produced VOCs with anti-P. destructans activity indicates disease-suppressive and fungistatic soils as a potentially significant reservoir of biological and chemical control options for WNS and provides wildlife management personnel with tools to combat this devastating disease.

Differential DNA methylation may contribute to temporal and spatial regulation of gene expression and the development of mycelia and conidia in entomopathogenic fungus Metarhizium robertsii.

Science.gov (United States)

Li, Wanzhen; Wang, Yulong; Zhu, Jianyu; Wang, Zhangxun; Tang, Guiliang; Huang, Bo

2017-03-01

Conidia and mycelia are two important developmental stages in the asexual life cycle of entomopathogenic fungus Metarhizium. Despite the crucial role that DNA methylation plays in many biological processes, its role in regulation of gene expression and development in fungi is not yet fully understood. We performed genome-wide analysis of DNA methylation patterns of an M. robertsii strain with single base pair resolution. Specifically, we examined for changes in methylation patterns between the conidia and mycelia stages. The results showed that approximately 0.38 % of cytosines are methylated in conidia, which is lower than the DNA methylation level (0.42 %) in mycelia. We found that DNA methylation undergoes genome-wide reprogramming during fungal development in M. robertsii. 132 differentially methylated regions (DMRs), which were mostly distributed in gene regions, were identified. KEGG analysis revealed that the DMR-associated genes belong to metabolic pathways. Intriguingly, in contrast to most other eukaryotes, promoter activities in M. robertsii seemed differentially modulated by DNA methylation levels. We found that transcription tended to be enhanced in genes with moderate promoter methylation, while gene expression was decreased in genes with high or low promoter methylation. Copyright © 2017 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Adherencia de las conidias de Paracoccidioides brasiliensis a proteínas de matriz extracelular y líneas celulares de mamífero

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Luz Elena Cano

2001-04-01

Full Text Available Durante la colonización exitosa y la producción de enfermedad
por un patógeno es crítica su habilidad para adherirse a las superficies del hospedero ya que la invasión de los tejidos es estimulada por la unión a proteínas séricas o de matriz extracelular (MEC; éstas podrían funcionar como un puente entre el microorganismo y los receptores celulares o como un ligando opsónico (1. Poco se conoce sobre los mecanismos de patogenicidad empleados por Paracoccidioides brasiliensis durante los estados iniciales de la infección; hallazgos experimentales han mostrado que las propágulas infectantes del hongo (conidias una vez inhaladas, alcanzan los alvéolos pulmonares, se transforman en levaduras para finalmente diseminarse a órganos distantes (2. Aunque no existen datos concluyentes, la interacción inicial de las conidias con los tejidos del hospedero probablemente involucra algún proceso de reconocimiento específico de componentes de la MEC, así como de células epiteliales y/o endoteliales del pulmón (3. El
presente trabajo pretende identificar las proteínas de matriz extracelular que representan un blanco potencial para la unión de conidias de P. brasiliensis, así como definir la naturaleza de los procesos de interacción con proteínas de MEC y líneas celulares de mamíferos.

Phytopharmacological studies on the rice diseases. IV. Protoplasmic alteration of conidia of Cochliobolus miyabeanus in the organomercurial solutions

Energy Technology Data Exchange (ETDEWEB)

Hashioka, Y; Takamura, Y

1956-01-01

Conidia germinated readily in solutions lower than 5 x 10/sup -7/ Hg(Cu) g/litre, while coagulation of protoplasm occurred at higher concentrations irrespective of the kinds of fungicides employed. There were considerable differences in the percentage of protoplasm coagulation between the various types and concentrations of fungicides. This suggests that fungicidal action on spore germination maybe due to a rather simple and unusual process such as inactivation of the SH system.

Repellent activity of desiccant dusts and conidia of the entomopathogenic fungus Beauveria bassiana when tested against poultry red mites (Dermanyssus gallinae) in laboratory experiments.

Science.gov (United States)

Kilpinen, Ole; Steenberg, Tove

2016-11-01

Desiccant dusts and entomopathogenic fungi have previously been found to hold potential against the poultry red mite, which is an important pest in egg production and notoriously difficult to control. Both control agents may cause repellence in other arthropods and potentially also influence control levels adversely when used against the poultry red mite. Five desiccant dust products with good efficacy against the poultry red mite Dermanyssus gallinae caused avoidance behavior in mites when tested in bioassays. The repellent activity was correlated with efficacy, which was found to depend on both dose and relative humidity (RH). However, one desiccant dust was significantly less repellent compared to other dusts with similar levels of efficacy. Further, dry conidia of the fungus Beauveria bassiana were also shown to be repellent to poultry red mites, both when applied on its own and when admixed with a low dose of the desiccant dust Diamol. The pick-up of desiccant dust particles and fungus conidia from treated surfaces by mites did not differ depending on RH, whereas the overall efficacy of the two control agents were significantly higher at 75 than at 85 % RH. In addition, the combined effect of the two substances was synergistic when tested in a bioassay where mites could choose whether to cross a treated surface. This is the first time a member of Acari has been shown to be repelled by desiccant dusts and by conidia of an entomopathogenic fungus.

Visual expression analysis of the responses of the alternative oxidase gene (aox1) to heat shock, oxidative, and osmotic stresses in conidia of citric acid-producing Aspergillus niger.

Science.gov (United States)

Honda, Yuki; Hattori, Takasumi; Kirimura, Kohtaro

2012-03-01

The citric acid-producing filamentous fungus Aspergillus niger WU-2223L shows cyanide-insensitive respiration catalyzed by alternative oxidase in addition to the cytochrome pathway. Sequence analysis of the 5' flanking region of the alternative oxidase gene (aox1) revealed a potential heat shock element (HSE) and a stress response element (STRE). We have previously confirmed aox1 expression in conidia. In this study, to confirm whether the upstream region of aox1 responds to various stresses, we used a visual expression analysis system for single-cell conidia of the A. niger strain AOXEGFP-1. This strain harbored a fusion gene comprising aox1 and egfp, which encodes the enhanced green fluorescent protein (EGFP). The fluorescence intensity of EGFP increased in conidia of A. niger AOXEGFP-1 that were subjected to heat shock at 35-45 °C, oxidative stress by exposure to 5mM paraquat or 1 mM t-butylhydroperoxide, or osmotic stresses by exposure to 0.5 M KCl or 1.0 M mannitol. These results indicate that the putative HSE and STRE in the upstream region of aox1 directly or indirectly respond to heat shock, oxidative, and osmotic stresses. Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Repellent activity of desiccant dusts and conidia of the entomopathogenic fungus Beauveria bassiana when tested against poultry red mites (Dermanyssus gallinae) in laboratory experiments

DEFF Research Database (Denmark)

Kilpinen, Ole; Steenberg, Tove

2016-01-01

on both dose and relative humidity (RH). However, one desiccant dust was significantly less repellent compared to other dusts with similar levels of efficacy. Further, dry conidia of the fungus Beauveria bassiana were also shown to be repellent to poultry red mites, both when applied on its own and when...

Evaluación molecular y fenotípica de la sensibilidad al fungicida fenamidone en aislamientos de Peronospora sparsa

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Argel Luz Edith

2007-06-01

Full Text Available

El Mildeo velloso es uno de los mayores limitantes del cultivo de la rosa en Colombia. Esta enfermedad es causada por el Oomycete holobiótrofo Peronospora sparsa que fue detectado en la década de 1970 en la Sabana de Bogotá. En los últimos años su efecto negativo sobre la floricultura aumentó conduciendo a los cultivadores al empleo excesivo de fungicidas con acción sistémica, muchos de los cuales no cuentan con estudios de líneas base de sensibilidad ni con el diseño de estrategias anti-resistencia apropiadas. A fin de determinar el grado de sensibilidad de diez aislamientos de P. sparsa al fungicida fenamidone, representante del grupo de resistencia cruzada QoI, se realizaron pruebas de sensibilidad in vitro y evaluaciones moleculares. Los resultados muestran que este patógeno presenta un alto grado de sensibilidad a dicha molécula (EC₅₀ promedio: 0,51 mg· L^-1 y que no hay evidencia de la presencia de las mutaciones puntuales G143A y F129L asociadas con fenotipos resistentes a QoI. Este estudio aporta las bases para el desarrollo de una metodología de detección molecular de individuos con resistencia a fungicidas QoI que puede ser utilizada para monitorear las poblaciones del patógeno en nuestro medio.

Kale BoRACK1 is involved in the plant response to salt stress and Peronospora brassicae Gaumann.

Science.gov (United States)

Li, Da-Hong; Shen, Fu-Jia; Li, Hong-Yan; Li, Wei

2017-06-01

The receptor for activated C kinase 1 (RACK1) belongs to a protein subfamily containing a tryptophan-aspartic acid-domain (WD) repeat structure. Compelling evidence indicates that RACK1 can interact with many signal molecules and affect different signal transduction pathways. In this study, a kale (Brassica oleracea var. acephala f.tricolor) RACK1 gene (BoRACK1) was cloned by RT-PCR. The amino acid sequence of BoRACK1 had seven WD repeats in which there were typical GH (glycine-histidine) and WD dipeptides. Comparison with AtRACK1 from Arabidopsis revealed 87.1% identity at the amino acid level. Expression pattern analysis by RT-PCR showed that BoRACK1 was expressed in all analyzed tissues of kale and that its transcription in leaves was down-regulated by salt, abscisic acid, and H 2 O 2 at a high concentration. Overexpression of BoRACK1 in kale led to a reduction in symptoms caused by Peronospora brassicae Gaumann on kale leaves. The expression levels of the pathogenesis-related protein genes, PR-1 and PRB-1, increased 2.5-4-fold in transgenic kale, and reactive oxygen species production was more active than in the wild-type. They also exhibited increased tolerance to salt stress in seed germination. H 2 O 2 may also be involved in the regulation of BoRACK1 during seed germination under salt stress. Quantitative real-time PCR analyses showed that the transcript levels of BoRbohs genes were significantly higher in overexpression of BoRACK1 transgenic lines. Yeast two-hybrid assays showed that BoRACK1 could interact with WNK8, eIF6, RAR1, and SGT1. This study and previous work lead us to believe that BoRACK1 may form a complex with regulators of plant salt and disease resistance to coordinate kale reactions to pathogens. Copyright © 2017 Elsevier GmbH. All rights reserved.

Inactivation of normal and mutant Neurospora crassa conidia by visible light and near-UV: role of 1O2, carotenoid composition and sensitizer location

International Nuclear Information System (INIS)

Thomas, S.A.; Sargent, M.L.; Tuveson, R.W.

1981-01-01

Inactivation of Neurospora crassa conidia from wild-type and mutant strains by visible and near-ultraviolet light was investigated in the presence and absence of photosensitizing dyes. Inactivation by near-UV was virtually unchanged by the presence of deuterium oxide or azide suggesting that, contrary to the situation with visible light and photosensitizing dyes, 1 O 2 is not involved in any substantial way in the formation of lethal lesions. Carotenoid deficient strains were similar to wild-type strains in sensitivity to near-UV inactivation which is consistent with 1 O 2 not being involved. Photodynamic inactivation of conidia by visible light occurred in the presence of methylene blue (MB), toluidine blue O (TB), or acridine orange (AO). Carotenoid deficient strains were more sensitive to such inactivation only when MB and TB were used. This suggests that MB and TB mediated damage involves the cell membrane where carotenoids are available for quenching, whereas AO mediated damage occurs in the nucleus sequestered from the protective influence of carotenoids. A newly isolated, lemon-yellow mutant exhibited sensitivities to photodynamic inactivation similar to other pure-white mutants. The sensitivity of this pigmented mutant is apparently related to insufficient unsaturation of the two coloured carotenoids produced by the mutant. (author)

Temperature and light intensity interaction on Cercospora coffeicola sporulation and conidia germination

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Marília Goulart da Silva

2016-04-01

Full Text Available ABSTRACT Difficulty in obtaining abundant sporulation in culture of many species of Cercospora may be the limiting factor for studies of biology, systematics, and inoculation of the genus. Therefore, it is necessary to understand the nutritional and environmental requirements that influence mycelial growth, sporulation and germination. As it is difficult to obtain conidia of Cercospora coffeicola in vitro, different temperatures (17, 22, 27, and 32 °C and light intensities (80, 160, 240, and 320 μmol m-2 s-1 were evaluated to optimize pathogen sporulation and assess favorable conditions for spore germination, aiming for a strategy of disease control. The dark treatment (0 μmol m-2 s-1 was added for sporulation. A significant interaction was found between temperature and light intensity for both variables. The highest sporulation rate of C. coffeicola occurred at a light intensity of 240 μmol m-2 s-1 and air temperature of 22 °C, reaching 5.9x106 con mL-1. Germination was higher at temperature 17 °C and light intensity of 320 μmol m-2 s-1, reaching 52%. Interaction between light intensity and temperature proved to influence the processes of sporulation and germination of C. coffeicola.

Yield and cold storage of Trichoderma conidia is influenced by substrate pH and storage temperature.

Science.gov (United States)

Steyaert, Johanna M; Chomic, Anastasia; Nieto-Jacobo, Maria; Mendoza-Mendoza, Artemio; Hay, Amanda J; Braithwaite, Mark; Stewart, Alison

2017-05-01

In this study we examined the influence of the ambient pH during morphogenesis on conidial yield of Trichoderma sp. "atroviride B" LU132 and T. hamatum LU593 and storage at low temperatures. The ambient pH of the growth media had a dramatic influence on the level of Trichoderma conidiation and this was dependent on the strain and growth media. On malt-extract agar, LU593 yield decreased with increasing pH (3-6), whereas yield increased with increasing pH for LU132. During solid substrate production the reverse was true for LU132 whereby yield decreased with increasing pH. The germination potential of the conidia decreased significantly over time in cold storage and the rate of decline was a factor of the strain, pH during morphogenesis, growth media, and storage temperature. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Efecto del fotoperiodo y la intensidad lumínica sobre la esporulación de Peronospora sparsa Berkeley, bajo condiciones controladas

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Soto S. Johana Carolina

2009-08-01

Full Text Available

El mildeo velloso de la rosa, producido por Peronospora sparsa, es una de las enfermedades más limitantes y costosas en los cultivos comerciales de rosa en Colombia. La estrecha relación de la enfermedad con las condiciones climáticas ha llevado a realizar estudios del parásito en condiciones microclimáticas controladas. Esta investigación buscó ahondar en el conocimiento de la biología del mildeo velloso de la rosa, evaluando para ello el efecto del fotoperiodo y la intensidad lumínica sobre la esporulación del parásito cultivado en folíolos de rosa en cámara húmeda bajo condiciones ambientales controladas. Se evaluaron los siguientes fotoperiodos: 24 horas bajo una intensidad lumínica de 1.000 lux; 8, 12 y 24 horas luz a 790 lux; 8, 16 y 24 horas a 560 lux; 12 horas a 120 lux, y finalmente 0 horas luz (oscuridad total. La esporulación se vio favorecida por fotoperiodos de 8 a 16 horas a 560 lux, y presentó un alto desempeño con periodos de 12 horas a 790 lux. La exposición continua a la luz demostró tener un efecto inhibidor sobre la esporulación de P. sparsa. Así mismo, determinó la producción de esporangios con características atípicas, como bajo contenido citoplasmático y menor tamaño.

A spindle pole antigen gene MoSPA2 is important for polar cell growth of vegetative hyphae and conidia, but is dispensable for pathogenicity in Magnaporthe oryzae.

Science.gov (United States)

Li, Chao; Yang, Jun; Zhou, Wei; Chen, Xiao-Lin; Huang, Jin-Guang; Cheng, Zhi-Hua; Zhao, Wen-Sheng; Zhang, Yan; Peng, You-Liang

2014-11-01

Spa2 is an important component of the multiprotein complex polarisome, which is involved in the establishment, maintenance, termination of polarized cell growth and is important for defining tip growth of filamentous fungi. In this study, we isolated an insertional mutant of the rice blast fungus Magnaporthe oryzae that formed smaller colony and conidia compared with the wild type. In the mutant, a spindle pole antigen gene MoSPA2 was disrupted by the integration of an exogenous plasmid. Targeted gene deletion and complementation assays demonstrated the gene disruption was responsible for the defects of the insertional mutant. Interestingly, the MoSpa2-GFP fusion protein was found to accumulate as a spot at hyphal tips, septa of hyphae and conidial tip cells where germ tubes are usually produced, but not in appressoria, infection hyphae or at the septa of conidia. Furthermore, the deletion mutants of MoSPA2 exhibited slower hyphal tip growth, more hyphal branches, and smaller size of conidial tip cells. However, MoSPA2 is not required for plant infection. These results indicate that MoSPA2 is required for vegetative hyphal growth and maintaining conidium morphology and that spotted accumulation of MoSpa2 is important for its functions during cell polar growth.

Distinct innate immune phagocyte responses to Aspergillus fumigatus conidia and hyphae in zebrafish larvae.

Science.gov (United States)

Knox, Benjamin P; Deng, Qing; Rood, Mary; Eickhoff, Jens C; Keller, Nancy P; Huttenlocher, Anna

2014-10-01

Aspergillus fumigatus is the most common filamentous fungal pathogen of immunocompromised hosts, resulting in invasive aspergillosis (IA) and high mortality rates. Innate immunity is known to be the predominant host defense against A. fumigatus; however, innate phagocyte responses to A. fumigatus in an intact host and their contributions to host survival remain unclear. Here, we describe a larval zebrafish A. fumigatus infection model amenable to real-time imaging of host-fungal interactions in live animals. Following infection with A. fumigatus, innate phagocyte populations exhibit clear preferences for different fungal morphologies: macrophages rapidly phagocytose conidia and form aggregates around hyphae, while the neutrophil response is dependent upon the presence of hyphae. Depletion of macrophages rendered host larvae susceptible to invasive disease. Moreover, a zebrafish model of human leukocyte adhesion deficiency with impaired neutrophil function also resulted in invasive disease and impaired host survival. In contrast, macrophage-deficient but not neutrophil-deficient larvae exhibited attenuated disease following challenge with a less virulent (ΔlaeA) strain of A. fumigatus, which has defects in secondary metabolite production. Taking these results together, we have established a new vertebrate model for studying innate immune responses to A. fumigatus that reveals distinct roles for neutrophils and macrophages in mediating host defense against IA. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Efecto del Fosfito de Potasio en Combinación con el Fungicida Metalaxyl + Mancozeb en el Control de Mildeo Velloso (Peronospora destructor Berk en Cebolla de Bulbo (Allium cepa L. Effect of the Potasium Phosphite in Combination with the Fungicide Metalaxyl plus Mancozeb on the Control of Downy Mildew (Peronospora destructor Berk in Onion Bulb (Allium cepa L.

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Jorge Velandia Monsalve

2012-06-01

Full Text Available Resumen. El hongo Peronospora destructor causa considerables pérdidas en la producción de cebolla de bulbo en el mundo. Fosfito de potasio en combinación con el fungicida Metalaxyl + Mancozeb, fue evaluado por su eficiencia en el control de este patógeno en el híbrido de cebolla de bulbo Yellow Granex. Los tratamientos consistieron en una aplicación de fosfito (5 mL L-1 alternada con una de fungicida (2 g L-1; dos y tres aplicaciones consecutivas de fosfito alternadas con una de fungicida; una aplicación de fungicida alternada con una de fosfito; dos y tres aplicaciones consecutivas de fungicida alternadas con una de fosfito; aplicaciones de sólo fosfito y sólo fungicida y un testigo. Se utilizó el diseño experimental de bloques completos al azar con tres repeticiones. Se evaluó la incidencia y severidad de la enfermedad, el número, tamaño y peso de bulbos de primera, segunda, tercera y peso total de bulbos. Los resultados mostraron que las aplicaciones de fosfito de potasio durante dos semanas consecutivas alternadas en la tercera semana con una de fungicida y las aplicaciones semanales de sólo fosfito, tuvieron efectos altamente significativos en el control de la incidencia y severidad de P. destructor y diferencias significativas en el peso de los bulbos de primera (28,1 y 27,9 t ha-1 y peso total de bulbos (55,5 y 52,1 t ha-1, representando en relación al testigo, un incremento de 200 y 196% en el peso de los bulbos de primera y de 55 y 45,7% en el peso total de bulbos, respectivamente. Se concluye que con el fosfito de potasio se reduce la aplicación de fungicidas y ello es una alternativa viable para el manejo ecológico de P. destructor en la producción de cebolla de bulbo.Abstract. Peronospora destructor fungi causes considerable loses in the production of onion bulb in the world. Potassium phosphite in combination with the fungicide Metalaxyl + Mancozeb was evaluated for its efficiency in the control of this pathogen

OBSERVATIONS ON THE SUSCEPTIBILITY OF SOME WILD SPECIES OF THE GENUS NICOTINA AND OF SOME VARIETIES OF NICOTIANA TABACUM L. AND N. RUSTICA L. TO BLUE MOULD (PERONOSPORA TABACINA ADAM) -- PULAWY 1962

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Nicotiana , no manifestations of the mould were observed on the species of N. debneyi and N. exigua. Very weak symptoms appeared in N. paniculata and N... plumbaginifolia . In the group of cigarette-tobacco varieties only the Hicks Resistant and Hicks fixed A2 (varieties of Australian origin, obtained by

Flutuação populacional de esporângios de Peronospora destructor no ar e sua relação com a severidade do míldio da cebola

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Leandro Luiz Marcuzzo

2015-03-01

Full Text Available Em experimento de campo nos anos de 2011, 2012 e 2013 quantificou-se a flutuação de esporângios de Peronospora destructor e sua relação com a severidade de míldio em cebola. Mudas de cebola do cultivar Bola Precoce foram transplantadas em quatro repetições de 60 plantas. Os esporângios foram coletados através de um coletor de esporos tipo "cata-vento", contendo uma lâmina de microscópio untada com vaselina, a qual era quantificada semanalmente com auxílio de microscópio. A severidade da doença foi analisada através da porcentagem de área foliar afetada pela doença. O número de esporângios e a severidade do míldio da cebola foram submetidos ao cálculo do coeficiente de correlação linear de Pearson (r. Nos anos de 2011 e 2012 a coleta de esporângios ocorreu posteriormente à constatação da doença, provavelmente devido haver baixo inóculo no ar. Já no ano de 2013 a coleta de esporângios ocorreu anteriormente à ocorrência da doença, porém de forma esporádica e em pequena quantidade, presupondo que esses estavam no ar e foram capturados pelo coletor. A correlação entre as variáveis flutuações de esporângios e a severidade da doença foi significativa, r=0,90, r=0,94 e r=0,80; para os anos de 2011, 2012 e 2013 respectivamente. A severidade da doença em cebola é influenciada pela presença dos esporângios coletadas do ar.

Survival of Penicillium spp. conidia during deep-frying and baking steps of frozen chicken nuggets processing.

Science.gov (United States)

Wigmann, Évelin Francine; Moreira, Rafael Chelala; Alvarenga, Verônica Ortiz; Sant'Ana, Anderson S; Copetti, Marina Venturini

2016-05-01

This study aimed at determining whether Penicillium spp. strains could survive through the heat treatment applied during the processing of frozen chicken nuggets. Firstly, it was found that the conidia of Penicillium were not able to survive the heat shock in phosphate buffer at pH 7.2 in thermal death tubes (TDT) at 80 °C/30 min. Subsequently, each Penicillium strain was inoculated in frozen chicken nuggets, which were subjected to the following treatments: i) only deep frying (frying oil at 195-200 °C), ii) only baking (120-130 °C until the internal temperature reached 70 °C) and iii) deep frying followed by baking (frying oil temperature of 195-200 °C and baking temperature of 120-130 °C, until the internal temperature reached 70 °C). The results indicated that Penicillium polonicum NGT 23/12, Penicillium commune NGT 16/12, Penicillium solitum NGT 30/12 and Penicillium crustosum NGT 51/12 were able to survive after the combined treatment (deep frying followed by baking) when inoculated in chicken nuggets. P. polonicum NGT 23/12 was the most resistant strain to the combined treatment (deep frying and baking), as its population was reduced by 3 log cycles CFU/g, when the internal temperature reached 78 °C after 10 min and 30 s of baking. The present data show that if Penicillium spp. is present in high numbers in raw materials, such as breading flours, it will survive the thermal processing applied during chicken nuggets production. Copyright © 2015 Elsevier Ltd. All rights reserved.

Journal of Genetics | Indian Academy of Sciences

Indian Academy of Sciences (India)

mildew-susceptible genotypes of opium poppy · Mukesh K. Dubey Ajit K. Shasany Om P. Dhawan Ashutosh K. Shukla Suman P. S. Khanuja · More Details Abstract Fulltext PDF. Downy mildew (DM) caused by Peronospora arborescens, is a ...

Microcycle Conidiation in Cercospora zeae-maydis.

Science.gov (United States)

Lapaire, Carrie L; Dunkle, Larry D

2003-02-01

ABSTRACT Conidia of Cercospora zeae-maydis are the primary inoculum causing gray leaf spot of maize. On nutrient-deficient substrates, but not on water on the leaf surface, conidia germinate and develop secondary conidia on conidiophores produced from germ tubes or conidial cells. A population of conidia increases its numbers more than twofold by 2 days on the surface of a water droplet and by fourfold on trichomes. This microcycle conidiation is suppressed by hydrogen peroxide and ammonium compounds but not by nitrate compounds, amino acids, or simple sugars. Microcycle conidiation is sensitive to alpha-amanitin and cycloheximide, suggesting that new RNA and proteins must be synthesized. Upon transfer from a humid to a dry atmosphere, secondary conidia and conidiophores dehydrate and collapse. Mature, dehydrated, secondary conidia are liberated by wind speeds approximately one-third those required to liberate hydrated conidia. The dispersed secondary conidia can rehydrate and germinate normally. Because this microcycle conidiation occurs at the expense of endogenous reserves, the ability to produce secondary conidia is lost after four successive cycles without a period of growth on nutrient media. This alternative method of maintaining inoculum potential during periods of fluctuating relative humidity may have epidemiological consequences when primary conidia fail to infect.

REARING TOMATO WHITEFLY AND FIELD EVALUATION OF ...

African Journals Online (AJOL)

ACSS

2015-05-27

May 27, 2015 ... important agricultural and ornamental crops in all agro-ecological regions in the world. This study evaluated the ... conidia: (a) modified conidia (conidia with improved ecological competence), and (b) unmodified conidia of the three isolates were subjected ..... control of cotton spider mites. There was a wide ...

In vitro attachment of phylloplane yeasts to Botrytis cinerea, Rhizoctonia solani, and Sclerotinia homoeocarpa.

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Allen, Tom W; Burpee, Leon L; Buck, James W

2004-12-01

The ability of yeasts to attach to hyphae or conidia of phytopathogenic fungi has been speculated to contribute to biocontrol activity on plant surfaces. Attachment of phylloplane yeasts to Botrytis cinerea, Rhizoctonia solani, and Sclerotinia homoeocarpa was determined using in vitro attachment assays. Yeasts were incubated for 2 d on potato dextrose agar (PDA) prior to experimentation. A total of 292 yeasts cultured on PDA were screened for their ability to attach to conidia of B. cinerea; 260 isolates (89.1%) attached to conidia forming large aggregates of cells, and 22 isolates (7.5%) weakly attached to conidia with 1 or 2 yeast cells attached to a few conidia. Ten yeasts (3.4%), including 8 isolates of Cryptococcus laurentii, 1 isolate of Cryptococcus flavescens, and an unidentified species of Cryptococcus, failed to attach to conidia. All non-attaching yeasts produced copious extracellular polysaccharide (EPS) on PDA. Seventeen yeast isolates did not attach to hyphal fragments of B. cinerea, R. solani, and S. homoeocarpa after a 1 h incubation, but attachment was observed after 24 h. Culture medium, but not culture age, significantly affected the attachment of yeast cells to conidia of B. cinerea. The 10 yeast isolates that did not attach to conidia when grown on agar did attach to conidia (20%-57% of conidia with attached yeast cells) when cultured in liquid medium. Attachment of the biocontrol yeast Rhodotorula glutinis PM4 to conidia of B. cinerea was significantly greater at 1 x 10(7) yeast cells x mL(-1) than at lower concentrations of yeast cells. The ability of yeast cells to attach to fungal conidia or hyphae appears to be a common phenotype among phylloplane yeasts.

Downy mildews on ornamental plants and their control.

Science.gov (United States)

Skrzypczak, C

2006-01-01

Among downy mildews occurring on ornamentals in Poland the most dangerous are downy mildew of rose and downy mildew of German statice (Tartarian statice). Downy mildew of rose caused by Peronospora sparsa Berk. is a serious threat to commercial cultivation of cut roses, especially grown under plastic tunnels. Peronospora statices Lobik casual agent of German statice downy mildew can causes the total losses in the second year of statice cultivation more than 70%. Both pathogens are very difficult to control. Effectiveness of azoxystrobine, cymoxanil + famoxate, mancozeb, phosethyl aluminium, phosethyl aluminium + fenamidone, propamocarb in the control of P. sparsa and P. statices was presented. In the control of statice downy mildew none of tested compounds was able to control satisfactory the pathogen. Relatively the best results were obtained with mixture of fenamidone (88 microg/cm3) and phosethyl AL (1334 microg/cm3). In the control of rose downy mildew, the best results were obtained with phosethyl Al at concentration 1600 microg/cm3.

Induced variation in potato dry rot fusarium roseum by ultraviolet irradiation

International Nuclear Information System (INIS)

Lee, C.U.

1979-01-01

Variations were readily induced in Fusarium roseum isolated from decayed potato tubers by subjecting its conidia to ultraviolet irradiation. The dosage that resulted in complete absence of developing colonies in five minutes of irradiation did not cause the mortality of conidia for the initial one minute. Eighty eight precent of conidia were killed in three minutes. Approximately 90% of the conidia showed resistance to lethality for the initial two minutes whereas 10% began to show resistance after three minutes of the irradiation. The maximum 42% of color variation was obtained among the surviving conidia. Varients in both decreased and increased pathogenicity were induced in 8% among the color variants. Variations occurred in such other characteristics as growth rate, rigidity of mycelium, and conidia formation. (author)

Downy mildew: a serious disease threat to rose health worldwide

Science.gov (United States)

Peronospora sparsa is a downy mildew-causing oomycete that can infect roses, blackberries and other members of the rose family. During the last 20 years, this disease has become a serious problem for rose growers in the U.S. and worldwide. While much is known about the disease and its treatment, inc...

Journal of Genetics | Indian Academy of Sciences

Indian Academy of Sciences (India)

Downy mildew (DM) caused by Peronospora arborescens, is a serious disease in opium poppy (Papaver somniferum), which has a world-wide spread. The establishment of DM-resistant cultivars appears to be a sustainable way to control the disease. In this paper, we present the results of a study aimed at the identification ...

Conidiation of Penicillium camemberti in submerged liquid cultures is dependent on the nitrogen source.

Science.gov (United States)

Boualem, Khadidja; Labrie, Steve; Gervais, Patrick; Waché, Yves; Cavin, Jean-François

2016-02-01

To study the ability of a commercial Penicillium camemberti strain, used for Camembert type cheese ripening, to produce conidia during growth in liquid culture (LC), in media containing different sources of nitrogen as, industrially, conidia are produced by growth at the surface of a solid state culture because conidiation in stirred submerged aerobic LC is not known. In complex media containing peptic digest of meat, hyphae ends did not differentiate into phialides and conidia. Contrarily, in a synthetic media containing KNO3 as sole nitrogen source, hyphae ends differentiated into phialides producing 0.5 × 10(7) conidia/ml. Conidia produced in LC were 25 % less hydrophobic than conidia produced in solid culture, and this correlates with a seven-times-lower expression of the gene rodA encoding hydrophobin RodA in the mycelium grown in LC. Conidiation of P. camembertii is stimulated in iquid medium containing KNO3 as sole source of nitrogen and therefore opens up opportunities for using liquid medium in commercial productions.

Evaluation of Metarhizium anisopliae var. anisopliae Qu-M845 Isolate to Control Varroa destructor (Acari: Varroidae in Laboratory and Field Trials Evaluación del Aislamiento Qu-M845 de Metarhizium anisopliae var. anisopliae para el Control de Varroa destructor (Acari: Varroidae en Ensayos de Laboratorio y Terreno

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Marta Rodríguez

2009-12-01

Full Text Available The effectiveness of the Metarhizium anisopliae (Metschinkoff Qu-M845 isolate was evaluated in laboratory and field trials. It was previously selected for thermal resistance (at 30 and 35 ºC and pathogenicity on Varroa destructor (Anderson and Trueman. In the laboratory, the first evaluations were carried out by spraying and increasing the concentration from zero to 10(8 conidia mL-1 on varroa adults. Lethal concentrations required for 50 and 90% mite mortality (LC50 and LC90 were 3.8 x 10(5 and 8 x 10(7 conidia mL-1, respectively (χ2 = 2.03. In the autumn field trials, three application methods (doses of 5 x 10(10 conidia per hive were evaluated. The treatments were: a conidia stamped on filter paper, located on every second frame inside the hive; b dry conidia sprinkled on and between frames; and c dry conidia in a dispenser path at the entrance of the hive. Furthermore, untreated hives were included as controls. After 21 days of treatment, the dry conidia sprinkled on and between frames showed 67% less bees infested by the mite than the control (p La efectividad del aislamiento Qu-M845 de Metarhizium anisopliae (Metschinkoff, seleccionado previamente por su resistencia a temperaturas de 30 y 35 ºC, y patogenicidad sobre Varroa destructor Anderson y Trueman fue evaluada en laboratorio y en ensayos de terreno. Las primeras pruebas consistieron en pulverizar concentraciones crecientes de 0 a 10(8 conidias mL-1 sobre varroas adultas. La concentración letal para matar el 50 y 90% de la población (CL50 y CL90 fueron de 3,8 x 10(5 y 8 x 10(7 conidias mL-1, respectivamente (χ² = 2,03. En otoño se evaluaron en terreno tres métodos de aplicación de una dosis de 5 x 10(10 conidias por colmena. Los tratamientos fueron: a conidias estampadas en papel filtro ubicado cada dos panales móviles al interior de la colmena; b conidias espolvoreadas sobre y entre los panales; y c dispensador de conidias ubicado en la piquera de las colmenas. Además se

Scorpion neurotoxin AaIT-expressing Beauveria bassiana enhances the virulence against Aedes albopictus mosquitoes.

Science.gov (United States)

Deng, Sheng-Qun; Cai, Qun-Di; Deng, Ming-Zhi; Huang, Qiang; Peng, Hong-Juan

2017-12-01

To improve the insecticidal efficacy of this entomopathogen Beauveria bassiana, the fungus was genetically modified to express an insect-specific scorpion neurotoxin AaIT. The virulence of the recombinant B. bassiana strain (Bb-AaIT) against Aedes albopictus adults (which occurs via penetration through the cuticle during spore germination or by conidia ingestion), and the larvae (by conidia ingestion) was measured with bioassays. The median lethal concentration (LC 50 ) of Bb-AaIT against A. albopictus larvae was 313.3-fold lower on day 4 and 11.3-fold lower on day 10 than that of the wild type (WT). Through conidia feeding or body contact, Bb-AaIT killed 50% of adult female mosquitoes at 3.9- or 1.9-fold reduced concentrations on day 4 and at 2.1- or 2.4-fold reduced concentrations on day 10. Compared with the results for the WT, the median lethal time (LT 50 ) of Bb-AaIT was reduced by 28.6% at 1 × 10 7 conidia ml -1 and 34.3% at 1 × 10 6 conidia ml -1 in the larvae bioassay by conidia ingestion, while it decreased 32.3% at 1 × 10 7 conidia ml -1 by conidia ingestion and 24.2% at 1 × 10 8 conidia ml -1 by penetrating through the cuticle in the adult bioassay. All the differences were significant. Our findings indicated that Bb-AaIT had higher virulence and faster action than the WT in killing the larval and adult mosquitoes, and therefore, it is valuable for development as a commercial mosquito pesticide.

Production, Survival, and Evaluation of Solid-Substrate Inocula of Penicillium oxalicum, a Biocontrol Agent Against Fusarium Wilt of Tomato.

Science.gov (United States)

Larena, I; Melgarejo, P; De Cal, A

2002-08-01

ABSTRACT Production of conidia of Penicillium oxalicum (ATCC number pending), a biocontrol agent of Fusarium oxysporum f. sp. lycopersici, was tested in liquid and solid fermentation. P. oxalicum produced 250-fold more conidia in solid than in liquid fermentation at 30 days after inoculation of substrate. Solid fermentation was carried out in plastic bags (600 cm(3)) especially designed for solid fermentation (VALMIC) containing 50 g of peat/vermiculite (PV) (1:1, wt/wt) with 40% moisture, sealed, sterilized, and then inoculated with 1 ml of a conidial suspension of P. oxalicum (10(5) conidia g(-1) dry substrate), sealed again, and incubated in darkness at 20 to 25 degrees C for 30 days. Addition of amendments to PV in a proportion of 0.5 (wt/wt) significantly increased conidial production of P. oxalicum. The best production was obtained on PV plus meal of cereal grains (barley) or leguminous seeds (lentil) (100-fold higher). Conidial production obtained after 5 days of inoculation was similar to that obtained at 30 days. However, viability of conidia produced in PV plus lentil meal was 35% higher than that of conidia produced in PV plus barley meal. Changes in proportions (1:1:0.5, wt/wt/wt; 1:1:1, wt/wt/wt; 1:0.5:0.5, wt/wt/wt; 1:1:0.5, vol/vol/vol) of components of the substrate (peat/vermiculite/lentil meal) did not enhance production or viability of conidia. Optimal initial moisture in the substrate was 30 to 40%. At lower moistures, significant reductions of production of conidia were observed, particularly at 10%. There was a general decline in the number of conidia in bags with time of storage at -80, -20, 4, and 25 degrees C, or at room temperature (range from 30 to 15 degrees C), with the highest decline occurring from 60 to 180 days. Conidial viability also was reduced with time, except for conidia stored at -20 degrees C. Fresh conidia produced in solid fermentation system or those conidia stored at -20 degrees C for 180 days reduced Fusarium wilt of

Leveillula taurica ON Ficus carica ABSTRACT RÉSUMÉ

African Journals Online (AJOL)

Prof. Adipala Ekwamu

Les conidiophores avaient des pattes à cellules cylindriques, munis d'une seule de conidium ou occasionnellement avec des chaînes courtes de 2-3 conidia. Les champignons avaient produit des conidia primaries à bout en forme de lance, ainsi que des conidia secondaires ellipsoides voire cylindriques. Les cléistothèces ...

Conidial germination in Scedosporium apiospermum, S. aurantiacum, S. minutisporum and Lomentospora prolificans: influence of growth conditions and antifungal susceptibility profiles

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Thaís Pereira de Mello

2016-01-01

Full Text Available In the present study, we have investigated some growth conditions capable of inducing the conidial germination in Scedosporium apiospermum, S. aurantiacum, S. minutisporum and Lomentospora prolificans. Germination in Sabouraud medium (pH 7.0, 37ºC, 5% CO2 showed to be a typically time-dependent event, reaching ~75% in S. minutisporum and > 90% in S. apiospermum, S. aurantiacum and L. prolificans after 4 h. Similar germination rate was observed when conidia were incubated under different media and pHs. Contrarily, temperature and CO2 tension modulated the germination. The isotropic conidial growth (swelling and germ tube-like projection were evidenced by microscopy and cytometry. Morphometric parameters augmented in a time-dependent fashion, evidencing changes in size and granularity of fungal cells compared with dormant 0 h conidia. In parallel, a clear increase in the mitochondrial activity was measured during the transformation of conidia-into-germinated conidia. Susceptibility profiles to itraconazole, fluconazole, voriconazole, amphotericin B and caspofungin varied regarding each morphotype and each fungal species. Overall, the minimal inhibitory concentrations for hyphae were higher than conidia and germinated conidia, except for caspofungin. Collectively, our study add new data about the conidia-into-hyphae transformation in Scedosporium and Lomentospora species, which is a relevant biological process of these molds directly connected to their antifungal resistance and pathogenicity mechanisms.

Pengendalian Diaphorina citri (Vektor Penyakit CVPD dengan Metarrhizium anisopliae

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Kardi Raharjo

2000-07-01

and after insect infestation. The first phase of the research phase has been conducted in Temanggung, Completely Randomize Design (CRD factorial with three time replication. Factor I: sterile water without fructose, concentration 10^6 conidia/ml without fructose, concentration 10^8 conidia/ml without fructose, concentration 10^10 conidia/ml: without fructose, sterile water + fructose 5 mg/ml, concentration 10^6 conidia/ml without fructose 5 mg/ml, concentration 10^8 conidia/ml + fructose (fungi application before insect infestation and W1 (fungi application after insect infestation. Research phase II was carried out with the best treatment combination compare with control treatment in Temanggung and Bantul. The results of experiment showed that the initial die of D. citri caused by M. anisopliae infection are on 4-6 days after application. The application of M. anisopliae at concentration 10^10 conidia/ml without fructose, applied after insect infestation was most effective. The application after insect infestation was more effective compare with application before insect infestation especially on 4 days after application, but on 35th days after application there was no significant difference. Fructose has no effect to mortality of D. citri.

Aspergillus fumigatus spore proteomics and genetics reveal that VeA represses DefA-mediated DNA damage response.

Science.gov (United States)

Shin, Kwang-Soo; Park, Hee-Soo; Kim, Young; Heo, In-Beom; Kim, Young Hwan; Yu, Jae-Hyuk

2016-10-04

Aspergillus fumigatus reproduces and infects host by forming a high number of small asexual spores (conidia). The velvet proteins are global transcriptional regulators governing the complex process of conidiogenesis in this fungus. Here, to further understand the velvet-mediated regulation, we carried out comparative proteomic analyses of conidia of wild type (WT) and three velvet mutants (ΔveA, ΔvelB and ΔvosA). Cluster analysis of 184 protein spots showing at least 1.5-fold differential accumulation between WT and mutants reveal the clustering of WT- ΔveA and ΔvelB-ΔvosA. Among 43 proteins identified by Nano-LC-ESI-MS/MS, 23 including several heat shock proteins showed more than two-fold reduction in both the ∆velB and ∆vosA conidia. On the contrary, three proteins exhibited more than five-fold increase in ∆veA only, including the putative RNA polymerase II degradation factor DefA. The deletion of defA resulted in a reduced number of conidia and restricted colony growth. In addition, the defA deletion mutant conidia showed hypersensitivity against the DNA damaging agents NQO and MMS, while the ΔveA mutant conidia were more resistant against to NQO. Taken together, we propose that VeA controls protein level of DefA in conidia, which are dormant and equipped with multiple layers of protection against environmental cues. Copyright © 2016 Elsevier B.V. All rights reserved.

Modifying action of DNA synthesis precursors on Aspergillus nidulans conidium irradiated by ultraviolet and X-rays

International Nuclear Information System (INIS)

Muronets, E.M.; Kameneva, S.V.

1975-01-01

Modification of inactivation action of radiation on conidia Aspergillus nidulans, UVS + and UVS strains, by desoxynucleosides, purine and pyrimidine bases is shown. The modification manifested in increased conidia survival is revealed when the precursor of DNA synthesis is added to the suspension before exposure to ultraviolet or X-rays. In the case of postradiation application of the substance no modification is observed. The modifying effect of different precursors becomes equally apparent with equimolar solutions and increases at higher concentration of the latter. An increase in thymidine endogenic pool in the exposed conidia does not affect their survival. When conidia are exposed to ultraviolet rays through a thymidine filter the survival rate increases to the same extent as in the case when they are exposed to irradiation in thymidine solution. The authors suggest that modification of the inactivating radiation action by DNA precursors at exposure of conidia Aspergillus nidulans is caused by the radioprotective effect of precursors not related to reparation [ru

Effects of temperature, light and incubation period on production, germination and bioactivity of Trichoderma atroviride.

Science.gov (United States)

Daryaei, A; Jones, E E; Ghazalibiglar, H; Glare, T R; Falloon, R E

2016-04-01

The goal was to determine the effect of temperature, light and incubation period on production, germination and bioactivity of Trichoderma atroviride LU132 against Rhizoctonia solani. The incubation temperatures of 20, 25 or 30°C were assessed on the production of T. atroviride conidia under constant light over a 25 and 50 days periods. The resulting conidia were also studied for germination and bioactivity. Conidium production was maximum at 25°C after 20 days. The second peak of conidium production occurred at 45-50 days. Incubation at 25°C after 15 days showed optimum production of T. atroviride LU132. Conidia produced at 30°C gave the greatest germination and bioactivity in comparison with incubation at 20 or 25°C. This study indicates that the temperature at which conidia of T. atroviride are produced affects germination and bioactivity. Formulations based on production of the high conidia yield may not result in optimal bioactivity and there is a trade-off between quantity and quality of T. atroviride LU132 conidia. Conidium production was shown to be a continuous process, and increased under a dark/light regime. This is the first report of bimodal conidium production in a Trichoderma biological control agent (BCA), which is likely to be on 20 days cycle, and is dependent on colony age rather than abiotic factors. Conidia produced after 15 days are likely to be the most suitable for use in commercial production of this strain as a BCA. Most studies on Trichoderma-based BCA have only shown the effect of culture conditions on the high conidia yield regardless of conidium quality. This study is the first report on conidium quality affected by principal culture conditions for Trichoderma biological control formulations. © 2016 The Society for Applied Microbiology.

The study on the infection of apple fruits by Botrytis cinerea Pers. after harvest

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Henryk Bryk

2013-12-01

Full Text Available The aim of this studv was to determine the possibility to infection of apples after harvest by conidia and/or mycelium of Botrytis cinerea Pers. Conidia were unable to infect uninjured apple skin regardless of inoculum density and presence of nutrients. The infection of apples by conidia occurred after the surface wax had been removed by washing of apples with chloroform. Injuries of skin appeared to be a favourable entry point for conidia and mycelium of B.cinerea. Only the mycelium of B.cinerea developed on the apple but not that grown on the artificial medium (PDA was able to directly penetration uninjured apple skin. It was observed that sometimes rotted spots develo ped arround the lenticels.

Entomopathogenic Fungus as a Biological Control for an Important Vector of Livestock Disease: The Culicoides Biting Midge

Science.gov (United States)

Ansari, Minshad Ali; Pope, Edward C.; Carpenter, Simon; Scholte, Ernst-Jan; Butt, Tariq M.

2011-01-01

Background The recent outbreak of bluetongue virus in northern Europe has led to an urgent need to identify control measures for the Culicoides (Diptera: Ceratopogonidae) biting midges that transmit it. Following successful use of the entomopathogenic fungus Metarhizium anisopliae against larval stages of biting midge Culicoides nubeculosus Meigen, we investigated the efficacy of this strain and other fungi (Beauveria bassiana, Isaria fumosorosea and Lecanicillium longisporum) as biocontrol agents against adult C. nubeculosus in laboratory and greenhouse studies. Methodology/Findings Exposure of midges to ‘dry’ conidia of all fungal isolates caused significant reductions in survival compared to untreated controls. Metarhizium anisopliae strain V275 was the most virulent, causing a significantly decrease in midge survival compared to all other fungal strains tested. The LT50 value for strain V275 was 1.42 days compared to 2.21–3.22 days for the other isolates. The virulence of this strain was then further evaluated by exposing C. nubeculosus to varying doses (108–1011 conidia m−2) using different substrates (horse manure, damp peat, leaf litter) as a resting site. All exposed adults were found to be infected with the strain V275 four days after exposure. A further study exposed C. nubeculosus adults to ‘dry’ conidia and ‘wet’ conidia (conidia suspended in 0.03% aq. Tween 80) of strain V275 applied to damp peat and leaf litter in cages within a greenhouse. ‘Dry’ conidia were more effective than ‘wet’ conidia, causing 100% mortality after 5 days. Conclusion/Significance This is the first study to demonstrate that entomopathogenic fungi are potential biocontrol agents against adult Culicoides, through the application of ‘dry’ conidia on surfaces (e.g., manure, leaf litter, livestock) where the midges tend to rest. Subsequent conidial transmission between males and females may cause an increased level of fungi-induced mortality in midges

The pmr gene, encoding a Ca2+-ATPase, is required for calcium and manganese homeostasis and normal development of hyphae and conidia in Neurospora crassa.

Science.gov (United States)

Bowman, Barry J; Abreu, Stephen; Johl, Jessica K; Bowman, Emma Jean

2012-11-01

The pmr gene is predicted to encode a Ca(2+)-ATPase in the secretory pathway. We examined two strains of Neurospora crassa that lacked PMR: the Δpmr strain, in which pmr was completely deleted, and pmr(RIP), in which the gene was extensively mutated. Both strains had identical, complex phenotypes. Compared to the wild type, these strains required high concentrations of calcium or manganese for optimal growth and had highly branched, slow-growing hyphae. They conidiated poorly, and the shape and size of the conidia were abnormal. Calcium accumulated in the Δpmr strains to only 20% of the wild-type level. High concentrations of MnCl(2) (1 to 5 mM) in growth medium partially suppressed the morphological defects but did not alter the defect in calcium accumulation. The Δpmr Δnca-2 double mutant (nca-2 encodes a Ca(2+)-ATPase in the plasma membrane) accumulated 8-fold more calcium than the wild type, and the morphology of the hyphae was more similar to that of wild-type hyphae. Previous experiments failed to show a function for nca-1, which encodes a SERCA-type Ca(2+)-ATPase in the endoplasmic reticulum (B. J. Bowman, S. Abreu, E. Margolles-Clark, M. Draskovic, and E. J. Bowman, Eukaryot. Cell 10:654-661, 2011). The pmr(RIP) Δnca-1 double mutant accumulated small amounts of calcium, like the Δpmr strain, but exhibited even more extreme morphological defects. Thus, PMR can apparently replace NCA-1 in the endoplasmic reticulum, but NCA-1 cannot replace PMR. The morphological defects in the Δpmr strain are likely caused, in part, by insufficient concentrations of calcium and manganese in the Golgi compartment; however, PMR is also needed to accumulate normal levels of calcium in the whole cell.

Influence of Cultivars and Seed Thermal Treatment on the Development of Fungal Pathogens in Carrot and Onion Plants

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Martin Koudela

2016-01-01

Full Text Available Carrot and onion are vegetables representing an important segment of fresh market. They suffer from serious fungal diseases that can inflict great damage on crops, i.e. alternaria leaf blight, peronospora downy mildew, and botrytis neck rot. The resistance of selected carrot and onion cultivars important for the production of vegetables in the Czech Republic was tested by exposure to targeted infection by the above fungal pathogens. The exposure of eleven carrot cultivars to spores of Alternaria dauci showed that the most resistant and sensitive cultivars were Katrin, Cortina F1, Afalon F1 and Favorit, Tinga, Berlika F1, respectively. A targeted infection of onion cultivars with Botrytis aclada clustered them into three groups: Amfora F1, Bolero, Tosca, Triumf F1 (strong resistance, Avalon, Grenada (medium resistance, Alice, Karmen, Všetana (low resistance. Similar groups were distinguished also after the infection with Peronospora destructor: Avalon, Bolero, Tosca (strong resistance, Alice, Amfora F1, Grenada, Karmen, Triumf F1 (medium resistance,Všetana (low resistance. Hot water treatment of carrot seeds applied after the inoculation with A. dauci decreased the development of the infection 1.3-2.3-fold, whereas the protective effect observed with onion seeds against the infection by P. destructor and B. aclada was lower.

Soybean diseases in Poland

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J. Marcinkowska

2013-12-01

Full Text Available Field observations on the occurrence of soybean diseases were undertaken in the southern and central regions of Poland in the period 1976-1980. Most prevalent were foliage diseases caused by Peronospora manshurica, Pseudomonas syrinqae pv. glycinea and soybean mosaic virus (SMV. Sclerotinia sclerotiorum and Ascochyta sojaecola were reported as pathogens of local importance. The following pathogenic fungi: Botrytis cinerea, Fusarium culmorum, F. oxysporum and Rhizoctonia solani were also isolated from soybean.

Infection of the malaria mosquito, Anopheles gambiae, with two species of entomopathogenic fungi: effects of concentration, co-formulation, exposure time and persistence

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Lwetoijera Dickson W

2009-12-01

Full Text Available Abstract Background Entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana isolates have been shown to infect and reduce the survival of mosquito vectors. Methods Here four different bioassays were conducted to study the effect of conidia concentration, co-formulation, exposure time and persistence of the isolates M. anisopliae ICIPE-30 and B. bassiana I93-925 on infection and survival rates of female Anopheles gambiae sensu stricto. Test concentrations and exposure times ranged between 1 × 107 - 4 × 1010 conidia m-2 and 15 min - 6 h. In co-formulations, 2 × 1010 conidia m-2 of both fungus isolates were mixed at ratios of 4:1, 2:1, 1:1,1:0, 0:1, 1:2 and 1:4. To determine persistence, mosquitoes were exposed to surfaces treated 1, 14 or 28 d previously, with conidia concentrations of 2 × 109, 2 × 1010 or 4 × 1010. Results Mosquito survival varied with conidia concentration; 2 × 1010 conidia m-2 was the concentration above which no further reductions in survival were detectable for both isolates of fungus. The survival of mosquitoes exposed to single and co-formulated treatments was similar and no synergistic or additive effects were observed. Mosquitoes were infected within 30 min and longer exposure times did not result in a more rapid killing effect. Fifteen min exposure still achieved considerable mortality rates (100% mortality by 14 d of mosquitoes, but at lower speed than with 30 min exposure (100% mortality by 9 d. Conidia remained infective up to 28 d post-application but higher concentrations did not increase persistence. Conclusion Both fungus isolates are effective and persistent at low concentrations and short exposure times.

Effects of ozone on the sporulation, germination, and pathogenicity of Botrytis cinerea

Energy Technology Data Exchange (ETDEWEB)

Krause, C.R.; Weidensaul, T.C.

1978-02-01

Studies were initiated to determine if Botrytis cinerea conidia remain viable when grown in vivo and in vitro in the presence of ambient ozone levels and whether ozonized conidia retain pathogenicity. Experimental materials and methods used are described.

Inactivation of carotenoid-producing and albino strains of Neurospora crassa by visible light, blacklight, and ultraviolet radiation

International Nuclear Information System (INIS)

Blanc, P.L.; Tuveson, R.W.; Sargent, M.L.

1976-01-01

Suspensions of Neurospora crassa conidia were inactivated by blacklight (BL) radiation (300 to 425 nm) in the absence of exogenous photosensitizing compounds. Carotenoid-containing wild-type conidia were less sensitive to BL radiation than albino conidia, showing a dose enhancement factor (DEF) of 1.2 for dose levels resulting in less than 10 percent survival. The same strains were about equally sensitive to shortwave ultraviolet (uv) inactivation. The kinetics of BL inactivation are similar to those of photodynamic inactivation by visible light in the presence of a photosensitizing dye (methylene blue). Only limited inactivation by visible light in the absence of exogenous photosensitizers was observed. BL and UV inactivations are probably caused by different mechanisms since wild-type conidia are only slightly more resistant to BL radiation (DEF = 1.2 at 1.0 percent survival) than are conidia from a uv-sensitive strain (upr-1, uvs-3). The BL-induced lethal lesions are probably not cyclobutyl pyrimidine dimers since BL-inactivated Haemophilus influenzae transforming deoxyribonucleic acid is not photoreactivated by N. crassa wild-type enzyme extracts, whereas uv-inactivated transforming deoxyribonucleic acid is photoreactivable with this treatment

Studies on Aspergillus flavus

International Nuclear Information System (INIS)

Padwal-Desai, S.R.; Ghanekar, A.S.; Sreenivasan, A.

1976-01-01

The response of germinating conidia of Asphergillus flavus (aflatoxin producing) and A.flavus oryzae (non-toxigenic) to heat, radiation and combination treatments has been studied. Germinating conidia of both strains of fungi were more susceptible to heat injury than non-germinating conidia and the susceptibility was enhanced with increased incubation period before treatment. Susceptibility to radiation injury, however, increased only for a short period followed by development of resistance. The germinating stage which was highly resistant to radiation was very sensitive to heat (50 0 /5 min). With combinations of heat and radiation, an irradiation-heat sequence was more effective than the reverse sequence for all germinating stages of A.flavus (toxigenic). However, in the case of the non-toxigenic strain, irrespective of germinating stage, irradiation of conidia before heating proved to be less effective than the reverse sequence. (author)

Studies on Aspergillus flavus

International Nuclear Information System (INIS)

Padwal-Desai, S.R.; Ghanekar, A.S.; Sreenivasan, A.

1976-01-01

In vitro studies were conducted on conidia of Aspergillus flavus Link (aflatoxin producing) and Aspergillus flavus oryzae (non-toxigenic) strains isolated and identified in this laboratory. These strains differed in resistance to heat and gamma radiation, the toxigenic strain being more resistant to both treatments. Results of tests on dose-modifying factors indicated that composition, temperature and pH of suspending media affected radiation resistance. On the other hand, the size of the initial population and the age of the conidia did not influence the radiation resistance of either strain. Studies on thermal inactivation of the conidia suggested that the temperature employed was more important than the time of heat treatment. Conidia of both strains showed a synergistic effect of combined heat and radiation treatments, although a heat-radiation sequence was more effective than a radiation-heat sequence. (author)

Comparison of the pathogenicity, growth, sporulation and morphology of Pezicula alba Guthrie (Gloeosporium album Osterw.

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H. Borecka

2013-12-01

Full Text Available Sixteen isolates of Pezicula alba Guthr. were examined. The intensity of growth on various media, pathogenicity to apple fruits and twigs, colour of cultures and size of conidia were measured. Some isolates are pathogenic to fruits, some others to twigs; one isolate (only no. 19 is pathogenic to both twigs and fruits; many isolates are not - pathogenic at all. Culture growth, sporulation and size of conidia are not correlated with the pathogenicity of the isolate. The mean size of conidia is 21.29µm x 3.48µm.

Aspergillus fumigatus conidial melanin modulates host cytokine response

NARCIS (Netherlands)

L.Y.A. Chai (Louis); M.G. Netea (Mihai); J. Sugui (Janyce); A.G. Vonk (Alieke); W.W.J. van de Sande (Wendy); A. Warris (Adilia); K.J. Kwon-Chung (Kyung); B. Jan Kullberg (Bart)

2010-01-01

textabstractMelanin biopigments have been linked to fungal virulence. Aspergillus fumigatus conidia are melanised and are weakly immunogenic. We show that melanin pigments on the surface of resting Aspergillus fumigatus conidia may serve to mask pathogen-associated molecular patterns (PAMPs)-induced

Aspergillus fumigatus conidial melanin modulates host cytokine response.

NARCIS (Netherlands)

Chai, L.; Netea, M.G.; Sugui, J.; Vonk, A.G.; Sande, W.W. van de; Warris, A.; Kwon-Chung, K.J.; Kullberg, B.J.

2010-01-01

Melanin biopigments have been linked to fungal virulence. Aspergillus fumigatus conidia are melanised and are weakly immunogenic. We show that melanin pigments on the surface of resting Aspergillus fumigatus conidia may serve to mask pathogen-associated molecular patterns (PAMPs)-induced cytokine

Differentiation of the entomopathogenic fungus Metarhizium flavoviride (Hyphomycetes

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Xavier-Santos Solange

1999-01-01

Full Text Available The differentiation of a Brazilian isolate of Metarhizium flavoviride (CG 423, a promising candidate for the biocontrol of grasshoppers, was investigated. Conidia were spread onto solid medium (1% yeast extract, 2.8% agar, 96.2% distilled water, incubated at 28°C and observed during 26 h. Germination initiated as conidia size increased from 5.3 (±0.6 x 3.1 (±0.3 µm (0 h incubation to 8.1 (±0.2 x 6.1 (±0.2 µm (8 h incubation. Germ tubes started to appear after 10 h incubation showing a high degree of multipolarity. Twenty six hours after inoculation, hyphal differentiation and anastomosis among hyphae from adjacent conidia were recorded. Appressoria were formed only from conidia incubated in liquid medium containing minimum concentration of yeast extract (0.06%; w/v. Appressoria were firmly adhered to the bottom of plastic dishes.

Tools for delivering entomopathogenic fungi to malaria mosquitoes: effects of delivery surfaces on fungal efficacy and persistence

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Mnyone Ladslaus L

2010-08-01

Full Text Available Abstract Background Entomopathogenic fungi infection on malaria vectors increases daily mortality rates and thus represents a control measure that could be used in integrated programmes alongside insecticide-treated bed nets (ITNs and indoor residual spraying (IRS. Before entomopathogenic fungi can be integrated into control programmes, an effective delivery system must be developed. Methods The efficacy of Metarhizium anisopliae ICIPE-30 and Beauveria bassiana I93-825 (IMI 391510 (2 × 1010 conidia m-2 applied on mud panels (simulating walls of traditional Tanzanian houses, black cotton cloth and polyester netting was evaluated against adult Anopheles gambiae sensu stricto. Mosquitoes were exposed to the treated surfaces 2, 14 and 28 d after conidia were applied. Survival of mosquitoes was monitored daily. Results All fungal treatments caused a significantly increased mortality in the exposed mosquitoes, descending with time since fungal application. Mosquitoes exposed to M. anisopliae conidia on mud panels had a greater daily risk of dying compared to those exposed to conidia on either netting or cotton cloth (p B. bassiana conidia on mud panels or cotton cloth had similar daily risk of death (p = 0.14, and a higher risk than those exposed to treated polyester netting (p Conclusion Both fungal isolates reduced mosquito survival on immediate exposure and up to 28 d after application. Conidia were more effective when applied on mud panels and cotton cloth compared with polyester netting. Cotton cloth and mud, therefore, represent potential substrates for delivering fungi to mosquitoes in the field.

Temperature effect on rose downy mildew development under environmental controlled conditions

OpenAIRE

Filgueira D., Juan José; Zambrano, Angélica

2014-01-01

The rose downy mildew disease, caused by Peronospora sparsa Berkeley, is one of the most important that affect rose crops in Colombia. To manage this disease, flower growers must deal with high-costs due to the excessive application of fungicides, but without good results. Studies on P. sparsa behavior have shown its narrow relationship with environmental conditions. In this study, the temperature effect was evaluated during the infection and sporulation of P. sparsa in Charlotte leaflets, a ...

Efecto del sustrato y la exposición a la luz en la producción de una cepa de Trichoderma sp.

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Mónica Chávez-Garcia

2008-12-01

Full Text Available Effects of substrate and light exposition in the production of Trichoderma sp. In order to evaluate the effect of temperature and lightconditions on biomass production of a Trichoderma sp. strain, three culture media were tested: rice 53% (w/w, rice 53% (w/w -molasses3% (w/w and rice 53% (w/w-molasses 10% (w/w in distilled water. Incubation conditions were: 25°C, constant light and a photoperiodof 24 h light/24 h darkness during 8 days. The evaluated parameters were population density (conidia/mL, spore germination after 24 hoursand purity percentage. The results showed that solid fermentation using rice - distilled water as substrate at 25°C and constant light, allowedthe highest conidia yield (45x1018 conidia/mL, 96% germination after 24 hours, and 92.1% purity. The liquid fermentation rendered a purityof 76.8% and conidia germination of 91.2% after 24 hours, showing a disadvantageous lower purity percentage compared to solidfermentation.

Does methyl salicylate, a component of herbivore-induced plant odour, promote sporulation of the mite-pathogenic fungus Neozygites tanajoae?

Science.gov (United States)

Hountondji, Fabien C C; Hanna, Rachid; Sabelis, Maurice W

2006-01-01

Blends of volatile chemicals emanating from cassava leaves infested by the cassava green mite were found to promote conidiation of Neozygites tanajoae, an entomopathogenic fungus specific to this mite. Methyl salicylate (MeSA) is one compound frequently present in blends of herbivore-induced plant volatiles (HIPV) as well as that of mite-infested cassava. Here, we investigated the effect of methyl salicylate in its pure form on the production of pre-infective spores (conidia), and the germination of these spores into infective spores (capilliconidia), by a Brazilian isolate and a Beninese isolate of N. tanajoae. Mummified mites previously infected by the fungal isolates were screened under optimal abiotic conditions for sporulation inside tightly closed boxes with or without methyl salicylate diffusing from a capillary tube. Production of conidia was consistently higher (37%) when the Beninese isolate was exposed to MeSA than when not exposed to it (305.5 +/- 52.62 and 223.2 +/- 38.13 conidia per mummy with and without MeSA, respectively). MeSA, however, did not promote conidia production by the Brazilian isolate (387.4 +/- 44.74 and 415.8 +/- 57.95 conidia per mummy with and without MeSA, respectively). Germination of the conidia into capilliconidia was not affected by MeSA for either isolate (0.2%, 252.6 +/- 31.80 vs. 253.0 +/- 36.65 for the Beninese isolate and 4.2%, 268.5 +/- 37.90 vs. 280.2 +/- 29.43 for the Brazilian isolate). The effects of MeSA on the production of conidia were similar to those obtained under exposure to the complete blends of HIPV for the case of the Beninese isolate, but dissimilar (no promoting effect of MeSA) for the case of the Brazilian isolate. This shows that MeSA, being one compound out of many HIPV, can be a factor promoting sporulation of N. tanajoae, but it may not be the only factor as its effect varies with the fungal isolate under study.

Conidiation of Neurospora crassa induced by treatment with natrium fluoride in submerged culture

Energy Technology Data Exchange (ETDEWEB)

Timberlake, W E; Turian, G

1975-01-01

A transient treatment of pregerminated conidia of Neurospora crassa with NaF induced young, submerged cultures to prematurely differentiate conidia. The inductive treatment decreased the rate of respiration (with lower RQ), reduced the relative concentration of nucleoside triphosphates, and inhibited leucine incorporation into protein and adenosine incorporation into RNA.

VeA of Aspergillus niger increases spore dispersing capacity by impacting conidiophore architecture

NARCIS (Netherlands)

Wang, Fengfeng; Dijksterhuis, Jan; Wyatt, Timon; Wösten, Han A B; Bleichrodt, Robert-Jan

Aspergillus species are highly abundant fungi worldwide. Their conidia are among the most dominant fungal spores in the air. Conidia are formed in chains on the vesicle of the asexual reproductive structure called the conidiophore. Here, it is shown that the velvet protein VeA of Aspergillus niger

Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis

DEFF Research Database (Denmark)

Chen, Chun; Xie, Tingna; Ye, Sudan

2016-01-01

candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongationfactor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reac-tion at different developmental stages (conidia, conidia with germ tubes, short hyphae andelongated hyphae), and under different nutritional conditions...

A preliminary report on the contact-independent antagonism of Pseudogymnoascus destructans by Rhodococcus rhodochrous strain DAP96253.

Science.gov (United States)

Cornelison, Christopher T; Keel, M Kevin; Gabriel, Kyle T; Barlament, Courtney K; Tucker, Trudy A; Pierce, George E; Crow, Sidney A

2014-09-26

The recently-identified causative agent of White-Nose Syndrome (WNS), Pseudogymnoascus destructans, has been responsible for the mortality of an estimated 5.5 million North American bats since its emergence in 2006. A primary focus of the National Response Plan, established by multiple state, federal and tribal agencies in 2011, was the identification of biological control options for WNS. In an effort to identify potential biological control options for WNS, multiply induced cells of Rhodococcus rhodochrous strain DAP96253 was screened for anti-P. destructans activity. Conidia and mycelial plugs of P. destructans were exposed to induced R. rhodochrous in a closed air-space at 15°C, 7°C and 4°C and were evaluated for contact-independent inhibition of conidia germination and mycelial extension with positive results. Additionally, in situ application methods for induced R. rhodochrous, such as fixed-cell catalyst and fermentation cell-paste in non-growth conditions, were screened with positive results. R. rhodochrous was assayed for ex vivo activity via exposure to bat tissue explants inoculated with P. destructans conidia. Induced R. rhodochrous completely inhibited growth from conidia at 15°C and had a strong fungistatic effect at 4°C. Induced R. rhodochrous inhibited P. destructans growth from conidia when cultured in a shared air-space with bat tissue explants inoculated with P. destructans conidia. The identification of inducible biological agents with contact-independent anti- P. destructans activity is a major milestone in the development of viable biological control options for in situ application and provides the first example of contact-independent antagonism of this devastating wildlife pathogen.

Quantitative proteomics revealed partial fungistatic mechanism of ammonia against conidial germination of nematode-trapping fungus Arthrobotrys oligospora ATCC24927.

Science.gov (United States)

Liu, Tong; Tian, Dong-Wei; Zou, Li-Juan; Liu, Fang-Yu; Can, Qi-Yan; Yang, Jin-Kui; Xu, Jian-Ping; Huang, Xiao-Wei; Xi, Jia-Qin; Zhu, Ming-Liang; Mo, Ming-He; Zhang, Ke-Qin

2018-05-01

Ammonia is one of the fungistatic factors in soil that can suppress conidial germination, but the molecular mechanism underlying the suppression is unknown. In this study, the proteomes of fungistatic conidia, fresh conidia and germinated conidia of Arthrobotrys oligospora ATCC24927 were determined and quantified. The protein expression profile of fungistatic conidia was significantly different from those in the other two conditions. 281 proteins were down expressed in fungistatic conidia and characterized by GO annotation. Gene transcription analysis and inhibition of puromycin (a protein translation inhibitor) on conidial germination suggested that down expression of 33 protein translation related proteins might well result in repression of protein synthesis and inhibition of conidial germination. In addition, 16 down-expressed proteins were mapped to the Ras/mitogen-activated protein (Ras/MAP) regulatory networks which regulate conidial DNA synthesis. The conidial DNA synthesis was found to be definitely inhibited under by ammonia, and function studies of two Ras/MAP proteins by using knock-out strains provided partial evidence that Ras/MAP pathway regulate the conidial germination. These results suggested that down-expression of Ras/MAP related proteins might result in inhibition of DNA synthesis and finally result in inhibition conidial germination. This study revealed partial fungistatic mechanism of ammonia against conidial germination. Copyright © 2018 Elsevier Ltd. All rights reserved.

Studying of cellular interaction of hairpin-like peptide EcAMP1 from barnyard grass (Echinochloa crusgalli L.) seeds with plant pathogenic fungus Fusarium solani using microscopy techniques.

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Vasilchenko, Alexey S; Yuryev, Mikhail; Ryazantsev, Dmitry Yu; Zavriev, Sergey K; Feofanov, Alexey V; Grishin, Eugene V; Rogozhin, Eugene A

2016-11-01

An interaction of recombinant hairpin-like cationic peptide EcAMP1 with conidia of plant pathogenic fungus Fusarium solani at the cellular level was studied by a combination of microscopic methods. EcAMP1 is from barnyard grass (Echinochloa crusgalli L.), and obtained by heterologous expression in Escherichia coli system. As a result, a direct relationship between hyphal growth inhibition and increasing active peptide concentration, time of incubation and fungal physiological condition has been determined. Dynamics of accumulation and redistribution of the peptide studied on fungal cellular cover and inside the conidia cells has been shown. The dynamics are dependent on time of coupling, as well as, a dissimilarity of EcAMP1 binding with cover of fungal conidia and its stepwise accumulation and diffuse localization in the cytoplasm. Correlation between structural disruption of fungal conidia and the presence of morphological changes has also been found. The correlation was found under the influence of peptide high concentrations at concentrations above 32 μM. The results indicate the presence of a binding of EcAMP1 with the surface of fungal conidia, thus, demonstrating a main specificity for its antifungal action at the cellular level. These results, however, cannot exclude the existence of attendant EcAMP1 action based on its intracellular localization on some specific targets. SCANNING 38:591-598, 2016. © 2016 Wiley Periodicals, Inc. © Wiley Periodicals, Inc.

Colonization by Cladosporium spp. of painted metal surfaces associated with heating and air conditioning systems

Science.gov (United States)

Ahearn, D. G.; Simmons, R. B.; Switzer, K. F.; Ajello, L.; Pierson, D. L.

1991-01-01

Cladosporium cladosporioides and C. hebarum colonized painted metal surfaces of covering panels and register vents of heating, air conditioning and ventilation systems. Hyphae penetrated the paint film and developed characteristic conidiophores and conidia. The colonies were tightly appressed to the metal surface and conidia were not readily detectable via standard air sampling procedures.

Pengendalian hama penggerek ubi jalar Cylas formicarius (Fabricus (Coleoptera: Curculionidae menggunakan cendawan entomopatogen Beauveria bassiana (Balsamo Vuillemen

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Marida Santi Yudha Ika Bayu

2016-12-01

Full Text Available Cylas formicarius (Fabricus (Coleoptera: Curculionidae is one of the major pest on sweetpotato which can cause yield lost up to 100%. The objective of this study was to obtain the effective and efficiency control technique of sweetpotato weevils. The research was conducted on June–October 2014 at Muneng Research Station, Probolinggo, East Java, using randomized block design, five treatments and three replicates. The treatments consisted of immersion of sweetpotato cuttings in the suspension of conidia Beauveria bassiana (Bals. Vuill. for 30 minutes before planting; application of suspension of conidia B. bassiana on soil at 2, 4, 6, 8, 10, and 12 weeks after sowing (WAS; immersion of sweetpotato cuttings in the suspension of conidia B. bassiana for 30 minutes before planting combined with application of suspension of conidia B. bassiana on soil at 2, 4, 6, 8, 10, and 12 WAS; application of chemical insecticide every week; and control. Analysis of variance showed that the application of various control technique significantly lowered the population of larvae and imago of C. formicarius and the level of tuber damage. The population level found on all treatments ranged from 1–4 eggs/plant, 1–19 larvae/plant, 0–0.2 pupae/plant and 0–4 imago/plant. The weight of tuber was 571–700 g/plant with the level of damage 6–53%. Low level of damage was found in the aplication of suspension of conidia B. bassiana on soil at 2, 4, 6, 8, 10, and 12 WAS which were 6%. Application of suspension of conidia B. bassiana on soil at 2, 4, 6, 8, 10, and 12 WAS can suppress the population of C. formicarius on tuber and reduce the level of damage up to 48%.

Achieving mutations of glycogenic strain Aspergillus awamori using ultraviolet radiation

International Nuclear Information System (INIS)

Rykala-Ziobro, M.; Dluzniewska, J.; Jedrzejowska, A.

1994-01-01

It was attempted to increase glucoamylase productivity of strain Aspergillus awamori, using UV light as a mutagenic agent. To this end, the conidias were twice irradiated, what resulted in obtaining the mutant with about 27% higher glucoamylase activity followed first irradiation of the conidias. Mutant form did not differ morphologically from the initial material (author)

Variability in morphology of Microsphaera alphitoides Griffon et Maubl. in Poland

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Krzysztof Ufnalski

2011-01-01

Full Text Available The aim of this study was to verify descriptions of the morphology of the fungus responsible for powdery mildew on oak leaves of nursery seedlings and of young trees in forest stands. Infected, fully developed leaves from apical parts of stems of Quercus robur or Q. petraea were collected in two national parks and in six forest districts in Poland. The taxonomic characteres of all specimens were generally consistent with those given in keys for Microsphaera alphitoides. However, the limoniform shape of conidia, characteristic of two sites (nurseries, has not been found in available synopses. Limoniform conidia were significantly longer and wider than others. Agreat variety of forms of mycelium on leaves were observed. The mycelium appeared in three forms: (1 floccose or felted oval patches (always with yellow or brown discoloration, (2 faveolate oval patches (often with yellow or brown discoloration, and (3 felted or mealy mycelium along veins (rarely with discoloration. Mycelium without limoniform conidia appeared in all forms, while mycelium with limoniform conidia appeared only in the form of patches, always causing leaf discoloration.

Evaluación del desarrollo de Salíx humboldtiana, Erythrina edulis y Trichantera gigantea asociados al cultivo de mora sin espina (Rubus glaucus Benth)

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Guapacha, Cristiam Camilo; Mesa, Jhon Jairo; Patiño, Andrés Alfonso

2015-01-01

En el Municipio de Quinchía, Departamento de Risaralda (Colombia), se evaluó el desarrollo de los tutores vivos Salíx humboldtiana, Erythrina edulis y Trichantera gigantea asociados al cultivo de mora sin espina (Rubus glaucus Benth), también la incidencia de Peronospora sparsa y Oidium sp en plantas de mora asociadas a estos sistemas agroforestales. Se utilizo un diseño de bloques al azar con tres repeticiones cada uno. Los resultados indican que Erythrina edulis es el tutor con mejor adapta...

Posaconazole Enhances the Activity of Amphotericin B against Aspergillus Hyphae In Vitro▿

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Perkhofer, Susanne; Lugger, Helene; Dierich, Manfred P.; Lass-Flörl, Cornelia

2006-01-01

The MICs and fractional inhibitory concentrations of posaconazole (POS) and voriconazole (VRZ), alone and in combination with amphotericin B (AMB), for the conidia and hyphae of 100 Aspergillus isolates were evaluated. POS-AMB had more synergistic activity against hyphae (75% of isolates) than VRZ-AMB (37%) and significantly more synergistic activity against hyphae than against conidia (12%).

First report of the infection of insecticide-resistant malaria vector mosquitoes with an entomopathogenic fungus under field conditions

Science.gov (United States)

2011-01-01

Background Insecticide-resistant mosquitoes are compromising the ability of current mosquito control tools to control malaria vectors. A proposed new approach for mosquito control is to use entomopathogenic fungi. These fungi have been shown to be lethal to both insecticide-susceptible and insecticide-resistant mosquitoes under laboratory conditions. The goal of this study was to see whether entomopathogenic fungi could be used to infect insecticide-resistant malaria vectors under field conditions, and to see whether the virulence and viability of the fungal conidia decreased after exposure to ambient African field conditions. Methods This study used the fungus Beauveria bassiana to infect the insecticide-resistant malaria vector Anopheles gambiae s.s (Diptera: Culicidae) VKPER laboratory colony strain. Fungal conidia were applied to polyester netting and kept under West African field conditions for varying periods of time. The virulence of the fungal-treated netting was tested 1, 3 and 5 days after net application by exposing An. gambiae s.s. VKPER mosquitoes in WHO cone bioassays carried out under field conditions. In addition, the viability of B. bassiana conidia was measured after up to 20 days exposure to field conditions. Results The results show that B. bassiana infection caused significantly increased mortality with the daily risk of dying being increased by 2.5× for the fungus-exposed mosquitoes compared to the control mosquitoes. However, the virulence of the B. bassiana conidia decreased with increasing time spent exposed to the field conditions, the older the treatment on the net, the lower the fungus-induced mortality rate. This is likely to be due to the climate because laboratory trials found no such decline within the same trial time period. Conidial viability also decreased with increasing exposure to the net and natural abiotic environmental conditions. After 20 days field exposure the conidial viability was 30%, but the viability of control

Resting spore formation of aphid-pathogenic fungus Pandora nouryi depends on the concentration of infective inoculum.

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Huang, Zhi-Hong; Feng, Ming-Guang

2008-07-01

Resting spore formation of some aphid-pathogenic Entomophthorales is important for the seasonal pattern of their prevalence and survival but this process is poorly understood. To explore the possible mechanism involved in the process, Pandora nouryi (obligate aphid pathogen) interacted with green peach aphid Myzus persicae on cabbage leaves under favourable conditions. Host nymphs showered with primary conidia of an isolate (LC(50): 0.9-6.7 conidia mm(-2) 4-7 days post shower) from air captures in the low-latitude plateau of China produced resting spores (azygospores), primary conidia or both spore types. Surprisingly, the proportion of mycosed cadavers forming resting spores (P(CFRS)) increased sharply within the concentrations (C) of 28-240 conidia mm(-2), retained high levels at 240-1760, but was zero or extremely low at 0.3-16. The P(CFRS)-C relationship fit well the logistic equation P(CFRS) = 0.6774/[1 + exp(3.1229-0.0270C)] (r(2) = 0.975). This clarified for the first time the dependence of in vivo resting spore formation of P. nouryi upon the concentration of infective inoculum. A hypothesis is thus proposed that some sort of biochemical signals may exist in the host-pathogen interaction so that the fungal pathogen perceives the signals for prompt response to forthcoming host-density changes by either producing conidia for infecting available hosts or forming resting spores for surviving host absence in situ.

Gray Mold on Saintpaulia ionantha Caused by Botrytis cinerea in Korea

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Hyung-Moo Kim

2011-04-01

Full Text Available Gray mold caused by Botrytis cinerea occurred on Saintpaulia ionantha in flower shop of the Jeonju city in Korea. Typical symptoms with brown water-soaked and rotting lesions were appeared on the flowers, leaves and petiole of infected plants. Many conidia spores appeared on the lesions under humid conditions. Colonies were grayish brown and sclerotial formation on potato dextrose agar. Conidia were one celled, mostly ellipsoidal or ovoid in shape, and were colorless to pale brown in color. The conidia were 7~14×5~9 μm in size. Based on pathogenicity and morphological characteristics of the isolated fungus, the causal fungus was identified as B. cinerea Persoon: Fries. Gray mold of S. ionantha was proposed to the name of this disease.

Use of high throughput sequencing to study oomycete communities in soil and roots

DEFF Research Database (Denmark)

Sapkota, Rumakanta; Nicolaisen, Mogens

2015-01-01

taxonomic units from symptomatic lesions in carrot resulted in 94% of the reads belonging to oomycetes with a dominance of species of Pythium that are known to be involved in causing cavity spot. Moreover, soil samples showed that 95% of the sequences could be assigned to oomycetes including Pythium......, Aphanomyces, Peronospora, Saprolegnia and Phytophthora. A high proportion of oomycete reads was consistently present in all symptomatic lesions and soil samples showing the versatility of the strategy and thus demonstrating the usefulness of the method in plant and soil DNA background....

Characterization of Emericella nidulans RodA and DewA hydrophobin mutants

DEFF Research Database (Denmark)

Jensen, Britt Guillaume; Nielsen, Jakob Blæsbjerg; Pedersen, Mona Højgaard

hydrophobins RodA and DewA. Individual knock-out mutants rodAΔ, dewAΔ and the double deletion strain rodAΔdewAΔ were constructed. Furthermore, two strains containing a point mutation in the first of the cysteines of RodA (rodA-C57G), where one was coupled to the dewA deletion, were included. The reference...... strain (NID1) and dewAΔ displayed green conidia. However, rodAΔ and rodAΔdewAΔ showed a dark green/brown conidial pigmentation, while rodA-C57G and rodAC57G dewAΔ displayed lighter brown conidia. rodAΔ and rodAΔdewAΔ displayed a higher degree of hülle cells compared to the moderate amount observed...... for NID1 and dewAΔ, while rodA-C57G and rodA-C57G dewAΔ displayed a low number of hülle cells. NID1 and dewAΔ conidia were dispersed as spore chains. rodAΔ, rodAΔdewAΔ, rodA-C57G and rodA-C57G dewAΔ spores were associated in large clumps, where the conidia seemed to adhere to one another. The largest...

Bioassay and Scanning Electron Microscopic Observations Reveal High Virulence of Entomopathogenic Fungus, Beauveria bassiana, on the Onion Maggot (Diptera: Anthomyiidae) Adults.

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Zhang, Hui; Wu, Shengyong; Xing, Zhenlong; Wang, Xiaoqing; Lei, Zhongren

2016-12-01

When flies were dipped in 1 × 10 8 conidia/ml conidia suspensions and then kept in the incubator (22 ± 1 °C, 70 ± 5% RH), scanning electron microscope observations revealed that, at 2 h, the majority of adhering Beauveria bassiana conidia were attached to either the wing surface or the interstitial area between the macrochaetae on the thorax and abdomen of the onion maggot adults. Germ tubes were being produced and had oriented toward the cuticle by 18 h. Penetration of the insect cuticle had occurred by 36 h, and by 48 h, germ tubes had completely penetrated the cuticle. Fungal mycelia had emerged from the insect body and were proliferating after 72 h. The superficial area and structure of the wings and macrochaetae may facilitate the attachment of conidia and enable effective penetration. The susceptibility of adults to 12 isolates, at a concentration of 1 × 10 7 conidia/ml, was tested in laboratory experiments. Eight of the more potent strains caused in excess of 85% adult mortality 8 d post inoculation, while the median lethal time (LT 50 ) of these strains was bassiana strains are highly virulent to onion maggot adults and should be considered as potential biocontrol agents against the adult flies. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Aspergillus niger mutants affected in conidial pigmentation do not have an increased susceptibility to water stress during growth at low water activity.

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Segers, F J J; Wösten, H A B; Dijksterhuis, J

2018-03-01

Aspergillus niger forms conidia that contain melanin in their cell wall. This black pigment has been shown to protect fungi against UV radiation, and experimental evidence has indicated that it also protects against drought and high salt concentrations. In this study, growth of A. niger was evaluated at low water activity (a w ) and after changes in relative humidity (RH). In addition, deletion strains of A. niger affected in the melanin synthesis pathway were compared. Germination of conidia of the wild-type and deletion strains was observed at 0·81 a w and germ tubes continued growth at a w  ≥ 0·83. Conidia and microcolonies of the different strains were incubated for 1 week at lowered RH (33-84%). Conidia of all strains germinated and formed colonies after exposure to RH ≥33% when transferred back to malt extract medium at a w 0·98. Conidia germinated and showed limited growth at 84% RH. Microcolonies of all strains did not survive an incubation of 1 week at RH ≤75%, but continued growth after exposure to 84% RH. Together, this is the first genetic evidence that melanin does not play a role during germination and radial extension of fungi at low water conditions. Aspergillus niger, a cosmopolitan fungus with melanized conidia, is used here as a model system for fungal growth at low water activity (a w ) and humidity dynamics. From this study it becomes clear that melanin, contrary to what has been suggested before, is not a key factor in survival and growth during situations that mimic indoor conditions. Indoor fungal growth can lead to cosmetic damage to building materials and health problems. This knowledge makes clear that novel ways to limit indoor fungal growth have to be based on interference with other cellular traits of fungi. © 2018 The Society for Applied Microbiology.

New use of broomcorn millets for production of granular cultures of aphid-pathogenic fungus Pandora neoaphidis for high sporulation potential and infectivity to Myzus persicae.

Science.gov (United States)

Hua, Li; Feng, Ming-Guang

2003-10-24

Glutinous broomcorn millets from the crop Panicum miliaceum were first used as substrate to produce granular cultures of Pandora neoaphidis, an obligate fungal pathogen specific to aphids. Carrying a water content of 36.5% after being steamed in a regular autoclaving procedure, millet grains of each 15 g (dry weight) in a 100-ml flask were mixed with 3 ml modified Sabouraud dextrose broth containing half a mashed colony of P. neoaphidis grown on egg yolk milk agar and then incubated at 20 degrees C and a light/dark cycle of 12 h/12 h for 21 days. Based on individually monitoring conidial production potential of 20 millet grains sampled from an arbitrarily taken flask at 3-day intervals, the millet cultures incubated for 6-15 days were capable of producing 16.8-23.4 x 10(4) conidia per millet grain with conidial ejection lasting for up to 6 days. The cultured millet grains individually produced significantly more conidia than apterous adults of Myzus persicae killed by P. neoaphidis (8.4 x 10(4) conidia per cadaver) and sporulated twice longer. The modeling of time-dose-mortality data from bioassays on M. persicae apterae exposed to conidial showers from the cultured millet grains and the mycelial mats produced in liquid culture resulted in similar estimates of LC(50) (millets: 21.4, 7.3, and 4.9 conidia mm(-2) on days 5-7 after exposure; mycelial mats: 22.1, 10.6, and 7.7 conidia mm(-2)) although the LT(50) estimated at a given conidial concentration was slightly smaller for the millet cultures than for the mycelial mats. This indicates that the millet grains cultured with P. neoaphidis produced conidia as infective as or slightly more infective to M. persicae than those from the mycelial mats. Based on the sporulation potential, infectivity, and ease and cost of the millet cultures, the method developed in this study highly improved in vitro cultures of P. neoaphidis and may adapt to culturing other entomophthoralean fungi for microbial control of insect pests.

Neem oil increases the efficiency of the entomopathogenic fungus Metarhizium anisopliae for the control of Aedes aegypti (Diptera: Culicidae) larvae.

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Gomes, Simone A; Paula, Adriano R; Ribeiro, Anderson; Moraes, Catia O P; Santos, Jonathan W A B; Silva, Carlos P; Samuels, Richard I

2015-12-30

Entomopathogenic fungi are potential candidates for use in integrated vector management and many isolates are compatible with synthetic and natural insecticides. Neem oil was tested separately and in combination with the entomopathogenic fungus Metarhizium anisopliae against larvae of the dengue vector Aedes aegypti. Our aim was to increase the effectiveness of the fungus for the control of larval mosquito populations. Commercially available neem oil was used at concentrations ranging from 0.0001 to 1%. Larval survival rates were monitored over a 7 day period following exposure to neem. The virulence of the fungus M. anisopliae was confirmed using five conidial concentrations (1 × 10(5) to 1 × 10(9) conidia mL(-1)) and survival monitored over 7 days. Two concentrations of fungal conidia were then tested together with neem (0.001%). Survival curve comparisons were carried out using the Log-rank test and end-point survival rates were compared using one-way ANOVA. 1% neem was toxic to A. aegypti larvae reducing survival to 18% with S50 of 2 days. Neem had no effect on conidial germination or fungal vegetative growth in vitro. Larval survival rates were reduced to 24% (S50 = 3 days) when using 1 × 10(9) conidia mL(-1). Using 1 × 10(8) conidia mL(-1), 30% survival (S50 = 3 days) was observed. We tested a "sub-lethal" neem concentration (0.001%) together with these concentrations of conidia. For combinations of neem + fungus, the survival rates were significantly lower than the survival rates seen for fungus alone or for neem alone. Using a combination of 1 × 10(7) conidia mL(-1) + neem (0.001%), the survival rates were 36%, whereas exposure to the fungus alone resulted in 74% survival and exposure to neem alone resulted in 78% survival. When using 1 × 10(8) conidia mL(-1), the survival curves were modified, with a combination of the fungus + neem resulting in 12% survival, whilst the fungus alone at this concentration also

Entomophthora muscae — moisture as a factor affecting its transmission and conidial germination

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John P. Kramer

2014-08-01

Full Text Available The role played by moisture in the transmission of Entomophthora muscae and in the germination of its conidia was investigated. A majority of adult house flies exposed to conidial showers that fell upon surfaces covered with droplets of condensation acquired the parasite, while no flies exposed to conidial showers that fell upon dry surfaces did so. A microscopical study of conidial showers showed that germination was practically non-existent on dry surfaces while a vast majority of conidia that fell upon a droplet-covered surface germinated. A method for the in vivo culture of E. muscae was developpd and 11 serial passages of the fungus were achieved. Resting spores rather than conidia became the dominant form produced in the cadavers, and flies in a twelfth group remained unifected.

Localization of Cladosporium fulvum hydrophobins reveals a role for HCf-6 in adhesion

DEFF Research Database (Denmark)

Lacroix, Hélène; Whiteford, James; Spanu, Pietro D

2008-01-01

for establishing hydrophobin localization during growth in culture and in plants. In this paper we localize HCf-2, -3, -4 and -5 and compare the data to our previous observations for HCf-1 and -6. In culture, HCf-1, -2, -3 and 4 localize to conidia and also appear on aerial hyphae. HCf-4 is unique...... in that it appears on submerged hyphae. HCf-5 expression is tightly regulated and appears on aerial hyphae early on during growth. Only HCf-1, -3 and -6 were observed during infection; HCf-3 appears on both conidia and emerging germ tubes. We also show that HCf-6 is secreted and coats surfaces under and around...... growing hyphae and demonstrate the effect of deleting HCf-6 on the adhesion of germinating C. fulvum conidia to glass slides....

Identification and validation of reference genes for qRT-PCR studies of the obligate aphid pathogenic fungus Pandora neoaphidis during different developmental stages

OpenAIRE

Zhang, Shutao; Chen, Chun; Xie, Tingna; Ye, Sudan

2017-01-01

The selection of stable reference genes is a critical step for the accurate quantification of gene expression. To identify and validate the reference genes in Pandora neoaphidis-an obligate aphid pathogenic fungus-the expression of 13classical candidate reference genes were evaluated by quantitative real-time reverse transcriptase polymerase chain reaction(qPCR) at four developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae). Four statistical algorithms, inc...

Biocontrol of the Brown-banded Cockroach, Supella longipalpa F. (Blattaria: Blattellidae, with Entomopathogenic Fungus, Metharhizium anisopliae

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Mona Sharififard

2016-01-01

Full Text Available Background: Considering to the high distribution of cockroaches as urban pests, the efficacy of different formula­tions of Metarhizium anisopliae strain Iran 437C were assessed against the brown-banded cockroach, Supella longi­palpa F. under laboratory and field conditions.Methods: Metarhizium anisopliae isolates were screened with immersing adults of the brown-banded cockroachs in aqueous suspension of 108 conidia ml-1 followed by surface or bait treated with different doses of the most virulent isolate against the nymphs. Then formulations of conidia oil-in-water were examined versus cockroach nymphs us­ing different plant oils and paraffin. Then they were evaluated and compared with aqueous suspension and control group. On a large-scale, the sunflower oil-in-water formulation of conidia was sprayed at houses using a hand sprayer.Results: Metarhizium anisopliae IRAN 437C was the most virulent isolate against the brown-banded cockroach, causing 100% mortality in adults at seven days post-exposure. Inoculated bait with this isolate was not enough path­ogenic against the cockroach even at two weeks after treatment. Treated surface with conidia as aqueous suspension or oil-in- water formulation was more effective than the bait formulation against the cockroach caused 39.4–97.2% mortality compared with 2.5% mortality in control group after two days. Spraying the conidia formulated with sun­flower oil was an effective formulation causing 76.1% reduction in the cockroach density on the third day post treatment in the houses.Conclusion: The oil-in-water formulation of M. anisopliae IRAN 437C could be recommended as a promising al­ternative for cockroach control.  

Patogenisitas Isolat Beauveria bassiana dan Metarhizium anisopliae asal Tanah Lebak dan Pasang Surut Sumatera Selatan untuk Agens Hayati Scirpophaga incertulas

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Rosdah Thalib

2014-08-01

Full Text Available Pathogeicity of Beauveria bassiana and  Metarhizium anisopliae Isolates from Fresh Swamp and  Tidal Lowland, South Sumatra for Scirpophaga incertulas Biological Agents.  The objectives of the research weret o explore and to determine the pathogenicity of entomopathogenic  fungi againts the larvae of Scirpophaga incertulas, and to measure conidial viability and density of the fungi.  The method for fungi exploration used larvae of Tenebrio molitor baiting submerged in the soil.  The soil was taken from fresh swampand tidal lowland rice in South  Sumatra.  From the exploration study, we found two species of entomopathogenic fungi: Beauveria bassiana and Metarhizium anisopliae. Mortality of S. incertulas larvae that had been treated topically with fungal conidia (1x106 conidia ml-1 varied among the isolates. The highest mortality (98.33% caused by BPlus isolate of B. Bassiana and the lowest by MtmIn  isolate of M. anisopliae (57.50% and BTmTr  isolate of B. bassiana (57.50%.  The fungal colonies grew fast from the second day up to the fourth day after incubation but the growth became slow after the fifth day.  The highest conidial density was resulted by   BPcMs of B. bassiana isolate (63.33x106 conidia ml-1 but  this density was not significantly different from that of the BPlus  of B. bassiana isolate (63.11x106 conidia ml-1.  The lowest conidial density found in BTmTr of B. bassiana isolate (20.97x106 conidia ml-1 .   The isolate B. bassiana was more effective than M. anisopliae againt the larvae of S.incertulas.

Biochemical analysis of plant protection afforded by a nonpathogenic endophytic mutant of Colletotrichum magna

Energy Technology Data Exchange (ETDEWEB)

Redman, R.S.; Rodriguez, R.J. (Geological Survey, Seattle, WA (United States) Univ. of Washington, Seattle, WA (United States). Dept. of Botany); Clifton, D.R.; Morrel, J.; Brown, G. (Geological Survey, Seattle, WA (United States)); Freeman, S. (Volcani Center, Bet Dagan (Israel). Dept. of Plant Pathology)

1999-02-01

A nonpathogenic mutant of Colletotrichum magna (path-1) was previously shown to protect watermelon (Citrullus lanatus) and cucumber (Cucumis sativus) seedlings from anthracnose disease elicited by wild-type C. magna. Disease protection was observed in stems of path-1-colonized cucurbits but not in cotyledons, indicating that path-1 conferred tissue-specific and/or localized protection. Plant biochemical indicators of a localized and systemic (peroxidase, phenylalanine ammonia-lyase, lignin, and salicylic acid) plant-defense response were investigated in anthracnose-resistant and-susceptible cultivars of cucurbit seedlings exposed to four treatments: (1) water (control), (2) path-1 conidia, (3) wild-type conidia, and (4) challenge conditions (inoculation into path-1 conidia for 48 h and then exposure to wild-type conidia). Collectively, these analyses indicated that disease protection in path-1-colonized plants was correlated with the ability of these plants to mount a defense response more rapidly and to equal or greater levels than plants exposed to wild-type C. magna alone. Watermelon plants colonized with path-1 were also protected against disease caused by Colletotrichum orbiculare and Fusarium oxysporum. A model based on the kinetics of plant-defense activation is presented to explain the mechanism of path-1-conferred disease protection.

Beauveria medogensis sp. nov., a new fungus of the entomopathogenic genus from China.

Science.gov (United States)

Imoulan, Abdessamad; Wu, Hai-Jun; Lu, Wei-Lai; Li, Yi; Li, Bin-Bin; Yang, Rei-Heng; Wang, Wen-Jing; Wang, Xiao-Liang; Kirk, Paul M; Yao, Yi-Jian

2016-09-01

Beauveria is among the most ubiquitous genera of entomopathogenic fungi throughout the world. A previously unknown species of the genus was recently discovered from a soil sample collected from Tibetan Plateau, China and is here described as new to science, B. medogensis sp. nov. The new species is distinguished from its closest relatives based on both morphological characterization and molecular phylogenetic analyses. Beauveria medogensis is characterized by globose to subglobose conidia, morphologically similar to some other species of in the genus, but was conclusively separated from those species in the phylogenetic analyses including sequences of four nuclear genes (RPB1, RPB2, TEF1 and Bloc). The new species was clustered in the analyses in a single terminal lineage which was grouped with B. australis sequences together as a sister clade to the B. brongniartii terminal clade. Although molecularly closely related, the new species is distinct morphologically from its closest sisters, B. australis and B. brongniartii, in producing globose to subglobose conidia rather than subglobose, broadly ellipsoid to ellipsoid conidia or ellipsoidal to cylindrical conidia. As isolated from a soil sample, the entomopathogenicity of the new species has been confirmed using Helicoverpa armigera and Tenebrio molitor larvae. Copyright © 2016 Elsevier Inc. All rights reserved.

Food consumption by Chilo partellus (Lepidoptera: Pyralidae) larvae infected with Beauveria bassiana and Metarhizium anisopliae and effects of feeding natural versus artificial diets on mortality and mycosis.

Science.gov (United States)

Tefera, Tadele; Pringle, K L

2003-11-01

Second and third instar Chilo partellus larvae were infected with Beauveria bassiana and Metarhizium anisopliae (both at 1x10(8)conidia/ml) and daily consumption of maize leaves was measured. Infection by the fungi was associated with reduced mean daily food consumption. Reduction in food consumption became evident 3-4 days after treatment with the fungi for second instar larvae and 4-5 days for third instar larvae. Four conidial concentrations, 1x10(5), 1x10(6), 1x10(7), and 1x10(8)conidia/ml, were tested against second instar larvae. Food consumption dropped by 70-85% when the second instar larvae were treated with the fungi at 1x10(8)conidia/ml. Reduction in food consumption by C. partellus larvae infected with B. bassiana and M. anisopliae may offset the slow speed of kill of the fungi. The effect of artificial versus natural diets on mortality and mycoses of second instar larvae treated with the fungi at 1x10(8)conidia/ml was determined. Larvae provided with artificial diet suffered little mortality and mycoses than larvae provided with maize leaves. The LT(50) was longer for larvae provided with artificial diet.

Sporulation of Metarhizium anisopliae var. Acridum and Beauveria bassiana on Rhammatocerus schistocercoides under humid and dry conditions

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Magalhães Bonifácio Peixoto

2000-01-01

Full Text Available The sporulation of the fungi Metarhizium anisopliae var. acridum and Beauveria bassiana in cadavers of the grasshopper Rhammatocerus schistocercoides was studied in dry and humid environments. Both fungi were equally virulent against R. schistocercoides. However, internally, M. anisopliae produced more conidia than B. bassiana at 53% and 75% relative humidity. Externally, there was no sporulation at 53% and 75% RH, and M. anisopliae produced more conidia than B. bassiana at 100% RH.

Development stage-specific proteomic profiling uncovers small, lineage specific proteins most abundant in the Aspergillus Fumigatus conidial proteome

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Suh Moo-Jin

2012-04-01

Full Text Available Abstract Background The pathogenic mold Aspergillus fumigatus is the most frequent infectious cause of death in severely immunocompromised individuals such as leukemia and bone marrow transplant patients. Germination of inhaled conidia (asexual spores in the host is critical for the initiation of infection, but little is known about the underlying mechanisms of this process. Results To gain insights into early germination events and facilitate the identification of potential stage-specific biomarkers and vaccine candidates, we have used quantitative shotgun proteomics to elucidate patterns of protein abundance changes during early fungal development. Four different stages were examined: dormant conidia, isotropically expanding conidia, hyphae in which germ tube emergence has just begun, and pre-septation hyphae. To enrich for glycan-linked cell wall proteins we used an alkaline cell extraction method. Shotgun proteomic resulted in the identification of 375 unique gene products with high confidence, with no evidence for enrichment of cell wall-immobilized and secreted proteins. The most interesting discovery was the identification of 52 proteins enriched in dormant conidia including 28 proteins that have never been detected in the A. fumigatus conidial proteome such as signaling protein Pil1, chaperones BipA and calnexin, and transcription factor HapB. Additionally we found many small, Aspergillus specific proteins of unknown function including 17 hypothetical proteins. Thus, the most abundant protein, Grg1 (AFUA_5G14210, was also one of the smallest proteins detected in this study (M.W. 7,367. Among previously characterized proteins were melanin pigment and pseurotin A biosynthesis enzymes, histones H3 and H4.1, and other proteins involved in conidiation and response to oxidative or hypoxic stress. In contrast, expanding conidia, hyphae with early germ tubes, and pre-septation hyphae samples were enriched for proteins responsible for

Metarhizium brunneum (Ascomycota; Hypocreales) Treatments Targeting Olive Fly in the Soil for Sustainable Crop Production.

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Yousef, Meelad; Alba-Ramírez, Carmen; Garrido Jurado, Inmaculada; Mateu, Jordi; Raya Díaz, Silvia; Valverde-García, Pablo; Quesada-Moraga, Enrique

2018-01-01

Soil treatments with Metarhizium brunneum EAMa 01/58-Su strain conducted in both Northern and Southern Spain reduced the olive fly ( Bactrocera oleae ) population density emerging from the soil during spring up to 70% in treated plots compared with controls. A model to determine the influence of rainfall on the conidial wash into different soil types was developed, with most of the conidia retained at the first 5 cm, regardless of soil type, with relative percentages of conidia recovered ranging between 56 and 95%. Furthermore, the possible effect of UV-B exposure time on the pathogenicity of this strain against B. oleae adults coming from surviving preimaginals and carrying conidia from the soil at adult emergence was also evaluated. The UV-B irradiance has no significant effect on M. brunneum EAMa 01/58-Su pathogenicity with B. oleae adult mortalities of 93, 90, 79, and 77% after 0, 2, 4, and 6 of UV-B irradiance exposure, respectively. In a next step for the use of these M. brunneum EAMa 01/58-Sun soil treatments within a B. oleae IPM strategy, its possible effect of on the B. oleae cosmopolitan parasitoid Psyttalia concolor , its compatibility with the herbicide oxyfluorfen 24% commonly used in olive orchards and the possible presence of the fungus in the olive oil resulting from olives previously placed in contact with the fungus were investigated. Only the highest conidial concentration (1 × 10 8 conidia ml - ) caused significant P. concolor adult mortality (22%) with enduing mycosis in 13% of the cadavers. There were no fungal propagules in olive oil samples resulting from olives previously contaminated by EAMa 01/58-Su conidia. Finally, the strain was demonstrated to be compatible with herbicide since the soil application of the fungus reduced the B. oleae population density up to 50% even when it was mixed with the herbicide in the same tank. The fungal inoculum reached basal levels 4 months after treatments (1.6 × 10 3 conidia g soil -1 ). These results

Metarhizium brunneum (Ascomycota; Hypocreales Treatments Targeting Olive Fly in the Soil for Sustainable Crop Production

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Meelad Yousef

2018-01-01

Full Text Available Soil treatments with Metarhizium brunneum EAMa 01/58-Su strain conducted in both Northern and Southern Spain reduced the olive fly (Bactrocera oleae population density emerging from the soil during spring up to 70% in treated plots compared with controls. A model to determine the influence of rainfall on the conidial wash into different soil types was developed, with most of the conidia retained at the first 5 cm, regardless of soil type, with relative percentages of conidia recovered ranging between 56 and 95%. Furthermore, the possible effect of UV-B exposure time on the pathogenicity of this strain against B. oleae adults coming from surviving preimaginals and carrying conidia from the soil at adult emergence was also evaluated. The UV-B irradiance has no significant effect on M. brunneum EAMa 01/58-Su pathogenicity with B. oleae adult mortalities of 93, 90, 79, and 77% after 0, 2, 4, and 6 of UV-B irradiance exposure, respectively. In a next step for the use of these M. brunneum EAMa 01/58-Sun soil treatments within a B. oleae IPM strategy, its possible effect of on the B. oleae cosmopolitan parasitoid Psyttalia concolor, its compatibility with the herbicide oxyfluorfen 24% commonly used in olive orchards and the possible presence of the fungus in the olive oil resulting from olives previously placed in contact with the fungus were investigated. Only the highest conidial concentration (1 × 108 conidia ml− caused significant P. concolor adult mortality (22% with enduing mycosis in 13% of the cadavers. There were no fungal propagules in olive oil samples resulting from olives previously contaminated by EAMa 01/58-Su conidia. Finally, the strain was demonstrated to be compatible with herbicide since the soil application of the fungus reduced the B. oleae population density up to 50% even when it was mixed with the herbicide in the same tank. The fungal inoculum reached basal levels 4 months after treatments (1.6 × 103 conidia g soil−1

Efficacy of the entomopathogenic fungus Metarhizium brunneum in controlling the tick Rhipicephalus annulatus under field conditions.

Science.gov (United States)

Samish, M; Rot, A; Ment, D; Barel, S; Glazer, I; Gindin, G

2014-12-15

High infectivity of entomopathogenic fungi to ticks under laboratory conditions has been demonstrated in many studies. However, the few reports on their use under field conditions demonstrate large variations in their success, often with no clear explanation. The present study evaluated the factors affecting the efficacy of the fungus Metarhizium brunneum against the tick Rhipicephalus (Boophilus) annulatus. It demonstrates how environmental conditions and ground cover affect the efficiency of the fungus under field conditions. During the summer, 93% of tick females exposed to fungus-contaminated ground died within 1 week, whereas during the winter, only 62.2% died within 6 weeks. Nevertheless, the hatchability of their eggs was only 6.1% during the summer and 0.0% during winter. Covering the ground with grass, leaves or gravel improved fungal performance. Aside from killing female ticks, the fungus had a substantial effect on tick fecundity. Fungal infection reduced the proportion of female ticks laying full-size egg masses by up to 91%, and reduced egg hatchability by up to 100%. To reduce the negative effect of outdoor factors on fungal activity, its conidia were mixed with different oils (olive, canola, mineral or paraffin at 10% v/v) and evaluated in both laboratory and field tests for efficacy. All tested oils without conidia sprayed on the sand did not influence tick survival or weight of the laid eggs but significantly reduced egghatchability. Conidia in water with canola or mineral oil spread on agarose and incubated for 18 h showed 57% and 0% germination, respectively. Comparing, under laboratory conditions, the effects of adding each of the four oils to conidia in water on ticks demonstrated no effect on female mortality or weight of the laid egg mass, but the percentage of hatched eggs was reduced. In outdoor trials, female ticks placed on the ground sprayed with conidia in water yielded an average of 175 larvae per female and there was no hatching of

Pathogenicity gene variations within the order Entomophthorales

DEFF Research Database (Denmark)

Grell, Morten Nedergaard; Jensen, Annette Bruun; Lange, Lene

Fungi within the order Entomophthorales (subphylum Entomophthoromycotina) are obligate biotrophic pathogens of arthropods with a remarkable narrow host range. Infection takes place through the cuticle when conidia hit a susceptible host, facilitated by enzymatic and mechanical mechanisms. In the ...... pathogenicity genes within genera Entomophthora and Pandora, using fungal genomic DNA originating from field-collected, infected insect host species of dipteran (flies, mosquitoes) or hemipteran (aphid) origin.......Fungi within the order Entomophthorales (subphylum Entomophthoromycotina) are obligate biotrophic pathogens of arthropods with a remarkable narrow host range. Infection takes place through the cuticle when conidia hit a susceptible host, facilitated by enzymatic and mechanical mechanisms......, conidia are produced and discharged when humidity gets high—usually during night. In an earlier secretome study of field-collected grain aphids (Sitobion avenae) infected with entomophthoralean fungi, a number of pathogenesis-related, secreted enzymes were discovered (Fungal Genetics and Biology 2011, vol...

Improvement of Aspergillus niger 55, a raw corn meal saccharifying enzyme hyperproducer, through mutation and selective screening techniques

International Nuclear Information System (INIS)

Oh, S.H.; O, P.S.

1991-01-01

Mutation experiments were performed to select the mutant of Aspergillus niger 55, which had lost almost all the ability to produce transglucosidases but retained that of high productivity of raw meal saccharifying enzyme, by means of successive induction with N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), ultraviolet(UV) light, and γ-rays. Also, we used the mutant enrichment techniques, such as liquid culture-filtration procedure and differential heat sensitivity of conidia, in order to increase the possibility of obtaining a mutant. The glucoamylase productivity of mutant PFST-38 was 11 times higher than that of the parent strain. The mutant PFST-38 was morphologically identical to the parent strain, except for the size of conidia, the tendency to form conidia and the length of conidiophore. Asp. niger mutant PFST-38 appeared to be useful for the submerged production of the raw corn meal saccharifying enzyme

Inheritance of resistance to powdery mildew in pea and pathogenesis-related aspects

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Ricardo Lima dos Santos

2012-06-01

Full Text Available The inheritance of resistance to powdery mildew in the pea cultivar MK-10 and some histological aspects of infection were assessed. For the inheritance study, F1, F2, backcrosses and F3 generations of MK-10 crossed with two susceptible populations were evaluated. Histological evaluations included percentage of germinated conidia, percentage of conidia that formed appresoria, percentage of conidia that established colonies, and number of haustoria per colony. Segregation ratios obtained in the resistance inheritance study were compared by Chi-square (ײ test and the histological data were analyzed by Tukey's test at 5% probability. It was concluded that resistance of MK-10 to powdery mildew is due to a pair of recessive alleles since it is expressed in the pre-penetration stage and completed by post-penetration localized cellular death, characteristic of the presence of the pair of recessive alleles er1er1.

Identification and functional analysis of endogenous nitric oxide in a filamentous fungus.

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Pengkit, Anchalee; Jeon, Seong Sil; Son, Soo Ji; Shin, Jae Ho; Baik, Ku Yeon; Choi, Eun Ha; Park, Gyungsoon

2016-07-18

In spite of its prevalence in animals and plants, endogenous nitric oxide (NO) has been rarely reported in fungi. We present here our observations on production of intracellular NO and its possible roles during development of Neurospora crassa, a model filamentous fungus. Intracellular NO was detected in hypha 8-16 hours after incubation in Vogel's minimal liquid media and conidiophores during conidiation using a fluorescent indicator (DAF-FM diacetate). Treatment with cPTIO, an NO scavenger, significantly reduced fluorescence levels and hindered hyphal growth in liquid media and conidiation, whereas exogenous NO enhanced hyphal extension on VM agar media and conidia formation. NO scavenging also dramatically diminished transcription of con-10 and con-13, genes preferentially expressed during conidiation. Our results suggest that intracellular NO is generated in young hypha growing in submerged culture and during conidia development and regulate mycelial development and conidia formation.

Toxic reagents and expensive equipment: are they really necessary for the extraction of good quality fungal DNA?

Science.gov (United States)

Rodrigues, P; Venâncio, A; Lima, N

2018-01-01

The aim of this work was to evaluate a fungal DNA extraction procedure with the lowest inputs in terms of time as well as of expensive and toxic chemicals, but able to consistently produce genomic DNA of good quality for PCR purposes. Two types of fungal biological material were tested - mycelium and conidia - combined with two protocols for DNA extraction using Sodium Dodecyl Sulphate (SDS) and Cetyl Trimethyl Ammonium Bromide as extraction buffers and glass beads for mechanical disruption of cell walls. Our results showed that conidia and SDS buffer was the combination that lead to the best DNA quality and yield, with the lowest variation between samples. This study clearly demonstrates that it is possible to obtain high yield and pure DNA from pigmented conidia without the use of strong cell disrupting procedures and of toxic reagents. There are numerous methods for DNA extraction from fungi. Some rely on expensive commercial kits and/or equipments, unavailable for many laboratories, or make use of toxic chemicals such as chloroform, phenol and mercaptoethanol. This study clearly demonstrates that it is possible to obtain high yields of pure DNA from pigmented conidia without the use of strong and expensive cell disrupting procedures and of toxic reagents. The method herein described is simultaneously inexpensive and adequate to DNA extraction from several different types of fungi. © 2017 The Society for Applied Microbiology.

Chitin Synthases with a Myosin Motor-Like Domain Control the Resistance of Aspergillus fumigatus to Echinocandins

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Jiménez-Ortigosa, Cristina; Aimanianda, Vishukumar; Muszkieta, Laetitia; Mouyna, Isabelle; Alsteens, David; Pire, Stéphane; Beau, Remi; Krappmann, Sven; Beauvais, Anne; Dufrêne, Yves F.

2012-01-01

Aspergillus fumigatus has two chitin synthases (CSMA and CSMB) with a myosin motor-like domain (MMD) arranged in a head-to-head configuration. To understand the function of these chitin synthases, single and double csm mutant strains were constructed and analyzed. Although there was a slight reduction in mycelial growth of the mutants, the total chitin synthase activity and the cell wall chitin content were similar in the mycelium of all of the mutants and the parental strain. In the conidia, chitin content in the ΔcsmA strain cell wall was less than half the amount found in the parental strain. In contrast, the ΔcsmB mutant strain and, unexpectedly, the ΔcsmA/ΔcsmB mutant strain did not show any modification of chitin content in their conidial cell walls. In contrast to the hydrophobic conidia of the parental strain, conidia of all of the csm mutants were hydrophilic due to the presence of an amorphous material covering the hydrophobic surface-rodlet layer. The deletion of CSM genes also resulted in an increased susceptibility of resting and germinating conidia to echinocandins. These results show that the deletion of the CSMA and CSMB genes induced a significant disorganization of the cell wall structure, even though they contribute only weakly to the overall cell wall chitin synthesis. PMID:22964252

Geomyces destructans sp. nov. associated with bat white-nose syndrome

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Gargas, Andrea; Trest, M.T.; Christensen, M.; Volk, T.J.; Blehert, David S.

2009-01-01

We describe and illustrate the new species Geomyces destructans. Bats infected with this fungus present with powdery conidia and hyphae on their muzzles, wing membranes, and/or pinnae, leading to description of the accompanying disease as white-nose syndrome, a cause of widespread mortality among hibernating bats in the northeastern US. Based on rRNA gene sequence (ITS and SSU) characters the fungus is placed in the genus Geomyces, yet its distinctive asymmetrically curved conidia are unlike those of any described Geomyces species.

Analysis of mitotic nondisjunction with Aspergillus nidulans.

Science.gov (United States)

Morpurgo, G; Bellincampi, D; Gualandi, G; Baldinelli, L; Crescenzi, O S

1979-08-01

Two methods to detect the induction of nondisjunction with a diploid stable strain of A. nidulans are described. The first method gives only qualitative results, while the second method is quantitative and dose-effect curves can be done. Some physiological parameters affecting the induction of nondisjunction can also be studied, because either quiescent or germinating conidia can be treated with the drug under test. Some agents inducing nondisjunction were also tested for the induction of point mutation and somatic crossing-over with these comparative analysis. Two classes of agents inducing nondisjunction may be detected: the first causes all possible types of genetic damage either on quiescent or germinating conidia (a representative of this class is MMS) and acts presumably on the DNA level; the second acts only on germinating conidia and does not produce point mutation or crossing over. A representative of this class is Benomyl which interferes with spindle microtubules. A list of compounds tests is included.

Conidioma production of the white root rot fungus [Rosellinia] in axenic culture under near-ultraviolet light radiation

International Nuclear Information System (INIS)

Nakamura, H.; Ikeda, K.; Arakawa, M.; Matsumoto, N.

2002-01-01

Conidiomata of the white root rot fungus were produced in axenic culture under near-ultraviolet light radiation. Pieces of sterilized Japanese pear twigs were placed on 7-day-old oatmeal agar culture in plates. The plates were further incubated for 5 days and then illuminated by near-ultraviolet light. Synnemata developed on the twigs within 5 weeks in 19 of 20 isolates tested, and conidia were observed in 12 of the 19 isolates. The synnemata and conidia produced were morphologically identical to those of Dematophora necatrix

PHENOTYPIC CHARACTERISATION AND GENETIC ANALYSIS OF MUTANTS OF ASPERGILLUS NIDULANS RESISTANT TO THE FUNGICIDE TOLCLOFOS-METHYL

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A CHIBANI

2000-12-01

Full Text Available Spontaneous mutants of Aspergillus nidulans were recovered from 0,55.10+7  conidia incubated on synthetic medium supplemented with 100 mg tolclofos-methyl/ml. They differed considerably in morphology, growth rate, and level of resistance to two other fungicides. All mutants tested were cross-resistant to quintozene and vinclozolin; they produced fewer conidia than their wild-type parent. Some mutants required fungicides for maximum growth. Genetic analysis revealed that the mutants carried mutations in one gene located on linkage group III.

An efficient Agrobacterium-mediated transformation method for aflatoxin generation fungus Aspergillus flavus.

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Han, Guomin; Shao, Qian; Li, Cuiping; Zhao, Kai; Jiang, Li; Fan, Jun; Jiang, Haiyang; Tao, Fang

2018-05-01

Aspergillus flavus often invade many important corps and produce harmful aflatoxins both in preharvest and during storage stages. The regulation mechanism of aflatoxin biosynthesis in this fungus has not been well explored mainly due to the lack of an efficient transformation method for constructing a genome-wide gene mutant library. This challenge was resolved in this study, where a reliable and efficient Agrobacterium tumefaciens-mediated transformation (ATMT) protocol for A. flavus NRRL 3357 was established. The results showed that removal of multinucleate conidia, to collect a homogenous sample of uninucleate conidia for use as the transformation material, is the key step in this procedure. A. tumefaciens strain AGL-1 harboring the ble gene for zeocin resistance under the control of the gpdA promoter from A. nidulans is suitable for genetic transformation of this fungus. We successfully generated A. flavus transformants with an efficiency of ∼ 60 positive transformants per 10 6 conidia using our protocol. A small-scale insertional mutant library (∼ 1,000 mutants) was constructed using this method and the resulting several mutants lacked both production of conidia and aflatoxin biosynthesis capacity. Southern blotting analysis demonstrated that the majority of the transformants contained a single T-DNA insert on the genome. To the best of our knowledge, this is the first report of genetic transformation of A. flavus via ATMT and our protocol provides an effective tool for construction of genome-wide gene mutant libraries for functional analysis of important genes in A. flavus.

Host, pathogen, and environmental characteristics predict white-nose syndrome mortality in captive little brown myotis (Myotis lucifugus).

Science.gov (United States)

Johnson, Joseph S; Reeder, DeeAnn M; McMichael, James W; Meierhofer, Melissa B; Stern, Daniel W F; Lumadue, Shayne S; Sigler, Lauren E; Winters, Harrison D; Vodzak, Megan E; Kurta, Allen; Kath, Joseph A; Field, Kenneth A

2014-01-01

An estimated 5.7 million or more bats died in North America between 2006 and 2012 due to infection with the fungus Pseudogymnoascus destructans (Pd) that causes white-nose syndrome (WNS) during hibernation. The behavioral and physiological changes associated with hibernation leave bats vulnerable to WNS, but the persistence of bats within the contaminated regions of North America suggests that survival might vary predictably among individuals or in relation to environmental conditions. To investigate variables influencing WNS mortality, we conducted a captive study of 147 little brown myotis (Myotis lucifugus) inoculated with 0, 500, 5000, 50,000, or 500,000 Pd conidia and hibernated for five months at either 4 or 10°C. We found that female bats were significantly more likely to survive hibernation, as were bats hibernated at 4°C, and bats with greater body condition at the start of hibernation. Although all bats inoculated with Pd exhibited shorter torpor bouts compared to controls, a characteristic of WNS, only bats inoculated with 500 conidia had significantly lower survival odds compared to controls. These data show that host and environmental characteristics are significant predictors of WNS mortality, and that exposure to up to 500 conidia is sufficient to cause a fatal infection. These results also illustrate a need to quantify dynamics of Pd exposure in free-ranging bats, as dynamics of WNS produced in captive studies inoculating bats with several hundred thousand conidia may differ from those in the wild.

Host, pathogen, and environmental characteristics predict white-nose syndrome mortality in captive little brown myotis (Myotis lucifugus.

Directory of Open Access Journals (Sweden)

Joseph S Johnson

Full Text Available An estimated 5.7 million or more bats died in North America between 2006 and 2012 due to infection with the fungus Pseudogymnoascus destructans (Pd that causes white-nose syndrome (WNS during hibernation. The behavioral and physiological changes associated with hibernation leave bats vulnerable to WNS, but the persistence of bats within the contaminated regions of North America suggests that survival might vary predictably among individuals or in relation to environmental conditions. To investigate variables influencing WNS mortality, we conducted a captive study of 147 little brown myotis (Myotis lucifugus inoculated with 0, 500, 5000, 50,000, or 500,000 Pd conidia and hibernated for five months at either 4 or 10°C. We found that female bats were significantly more likely to survive hibernation, as were bats hibernated at 4°C, and bats with greater body condition at the start of hibernation. Although all bats inoculated with Pd exhibited shorter torpor bouts compared to controls, a characteristic of WNS, only bats inoculated with 500 conidia had significantly lower survival odds compared to controls. These data show that host and environmental characteristics are significant predictors of WNS mortality, and that exposure to up to 500 conidia is sufficient to cause a fatal infection. These results also illustrate a need to quantify dynamics of Pd exposure in free-ranging bats, as dynamics of WNS produced in captive studies inoculating bats with several hundred thousand conidia may differ from those in the wild.

An Entomopathogenic Strain of Beauveria bassiana against Frankliniella occidentalis with no Detrimental Effect on the Predatory Mite Neoseiulus barkeri: Evidence from Laboratory Bioassay and Scanning Electron Microscopic Observation

Science.gov (United States)

Wu, Shengyong; Gao, Yulin; Zhang, Yaping; Wang, Endong; Xu, Xuenong; Lei, Zhongren

2014-01-01

Among 28 isolates of Beauveria bassiana tested for virulence against F. occidentalis in laboratory bioassays, we found strain SZ-26 as the most potent, causing 96% mortality in adults at 1×107 mL−1conidia after 4 days. The effect of the strain SZ-26 on survival, longevity and fecundity of the predatory mite Neoseiulus (Amblyseius) barkeri Hughes were studied under laboratory conditions. The bioassay results showed that the corrected mortalities were less than 4 and 8% at 10 days following inoculation of the adult and the larvae of the predator, respectively, with 1×107 conidia mL−1 of SZ-26. Furthermore, no fungal hyphae were found in dead predators. The oviposition and postoviposition durations, longevity, and fecundity displayed no significant differences after inoculation with SZ-26 using first-instar larvae of F. occidentalis as prey in comparison with untreated predator. In contrast, the preoviposition durations were significantly longer. Observations with a scanning electron microscope, revealed that many conidia were attached to the cuticles of F. occidentalis at 2 h after treatment with germ tubes oriented toward cuticle at 24 h, penetration of the insect cuticle at 36 h, and finally, fungal colonization of the whole insect body at 60 h. In contrast, we never observed penetration of the predator's cuticle and conidia were shed gradually from the body, further demonstrating that B. bassiana strain SZ-26 show high toxicity against F. occidentalis but no pathogenicity to predatory mite. PMID:24454744

Pengaruh Kemasaman, Suhu, dan Cahaya terhadap Golovinomyces sordidus Penyebab Penyakit Embun Tepung pada Plantago major

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Dini Florina

2014-11-01

Full Text Available Unidentified powdery mildew was found on leaves of a medicinal plant, Plantago major in Indonesia. The present studies were aimed to identify the causal fungal species of powdery mildew of  P. major and study the effect of acidity, temperature and light on the causal fungus. Identification was conducted by observing morphological characteristics of the fungus scraped from diseased leaves under light microscope. The infection process was observed by staining the inoculated leaves followed observation under light microscope. Conidia were suspended in various pH solutions to examine effect of pH on conidial germination and hyphal length. For testing effects of temperature on conidial germination and hyphal length, conidial suspensions were dropped onto glass slides then incubated in temperature ranges from 20–35 °C, the germinating conidia and length of the existing hypha were counted and measured 24 hours later. With the same method as above the glass slides were incubated in 25 °C either in the dark or exposed under illuminated white light tube (400 Lux for study effect of light on conidial germination. The causal fungus was identified as Golovinomyces sordidus (syn. Erysiphe sordida with its anamorph state as Oidium. The germinating conidia penetrate directly into leaf tissue within 24 hours. The optimal conditions for the conidia to germinate are pH between 4 and 7, temperature between 25 °C and 30 °C, and dark condition.  

CCR4-Not Complex Subunit Not2 Plays Critical Roles in Vegetative Growth, Conidiation and Virulence in Watermelon Fusarium Wilt Pathogen Fusarium oxysporum f. sp. niveum

Science.gov (United States)

Dai, Yi; Cao, Zhongye; Huang, Lihong; Liu, Shixia; Shen, Zhihui; Wang, Yuyan; Wang, Hui; Zhang, Huijuan; Li, Dayong; Song, Fengming

2016-01-01

CCR4-Not complex is a multifunctional regulator that plays important roles in multiple cellular processes in eukaryotes. In the present study, the biological function of FonNot2, a core subunit of the CCR4-Not complex, was explored in Fusarium oxysporum f. sp. niveum (Fon), the causal agent of watermelon wilt disease. FonNot2 was expressed at higher levels in conidia and germinating conidia and during infection in Fon-inoculated watermelon roots than in mycelia. Targeted disruption of FonNot2 resulted in retarded vegetative growth, reduced conidia production, abnormal conidial morphology, and reduced virulence on watermelon. Scanning electron microscopy observation of infection behaviors and qRT-PCR analysis of in planta fungal growth revealed that the ΔFonNot2 mutant was defective in the ability to penetrate watermelon roots and showed reduced fungal biomass in root and stem of the inoculated plants. Phenotypic and biochemical analyses indicated that the ΔFonNot2 mutant displayed hypersensitivity to cell wall perturbing agents (e.g., Congo Red and Calcofluor White) and oxidative stress (e.g., H2O2 and paraquat), decreased fusaric acid content, and reduced reactive oxygen species (ROS) production during spore germination. Our data demonstrate that FonNot2 plays critical roles in regulating vegetable growth, conidiogenesis and conidia morphology, and virulence on watermelon via modulating cell wall integrity, oxidative stress response, ROS production and FA biosynthesis through the regulation of transcription of genes involved in multiple pathways. PMID:27695445

Laboratory and field evaluation of an entomopathogenic fungus, Isaria cateniannulata strain 08XS-1, against Tetranychus urticae (Koch).

Science.gov (United States)

Zhang, Xiaona; Jin, Daochao; Zou, Xiao; Guo, Jianjun

2016-05-01

The two-spotted mite, Tetranychus urticae Koch, is one of the most serious mite pests of crops throughout the world. Biocontrol of the mite with fungal agents has long been paid much attention because of the development of insecticide resistance and the severe restriction of chemical pesticides. In this study, the efficacy of submerged conidia of the entomopathogenic fungus Isaria cateniannulata strain 08XS-1 against T. urticae eggs, larvae and female adults was evaluated at different temperatures and humidity in the laboratory and under field conditions. The results showed that a suspension of 2 × 10(7) submerged conidia mL(-1) caused the highest mortalities of mite eggs, larvae and females (100, 100 and 70% respectively) at 100% relative humidity and 25 °C in the laboratory. In the field experiments against the mites, a suspension of 2 × 10(8) submerged conidia mL(-1) achieved significant efficiency - the relative control effects were 88.6, 83.8 and 83%, respectively, in cucumber, eggplant and bean fields after 10 days of treatment. The results suggest that the I. cateniannulata strain 08XS-1 is a potential fungal agent, with acceptable production cost of conidia, against T. urticae in the field in an area such as southwestern China with higher air humidity. © 2016 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Predatory activity of Butlerius nematodes and nematophagous fungi against Haemonchus contortus infective larvae

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Manoel Eduardo da Silva

Full Text Available Abstract The purpose of this study was to evaluate the predatory activity of the nematode Butlerius spp. and fungal isolates of Duddingtonia flagrans, Clonostachys rosea, Arthrobotrys musiformis and Trichoderma esau against H. contortus infective larvae (L3 in grass pots. Forty-eight plastic gardening pots containing 140 g of sterile soil were used. Panicum spp. grass seeds (200 mg were sown into each pot and individually watered with 10 mL of tap water. Twelve days after seeding, the pots were randomly divided into 6 groups (n=8. Two thousand H. contortus infective larvae (L3 were added to each group. Additionally, the following treatments were established: Group 1 – 2000 Butlerius spp. larvae; group 2 – A. musiformis (1x107 conidia; group 3 – T. esau (1x107 conidia; group 4 – C. rosea (1x107 conidia, group 5 – D. flagrans (1x107conidia and Group 6 – no biological controller (control group. The larval population of H. contortus exposed to Butlerius spp. was reduced by 61.9%. Population reductions of 90.4, 66.7, 61.9 and 85.7% were recorded in the pots containing A. musiformis, T. esau, C. rosea and D. flagrans, respectively. The results of this study indicate that the predatory nematode Butlerius spp. and the assessed fungi display an important predatory activity can be considered suitable potential biological control agents.

The changeability of Pyricularia oryzae Cav. 1. The action of some mutagenous factors

International Nuclear Information System (INIS)

Voinova, T.M.; Terekhova, V.A.; D'yakov, Yu.T.

1983-01-01

The lethal and mutagenous actions of UV rays, nitrozomethylurea, and nitrosoguanidine in respect to Conidia of rice Pyricularia oryzae Cav. agent have been investigated. It has been found out that low doses of UV-radiation, which are not lethal for a three-cell conidia, increase the intensity of two-cell vegetation. All the investigated mutagens cause a formation of mutants which are deficient according to pigment synthesis white and pink colonies and differ by their reduced growth. Auxotrophic mutants were mainly obtained under the action of nitroso compounds

EVALUACIÓN DEL ANTAGONISMO Y MULTIPLICACIÓN DE Trichoderma sp. EN SUSTRATO DE PLÁTANO EN MEDIO LÍQUIDO ESTÁTICO

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Elkin Agamez Ramos

2009-09-01

Full Text Available Se evaluó la capacidad antagónica de nueve aislados del hongo Trichoderma sp. proveniente de la colección de cepas del Laboratorio de Fitopatología de la Universidad de Córdoba, Colombia y su producción de esporas en medios de cultivo líquido estático, a través de pruebas in vitro de antagonismo, utilizando dos hongos fitopatógenos: Fusarium sp. y Rhizoctonia sp.; esta prueba permitió seleccionar el aislado con la mayor actividad antagónica, para así ajustar la producción de esporas mediante un proceso de fermentación en estado líquido en condición estática mediante procesos de buferización y sin buferización con fosfato 0,2N, utilizándose como sustrato principal harina de plátano (HP 5%, además de complementos nutricionales que constituyeron los tratamientos: HPM (solución de melaza 5%, HPL (solución de levadura granulada al 2%, HPU (solución de urea 0,1%, y como testigo PGL (papa glucosa líquida. En los ensayos de antagonismo se seleccionó el aislado C2 de Trichoderma sp. por presentar la mayor capacidad antagónica frente al hongo Fusarium sp., con un porcentaje de inhibición del 78,30% y reportando un índice 4 de antagonismo, en los procesos de producción de esporas el tratamiento HPL (harina de plátano más levadura sin buffer se destacó por su favorable producción de conidias, con una concentración de 1,1 x 109 conidias/mL. Los procesos de buferización se diferenciaron significativamente, demostrando que la variable buffer influyó en la producción de conidias, los medios que se amortiguaron mostraron una producción de 1,01 x 109 conidias/mL, con respecto a los que no se amortiguaron 6,3 x 108 conidias/mL.

Pink Mold Rot on Unishiu Orange (Citrus unshiu Mac. Caused by Trichothecium roseum (Pers. Link ex Gray in Korea

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Jin-Hyeuk Kwon

2013-09-01

Full Text Available In 2012, a pink mold rot was observed on unishiu orange (Citrus unshiu Mac. fruits at the Wholesale Market for Agricultural Products, Jinju, Korea. The symptom on unishiu orange was a water-soaked lesion on the surface of fruit, which later on enlarged to form softened brown rot lesions. The diseased fruits were covered with pink-colored mold, consisting of conidia and conidiophores of the pathogen. Optimum temperature for mycelial growth was 25oC. Conidia were hyaline, smooth, 2-celled, and thick-walled conidia with truncate bases, ellipsoidal to pyriform, characteristically held together zig-zag chains and 12−26 × 8−12 μm in size. Conidiophore was erect, colorless, unbranched, and 4−5 μm wide. On the basis of mycological characteristics, pathogenicity test, and molecular analysis with complete ITS rDNA region, the causal fungus was identified as Trichothecium roseum (Pers. Link ex Gray. This is the first report of pink mold rot caused by T. roseum on unishiu orange in Korea.

Control of the Alphitobius diaperinus (Panzer (Coleoptera: Tenebrionidae with entomopathogenic fungi

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SRF Rezende

2009-06-01

Full Text Available The beetle Alphitobius diaperinus (Panzer, considered a worldwide pest in the poultry industry, is difficult to control and it is a vector for pathogens. The objective of this study was to evaluate the biological control of the lesser mealworm, by strains of fungi Beauveria bassiana, Cladosporium sp. and Trichoderma sp. Larvae and adults of the A. diaperinus were inoculated with suspensions of conidia in the concentration of 10(7 conídia.mL-1. The B. bassiana isolate caused higher insect mortality as compared to Cladosporium sp. and Trichoderma sp. isolates, with the larvae being more susceptible than adults. The entomopathogenicity of B. bassiana was further evaluated with 200 larvae and 200 adults of A. diaperinus inoculated with suspensions 10(6, 10(7, and 10(8 conidia.mL-1, and observed for ten days. Larvae mortality started at the fourth day at the lowest concentration, and the adult mortality was only observed on the sixth day at the concentration of 10(8 conidia.mL-1.

Morphological and ecological variation of Gremmeniella abietina var. abietina in Pinus sylvestris, Pinus contorta and Picea abies sapling stands in northern Finland and the Kola Peninsula

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Kaitera, Juha; Seitamaeki, Leena; Jalkanen, Risto [Finnish Forest Research Inst., Rovaniemi (Finland). Rovaniemi Research Station

2000-07-01

The morphological and ecological variation of two types of Gremmeniella abietina var. abietina causing scleroderris canker on conifers was investigated in Pinus spp. and Picea sp. sapling stands in northern Finland and the Kola Peninsula. Small-tree type (STT or B type) of G. abietina was detected alone in 13 Scots pine, three lodgepole pine and two Norway spruce sapling stands out of 26 stands investigated, both STT and large-tree type (LTT or A type) were observed in six Scots pine stands, and LTT was detected alone in two Scots pine stands. For the first time, G. abietina was found to injure Norway spruce saplings in a respective plantation in northern Fennoscandia. STT isolates produced statistically significantly more conidia in vitro than LTT isolates. Morphological variation in conidia septation revealed that STT produced conidia with more than five septa more frequently than did LTT. There was a greater range in variation in septation in STT than in LTT, with overlapping between the types. Isolates of both types were equally associated with cankers, coloured wood, pycnidia or apothecia in the infected saplings.

Evaluation in greenhouse of the response of two Musa cultivar to Black Sigatoka by artificial inoculation of conidial suspension of Pseudocercospora fijiensis

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Mayra Acosta Suárez

2004-04-01

Full Text Available For evaluating the resistance of Black sigatoka in genotypes from breeding programmes different methods have been used, the artificial inoculation of pathogens reproduction structures is one of them. Nevertheless, is necessary the standardization of condition for it in vitro production and to know the response of cultivars to the diseases in greenhouse. It will permit to establish methods for evaluating and selecting the new resistant genotypes. This work was carried out with the objectives to evaluate different culture media and incubation conditions for obtaining P. fijiensis conidia and for evaluating the response of two cultivars to the artificial inoculation with conidial suspensions of this fungus. Different culture media and incubation conditions for obtaining the conidia and the cultivars Grande naine (susceptible and FHIA-18 (partially resistant were used. The conidia of P. fijiensis could be obtained in vitro at 20ºC and constant light from 10 days of incubation in culture media Potato Carrot Agar, modified V-8 Agar and Potato Dextrose Agar. However, the greatest concentration was reached in Potato Dextrose Agar after 20 days of incubation at 20ºC under continuous white fluorescent light. The artificial inoculation of conidial suspensions of P. fijiensis allowed to evaluate the response of Grande Naine and FHIA-18 cultivars in greenhouse. By the use of qualitative scale of symptoms stages and the quantitative variables symptoms evolution time and disease development time was possible to differentiate both cultivars. They maintained in greenhouse similar response to the disease that in the field. Key words: conidia, culture medium, FHIA-18, Grande naine, sporulation

Rapid enumeration of low numbers of moulds in tea based drinks using an automated system.

Science.gov (United States)

Tanaka, Kouichi; Yamaguchi, Nobuyasu; Baba, Takashi; Amano, Norihide; Nasu, Masao

2011-01-31

Aseptically prepared cold drinks based on tea have become popular worldwide. Contamination of these drinks with harmful microbes is a potential health problem because such drinks are kept free from preservatives to maximize aroma and flavour. Heat-tolerant conidia and ascospores of fungi can survive pasteurization, and need to be detected as quickly as possible. We were able to rapidly and accurately detect low numbers of conidia and ascospores in tea-based drinks using fluorescent staining followed by an automated counting system. Conidia or ascospores were inoculated into green tea and oolong tea, and samples were immediately filtered through nitrocellulose membranes (pore size: 0.8 μm) to concentrate fungal propagules. These were transferred onto potato dextrose agar and incubated for 23 h at 28 °C. Fungi germinating on the membranes were fluorescently stained for 30 min. The stained mycelia were counted selectively within 90s using an automated counting system (MGS-10LD; Chuo Electric Works, Osaka, Japan). Very low numbers (1 CFU/100ml) of conidia or ascospores could be rapidly counted, in contrast to traditional labour intensive techniques. All tested mould strains were detected within 24h while conventional plate counting required 72 h for colony enumeration. Counts of slow-growing fungi (Cladosporium cladosporioides) obtained by automated counting and by conventional plate counting were close (r(2) = 0.986). Our combination of methods enables counting of both fast- and slow-growing fungi, and should be useful for microbiological quality control of tea-based and also other drinks. Copyright © 2011 Elsevier B.V. All rights reserved.

An entomopathogenic strain of Beauveria bassiana against Frankliniella occidentalis with no detrimental effect on the predatory mite Neoseiulus barkeri: evidence from laboratory bioassay and scanning electron microscopic observation.

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Shengyong Wu

Full Text Available Among 28 isolates of Beauveria bassiana tested for virulence against F. occidentalis in laboratory bioassays, we found strain SZ-26 as the most potent, causing 96% mortality in adults at 1×10(7 mL(-1conidia after 4 days. The effect of the strain SZ-26 on survival, longevity and fecundity of the predatory mite Neoseiulus (Amblyseius barkeri Hughes were studied under laboratory conditions. The bioassay results showed that the corrected mortalities were less than 4 and 8% at 10 days following inoculation of the adult and the larvae of the predator, respectively, with 1×10(7 conidia mL(-1 of SZ-26. Furthermore, no fungal hyphae were found in dead predators. The oviposition and postoviposition durations, longevity, and fecundity displayed no significant differences after inoculation with SZ-26 using first-instar larvae of F. occidentalis as prey in comparison with untreated predator. In contrast, the preoviposition durations were significantly longer. Observations with a scanning electron microscope, revealed that many conidia were attached to the cuticles of F. occidentalis at 2 h after treatment with germ tubes oriented toward cuticle at 24 h, penetration of the insect cuticle at 36 h, and finally, fungal colonization of the whole insect body at 60 h. In contrast, we never observed penetration of the predator's cuticle and conidia were shed gradually from the body, further demonstrating that B. bassiana strain SZ-26 show high toxicity against F. occidentalis but no pathogenicity to predatory mite.

Innate and adaptive immune response to chronic pulmonary infection of hyphae of Aspergillus fumigatus in a new murine model.

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Wang, Fengyuan; Zhang, Caiyun; Jiang, Yuan; Kou, Caixia; Kong, Qingtao; Long, Nanbiao; Lu, Ling; Sang, Hong

2017-10-01

The pathogenesis of chronic pulmonary aspergillosis (CPA) has seldom been studied due partly to a lack of animal models. Since hypha is the main morphology colonizing the airway in CPA, it's critical to study the immune reaction to chronic pulmonary infection of hyphae of Aspergillus fumigatus, which also has seldom been studied in vivo before. We established a novel murine model of chronic pulmonary infection of hyphae by challenging immunocompetent mice with tightly-structured hyphae balls intratracheally, and described the ensuing immunoreaction to hyphae and conidia, and the pathogenesis of CPA. Our experiment proved that the hyphae balls could induce a chronic pulmonary infection for 28 days with a considerable recrudescence at day 28 post-infection. Lungs infected with hyphae balls were remarkable for the many neutrophils and macrophages that flooded into airway lumens, with peribronchiolar infiltration of leukocytes. There was a transient increase of Th2 cells and Th17 cells at day 7 post-infection in the lung tissue. In contrast, lungs infected with conidia showed no peribronchiolar infiltration of leukocytes, but an influx of a great number of macrophages, and a much less number of neutrophils in the lumen. Besides, conidia activated the co-response of Th1, Th2 and Th17 cells with an increase of Treg cells in the lung tissue (quite different from most previous studies). We established a new murine model of chronic infection of hyphae to mimic the formation of CPA, and provide a new marker for different immune responses to hyphae and conidia.

Cultivation of Aschersonia placenta Berkeley and Broom and its efficacy for controlling Parlatoria ziziphi (Lucas (Hemiptera: Diaspididae

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Dokgluaymai Homrahud

2016-05-01

Full Text Available The entomopathogenic fungus genus Aschersonia (Deuteromycotina: Hyphomycetes is host specific to some aleyrodids and scale insects. In search of the Thai endemic species, fungus samples were isolated from cadavers of citrus whiteflies (Aleyrodes tabaci Gennadius found in citrus orchards in Trat province, Thailand. After morphological analysis and scanning electron microscopic examination, it was identified as Aschersonia placenta Berkeley and Broom. Seven synthetic media, namely: potato dextrose agar (PDA, PDA with pasteurized milk (Foremost® (PDA + M, Sabouraud dextrose agar with yeast extract (SDAY, SDA with pasteurized milk (Foremost® (SDA + M, corn meal agar (CMA, water agar with juice of eight vegetable species (V8® (WA + V8 and WA were explored as appropriate media for fungal cultivation. SDAY and SDA + M gave the best colony radial growth, producing 2.04 ± 0.13 cm and 2.09 ± 0.10 cm in 21 d, respectively. However, based on the ability of A. placenta to produce conidia, PDA and SDAY which produced 2.59 × 108 conidia/mL and 2.69 × 108 conidia/mL, respectively, were considered as the most suitable media for this fungal species. The efficiency assessment of A. placenta for controlling black parlatoria (Parlatoria ziziphi (Lucas, indicated that a conidial suspension at 1 × 109 conidia/mL gave 23.73% and 27.42% mortality at 14 and 21 d post inoculation, respectively.

Phylogenetic variation of the green muscadine fungus, Metarhizium anisopliae (Metchnikoff Sorokin, and its virulence to larvae of the sugarcane longhorn stem borer, Dorysthenes buqueti Guerin (Coleoptera: Cerambycidae

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Nichanun Kernasa

2016-11-01

Full Text Available The sugarcane longhorn stem borer (SLSB, Dorysthenes buqueti Guerin (Coleoptera: Cerambycidae has recently become a serious insect pest of sugarcane in Thailand and effective biological control agent must be evaluated. The green muscadine fungus (GMF, Metarhizium anisopliae (Metchnikoff Sorokin is a species complex of entomopathogenic fungi, which includes many cryptic subspecies and species. It has been reported that GMF infects and kills the sugarcane longhorn stem borer (SLSB, D. buqueti Guerin, so that GMF is a possible biological control agent of SLSB. Molecular analyses were conducted to gain a better understanding of the taxonomic position of GMF Thai strains. Virulence bioassays were carried out on four isolates of GMF to 5th–9th instars of SLSB. This study revealed that an isolate from Khon Kaen (KK showed the highest virulence to 5th–9th instars of SLSB. In biological control, an aqueous suspension containing 1 × 108 conidia/mL of KK isolate was best from the viewpoint of a tradeoff between the economic cost/benefit of the mass production cost and the consequent mortality after application. Comparing suspensions containing 1 × 108 conidia/mL with those containing 1 × 1013 conidia/mL, 100,000 times as much quantity of suspension can be obtained from the same quantity of conidia, though the difference in the D. buqueti mortality was relatively small. Six isolates of GMF from SLSB in Thailand were likely a cryptic species, although further molecular analysis using factor 1-alpha sequences is needed.

[Viability of nematophagous fungi Arthrobotrys robusta, Duddingtonia flagrans and Monacrosporium thaumasium after sporulation in different culture media].

Science.gov (United States)

Maciel, Alessandro S; de Araujo, Jackson V; Campos, Artur K

2006-01-01

Due to the shortage of studies that indicate the culture mediums that optimize the sporulation of namatophagous fungi for use in researche, the sporulation of the fungal isolates A. robusta (I31), D. flagrans (CG768) and M. thaumasium (NF34A) was evaluated in laboratorial conditions for 10 days in the means water-agar 2% (WA 2%), potato-dextrose-agar 2% (PDA 2%), corn-meal-agar 2% (CMA 2%) and yeast-phosphate-sulphate-sucrose-agar (YPSSA). The largest conidia production (P BDA 2% while in the isolates I31 and NF34A produced larger conidia number in YPSSA (P 0.05).

Influence of ultraviolet light on arising of induced mutants in Cercospora beticola sacc

Energy Technology Data Exchange (ETDEWEB)

Brillova, D [Institute of Experimental Phytopathology and Entomology of the Slovak Academy of Sciences, Ivanka pri Dunaji (Czechoslovakia)

1976-01-01

Ultraviolet radiation of wavelengths of 254 and 350 nm respectively, applied for 30 to 480 seconds to the conidia of Cercospora beticola, induced a large number of mutants. According to their appearance, the occurring mutants can be considered as visible with effect on morphology and colour. A considerable part of the mutants lost its ability to form reproductive organs in in vitro conditions, as well as on the host plant; they became avirulent. Moreover, mutants occurred with decreased virulence, with a weak forming of conidia and prolonged incubation period. In few cases, also reverse mutations were induced characterized by increased virulence.

Single application prophylaxis against gray mold in pot rose and pelargonium with Ulocladium atrum

DEFF Research Database (Denmark)

Yohalem, D S; Paaske, K; Kristensen, K

2007-01-01

Experiments were conducted to clarify the optimal dose and timing of single applications of Ulocladium atrum for control of gray mold on pot rose and pelargonium under conditions similar to those found in commercial greenhouses. In general, application of the antagonist 1-d prior to infestation...... with conidia of Botrytis cinerea was more effective than application after the infection period. For pot rose, the minimum effective dose when compared to the fungicide fenhexamid was 106 conidia ml-1 and persisted for up to 21-d in direct comparison to the fungicide. The effect of pre-emptive application...

Inactivation of certain insect pathogens by ultraviolet radiation

Energy Technology Data Exchange (ETDEWEB)

Krieg, A.; Groener, A.; Huber, J.; Zimmermann, G.

1981-01-01

The UV-sensitivity of two baculoviruses (granulosis virus, nuclear polyhedrosis virus) and two entomopathogenic microorganisms (Bacillus thuringiensis, Beauveria bassiana) was determined by radiation tests. In the far UV (254 nm) the stability, measured at an inactivation rate of 99%, was in declining order: nuclear polyhedra >= conidia of B. bassiana > granula > spores of B. thuringiensis >= vegetative cells of B. thuringiensis. In the near UV (285-380 nm) the following order could be found: conidia of B. bassiana >= nuclear polyhedra > spores of B. thuringiensis >= granula > vegetative cells of B. thuringiensis. Far UV had a much higher germicidal effect for all pathogens tested than near UV.

Microfungi of the Tatra Mts. 6. Fungus-like organisms: Albuginales, Peronosporales and Pythiales

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Wiesław Mułenko

2014-06-01

Full Text Available A list and the distribution of Oomycota species in the Tatra Mts (Western Carpathian Mts are presented. Revised herbarium vouchers and literature data were used for analysis. Thirty two species of oomycetes on fifty seven plant species were noted in the area, including two species of the order Albuginales (genera: Albugo and Pustula, on nine plant species, 29 species of the order Peronosporales (genera: Bremia, Hyaloperonospora, Peronospora and Plasmopara, on 49 plant species, and one species of the order Pythiales (genus: Myzocytium, on one species of algae. Twenty nine species were collected on the Polish side of the Tatra Mts and ten species were collected on the Slovak side. The oomycetes were collected at 185 localities.

New cost-effective bioconversion process of palm kernel cake into bioinsecticides based on Beauveria bassiana and Isaria javanica.

Science.gov (United States)

do Nascimento Silva, Jaqueline; Mascarin, Gabriel Moura; Dos Santos Gomes, Isabel Cristina; Tinôco, Ricardo Salles; Quintela, Eliane Dias; Dos Reis Castilho, Leda; Freire, Denise Maria Guimarães

2018-03-01

The present study aimed to add value to palm oil by-products as substrates to efficiently produce conidia of Beauveria bassiana and Isaria javanica (Hypocreales: Cordycipitaceae) for biological control of the whitefly Bemisia tabaci (Hemiptera: Aleyrodidae), through a solid-state fermentation process using palm kernel cake and palm fiber as nutrient source and solid matrix, respectively. The optimum culture conditions yielded high concentrations of viable conidia after air-drying, when the fungi were grown on palm kernel cake (B. bassiana 7.65 × 10 9 and I. javanica 2.91 × 10 9  conidia g -1 dry substrate) after 6 days under optimal growth conditions set to 60% substrate moisture and 32 °C. Both fungal strains exhibited high efficacy against third-instar whitefly nymphs, inducing mortality up to 62.9 and 56.6% by B. bassiana and I. javanica, respectively, assessed after 9 days post-application in a screenhouse. Furthermore, we noted that insect mortality was strongly correlated with high atmospheric moisture, while B. bassiana appeared to require shorter accumulative hours under high moisture to kill whitefly nymphs compared to I. javanica. Our results underpin a feasible and cost-effective mass production method for aerial conidia, using palm kernel as the main substrate in order to produce efficacious fungal bioinsecticides against an invasive whitefly species in Brazil. Finally, our fermentation process may offer a sustainable and cost-effective means to produce eco-friendly mycoinsecticides, using an abundant agro-industrial by-product from Brazil that will ultimately assist in the integrated management of agricultural insect pests.

Correlation of cell surface proteins of distinct Beauveria bassiana cell types and adaption to varied environment and interaction with the host insect.

Science.gov (United States)

Yang, Zhi; Jiang, Hongyan; Zhao, Xin; Lu, Zhuoyue; Luo, Zhibing; Li, Xuebing; Zhao, Jing; Zhang, Yongjun

2017-02-01

The insect fungal pathogen Beauveria bassiana produces a number of distinct cell types that include aerial conidia, blastospores and haemolymph-derived cells, termed hyphal bodies, to adapt varied environment niches and within the host insect. These cells display distinct biochemical properties and surface structures, and a highly ordered outermost brush-like structure uniquely present on hyphal bodies, but not on any in vitro cells. Here, we found that the outermost structure on the hyphal bodies mainly consisted of proteins associated to structural wall components in that most of it could be removed by dithiothreitol (DTT) or proteinase K. DTT-treatment also caused delayed germination, decreased tolerance to ultraviolet irradiation and virulence of conidia or blastospores, with decreased adherence and alternated carbohydrate epitopes, suggesting involvement in fungal development, stress responses and virulence. To characterize these cell surface molecules, proteins were released from the living cells using DTT, and identified and quantitated using label-free quantitative mass spectrometry. Thereafter, a series of bioinformatics programs were used to predict cell surface-associated proteins (CSAPs), and 96, 166 and 54 CSAPs were predicted from the identified protein pools of conidia, blastospores and hyphal bodies, respectively, which were involved in utilization of carbohydrate, nitrogen, and lipid, detoxification, pathogen-host interaction, and likely other cellular processes. Thirteen, sixty-nine and six CSAPs were exclusive in conidia, blastospores and hyphal bodies, respectively, which were verified by eGFP-tagged proteins at their N-terminus. Our data provide a crucial cue to understand mechanism of B. bassiana to adapt to varied environment and interaction with insect host. Copyright © 2016 Elsevier Inc. All rights reserved.

VeA of Aspergillus niger increases spore dispersing capacity by impacting conidiophore architecture.

Science.gov (United States)

Wang, Fengfeng; Dijksterhuis, Jan; Wyatt, Timon; Wösten, Han A B; Bleichrodt, Robert-Jan

2015-01-01

Aspergillus species are highly abundant fungi worldwide. Their conidia are among the most dominant fungal spores in the air. Conidia are formed in chains on the vesicle of the asexual reproductive structure called the conidiophore. Here, it is shown that the velvet protein VeA of Aspergillus niger maximizes the diameter of the vesicle and the spore chain length. The length and width of the conidiophore stalk and vesicle were reduced nearly twofold in a ΔveA strain. The latter implies a fourfold reduced surface area to develop chains of spores. Over and above this, the conidial chain length was approximately fivefold reduced. The calculated 20-fold reduction in formation of conidia by ΔveA fits the 8- to 17-fold decrease in counted spore numbers. Notably, morphology of the ΔveA conidiophores of A. niger was very similar to that of wild-type Aspergillus sydowii. This suggests that VeA is key in conidiophore architecture diversity in the fungal kingdom. The finding that biomass formation of the A. niger ΔveA strain was reduced twofold shows that VeA not only impacts dispersion capacity but also colonization capacity of A. niger.

OCCURRENCE IN THE SOIL AND DISPERSAL OF Lecanicillium lecanii, A FUNGAL PATHOGEN OF THE GREEN COFFEE SCALE (Coccus viridis AND COFFEE RUST (Hemileia vastatrix

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Doug William Jackson

2012-08-01

Full Text Available The fungus Lecanicillium lecanii attacks the green scale (Coccus viridis, a pest of coffee, and is also a hyperparasite of coffee rust (Hemileia vastatrix. Knowledge of the epizootiology of this fungus is potentially important for conservation biological control in coffee agroecosystems. The presence of viable propagules of L. lecanii in the soil, a possible environmental reservoir, was assessed using two baiting methods: the standard Galleria mellonella bait method and a C. viridis bait method. Infectious propagules of L. lecanii were detected in soil samples taken from a 45 ha study plot, both nearby and far from recent epizootics of L. lecanii. To test the potential for the transmission of L. lecanii conidia from the soil via rain splash or wind, coffee seedlings with populations of C. viridis were placed near L. lecanii-inoculated soil and then subjected to artificial rain and wind treatments. Rain splash was shown to be a potential transmission mechanism. Dispersal of L. lecanii conidia by the ant Azteca instabilis was tested using field and laboratory ant-exclusion experiments. Azteca instabilis was shown to transport conidia of L. lecanii; however, dispersal by A. instabilis may not be important under field conditions.

An extra-domiciliary method of delivering entomopathogenic fungus, Metharizium anisopliae IP 46 for controlling adult populations of the malaria vector, Anopheles arabiensis

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Mnyone Ladslaus L

2010-03-01

Full Text Available Abstract Fungal biopesticides have the potential to significantly reduce densities of malaria vectors as well as associated malaria transmission. In previous field trials, entomopathogenic fungus was delivered from within human dwellings, where its efficacy was limited by low infection rates of target mosquitoes, high costs of spraying fungus inside houses, and potential public health concerns associated with introducing fungal conidia inside houses. Here we have demonstrated that Metarhizium anisopliae IP 46, delivered within an extra-domiciliary odor-baited station (OBS, can infect and slowly-kill a high proportion of the wild adult malaria vector, Anopheles arabiensis which entered and exited the OBS. This study, carried out in rural Tanzania, showed that by using a concentration of 3.9 × 1010 conidia/m2, more than 95% of mosquitoes that flew in and out of the OBS died within 14 days post-exposure. At least 86% infection of mosquito cadavers was recorded with a significant reduction in the probability of daily survival of exposed An. arabiensis in both treatments tested: low quantity of conidia (eave baffles plus one cotton panel; HR = 2.65, P P

The Efficiency of Noni (Morinda citrifolia L.) Essential Oil on the Control of Leaf Spot Caused by Exserohilum turcicum in Maize Culture.

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Silva, Janaina Costa E; Mourão, Dalmarcia de Sousa Carlos; Lima, Fabia Silva de Oliveira; Sarmento, Renato de Almeida; Dalcin, Mateus Sunti; Aguiar, Raimundo Wagner de Souza; Santos, Gil Rodrigues Dos

2017-08-14

The objective of this work was to evaluate the efficiency of noni essential oil on the control of Exserohilum turcicum , a causative agent of Exserohilum spot in maize culture. In the sanitary test 400 seeds were incubated using the blotter test method. For the transmissibility test, the fragments of damaged leaves of seedlings were removed and put into a potato, dextrose and agar (PDA) culture environment. To verify the pathogenicity, Koch´s postulates were performed. In the phytotoxicity test different concentrations of noni oil were applied in maize seedlings. E. turcicum conidia were submitted to different concentrations of noni oil. In the preventive and curative tests noni essential oils were applied before and after the conidia inoculation, respectively. The results revealed the presence of fungi of the genres Aspergillus , Penicillium , Rhizopus , Fusarium , and Exserohilum in the maize seeds. The pathogenicity of E. turcicum and also the transmission of this fungus from the seeds to the maize seedlings was confirmed. The inhibition of conidia germination was proportional to the concentration increase. The preventive application of noni essential oil was the most efficient on the control of Exserohilum spot.

Baeuveria bassiana (BalsVuill and Metarhizium anisioplae (Metsch. Sorokin in the pupas control of Prodiplosis longifila Gagné on asparagus crop

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Carolina Cedano

2012-03-01

Full Text Available The objective of the present work was to evaluate the effect of the entomopathogen fungi Baeuveria bassiana and Metarhizium anisioplae of the Prodiplosis longifila pupas, in order to reduce the adult population of this insect. The treatments consisted on the application of two different propagel concentrations (mycel and conidia of each entomopathogen. One of these concentration was 1x106 propagels/ml (obtained from the total amount of conidia collected of 25 kg of rice colonized by the entomopathogen; and the other one, was 1x107 propagels/ml (obtained from the total amount of conidia collected of 40 kg of rice colonized by the entomopathogen. As a check a non application treatment was considered. The applications were trough the irrigation system and started 15 days after the end of harvest and were repeated each five days during a month, coincident with the period of most falls of pupas of the soil. As a result Baeuveria bassiana at 1x107 propagels/ml shown 53.4 % of the total pupas colonized by the entomopathogen, which allows indicating B. bassiana as a promissory biocontrol of this specie

Cellulose hydrolysis by fungi. 1. Screening of cellulolytic strains

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Roussos, S.; Raimbault, M. (Laboratoire de Microbiologie ORSTOM, Centre de Recherche IRCHA, 91 - Vert-le-Petit (France))

Trichoderma harzianum was selected from 30 strains of cellulolytic fungi with the aim of producing cellulases by solid state fermentation of lignocellulosic substrates. Special attention was paid to cellulase production (i. e. carboxymethylcellulase and filter paper activity), apical growth and conidia production. Under the conditions of our experiments, T. harzianum exhibited the highest cellulasic activities with 1,315 IU/I of carboxymethyl cellulose and 80 IU/l of filter paper activity. Apical growth (1 mm/h) and yield of conidial production (3.25 X 10/sup 10/ conidia/g of substrate dry weight) were also valuable characteristics of this strain in the use of solid state fermentation.

Real-time visualization of immune cell clearance of Aspergillus fumigatus spores and hyphae.

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Knox, Benjamin P; Huttenlocher, Anna; Keller, Nancy P

2017-08-01

Invasive aspergillosis (IA) is a disease of the immunocompromised host and generally caused by the opportunistic fungal pathogen Aspergillus fumigatus. While both host and fungal factors contribute to disease severity and outcome, there are fundamental features of IA development including fungal morphological transition from infectious conidia to tissue-penetrating hyphae as well as host defenses rooted in mechanisms of innate phagocyte function. Here we address recent advances in the field and use real-time in vivo imaging in the larval zebrafish to visually highlight conserved vertebrate innate immune behaviors including macrophage phagocytosis of conidia and neutrophil responses post-germination. Copyright © 2017 Elsevier Inc. All rights reserved.

Inactivation of certain insect pathogens by ultraviolet radiation

International Nuclear Information System (INIS)

Krieg, A.; Groener, A.; Huber, J.; Zimmermann, G.

1981-01-01

The UV-sensitivity of two baculoviruses (granulosis virus, nuclear polyhedrosis virus) and two entomopathogenic microorganisms (Bacillus thuringiensis, Beauveria bassiana) was determined by radiation tests. In the far UV (254 nm) the stability, measured at an inactivation rate of 99%, was in declining order: nuclear polyhedra >= conidia of B. bassiana > granula > spores of B. thuringiensis >= vegetative cells of B. thuringiensis. In the near UV (285-380 nm) the following order could be found: conidia of B. bassiana >= nuclear polyhedra > spores of B. thuringiensis >= granula > vegetative cells of B. thuringiensis. Far UV had a much higher germicidal effect for all pathogens tested than near UV. (orig.) [de

White blister rusts and downy mildews from bajaur agency fata, with some new records from pakistan

International Nuclear Information System (INIS)

Haq, M.A.; Shahzad, S.

2015-01-01

In a species diversity study of Oomycyctes of Bajaur Agency FATA, Pakistan, infection of white blister rusts and downy mildews recorded on three cultivated and four wild plants. Capsella bursa-pastoris showed mixed infection of Albugo candida and Hyaloperonospora parasitica (syn: Peronospora parasitica). Similarly, A. candida and H. brassicae (syn: P. brassicae) parasitized Brassica campestris. Wilsoniana portulacae (syn: Albugo portulacae) and W. occidentalis com. nov. (syn: Albugo occidentalis) recovered from Portulaca oleracea and Spinacia oleracea, respectively. Bremia taraxaci, B. sonchicola and B. saussureae recorded on Taraxicum officinale, Sonchus sp., and Saussurea sp., respectively. All these obligate parasites are new records from Bajaur Agency, while H. parasitica, W. occidentalis, B. taraxaci, and B. saussureae on the mentioned hosts are new records from Pakistan. (author)

A re-evaluation of the genus Myceliophthora (Sordariales, Ascomycota): its segregation into four genera and description of Corynascus fumimontanus sp. nov.

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Marin-Felix, Yasmina; Stchigel, Alberto M; Miller, Andrew N; Guarro, Josep; Cano-Lira, José F

2015-01-01

Based on a number of isolates of Myceliophthora (Chaetomiaceae, Sordariales, Ascomycota) recently isolated from soil samples collected in USA, the taxonomy of the genus was re-evaluated through phylogenetic analyses of sequences from the nuc rDNA internal transcribed spacer region and genes for the second largest subunit of RNA polymerase II and translation elongation factor 1α. Members of Myceliophthora were split into four monophyletic clades strongly supported by molecular and phenotypic data. Such clades correspond with Myceliophthora, now restricted only to the type species of the genus Corynascus, which is re-established with five species, the new monotypic genus Crassicarpon and also the new genus Thermothelomyces (comprising four species). Myceliophthora lutea is mesophilic and a permanently asexual morph compared to the members of the other three mentioned genera, which also are able to sexually reproduce morphs with experimentally proven links to their asexual morphs. The asexual morph of M. lutea is characterized by broadly ellipsoidal, smooth-walled conidia with a wide, truncate base. Crassicarpon thermophilum is thermophilic and heterothallic and produces spherical to cuneiform, smooth-walled conidia and cleistothecial ascomata of smooth-walled, angular cells and ascospores with a germ pore at each end. Corynascus spp. are homothallic and mesophilic and produce spherical, mostly ornamented conidia and cleistothecial ascomata with textura epidermoidea composed of ornamented wall cells, and ascospores with one germ pore at each end. Thermothelomyces spp. are thermophilic, heterothallic and characterized by similar ascomata and conidia as Corynascus spp., but its ascospores exhibit only a single germ pore. A dichotomous key to distinguish Myceliophthora from the other mentioned genera are provided, as well as dichotomous keys to identify the species of Corynascus and Thermothelomyces. A new species, namely Corynascus fumimontanus, characterized by

[Effects of Beauveria bassiana on Myzus persicae and its two predaceous natural enemies].

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Zhu, Hong; Luo, Xu-mei; Song, Jin-xin; Fan, Mei-zhen; Li, Zeng-zhi

2011-09-01

A Beauveria bassiana strain Bb21 was isolated from naturally infected green peach aphid Myzus persicae (Hemiptera: Aphididae). The effects of the strain on M. persicae and its two predaceous natural enemies Chrysoperla carnea (Neuroptera: Chrysopidae) and Harmonia axyridis (Coleoptera: Coccinellidae) were investigated under laboratory conditions. Bb21 had strong pathogenicity to M. persicae, with the LD50 of 97 conidia x mm(-2) (45-191, 95% confidence interval), but was less pathogenic to the second instar nymph of C. carnea, with the LD50 of 1089 conidia x mm(-2). The LD50 for C. carnea was 10.2 times higher than that for M. persicae. The pathogenicity of Bb21 to H. axyridis was very weak, with a low infection rate of 13% even at a high concentration 5 x 10(8) conidia x mL(-1). The Bb21 at low conidia concentration had less effect on the developmental period and fecundity of the two predaceous natural enemies. However, when applied at the high concentration 5 x 10(8) spores x mL(-1), Bb21 shortened the larval stage of H. axyridis averagely by 1.4 d and decreased the adult emergence rate and fecundity by 33% and 14%, respectively, and shortened the larval stage of C. carnea averagely by 0.7 d and decreased the adult emergence rate and fecundity by 24% and 11%, respectively. Since the LD50 for green peach aphid was much lower than that for the two predaceous natural enemies, and had very low effect on the adult emergence rate and fecundity of the two predators at the concentration recommended for field spray, Bb21 could be applied as a biocontrol agent of M. persicae in the integrated management of pernicious organisms.

Development of a population-based threshold model of conidial germination for analysing the effects of physiological manipulation on the stress tolerance and infectivity of insect pathogenic fungi.

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Andersen, M; Magan, N; Mead, A; Chandler, D

2006-09-01

Entomopathogenic fungi are being used as biocontrol agents of insect pests, but their efficacy can be poor in environments where water availability is reduced. In this study, the potential to improve biocontrol by physiologically manipulating fungal inoculum was investigated. Cultures of Beauveria bassiana, Lecanicillium muscarium, Lecanicillium longisporum, Metarhizium anisopliae and Paecilomyces fumosoroseus were manipulated by growing them under conditions of water stress, which produced conidia with increased concentrations of erythritol. The time-course of germination of conidia at different water activities (water activity, aw) was described using a generalized linear model, and in most cases reducing the water activity of the germination medium delayed the onset of germination without affecting the distribution of germination times. The germination of M. anisopliae, L. muscarium, L. longisporum and P. fumosoroseus was accelerated over a range of aw levels as a result of physiological manipulation. However, the relationship between the effect of physiological manipulation on germination and the osmolyte content of conidia varied according to fungal species. There was a linear relationship between germination rate, expressed as the reciprocal of germination time, and aw of the germination medium, but there was no significant effect of fungal species or physiological manipulation on the aw threshold for germination. In bioassays with M. anisopliae, physiologically manipulated conidia germinated more rapidly on the surface of an insect host, the melon cotton aphid Aphis gossypii, and fungal virulence was increased even when relative humidity was reduced after an initial high period. It is concluded that physiological manipulation may lead to improvements in biocontrol in the field, but choice of fungal species/isolate will be critical. In addition, the population-based threshold model used in this study, which considered germination in terms of physiological

Development of a Biofungicide Using a Mycoparasitic Fungus Simplicillium lamellicola BCP and Its Control Efficacy against Gray Mold Diseases of Tomato and Ginseng

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Teak Soo Shin

2017-06-01

Full Text Available To develop a commercial product using the mycoparasitic fungus Simplicillium lamellicola BCP, the scaleup of conidia production from a 5-l jar to a 5,000-l pilot bioreactor, optimization of the freeze-drying of the fermentation broth, and preparation of a wettable powder-type formulation were performed. Then, its disease control efficacy was evaluated against gray mold diseases of tomato and ginseng plants in field conditions. The final conidial yields of S. lamellicola BCP were 3.3 × 10⁸ conidia/ml for a 5-l jar, 3.5 × 10⁸ conidia/ml for a 500-l pilot vessel, and 3.1 × 10⁸ conidia/ml for a 5,000-l pilot bioreactor. The conidial yield in the 5,000-l pilot bioreactor was comparable to that in the 5-l jar and 500-l pilot vessel. On the other hand, the highest conidial viability of 86% was obtained by the freeze-drying method using an additive combination of lactose, trehalose, soybean meal, and glycerin. Using the freeze-dried sample, a wettable powder-type formulation (active ingredient 10%; BCP-WP10 was prepared. A conidial viability of more than 50% was maintained in BCP-WP10 until 22 weeks for storage at 40°C. BCP-WP10 effectively suppressed the development of gray mold disease on tomato with control efficacies of 64.7% and 82.6% at 500- and 250-fold dilutions, respectively. It also reduced the incidence of gray mold on ginseng by 65.6% and 81.3% at 500- and 250-fold dilutions, respectively. The results indicated that the new microbial fungicide BCP-WP10 can be used widely to control gray mold diseases of various crops including tomato and ginseng.

Efecto antifúngico de hidrodestilados de Zingiber officinale Roscoe sobre Moniliophthora roreri (Cif&Par

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José Gregorio Joya-Dávila

2015-07-01

Full Text Available La moniliasis, ocasionada por el hongo Moniliophthora roreri (Cif. & Par, es la principal enfermedad del cacao en Latinoamérica, y las medidas para su manejo sostenible son aún escasas. Con base en reportes previos, el objetivo de este trabajo fue optimizar el proceso de extracción por destilación de Zingiber officinale, evaluando in vitro su efecto antifúngico sobre conidias de M. roreri. Se elaboraron ocho hidrodestilados con material fresco (300 y 600 g/L y seco al sol (45 y 90 g/L, con dos solventes (agua-alcohol, relación 10:1 y 10:0, y en tubos de ensayo con conidias del hongo, mediante la técnica de cultivo en medio líquido (agua-extracto de cacao, se adicionó cada hidrodestilado de Z. officinale en relación 1:1 v/v, determinando su efecto sobre la formación y germinación de conidias del hongo en cinco momentos de observación (0, 24, 48, 72 y 96 horas. Los resultados muestran que todos los hidrodestilados de Z. officinale presentan metabolitos con efectos de inhibición sobre las variables evaluadas, siendo el tratamiento J5 (Z. officinale seco, 45 g/L, relación 10:1 agua: alcohol el mejor, pues alcanza el 88% y el 100% de control sobre la formación y germinación de conidias, respectivamente, con respecto al testigo absoluto a las 72 horas, siendo posible optimizar la extracción mediante destilación utilizando 45 g/L de material seco y el uso de solvente agua-alcohol en una relación 10:1.

The Mosquito Melanization Response Is Implicated in Defense against the Entomopathogenic Fungus Beauveria bassiana

Science.gov (United States)

Osta, Mike A.

2012-01-01

Mosquito immunity studies have focused mainly on characterizing immune effector mechanisms elicited against parasites, bacteria and more recently, viruses. However, those elicited against entomopathogenic fungi remain poorly understood, despite the ubiquitous nature of these microorganisms and their unique invasion route that bypasses the midgut epithelium, an important immune tissue and physical barrier. Here, we used the malaria vector Anopheles gambiae as a model to investigate the role of melanization, a potent immune effector mechanism of arthropods, in mosquito defense against the entomopathogenic fungus Beauveria bassiana, using in vivo functional genetic analysis and confocal microscopy. The temporal monitoring of fungal growth in mosquitoes injected with B. bassiana conidia showed that melanin eventually formed on all stages, including conidia, germ tubes and hyphae, except the single cell hyphal bodies. Nevertheless, melanin rarely aborted the growth of any of these stages and the mycelium continued growing despite being melanized. Silencing TEP1 and CLIPA8, key positive regulators of Plasmodium and bacterial melanization in A. gambiae, abolished completely melanin formation on hyphae but not on germinating conidia or germ tubes. The detection of a layer of hemocytes surrounding germinating conidia but not hyphae suggested that melanization of early fungal stages is cell-mediated while that of late stages is a humoral response dependent on TEP1 and CLIPA8. Microscopic analysis revealed specific association of TEP1 with surfaces of hyphae and the requirement of both, TEP1 and CLIPA8, for recruiting phenoloxidase to these surfaces. Finally, fungal proliferation was more rapid in TEP1 and CLIPA8 knockdown mosquitoes which exhibited increased sensitivity to natural B. bassiana infections than controls. In sum, the mosquito melanization response retards significantly B. bassiana growth and dissemination, a finding that may be exploited to design transgenic

Réponse des stades larvaires de Helicoverpa armigera (Hübner (Lepidoptera : Noctuidae à l'application de champignons entomopathogènes Metarhizium anisopliae et Beauveria bassiana

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Tamò, M.

2012-01-01

Full Text Available Response of the nymphs of Helicoverpa armigera (Hübner (Lepidoptera: Noctuidae to entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana. Two experiments on dose/mortality response between the instars of Helicoverpa armigera and two strains of entomopathogenic fungi, Metarhizium anisopliae (Met 31 and Beauveria bassiana (Bb 11 were carried out in laboratory conditions. In the first experiment, M. anisopliae Met 31 was tested on the third instar of H. armigera, while in the second experiment, both Met 31 and Bb 11 were tested on the fourth instar. In all the experiments, the following different doses of conidia per insect were used: 104, 105, 106, 107. The following parameters were measured: mortality and sporulation rates, the number of pupae formed and the number of adults that emerged. Abbott's formula was used to correct the treatment mortality rates. LD50 was determined using Cox-regression. For the third instar in experiment one, no significant difference was observed between high doses (106 and 107 conidia per insect. For instar L4, only the dose of 107 conidia per insect showed high mortality rates (74%. For the strain Bb 11, in spite of the variation observed between the mortality rates induced by high doses (106 and 107 conidia per insect, no significant difference was recorded at the 5% level. No mycosis was observed from cadavers resulting from lower doses when tested on L4. The control recorded the highest numbers of pupae and adults. These two parameters were related to the level of dosage: the higher the dose, the lower the numbers of pupae and adults that emerged. For all the strains of fungi used, whatever the larval stage of H. armigera, the dose/mortality response was significant.

The combination of the entomopathogenic fungus Metarhizium anisopliae with the insecticide Imidacloprid increases virulence against the dengue vector Aedes aegypti (Diptera: Culicidae

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Samuels Richard I

2011-01-01

Full Text Available Abstract Background Dengue fever transmitted by the mosquito Aedes aegypti, is one of the most rapidly spreading insect borne diseases, stimulating the search for alternatives to current control measures. The dengue vector A. aegypti has received less attention than anophelene species, although more than 2.5 billion people are at risk of infection worldwide. Entomopathogenic fungi are emerging as potential candidates for the control of mosquitoes. Here we continue our studies on the pathogenicity of the entomopathogenic fungus Metarhizium anisopliae against adult A. aegypti females. With the aim of further reducing mean survival times of A. aegypti exposed to fungus impregnated surfaces, a sub-lethal concentration of the neonicotinoid insecticide Imidacloprid (IMI was added to fungal suspensions. Results A sub-lethal concentration of IMI that did not significantly alter the daily survival rates or mean survival percentages of mosquitoes was identified to be 0.1 ppm. This sub-lethal concentration was combined with M. anisopliae conidia (1 × 109 conidia mL-1. Both the combined treatment and the conidia alone were able to reduce the survival of A. aegypti compared with untreated or IMI treated mosquitoes. Importantly, mosquito survival following exposure to the combined treatment for 6 and 12 hrs was significantly reduced when compared with mosquitoes exposed to conidia alone. Conclusions This is the first time that a combination of an insecticide and an entomopathogenic fungus has been tested against A. aegypti. Firstly, the study showed the potential of IMI as an alternative to the currently employed pyrethroid adulticides. Secondly, as an alternative to applications of high concentrations of chemical insecticides, we suggest that adult A. aegypti could be controlled by surface application of entomopathogenic fungi and that the efficiency of these fungi could be increased by combining the fungi with ultra-low concentrations of insecticides

The combination of the entomopathogenic fungus Metarhizium anisopliae with the insecticide Imidacloprid increases virulence against the dengue vector Aedes aegypti (Diptera: Culicidae).

Science.gov (United States)

Paula, Adriano R; Carolino, Aline T; Paula, Cátia O; Samuels, Richard I

2011-01-25

Dengue fever transmitted by the mosquito Aedes aegypti, is one of the most rapidly spreading insect borne diseases, stimulating the search for alternatives to current control measures. The dengue vector A. aegypti has received less attention than anophelene species, although more than 2.5 billion people are at risk of infection worldwide. Entomopathogenic fungi are emerging as potential candidates for the control of mosquitoes. Here we continue our studies on the pathogenicity of the entomopathogenic fungus Metarhizium anisopliae against adult A. aegypti females. With the aim of further reducing mean survival times of A. aegypti exposed to fungus impregnated surfaces, a sub-lethal concentration of the neonicotinoid insecticide Imidacloprid (IMI) was added to fungal suspensions. A sub-lethal concentration of IMI that did not significantly alter the daily survival rates or mean survival percentages of mosquitoes was identified to be 0.1 ppm. This sub-lethal concentration was combined with M. anisopliae conidia (1 × 10(9) conidia mL(-1)). Both the combined treatment and the conidia alone were able to reduce the survival of A. aegypti compared with untreated or IMI treated mosquitoes. Importantly, mosquito survival following exposure to the combined treatment for 6 and 12 hrs was significantly reduced when compared with mosquitoes exposed to conidia alone. This is the first time that a combination of an insecticide and an entomopathogenic fungus has been tested against A. aegypti. Firstly, the study showed the potential of IMI as an alternative to the currently employed pyrethroid adulticides. Secondly, as an alternative to applications of high concentrations of chemical insecticides, we suggest that adult A. aegypti could be controlled by surface application of entomopathogenic fungi and that the efficiency of these fungi could be increased by combining the fungi with ultra-low concentrations of insecticides, resulting in higher mortality following relatively short

STUDY OF BORER PEST CONTROL POTATO TUBER IN WAREHOUSES SEEDLINGS (With fungus Beauveria bassiana and Leaf of Tagetes erecta

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Loso Winarto

2013-01-01

Full Text Available Study of borer pest control potato tuber (Phthorimaea operculella Zell in warehouses seedlings with the fungus Beauveria bassiana (Balsamo and leaf Tagetes erecta was conducted in Kutagadung village, district of Brand, Karo District, starting in October until December 2010. The draft that is in use is completely randomized (CRD comprised of seven treatments with three replications. Treatment at Kaji is: Control (no treatment, the density of conidia 106 per ml, 105 per ml, 104 per ml, leaf thickness Tegetes two cm, three cm, and four cm. Observations were P. operculella and intensity of attacks healthy seedlings. In the final analysis, the intensity of attacks in 100 percent control treatment, the density of conidia.

Development of Colletotrichum gloeosporioides isolated from green pepper in different culture media, temperatures, and light regimes

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Mello Alexandre Furtado Silveira

2004-01-01

Full Text Available Control of anthracnose in green pepper involves the use of resistant varieties and/or fungicides. The selection of varieties and efficient products demands great amounts of conidia as inoculum. It is thus necessary to optimize the production of Colletotrichum gloeosporioides conidia in the laboratory, establishing the best conditions for fungus development. The present study aimed at determining the most favorable culture media, temperature, and light conditions for the production of fungus inoculum. The fungus was isolated from green pepper fruits (Capsicum annuum L. and transferred to four culture media (PDA, oat, filtered pepper extract, and autoclaved pepper extract, under different temperatures (15, 20, 25, 30, and 35ºC and light conditions (24h dark, and 24h light. Colony growth was evaluated after 7 and 12 days of incubation. No differences were found between the culture media. However, the greatest number of conidia was obtained from colonies grown in oat medium at 25ºC. Temperatures of 20 and 25ºC were the most favorable for colony growth and sporulation. Higher sporulation was obtained under incubation in constant light. Cultivation of C. gloeosporioides in oat medium, at 25ºC, and constant light is recommended.

Morphological and pathological characterisation of Colletotrichum sp. as casual agent of anthracnose in Dioscorea sp.

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Libia María Pérez Castro

2003-01-01

Full Text Available Colletotrichum gloeosporioides and Colletotrichum dematium strains were isolated from yam plants in the Sucre depart-ment. Macro- and microscopic descriptions were made, finding that the predominant colour for the colonies was light purple. Sporulation was higher than 58,000 conidia/ml for most colonies. Growth rate lay within a range of 4 to 5mm/day. C. gloeosporioide isolate conidial morphology revealed a cylindrical spore having one rounded end whilst the other was acute shaped. Conidia length and width averaged between 7-8 micras and 3-4 micras, respectively. C. dematium colonies, however, were characterised by being grey and having radial growth. The fálcate and fusiform conidia tapered at the acute apex. They had a length and width of 16-18 micras and 3-4 micras, respectively. C. gloeosporioides colonies showed different macroscopic morphology but similar microscopic characteristics. Vegeta-tive compatibility was evaluated amongst different geographical isolates resulting in 90% compatibility. A patho-genic assay was done for evaluating isolate virulence on tolerant and susceptible cultivars. The isolates showed great variability regarding their virulence. The Colletotrichum isolate collection represents a tool for use in molecular characterisation and evaluating yam germplasm tolerance to this pathogen. Keywords: Colletotrichum, yam, anthracnose, phytopathology.

Effect of Leaf Litters and Soils on Viability of Entomopathogenic Fungi Beauveria bassiana (Bals. Vuill

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LISDAR IDWAN SUDIRMAN

2008-09-01

Full Text Available Viability of Beauveria bassiana is extremely low due to toxic compounds in soils. This research was aimed to study the effect of four groups of media on viability of B. bassiana Bb-Pb2. The first group was leaf litters of onion, flowering white cabbage, cabbage, and chinese mustard, respectively; the second group was the soils containing decomposed residues of each plant of the first group; the third group was the mixtures of each media of both groups above (1:1, and the fourth group was natural top soil as a control. Each plastic bag filled with one kg of each medium was inoculated with ten ml of B. bassiana conidia (106/ml of concentration and incubated in open area for 8 weeks. The results showed that all leaf litters of those plants and their compost soils affected the fungal viability. The highest decreasing number of colony was found on onion's leaf litters, soil containing of decomposed onion, and the mixtures of both media. The treated B. bassiana showed significant reducing abilities of growth, conidia production and conidia germination on PDA media, except the one of control. It is suggested that the Bb-Pb2 isolate might not be effective as bioinsecticide in the soils containing either those leaf litters or composts.

Characterization of Peronospora belbahrii on basil under light and scanning electron microscopy

Science.gov (United States)

Basil (Ocimum spp.) downy mildew caused by Peronsoora belbahrii is a major yield-limiting disease of sweet basil (O. basilicum) production worldwide. In this study, sweet basil was grown in a soilless potting mix in plant growth chambers and inoculated with sporangia of P. belbahrii harvested from p...

Laboratory Assessment on Virulence of Nomuraea rileyi against Noctuid Larvae%莱氏野村菌对夜蛾科害虫防治潜力的室内评测

Institute of Scientific and Technical Information of China (English)

周立峰; 于士军; 刘元兵; 王滨

2012-01-01

本文用喷雾法测定了5株莱氏野村菌Nomuraea rileyi对棉铃虫Helicoverpa armigera、甜菜夜蛾Spodoptera exigua和斜纹夜蛾Spodoptera litura 3种夜蛾科害虫的致病力。配制孢子悬浮液对3种夜蛾科害虫进行雾化接种,孢子附着量为188.9～4048.3孢子.mm 2。接种后幼虫置于（24±1）℃和12L∶12D条件下人工饲料饲养,每天观察直至化蛹。时间-剂量-死亡率模型分析显示：5株莱氏野村菌菌株接种量为188.9～4048.3孢子.mm 2时,斜纹夜蛾第10 d的侵染率为60%～95%;菌株Nr15对斜纹夜蛾显示出极高的毒力水平（LC50=266.7孢子.mm 2,4048.3孢子.mm 2浓度下的LT50=5.81 d）,第10 d的侵染率达95%。供试菌株对棉铃虫和甜菜夜蛾的致病力相对较弱,侵染率均不足50%。研究表明,莱氏野村菌菌株对夜蛾科害虫体现出明显的寄主专化性,且有明显的量效关系。%In the current study,a laboratory bioassay was developed to measure the pathogenicities of five Nomuraea rileyi strains against Helicoverpa armigera,Spodoptera exigua and Spodoptera litura of noctuid larvae.The larvae were inoculated with conidia suspensions of N.rileyi at concentrations of 188.9 4048.3 conidia.mm 2 by a Potter Spray Tower,with 0.05% Tween-80 as control.After inoculation,the larvae were individually fed on artificial diet in plastic boxes at（24±1） ℃ and 12L：12D until pupated.Time-dose-mortality model was used to analyze the effect and indicated that the 5 fungal strains could kill 60% 95% of S.litura larvae at concentrations of 188.9 4048.3 conidia.mm 2within 10 d after conidia inoculation.Strain Nr15 demonstrated high virulence to S.litura larvae with a LC50value of 266.7 conidia.mm 2and a LT50value of 5.81 d at the concentration of 4048.3 conidia.mm 2,whereas mortalities of H.armigera and S.exigua didn’t reached 50%.The difference of mortality and fungal infection indicated that there is a host-specificity of N.rileyi strains to noctuid

Influence of culture media in viability test of conidia of entomopathogenic fungi Influência de meios de cultura em teste de viabilidade de fungos entomopatogênicos

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Edimara Aparecida Francisco

2006-08-01

Full Text Available This work aimed at investigatimg whether the culture medium used in viability tests affects the conidial germination of Lecanicillium lecanii, Beauveria bassiana and Paecilomyces fumosoroseus isolates. The tests were performed on microscope slides containing one of the culture media: agar-water (AW, minimal medium (MM, potato-dextrose agar (PDA, potato-dextrose-1% yeast extract agar (PDAY, Sabouraud-dextrose-yeast extract agar (SDAY, and complete medium (CM. Three areas per slide were delimited and 0.05ml of a 5.5 x 105 conidia ml-1 suspension was applied to each area. One bioassay was performed for each isolate. Germination was determined after 15 hours of incubation at 26±0.5°C. The culture media influenced the germination of the species studied, verifying within and inter specific variations. CM and PDA provided the highest germination of L. lecanii isolates and the lowest was obtained on SDAY and AW. The germination of B. bassiana isolates was favoured by CM, PDA and PDAY media, a fact not observed in AW and MM. P. fumosoroseus isolates showed the highest germination on CM and PDA media and the lowest on SDAY. However, some isolates presented high germination on nutrient-poor media (AW and MM.O presente trabalho objetivou investigar se meios de cultura utilizados em teste de viabilidade afetam a germinação de conídios de cinco isolados de Lecanicillium lecanii, cinco de Beauveria bassiana e quatro de Paecilomyces fumosoroseus. Os testes foram realizados em lâminas de microscopia contendo um dos seguintes meios de cultura: Ágar-água (AA, Meio Mínimo (MM, Batata, dextrose e ágar (BDA, Batata, dextrose, ágar e 1% de extrato de levedura (BDAL, Sabouraud, dextrose, ágar e extrato de levedura (SDAL e Meio Completo (MC. Delimitaram-se três áreas por lâmina e em cada uma aplicou-se 0,05mL de uma suspensão com concentração de 5,5 x 105 conídios ml-1. Para cada isolado foi realizado um bioensaio, com seis tratamentos e cinco repeti

Aplicação foliar de tratamentos para o controle do míldio e da podridão-de-escamas de bulbos de cebola Foliar spray of treatments in the control of downy mildew and bulb rot in onion

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João Américo Wordell Filho

2007-12-01

Full Text Available Em experimento de campo, avaliou-se o efeito da aplicação foliar de tratamentos para o controle do míldio (Peronospora destructor e da podridão de bulbos (Burkholderia cepacia de cebola: testemunha, clorotalonil/metalaxyl + clorotalonil, fosfito de potássio, fertilizante foliar (03-00-16, N-P-K, calda bordalesa, calda bordalesa/fosfito de potássio, acibenzolar-S-methyl, pulverizados semanalmente; extrato de alga (Ulva fasciata e ulvana, aplicados a cada 7, 14 e 21 dias. Somente a pulverização semanal com fungicidas sintéticos ou com o fertilizante (03-00-16; 400 mL de p.c./100 L foi capaz de reduzir significativamente a severidade do míldio, em 60 ou 23%, respectivamente, em relação à testemunha não pulverizada, sem aumentar o rendimento de bulbos. O tratamento com fertilizantes ricos em potássio resultou em maior incidência da podridão de bulbos armazenados por cinco meses. O conteúdo de açúcares solúveis e incidência da podridão de bulbos de cebola foram correlacionados significativamente (-0,629, p A field experiment was carried out to evaluate the effect of foliar sprays with the following treatments on the downy mildew (Peronospora destructor and bulb rot (Burkholderia cepacia in onions: non-treated control, fungicide chlorotalonil/metalaxyl + chlorotalonil, potassium phosphite, foliar fertilizer (03-00-16, N-P-K, bordeaux mixture, bordeaux mixture/potassium phosphite, acibenzolar-S-methyl weekly applied; extract of alga Ulva fasciata and ulvan sprayed every 7, 14 and 21 days. Only the weekly spraying of fungicides and fertilizer (03-00-16, 400 mL/100 L significantly reduced the mildew severity by 60 and 23%, respectively, but did not increase the bulb yield. The foliar application of potassium rich fertilizers resulted in a higher incidence of rotten bulbs after 5 months in storage. Soluble sugar content and rot incidence of onion bulbs were significantly correlated (-0,629, p < 0,05.

Blue and near ultraviolet reversible photoreaction in the induction of fungal conidiation

International Nuclear Information System (INIS)

Kumagai, T.

1982-01-01

The effects of blue light and near UV light on the induction of conidiation in the fungus, Alternaria tomato, were investigated. Induction of conidiation was repeatedly controlled by alternating doses of near UV light and blue light. When the final light was near UV, conidiation was induced and conidia developed in the following darkness; when it was blue, the induction of conidiation was suppressed. When conidiation was induced by irradiation with a light mixed with near UV and blue, not only the time lag for inducing conidiation but also the amount of conidia formed were regulated by the fluence rates of both those lights. Thus, 'mycochrome' is considered to function as a photoreceptor system in the induction of conidiation of this fungus. (author)

Inhibition of Blumeria graminis germination and germling development within colonies of oat mildew

DEFF Research Database (Denmark)

Carver, T.L.W.; Roberts, P.C.; Thomas, B.J.

2001-01-01

Germination by Blumeria graminis. DC Speer ff. spp. avenae, hordei and tritici, was greatly suppressed when conidia fell within colonies of ff. spp. avenae or hordei established on susceptible oat or barley, respectively. On healthy oat or barley, and when distant from powdery mildew, colonies. all...... ff. spp. formed normal appressoria. This was also true When conidia germinated within established barley mildew colonies. Within barley mildew colonies, appressoria of f. sp. hordei penetrated epidermal cells formed haustoria more frequently than appressoria distant from colonies. Similarly, ff. spp....... avenae and tritici, normally unable to infect barley. frequently penetrated epidermal cells subtending established barley mildew colonies. Thus, colony, establishment induced barley epidermal cell accessibility, even to non-pathogenic ff. spp, In contrast. when all three ff. spp. germinated within...

The nematophagous fungus Monacrosporium thaumasium and its nematicidal activity on Angiostrongylus vasorum.

Science.gov (United States)

Soares, Filippe Elias de Freitas; Braga, Fabio Ribeiro; de Araújo, Jackson Victor; Lima, Walter dos Santos; de Queiroz, José Humberto

2015-01-01

The dog acts as a reservoir and environmental disseminator of potentially zoonotic parasites. The objective of this work was to study the fungus Monacrosporium thaumasium regarding its nematicidal potential in laboratory trials and its proteolytic profile. The in vitro test was carried out through two assays (A and B). In assay A, conidia of the fungus N34a were added in positive coprocultures for Angiostrongylus vasorum. In assay B, crude extract (treated group) and distilled water (control group) were added to coprocultures. Next, the proteolytic profile of crude extract of the nematophagous fungus M. thaumasium (NF34a) was revealed by performing a zymogram. There was a reduction (p<0.01) in the averages of larvae recovered from the treated groups (conidia and crude extract) in relation to control groups. The zymogram suggested that the nematophagous fungus M. thaumasium produces a protease of approximately 40 kDa. The results of this work confirm that the conidia as well as the crude extract of the fungus M. thaumasium may be used to control A. vasorum L1. The proteolytic profile suggested the presence of one protease (Mt1) of approximately 40 kDa that in the future may be used in biological control of L1 of this nematode. Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

Aquatic hyphomycetes as environmental quality bioindicators in the Chirgua River (Bejuma, Venezuela

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Rafael Fernández

2017-01-01

Full Text Available At present, the evaluation of water resources uses bioindicators, such as additional diagnostic systems, because they indicate anthropogenic disturbances. Aquatic hyphomycetes,  part of the trophic chain of aquatic ecosystems, are indicators of water quality through their varying  presence and concentration of conidia based on environmental disturbance; therefore, this research assessed their bioindicator role for environmental quality in the Chirgua River in Venezuela. In 2011 in two areas: 1 Headwaters (unaffected and 2 downstream area (affected, physico-chemical parameters (O2, conductivity, pH, temperature and total solids and nutrients were determined weekly (SO4-2, PO4-3, NO2-2 and NO3-3, along with coliforms (total and fecal and water hyphomycetes (number of species and conidia/mL. We identified a total of 44 species, 42 (20 exclusive in zone 1 and 24 (2 exclusive in zone 2. In the affected sector, the number of species and conidia/mL of the coliforms (total and fecal decreased, as compared to the unaffected zone. Sensitive species to environmental disturbance for coliforms included Brachiosphaera tropicalis, Camposporium antenatum, Campylospora filicladia, Campylospora parvula, Clavatospora tentacula, Clavatospora stellata, Culicidospora gravida, Diplocladiella scalaroides, Flabellospora acuminata, Helicomyces colligatus, Helicomyces sp., Helicomyces torquatus, Phalangispora constricta, Tetracladium marchalianum and Triscelophorus monosporus, while tolerant species included Alatospora acuminata, Campylospora chaetocladia, Flabellospora crassa, Isthmotricladia gombakiensis, Tetraploa cf. Aristata and Triscelophorus acuminatus.

MMS induction of different types of genetic damage in Aspergillus nidulans: a comparative analysis in mutagenesis.

Science.gov (United States)

Gualandi, G; Bellincampi, D; Puppo, S

1979-09-01

Methyl methanesulphonate (MMS) was used to test the induction of gene mutation, somatic crossing-over and mitotic non-disjunction in A. nidulans. Gene mutation was tested by inducing mutants resistant to 8-azaguanine and revertants of methG1 in a haploid strain. Somatic crossing-over was tested in heterozygous diploids, both with a selective method, i.e. inducing homozygosis to FPA resistance in a heterozygous fpa A1/+ strain, and with a non-selective method, i.e. identifying the frequencies of colour sectors. This latter method was also used to estimate the induction of non-disjunction because additional markers were present which permitted us to distinguish the two types of colour segregant. Generally, 3 different experimental procedures were used, namely the "plate test", i.e. plating of conidia in agar media containing MMS, and two types of "liquid test", i.e. brief treatment of quiescent or pre-germinated conidia in MMS solution before they were plated on agar media. Point mutations were induced with about equal efficiency with each method, whereas crossing-over was induced preferentially when germinating conidia were exposed to MMS. On the other hand, non-disjunction was induced in germinating and quiescent spores with equal efficiency, but such segregants were not recovered with the selective (fpa) method. The results are discussed for both their practical use in the mutagenic testing procedure and their theoretical implication.

Effect of electron beam irradiation on conidial germination activity and pathogenicity of Botrytis cinerea

International Nuclear Information System (INIS)

Zhang Ting; Qiao Yongjin; Chen Zhaoliang

2011-01-01

Conidia of Botrytis cinerea were irradiated by electron beam at 0.5, 1.0, 2.0 and 3.0 kGy. The influence of electron beam on the activities of conidial germination and pathogenicity at the temperatures of 5 ℃ and 25 ℃ were tested, respectively. The results showed that the electron beam could inhibit germination of conidia and the length of germ tube of Botrytis cinerea, and delay the germination time. It could also decrease the pathogenicity obviously and higher irradiation dose showed stronger effects. Compared with control, the complete germination time of conidia extended to 5 and 9 d at the cultivate temperatures of 25 ℃ and 5 ℃, after 2 kGy of irradiation, and the germination rate was reduced 46.57% and 33.68%, respectively. The inhibition rates of germ tube were 25.12% and 74.29% when cultured 24 h. The pathogenicity of Botrytis cinerea to strawberry was reduced significantly. After 2.0 kGy irradiation and cultivate at 25 ℃ for 2 d, the disease index was 4.17 and it decreased to 15.28 after cultivation of 5 ℃ for 15 d. Electron beam treatment could inhibit the spore germination and germ tube elongation of Botrytis cinerea significantly, delayed the germination time, and reduced its pathogenicity, the higher the dose, the effect was more obvious. (authors)

Heated water and UV-C radiation to post harvest control of Cryptosporiopsis perennans on apples

International Nuclear Information System (INIS)

Bartnicki, Vinicius Adao; Amarante, Cassandro Vidal Talamini do; Castro, Luis Antonio Suita de; Rizzatti, Mara Regina; Souza, Joao Antonio Vargas de

2010-01-01

The objective of this work was to assess the colonization of Cryptosporiopsis perennans in the epidermis of apples and the efficiency of heated water and UV-C radiation application to control this pathogen. In apples inoculated with C. perennans, the colonization of lenticels and adjacent areas by the pathogen was observed by electronic scanning microscopy. The sensitivity of C. perennans conidia was evaluated in aqueous suspension, at temperatures of 28, 45, 50 and 55 deg C for 15 and 30 s, and at UV.C radiation doses of 0.018, 0.037, 0.075, 0.150, 0.375, 0.750, 1.500 and 3.000 kJ m.2. The effects of UV.C radiation doses at 0.375, 0.750 and 1.500 kJ m.2 and heated water at 50 deg C, sprayed during 15 and 30 s were evaluated for controlling C. perennans in apples inoculated with the pathogen. The fungus produced abundant mycelium and conidia in lenticels and adjacent areas on the epidermis of the apples. The heated water at 50 deg C during 15 s and a 0.750 kJ m.2 UV.C radiation dose reduced conidia survival in more than 99%. Heated water sprayed at 50 deg C during 15 s and a UV.C radiation dose of 0.375 kJ m.2 control C. perennans in apples. (author)

Diatomaceous earth and oil enhance effectiveness of Metarhizium anisopliae against Triatoma infestans.

Science.gov (United States)

Luz, Christian; Rodrigues, Juscelino; Rocha, Luiz F N

2012-04-01

Entomopathogenic fungi, especially Metarhizium anisopliae, have potential for integrated control of peridomestic triatomine bugs. However, the high susceptibility of these vectors to fungal infection at elevated ambient humidities decreases in the comparatively dry conditions that often prevail in their microhabitats. A formulation adapted to this target pest that induces high and quick mortality can help to overcome these drawbacks. In the present study diatomaceous earth, which is used against pests of stored grains or as an additive to mycoinsecticides, delayed but did not reduce in vitro germination of M. anisopliae s.l. IP 46 conidia after >24h agitation without affecting viability, and did not hamper the survival of Triatoma infestans nymphs exposed to treated surfaces. The settling behavior of nymphs on a treated surface in choice tests depended on the concentration of diatomaceous earth and ambient light level. Conidia formulated with diatomaceous earth and a vegetable oil synergized the insecticidal effect of the fungus in nymphs, and quickly killed all treated insects, even at 75% relative humidity (LT(90) 8.3 days) where unformulated conidia caused only 25% mortality after a 25 days exposure. The improved performance of a combined oil and desiccant dust formulation of this Metarhizium isolate raises the likelihood for its successful mycoinsecticidal use for triatomine control and, apparently, against other domestic insect pests. Copyright © 2011 Elsevier B.V. All rights reserved.

Heated water and UV-C radiation to post harvest control of Cryptosporiopsis perennans on apples; Agua aquecida e radiacao UV-C no controle pos-colheita de Cryptosporiopsis perennans em macas

Energy Technology Data Exchange (ETDEWEB)

Bartnicki, Vinicius Adao; Amarante, Cassandro Vidal Talamini do, E-mail: vinibart@hotmail.co, E-mail: amarante@cav.udesc.b [Universidade do Estado de Santa Catarina (UDESC), Lages, SC (Brazil). Centro de Ciencias Agroveterinarias. Dept. de Agronomia; Valdebenito-Sanhueza, Rosa Maria, E-mail: rosamaria@m2net.com.b [Proterra Engenharia Agronomica, Vacaria, RS (Brazil); Castro, Luis Antonio Suita de, E-mail: suita@cpact.embrapa.b [EMBRAPA Clima Temperado, Pelotas, RS (Brazil); Rizzatti, Mara Regina; Souza, Joao Antonio Vargas de, E-mail: marar@pucrs.b [Pontificia Universidade Catolica do Rio Grande do Sul (PUC-RS), Porto Alegre, RS (Brazil). Centro em Pesquisa e Desenvolvimento em Fisica. Grupo de Fisica das Radiacoes

2010-02-15

The objective of this work was to assess the colonization of Cryptosporiopsis perennans in the epidermis of apples and the efficiency of heated water and UV-C radiation application to control this pathogen. In apples inoculated with C. perennans, the colonization of lenticels and adjacent areas by the pathogen was observed by electronic scanning microscopy. The sensitivity of C. perennans conidia was evaluated in aqueous suspension, at temperatures of 28, 45, 50 and 55 deg C for 15 and 30 s, and at UV.C radiation doses of 0.018, 0.037, 0.075, 0.150, 0.375, 0.750, 1.500 and 3.000 kJ m.2. The effects of UV.C radiation doses at 0.375, 0.750 and 1.500 kJ m.2 and heated water at 50 deg C, sprayed during 15 and 30 s were evaluated for controlling C. perennans in apples inoculated with the pathogen. The fungus produced abundant mycelium and conidia in lenticels and adjacent areas on the epidermis of the apples. The heated water at 50 deg C during 15 s and a 0.750 kJ m.2 UV.C radiation dose reduced conidia survival in more than 99%. Heated water sprayed at 50 deg C during 15 s and a UV.C radiation dose of 0.375 kJ m.2 control C. perennans in apples. (author)

The homeobox BcHOX8 gene in Botrytis cinerea regulates vegetative growth and morphology.

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Zsuzsanna Antal

Full Text Available Filamentous growth and the capacity at producing conidia are two critical aspects of most fungal life cycles, including that of many plant or animal pathogens. Here, we report on the identification of a homeobox transcription factor encoding gene that plays a role in these two particular aspects of the development of the phytopathogenic fungus Botrytis cinerea. Deletion of the BcHOX8 gene in both the B. cinerea B05-10 and T4 strains causes similar phenotypes, among which a curved, arabesque-like, hyphal growth on hydrophobic surfaces; the mutants were hence named Arabesque. Expression of the BcHOX8 gene is higher in conidia and infection cushions than in developing appressorium or mycelium. In the Arabesque mutants, colony growth rate is reduced and abnormal infection cushions are produced. Asexual reproduction is also affected with abnormal conidiophore being formed, strongly reduced conidia production and dramatic changes in conidial morphology. Finally, the mutation affects the fungus ability to efficiently colonize different host plants. Analysis of the B. cinerea genome shows that BcHOX8 is one member of a nine putative homeobox genes family. Available gene expression data suggest that these genes are functional and sequence comparisons indicate that two of them would be specific to B. cinerea and its close relative Sclerotinia sclerotiorum.

Identification of fibrinogen-binding proteins of Aspergillus fumigatus using proteomic approach.

Science.gov (United States)

Upadhyay, Santosh Kumar; Gautam, Poonam; Pandit, Hrishikesh; Singh, Yogendra; Basir, Seemi Farhat; Madan, Taruna

2012-03-01

Aspergillus fumigatus, the main etiological agent for various forms of human aspergillosis, gets access to the respiratory system of human host by inhalation of airborne conidia. These conidia possibly adhere to extracellular matrix (ECM) proteins. Among the ECM proteins involved in adherence, fibrinogen is thought to be crucial. Here, we studied whether A. fumigatus three-week culture filtrate (3wcf) proteins promote binding of A. fumigatus to ECM proteins and promote fungal growth. We observed that incubation of ECM with 3wcf proteins led to dose- and time-dependent increase in adherence of conidia to the ECM. In order to identify the catalogue of fibrinogen-binding A. fumigatus proteins, we carried out fibrinogen affinity blotting using two-dimensional gel electrophoresed 3wcf proteins. A total of 15 fibrinogen-binding protein spots corresponding to 7 unique proteins were identified in 3wcf using matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF-TOF). Among these, 4 proteins, namely, beta-glucosidase, alpha-mannosidase, pectate lyase A and oryzin precursor were predicted to have cell wall or extracellular localization, whereas amidase family protein and two hypothetical proteins did not display the signal sequence. This study reports seven novel fibrinogen-binding proteins of A. fumigatus, some of which could be further explored for targeting the adhesion phenomenon as antifungal strategy.

Effect of Leaf Litters and Soils on Viability of Entomopathogenic Fungi Beauveria bassiana (Bals. Vuill

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LISDAR IDWAN SUDIRMAN

2008-09-01

Full Text Available Viability of Beauveria bassiana is extremely low due to toxic compounds in soils. This research was aimed to study the effect of four groups of media on viability of B. bassiana Bb-Pb2. The first group was leaf litters of onion, flowering white cabbage, cabbage, and chinese mustard, respectively; the second group was the soils containing decomposed residues of each plant of the first group; the third group was the mixtures of each media of both groups above (1:1, and the fourth group was natural top soil as a control. Each plastic bag filled with one kg of each medium was inoculated with ten ml of B. bassiana conidia (106/ml of concentration and incubated in open area for 8 weeks. The results showed that all leaf litters of those plants and their compost soils affected the fungal viability. The highest decreasing number of colony was found on onion’s leaf litters, soil containing of decomposed onion, and the mixtures of both media. The treated B. bassiana showed significant reducing abilities of growth, conidia production and conidia germination on PDA media, except the one of control. It is suggested that the Bb-Pb2 isolate might not be effective as bioinsecticide in the soils containing either those leaf litters or composts.

Effect of two dosages of Metarhizium anisopliae var. acridum against Rhammatocerus schistocercoides Rehn

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Faria Marcos Rodrigues de

2002-01-01

Full Text Available The fungus Metarhizium anisopliae var. acridum, strain CG 423, was tested under field conditions against the gregarious grasshopper Rhammatocerus schistocercoides (Rehn (Orthoptera: Acrididae. Conidia formulated in a racemic mixture of soybean oil and kerosene were sprayed under field conditions using an ultralow-volume hand-held atomizer Ulva Plus adjusted to deliver 2.9 L/ha. Bands composed of 2nd instar nymphs were treated with either 5.0x10(12 or 1.0x10(13 viable conidia/ha. The number of insects in each band was estimated at day one following spraying and by the end of the field trial (15 to 16 days post-treatment. Reductions in population size reached, in average, 65.8% and 80.4% for bands treated with the higher and lower dosage, respectively. For both dosages, total mortality rates of insects collected at two days post-application, and kept in cages for 14 days under lab conditions, showed no significant differences as compared to that obtained with insects collected immediately after spraying. Healthy insects were fed to native grasses sprayed on the field with 1.0x10(13 viable conidia/ha. Mortality levels of the nymphs fed on grasses collected two and four days post-application were not affected when compared to nymphs fed on grasses collected immediately following application.

Odor aversion and pathogen-removal efficiency in grooming behavior of the termite Coptotermes formosanus.

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Aya Yanagawa

Full Text Available The results of biocontrol with entomopathogens in termites have been discouraging because of the strong social hygiene behavior for removing pathogens from termite colonies. However, the mechanism of pathogen detection is still unclear. For the successful application of biopesticides to termites in nature, it would be beneficial to identify substances that could disrupt the termite's ability to perceive pathogens. We hypothesized that termites can perceive pathogens and this ability plays an important role in effective hygiene behavior. In this study, pathogen-detection in the subterranean termite Coptotermes formosanus was investigated. We performed quantitative assays on conidia removal by grooming behavior using epifluoresence microscopy and Y-maze tests to examine the perception of fungal odor by termites. Three species each of high- and low-virulence entomopathogenic fungi were used in each test. The results demonstrated that termites removed conidia more effectively from a nestmate's cuticle if its odor elicited stronger aversion. Highly virulent pathogens showed higher attachment rates to termite surfaces and their odors were more strongly avoided than those of low-virulence isolates in the same species. Moreover, termites appeared to groom each other more persistently when they had more conidia on their bodies. In brief, insect perception of pathogen-related odor seems to play a role in the mechanism of their hygiene behavior.

Molecular, morphological and pathogenic characterization of six strains of Metarhizium spp. (Deuteromycotina: Hyphomycetes for the control of Aegorhinus superciliosus (Coleoptera: Curculionidae

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María Sepúlveda

2016-03-01

Full Text Available Aegorhinus superciliosus is an important pest on blueberry (Vaccinium corymbosum L. and other fruit trees. The use of entomopathogenic fungi as Metarhizium spp. has been evaluated for the control of this insect, but variability has been observed among different strains. The aim of this study was to characterize six promising strains of Metarhizium spp. for the control of A. superciliosus. The studied strains were QuM173c, Qu-M363, Qu-M171a, Qu-M156a, Qu-M421, and Qu-M430, all of which belonged to the Chilean Collection of Microbial Genetic Resources (ChCMGR of the Institute de Investigaciones Agropecuarias (INIA, Chile. Molecular characterization was made by sequencing the ITS region (Internal Transcribed Spacers, ITS-5.8S rDNA. The morphology of conidia was evaluated through scanning electron microscopy and radial colony growth was evaluated in potato dextrose agar (PDA, Sabouraud dextrose agar (SDA, agar enriched with larvae of Galleria mellonella (Lepidoptera: Pyralidae (GA, and agar enriched with adults of A. superciliosus (AA. Pathogenicity was studied based on mortality of adults of A. superciliosus inoculated with conidia. Sequencing of the ITS-5.8S rDNA region indicates that the strains belong to the clade of M. anisopliae var. anisopliae, except for Qu-M171a, which was identified as M. anisopliae var. lepidiotum. Conidia average length for the six strains was 5.09 pm and average conidia width was 1.92 pm. Radial colony growth differences were observed between strains (p < 0.01 and between different growth media (p < 0.01. The strains exhibited the highest colony growth in the GA medium, while in the AA medium they showed the lowest (p < 0.01. Pathogenicity tests show that Qu-M430 reached a 90% mortality rate (p < 0.01. Results show that there is variability between the studied strains, which is expressed in their morphology, molecular characteristics and pathogenicity towards A. superciliosus.

Cellulase Activity in Solid State Fermentation of Palm Kernel Cake with Trichoderma sp.

Directory of Open Access Journals (Sweden)

Massaud, M. B. N.

2012-01-01

Full Text Available Aims: The effect of different types of fungal inocula to the cellulase activity measured on palm kernel cake (PKC was studied. Methodology and Results: Isolate Pro-A1 which was identified as Trichoderma sp. was selected as a potential producer of cellulase via solid state fermentation technique (SSF. Two types of PKCs were used; raw PKC (containing residual oil and defatted PKC. The PKCs were inoculated with different concentrations of conidia and varying amounts (g of solid mycelia plugs (SMP for SSF. The effect of ultrafiltered crude fungal filtrate (CFF as inocula was also being tested. The highest cellulase activity of 2.454 FPU/mL was detected with 60% (wt/wt SMP applied to the raw PKC. Conversely, 2.059 FPU/mL of cellulase activity was measured when 80% (wt/wt of SMP was applied to the defatted PKC which is 62.3% higher than the untreated defatted PKC; and more than 100% increase in enzymatic activity compared to raw PKC. The cellulase activity in the SSF inoculated with 8 x 106 conidia /mL and 12 x 106 conidia /mL were 1.704 FPU/mL for raw PKC and 1.856 FPU/mL for defatted PKC, an enhancement of about 46% from uninoculated batch. Inoculation with CFF bears corresponding maximum improvement of the cellulase activity on both PKCs of 13.58% (raw and 2.86% (defatted. Conclusion, significance and impact of study: The current study proves that Trichoderma sp. in the form of SMP can enhance the cellulase activity on PKCs effectively with more than 100% increment. Fungal conidia are also a better choice in enhancing cellulase activity of Trichoderma sp. permitted that the PKC used is devoid of oil. From this study, Trichoderma sp. holds the potential of converting lignocellulosic materials into products of commercial and industrial values such as glucose and other biofuels.

Monitoring persistence of the entomopathogenic fungus Metarhizium anisopliae under simulated field conditions with the aim of controlling adult Aedes aegypti (Diptera: Culicidae).

Science.gov (United States)

Carolino, Aline T; Paula, Adriano R; Silva, Carlos P; Butt, Tariq M; Samuels, Richard I

2014-04-25

Entomopathogenic fungi are potential candidates for use in integrated vector management, with recent emphasis aimed at developing adult mosquito control methods. Here we investigated the persistence of the fungus Metarhizium anisopliae when tested against female A. aegypti under field conditions. Black cotton cloths impregnated with M. anisopliae conidia, formulated in vegetable oil + isoparaffin, were maintained on a covered veranda for up to 30 days. At specific times, pieces of the cloths were removed, placed in Tween 80 and the resuspended conidia were sprayed directly onto mosquitoes. The persistence of conidia impregnated on black cloths using three different carriers was evaluated in test rooms. Fifty mosquitoes were released into each room and after a 5 day period, the surviving insects were captured. Another 50 insects were then released into each room. The capacity of the fungus at reducing mosquito survival was evaluated over a total of 35 days. Conidia extracted from cloths maintained on the veranda for 2 to 18 days remained virulent, with 28 to 60% mosquito survival observed. Mosquito survival following exposure to fungus impregnated cloths showed that fungus + Tween caused similar reductions to that of fungus + vegetable oil. Mosquitoes exposed to the formulation fungus + vegetable oil had survival rates of 36% over the first 5 days of the experiment. Following the release of the second cohort of mosquitoes (6-11days), survival increased to 50%. The survival of the 12-17 day cohort (78%) was statistically equal to that of the controls (84%). Formulation of the fungus in vegetable oil + isoparaffin increased the persistence of the fungus, with the 18-23 day cohort (64% survival) still showing statistical differences to that of the controls (87% survival). The potential of entomopathogenic fungi for the control of adult A. aegypti was confirmed under field conditions. Vegetable oil + isoparaffin formulations of M. anisopliae significantly increased the

Effects of Fungicides on the Development of the Entomopathogenic Fungus Metarhizium anisopliae var. anisopliae Efecto de los Fungicidas sobre el Desarrollo del Hongo Entomopatógeno Metarhizium anisopliae var. anisopliae

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Maribel Yáñez

2010-09-01

Full Text Available Metarhizium anisopliae (Metschnikoff Sorokin is an entomopathogenic fungus used for controlling different insect pests. It is most frequently applied to berry fruit crops, where fungicides are also used for disease control. Fungicides: azoxystrobin, benomyl, captan, chlorothalonil, fenhexamid, fludioxonil, iprodione, and metalaxyl in concentrations of 0.01, 0.1, 1.0, 10, and 100 mg L-1 were evaluated in this research study. Vegetative growth, conidia germination, and conidia germination tube length were measured on the Qu-M82, Qu-M151b, Qu-M253, Qu-M430, and Qu-M984 Metarhizium anisopliae var. anisopliae fungus strains. Those strains were selected because of their present use against different insect pest in bramble fruits. Vegetative growth was measured through the colony rate growth in agar media, and those reaching up to 50% of the check growth were considered compatible. Results indicate that the benomyl and fenhexamid fungicides were compatible with the five isolates whereas, azoxystrobin and fludioxonil were incompatible. Furthermore, benomyl and fludioxonil reduced conidia germination by 53 and 91%, and germination tube length by 18 and 37%, respectively.Metarhizium anisopliae (Metschnikoff Sorokin es un hongo entomopatógeno que se utiliza para el control de diferentes insectos, uno de sus usos más frecuentes es en frutales menores, donde también se utilizan fungicidas para el control de enfermedades. En este trabajo se evaluó el efecto de los fungicidas azoxystrobin, benomil, captan, chlorothalonil, fenhexamid, fludioxonil, iprodione y metalaxil, en concentraciones de 0,01; 0,1; 1; 10 y 100 mg L-1, sobre el crecimiento de la colonia, porcentaje de germinación de conidias y longitud de tubos germinativos de distintas cepas de M. anisopliae var. anisopliae. Las cepas utilizadas fueron Qu-M82, Qu-M151b, Qu-M253, Qu-M430 y Qu-M984, seleccionadas por su uso comercial para el control de diferentes insectos en frutales menores. El

Cellulose hydrolysis by fungi. 2. Cellulase production by Trichoderma harzianum in liquid medium fermentation

Energy Technology Data Exchange (ETDEWEB)

Roussos, S.; Raimbault, M. (Laboratoire de Microbiologie ORSTOM, Centre de Recherche IRCHA, 91 - Vert-le-Petit (France))

Microcrystalline cellulose (cellulose Avicel, Merck) supported growth of Trichoderma harzianum and induced production of cellulases in liquid cultures. After 50 h growth, the total cellulasic activities present in both the supernatant and the mycelium were 3,000 IU/l of carboxymethyl cellulose, 400 IU/l of filter paper activity, and 4 IU/l of cotton activity corresponding to 1.7 g/l of proteins. Cellulase production could be increased by a preliminary treatment of cellulose, and pH regulation during growth. The influence of inoculum concentration was studied and an optimum of 3 X 10/sup 7/ conidia/g dry weight of substrate was demonstrated. Using a synthetic culture medium, a soluble factor of germination was demonstrated which could be leached out by 3 successive washings of conidia.

ESTUDO DA TOXICIDADE DO MEDICAMENTO PERINDOPRIL ...

African Journals Online (AJOL)

Usuario

2013-08-28

Aug 28, 2013 ... is accomplished by anti-hypertensive drugs, among them, ... of perindopril on the conidia germination speed of the model fungus M. anisopliae, for detecting a possible ..... negative for tests with following parameters: bacterial.

Utilization of chitinolytic bacterial isolates to control anthracnose of ...

African Journals Online (AJOL)

SAM

2014-04-09

Apr 9, 2014 ... Assay of antagonistic bacterial chitinolytic to C. gloeosporioides was ... 1632 Afr. J. Biotechnol. ... microorganisms, by their interactions with various plant ... ristics such as colony, mycelia as well as shape and size of conidia.

Differential allergy responses to Metarhizium anisopliae fungal component extracts in BALB/c mice

Science.gov (United States)

Intratracheal aspiration (IA) exposure to Metarhizium anisopliae crude antigen (MACA), which is composed of equal protein amounts of mycelium (MYC), conidia (CON) and inducible proteases/chitinases (IND) extracts/filtrates, has resulted in responses characteristic of human allerg...

The MpkB MAP kinase plays a role in autolysis and conidiation of Aspergillus nidulans.

Science.gov (United States)

Kang, Ji Young; Chun, Jeesun; Jun, Sang-Cheol; Han, Dong-Min; Chae, Keon-Sang; Jahng, Kwang Yeop

2013-12-01

The mpkB gene of Aspergillus nidulans encodes a MAP kinase homologous to Fus3p of Saccharomyces cerevisiae which is involved in conjugation process. MpkB is required for completing the sexual development at the anastomosis and post-karyogamy stages. The mpkB deletion strain could produce conidia under the repression condition of conidiation such as sealing and even in the submerged culture concomitant with persistent brlA expression, implying that MpkB might have a role in timely regulation of brlA expression. The submerged culture of the deletion strain showed typical autolytic phenotypes including decrease in dry cell mass (DCM), disorganization of mycelial balls, and fragmentation of hyphae. The chiB, engA and pepJ genes which are encoding cell wall hydrolytic enzymes were transcribed highly in the submerged culture. Also, we observed that the enzyme activity of chitinase and glucanase in the submerged culture of mpkB deletion strain was much higher than that of wild type. The deletion of mpkB also caused a precocious germination of conidia and reduction of spore viability. The expression of the vosA gene, a member of velvet gene family, was not observed in the mpkB deletion strain. These results suggest that MpkB should have multiple roles in germination and viability of conidia, conidiation and autolysis through regulating the expression of vosA and brlA. Copyright © 2013 Elsevier Inc. All rights reserved.

WetA bridges cellular and chemical development in Aspergillus flavus.

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Ming-Yueh Wu

Full Text Available Bridging cellular reproduction and survival is essential for all life forms. Aspergillus fungi primarily reproduce by forming asexual spores called conidia, whose formation and maturation is governed by the central genetic regulatory circuit BrlA→AbaA→WetA. Here, we report that WetA is a multi-functional regulator that couples spore differentiation and survival, and governs proper chemical development in Aspergillus flavus. The deletion of wetA results in the formation of conidia with defective cell walls and no intra-cellular trehalose, leading to reduced stress tolerance, a rapid loss of viability, and disintegration of spores. WetA is also required for normal vegetative growth, hyphal branching, and production of aflatoxins. Targeted and genome-wide expression analyses reveal that WetA exerts feedback control of brlA and that 5,700 genes show altered mRNA levels in the mutant conidia. Functional category analyses of differentially expressed genes in ΔwetA RNA-seq data indicate that WetA contributes to spore integrity and maturity by properly regulating the metabolic pathways of trehalose, chitin, α-(1,3-glucan, β-(1,3-glucan, melanin, hydrophobins, and secondary metabolism more generally. Moreover, 160 genes predicted to encode transcription factors are differentially expressed by the absence of wetA, suggesting that WetA may play a global regulatory role in conidial development. Collectively, we present a comprehensive model for developmental control that bridges spore differentiation and survival in A. flavus.

Fusarium pathogenesis investigated using Galleria mellonella as a heterologous host

Science.gov (United States)

Coleman, Jeffrey J.; Muhammed, Maged; Kasperkovitz, Pia V.; Vyas, Jatin M.; Mylonakis, Eleftherios

2011-01-01

Members of the fungal genus Fusarium are capable of manifesting in a multitude of clinical infections, most commonly in immunocompromised patients. In order to better understand the interaction between the fungus and host, we have developed the larvae of the greater wax moth, Galleria mellonella, as a heterologous host for fusaria. When conidia are injected into the hemocoel of this Lepidopteran system, both clinical and environmental isolates of the fungus are able to kill the larvae at 37°C, although killing occurs more rapidly when incubated at 30°C. This killing was dependent on several other factors besides temperature, including the Fusarium strain, the number of conidia injected, and the conidia morphology, where macroconidia are more virulent than their microconidia counterpart. There was a correlation in the killing rate of Fusarium spp. when evaluated in G. mellonella and a murine model. In vivo studies indicated G. mellonella hemocytes were capable of initially phagocytosing both conidial morphologies. The G. mellonella system was also used to evaluate antifungal agents, and amphotericin B was able to confer a significant increase in survival to Fusarium infected-larvae. The G. mellonella-Fusarium pathogenicity system revealed that virulence of Fusarium spp. is similar, regardless of the origin of the isolate, and that mammalian endothermy is a major deterrent for Fusarium infection and therefore provides a suitable alternative to mammalian models to investigate the interaction between the host and this increasingly important fungal pathogen. PMID:22115447

Differential susceptibility of adults and nymphs of Blattella germanica (L.) (Blattodea: Blattellidae) to infection by Metarhizium anisopliae and assessment of delivery strategies

International Nuclear Information System (INIS)

Lopes, R.B.; Alves, S.B.

2011-01-01

Microbial insecticides for cockroach control, such as those containing entomopathogenic fungi, may be an alternative to reduce contamination by chemicals in housing and food storage environments. Virulence of isolate ESALQ1037 belonging to the Metarhizium anisopliae complex against nymphs and adults of Blattella germanica (L.), and its infectivity following exposure of insects to a contaminated surface or to M. anisopliae-bait were determined under laboratory conditions. Estimated LD50 15 d following topical inoculation was 2.69 x 105 conidia per adult, whereas for nymphs the maximum mortality was lower than 50%. Baits amended with M. anisopliae conidia had no repellent effect on targets; adult mortality was inferior to 25%, and nymphs were not susceptible. All conidia found in the digestive tract of M. anisopliae-bait fed cockroaches were unviable, and bait-treated insects that succumbed to fungal infection showed a typical mycelial growth on mouthparts and front legs, but not on the hind body parts. As opposed to baits, the use of a M. anisopliae powdery formulation for surface treatment was effective in attaining high mortality rates of B. germanica. Both nymphs and adults were infected when this delivery strategy was used, and mycelia growth occurred all over the body surface. Our results suggest that the development of powders or similar formulations of M. anisopliae to control B. germanica may provide faster and better results than some of the strategies based on baits currently available. (author)

Nest sanitation through defecation: antifungal properties of wood cockroach feces

Science.gov (United States)

Rosengaus, Rebeca B.; Mead, Kerry; Du Comb, William S.; Benson, Ryan W.; Godoy, Veronica G.

2013-11-01

The wood cockroach Cryptocercus punctulatus nests as family units inside decayed wood, a substrate known for its high microbial load. We tested the hypothesis that defecation within their nests, a common occurrence in this species, reduces the probability of fungal development. Conidia of the entomopathogenic fungus, Metarhizium anisopliae, were incubated with crushed feces and subsequently plated on potato dextrose agar. Relative to controls, the viability of fungal conidia was significantly reduced following incubation with feces and was negatively correlated with incubation time. Although the cockroach's hindgut contained abundant β-1,3-glucanase activity, its feces had no detectable enzymatic function. Hence, these enzymes are unlikely the source of the fungistasis. Instead, the antifungal compound(s) of the feces involved heat-sensitive factor(s) of potential microbial origin. When feces were boiled or when they were subjected to ultraviolet radiation and subsequently incubated with conidia, viability was "rescued" and germination rates were similar to those of controls. Filtration experiments indicate that the fungistatic activity of feces results from chemical interference. Because Cryptocercidae cockroaches have been considered appropriate models to make inferences about the factors fostering the evolution of termite sociality, we suggest that nesting in microbe-rich environments likely selected for the coupling of intranest defecation and feces fungistasis in the common ancestor of wood cockroaches and termites. This might in turn have served as a preadaptation that prevented mycosis as these phylogenetically related taxa diverged and evolved respectively into subsocial and eusocial organizations.

Isolate-specific conidiation in Trichoderma in response to different nitrogen sources.

Science.gov (United States)

Steyaert, Johanna M; Weld, Richard J; Stewart, Alison

2010-01-01

A characteristic feature of Trichoderma is the production of concentric rings of conidia in response to alternating light/dark conditions and a single ring of conidia in response to a single burst of light. In this study, conidiation was investigated in four biocontrol isolates (T. hamatum, T. atroviride, T. asperellum, T. virens) and one isolate from the mushroom pathogen species, T. pleuroticola. All five isolates produced concentric conidial rings under alternating light/dark conditions on potato-dextrose agar (PDA), however, in response to a 15min burst of blue light, only T. asperellum and T. virens produced a clearly defined conidial ring. Both T. pleuroticola and T. hamatum photoconidiated in a disk-like fashion and T. atroviride produced a broken ring with a partially filled in appearance. In the presence of primary nitrogen, T. asperellum and T. pleuroticola conidiated in a disk, whereas, when grown in the presence of secondary nitrogen, a ring of conidia was produced. Primary nitrogen promoted photoconidiation and competency to conidiate in response to light appeared dependent on the nitrogen catabolite repression state of the cell. Mycelial injury was also investigated in the same five isolates of Trichoderma on PDA and under different nitrogen statuses. For the first time, we report that conidiation in response to injury is differentially regulated in different isolates/species of Trichoderma. Copyright © 2009 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Differential susceptibility of adults and nymphs of Blattella germanica (L.) (Blattodea: Blattellidae) to infection by Metarhizium anisopliae and assessment of delivery strategies

Energy Technology Data Exchange (ETDEWEB)

Lopes, R.B., E-mail: rblopes@cenargen.embrapa.b [EMBRAPA Recursos Geneticos e Biotecnologia, Brasilia, DF (Brazil); Alves, S.B. [Escola Superior de Agricultura Luiz de Queiroz (ESALQ/USP), Piracicaba, SP (Brazil). Dept. de Entomologia e Acarologia

2011-05-15

Microbial insecticides for cockroach control, such as those containing entomopathogenic fungi, may be an alternative to reduce contamination by chemicals in housing and food storage environments. Virulence of isolate ESALQ1037 belonging to the Metarhizium anisopliae complex against nymphs and adults of Blattella germanica (L.), and its infectivity following exposure of insects to a contaminated surface or to M. anisopliae-bait were determined under laboratory conditions. Estimated LD50 15 d following topical inoculation was 2.69 x 105 conidia per adult, whereas for nymphs the maximum mortality was lower than 50%. Baits amended with M. anisopliae conidia had no repellent effect on targets; adult mortality was inferior to 25%, and nymphs were not susceptible. All conidia found in the digestive tract of M. anisopliae-bait fed cockroaches were unviable, and bait-treated insects that succumbed to fungal infection showed a typical mycelial growth on mouthparts and front legs, but not on the hind body parts. As opposed to baits, the use of a M. anisopliae powdery formulation for surface treatment was effective in attaining high mortality rates of B. germanica. Both nymphs and adults were infected when this delivery strategy was used, and mycelia growth occurred all over the body surface. Our results suggest that the development of powders or similar formulations of M. anisopliae to control B. germanica may provide faster and better results than some of the strategies based on baits currently available. (author)

Density and molecular epidemiology of Aspergillus in air and relationship to outbreaks of Aspergillus infection

NARCIS (Netherlands)

A.C.A.P. Leenders (Alexander); A.F. van Belkum (Alex); M.D. Behrendt (Myra); A. Luijendijk (Ad); H.A. Verbrugh (Henri)

1999-01-01

textabstractAfter five patients were diagnosed with nosocomial invasive aspergillosis caused by Aspergillus fumigatus and A. flavus, a 14-month surveillance program for pathogenic and nonpathogenic fungal conidia in the air within and outside the University Hospital in

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Indian Academy of Sciences (India)

PRAKASH KUMAR

natural populations of Neurospora are prevailingly sexual, not clonal (Perkins and Turner ... attesting to conidia being a highly palatable food for mites. .... Lauter F-R and Russo V E A 1989 Fast light regulated genes of Neurospora crassa;.

DIVERSIDAD GENÉTICA DE Peronospora sparsa (PERONOSPORACEAE) EN CULTIVOS DE ROSA DE COLOMBIA

OpenAIRE

MARILUZ AYALA-VÁSQUEZ; LUZ EDITH ARGEL-ROLDAN; MAURICIO MARÍN-MONTOYA

2008-01-01

El mildeo velloso de la rosa es la enfermedad más limitante de este cultivo en Colombia. Aunque algunos trabajos de investigación se han adelantado recientemente sobre la epidemiología de esta enfermedad, uno de los campos menos estudiados es el nivel de variabilidad de su agente causal. El conocimiento de este aspecto es fundamental para definir las estrategias de manejo, especialmente aquellas relacionadas con la resistencia genética y el control químico. Los objetivos de esta investigación...

DIVERSIDAD GENÉTICA DE Peronospora sparsa (PERONOSPORACEAE) EN CULTIVOS DE ROSA DE COLOMBIA

OpenAIRE

MARILUZ AYALA-VÁSQUEZ; LUZ EDITH ARGEL-ROLDAN; SONIA JARAMILLO-VILLEGAS; MAURICIO MARÍN-MONTOYA

2008-01-01

El mildeo velloso de la rosa es la enfermedad más limitante de este cultivo en Co- lombia. Aunque algunos trabajos de investigación se han adelantado recientemente sobre la epidemiología de esta enfermedad, uno de los campos menos estudiados es el nivel de variabilidad de su agente causal. El conocimiento de este aspecto es fun- damental para definir las estrategias de manejo, especialmente aquellas relacionadas con la resistencia genética y el control químico. Los objetivos de esta investiga...

Estado Actual de Peronospora sparsa, Causante del Mildiu Velloso en Rosa ( Rosa sp.)

OpenAIRE

Pablo Israel Álvarez Romero

2013-01-01

La rosa ( Rosa sp.) es una ornamental de importancia en México. Es afectada por un complejo de enfermedades, destacándose el Mildiu velloso ocasionado por el Chromista Per onospora sparsa que incide en la producti vidad, calidad, comercia lización y costos de producción, las pérdidas llegan hasta el 100 % de los ta llos florales. Entre los fungicidas empleados para el control del Mildiu velloso se destacan: fosetil-Al, dimetomorph, c y m o x a n i l , m e t a l a x y l . A l m o m e n t o e x...

Evaluating the virulence of Metarhizium anisopliae (Deuteromycotina: Hyphomycetes and Beauveria bassiana (Ascomycota: Hypocreales isolates to Arabian rhinoceros beetle, Oryctes agamemnon arabicus

Directory of Open Access Journals (Sweden)

M.W. Khudhair

2015-12-01

Full Text Available Virulence of entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana were tested against Arabian Rhinoceros Beetle, Oryctes agamemnon arabicus larvae. Four concentrations (1×105, 1×107, 1×109 and 1×1011 conidia/mL–1 of two locally isolated entomopathogenic fungi spore suspensions were used in this study via larval direct spraying. Results revealed that both isolates can cause high mortality rate reaching 100% after 29 days. However, Beauveria bassiana scored higher mortality rate in short time especially at the concentration of 1×1011 conidia/ mL–1 with lethal time (LT50 12.75 and LT90 20.00; while, Metarhizium anisopliae caused the higher percentage of malformed adults. Moreover, both isolates affected insect’s life cycle particularly in the pupal stage which was reduced remarkably by almost 50% in comparison with the control treatment.

Sem study on the invasion of Nomuraea rileyi (Farlow) on silkworm, Bombyx mori Linn. causing green muscardine.

Science.gov (United States)

Kumar, V; Singh, G P; Kumar, V; Babu, A M; Datta, R K

1997-01-01

The mature conidia of Nomuraea rileyi (Farlow) germinate on the larval integument of Bombyx mori within 24 h and penetrate the cuticle within 36 h after inoculation at 24.0 +/- 1.0 degrees C temperature and 80.0 +/- 5.0% relative humidity. The penetrating hyphae multiply by budding and septa formation in the hemocoel, and the larva succumbs to the infection 6-7 days post-treatment. The hyphal bodies elongate and become interwoven with other hyphae forming a mycelial complex across different tissues. The ramification of hyphae along the epidermal tissue results in larval mummification in 7-8 days. Numerous conidiophores emerge, producing a confluent white fungal mat over the entire surface of the host larva by 9-10 days. Pale green conidia develop, making the larval body green. Life cycle of the fungus on B. mori is completed in 10-11 days.

DIFFERENTIAL ALLERGIC AND NEUROTROPHIN RESPONSES TO FUNGAL COMPONENT EXTRACTS IN BALB/C MICE

Science.gov (United States)

Metarhizium anisopliae mycelium (MYC), conidia (CON) and inducible protease (IND) extracts were combined to produce the antigen MACA to screen for allergenic potential. Involuntary aspiration (IA) exposure to MACA in BALB/c mice has caused immune, inflammatory and physiological ...

GERMINATION, VIABILITY AND CLEARANCE OF STACHYBOTRYS CHARTARUM IN THE LUNGS OF INFANT RATS

Science.gov (United States)

The fungus Stachybotrys chartarum has been associated with many adverse health effects including the condition known as idiopathic pulmonary hemorrhage in infants. In order to gain some insight into possible mechanisms, viable conidia of S. chartarum were instilled into the lung...

Differentiation inside multicelled macroconidia of Fusarium culmorum during early germination

NARCIS (Netherlands)

Chitarra, Gilma S; Breeuwer, Pieter; Rombouts, Frans M; Abee, Tjakko; Dijksterhuis, Jan

Multicelled conidia are formed by many fungal species, but germination of these spores is scarcely studied. Here, the germination and the effects of antimicrobials on multicompartment macroconidia of Fusarium culmorum were investigated. Germ-tube formation was mostly from apical compartments. The

Differentiation inside multicelled macroconidia of Fusarium culmorum during early germination

NARCIS (Netherlands)

Chitarra, G.S.; Breeuwer, P.; Rombouts, F.M.; Abee, T.; Dijksterhuis, J.

2005-01-01

Multicelled conidia are formed by many fungal species, but germination of these spores is scarcely studied. Here, the germination and the effects of antimicrobials on multicompartment macroconidia of Fusarium culmorum were investigated. Germ-tube formation was mostly from apical compartments. The

Phylogeny and taxonomy of the genus Cylindrocladiella

NARCIS (Netherlands)

Lombard, L.; Shivas, R.G.; To-anun, C.; Crous, P.W.

2012-01-01

The genus Cylindrocladiella was established to accommodate Cylindrocladium-like fungi that have small, cylindrical conidia and aseptate stipe extensions. Contemporary taxonomic studies of these fungi have relied on morphology and to a lesser extent on DNA sequence comparisons of the internal

They seldom occur alone

NARCIS (Netherlands)

Crous, Pedro W.; Groenewald, Johannes Z.

2016-01-01

Species of Coleophoma have been reported as plant pathogenic, saprobic or endophytic on a wide host range. The genus is characterised by having pycnidial conidiomata, phialidic conidiogenous cells intermingled among paraphyses, and cylindrical conidia. Coleophoma has had a confusing taxonomic

Biotechnology of Aureobasidium pullulans: A phylogenetic perspective

Science.gov (United States)

Aureobasidium pullulans is a fungus historically included among the "black yeasts." Although many strains are predominantly yeast-like, the species is actually polymorphic, exhibiting complex forms ranging from blastic conidia and swollen cells to pseudophyphae, hyphae, and chlamydospores. A. pull...

Molecular systematics of the cotton root rot pathogen, Phymatotrichopsis omnivora

NARCIS (Netherlands)

Marek, S.M.; Hansen, K.; Romanish, M.; Thorn, R.G.

2009-01-01

Cotton root rot is an important soilborne disease of cotton and numerous dicot plants in the south-western United States and Mexico. The causal organism, Phymatotrichopsis omnivora (= Phymatotrichum omnivorum), is known only as an asexual, holoanamorphic (mitosporic) fungus, and produces conidia

A re-appraisal of Harknessia (Diaporthales), and the introduction of Harknessiaceae fam. nov.

NARCIS (Netherlands)

Crous, P.W.; Summerell, B.A.; Shivas, R.G.; Carnegie, A.J.; Groenewald, J.Z.

2012-01-01

Harknessiaceae is introduced as a new family in the ascomycete order Diaporthales to accommodate species of Harknessia with their Wuestneia-like teleomorphs. The family is distinguished by having pycnidial conidiomata with brown, furfuraceous margins, brown conidia with hyaline, tube-like basal

Exposure of bed bugs to metarhizium anisopliae, and the effect of defensive secretions on fungal growth in vitro

Science.gov (United States)

Bed bugs Cimex lectularius were treated with conidia of the entomopathogenic fungus Metarhizium anisopliae by topical, spray, and contact exposure. One week post-exposure, inconsistent mortalities were observed, averaging 30% across all treatment groups and replicates. Microscopic examination of top...

Eight novel Bipolaris species identified from John L. Alcorn’s collections at the Queensland Plant Pathology Herbarium (BRIP)

NARCIS (Netherlands)

Tan, Yu Pei; Crous, Pedro W.; Shivas, Roger G.

2016-01-01

Several unidentified specimens of Bipolaris deposited in the Queensland Plant Pathology Herbarium (BRIP) that were previously recognised by Dr. John L. Alcorn as taxonomically interesting were re-examined. The morphology of conidia and conidiophores, as well as phylogenetic inference from the

Sporothrix schenckii sensu stricto and Sporothrix brasiliensis Are Differentially Recognized by Human Peripheral Blood Mononuclear Cells

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José A. Martínez-Álvarez

2017-05-01

Full Text Available Sporothrix schenckii sensu stricto and S. brasiliensis are usually associated to sporotrichosis, a subcutaneous mycosis worldwide distributed. Comparative analyses between these two species indicate they contain genetic and physiological differences that are likely to impact the interaction with host cells. Here, we study the composition of the cell wall from conidia, yeast-like cells and germlings of both species and found they contained the same sugar composition. The carbohydrate proportion in the S. schenckii sensu stricto wall was similar across the three cell morphologies, with exception in the chitin content, which was significantly different in the three morphologies. The cell wall from germlings showed lower rhamnose content and higher glucose levels than other cell morphologies. In S. brasiliensis, the wall sugars were constant in the three morphologies, but glucose was lower in yeast-like cells. In S. schenckii sensu stricto cells most of chitin and β1,3-glucan were underneath wall components, but in S. brasiliensis germlings, chitin was exposed at the cell surface, and β1,3-glucan was found in the outer part of the conidia wall. We also compared the ability of these cells to stimulate cytokine production by human peripheral blood mononuclear cells. The three S. schenckii sensu stricto morphologies stimulated increased levels of pro-inflammatory cytokines, when compared to S. brasiliensis cells; while the latter, with exception of conidia, stimulated higher IL-10 levels. Dectin-1 was a key receptor for cytokine production during stimulation with the three morphologies of S. schenckii sensu stricto, but dispensable for cytokine production stimulated by S. brasiliensis germlings. TLR2 and TLR4 were also involved in the sensing of Sporothrix cells, with a major role for the former during cytokine stimulation. Mannose receptor had a minor contribution during cytokine stimulation by S. schenckii sensu stricto yeast-like cells and

Ocorrência de fungos em sementes de soja produzidas sob calagem e adubação potássica residuais The residual effect of liming and potassium fertilization on the occurrence of fungi in soybean seeds

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H.A.A. Mascarenhas

1995-12-01

Full Text Available Estudou-se o efeito residual da adubação potássica e da calagem sobre a ocorrência de fungos em sementes de soja. O experimento foi instalado com a cultivar IAC-17, no ano agrícola de 1991/92, aplicando doses a lanço de O, 3,5 e 7 t/ha de calcário dolomítico e de 0, 150, 300, 450 e 600 kg/ha de K2O. As sementes colhidas no terceiro ano de cultivo, (1993/94 foram submetidas ao teste de sanidade, pelo método do papel de filtro. Observou-se que a calagem reduziu significativamente a incidência de Phomopsis sp., favorecendo, no entanto, a presença de Aspergillus sp. e de Fusarium sp.. Embora não tenham sido detectadas diferenças significativas, nos tratamentos com calagem, verificou-se redução da incidência de Cercospora kikuchii, Colltotrichum dematium var. truncata e Peronospora manshurica. Apenas a incidência de Phomopsis sp. diminuiu significativamente devido à adubação potássica, sendo menor quando se utilizou a dose de 450 kg/ha de K2O.To study the residual effect of liming and potassium fertilizer on the occurrence of fungi in soybean seeds, an experiment was installed during the 1991/92 growing season, using 0, 3.5 and 7.0 t/ha of dolomitic lime and 0, 150, 300, 450 and 600 kg/ha of K2O in the form of potassium cloride. In 1993/94 (the third year, the harvested seeds were submitted to a seed health test (blotter method. The results showed that liming reduced significantly the incidence of Phomopsis sp. and increased the presence of Aspergillus sp. and Fusarium sp.. Although no significant differences were observed with liming, there was a reduction in the incidence of Cercospora kikuchii, Colletotrichum dematium var. truncala and Peronospora manshurica. Among the fungi found in the seeds, only Phomopsis sp. was significantly reduced by potassium fertilization, the least incidence being observed at the rate of 450 kg/ha.

Cladophialophora abundans, a novel environmental species in the Chaetothyriales isolated from the natural environment

NARCIS (Netherlands)

Feng, P.Y.; de Hoog, G.S.; Najafzadeh, M.J.; Gerrits van den Ende, A.H.G.; Stielow, J.B.; Badali, H.; Boeger, W.A.; Vicente, V.A.

2013-01-01

A novel species of Cladophialophora is described from the natural environment of the mangrove land crab (Ucides cordatus) along the coast, and from plant sources in Brazil. It is morphologically very similar to Cladophialophora bantiana, though with moderately melanized conidia produced in strongly

Borisade et al (10)

African Journals Online (AJOL)

DELL

Entomopathogenic fungi, Metarhizium anisopliae and Isaria farinosa are biocontrol agents (BCA) widely reported for the management of insect pests, and they are potential components of Integrated Pest Management (IPM) systems. Compatibility of their infective conidia with low rates of four agrochemicals; Champ-DP ...

Trehalose is required for the acquisition of tolerance to a variety of stresses in the filamentous fungus Aspergillus nidulans

NARCIS (Netherlands)

Fillinger, S.; Chaveroche, M.; Vries, de R.; Dijck, van P.; Ruijter, G.; Thevelien, J.; Enfert, d' C.

2001-01-01

Trehalose is a non-reducing disaccharide found at high concentrations in Aspergillus nidulans conidia and rapidly degraded upon induction of conidial germination. Furthermore, trehalose is accumulated in response to a heat shock or to an oxidative shock. The authors have characterized the A.

Penicilllium discolor, a new species from cheese, nuts and vegetables

DEFF Research Database (Denmark)

Frisvad, Jens Christian; Samson, Robert A.; Rassing, Birgitte A.

1997-01-01

The new species Penicillum discolor, frequently isolated from nuts, vegetables and cheese is described. It is characterised by rough, dark green conidia, synnemateous growth on malt agar and the production of the secondary metabolites chaetoglobosins A, B and C, palitantin, cyclopenin, cyclopenol...

Penicillium koreense sp. nov., Isolated from Various Soils in Korea

NARCIS (Netherlands)

You, Young-Hyun; Cho, Hye Sun; Song, Jaekyeong; Kim, Dae-Ho; Houbraken, Jos; Hong, Seung-Beom

2014-01-01

During an investigation of the fungal diversity of Korean soils, four Penicillium strains could not be assigned to any described species. The strains formed monoverticillate conidiophores with occasionally a divaricate branch. The conidia were smooth or finely rough-walled, globose to broadly

Epidemiology of Botrytis spotting on gerbera and rose flowers grown under glass

NARCIS (Netherlands)

Kerssies, A.

1994-01-01

Experiments described in this thesis were performed to improve the knowledge on the epidemiology of Botrytis cinerea in glasshouses, with gerbera and rose as model systems. B. cinerea is an airborne fungus, with conidia as the most important

Cladophialophora psammophila, a novel species of Chaetothyriales with a potential use in the bioremediation of volatile aromatic hydrocarbons

NARCIS (Netherlands)

Badali, H.; Prenafata - Boldu, F.X.; Guarro, J.; Klaasen, C.; Meis, J.F.; de Hoog, G.S.

2011-01-01

Cladophialophora is a genus of asexual black yeast-like fungi with one-celled, hydrophobic conidia which is predicted to have teleomorphs in the ascomycete genus Capronia, a member of the order Chaetothyriales. Cladophialophora species are relatively frequently involved in human disease ranging from

Transcriptome of Aspergillus flavus aswA (AFLA_085170) deletion strain related to sclerotial development and production of secondary metabolites

Science.gov (United States)

Aspergillus flavus produces many secondary metabolites including aflatoxins. Besides conidia, the fungus uses sclerotia as another type of propagule. We obtained transcriptomes from four growth conditions of the aswA mutant, a strain impaired in sclerotial development and production of sclerotium-sp...

Exact Molecular Typing of Aspergillus fumigatus. Methods and Applications.

NARCIS (Netherlands)

Valk-van Haren, J.A. de

2008-01-01

Aspergillus species are widely distributed fungi that release large amounts of airborne conidia that are dispersed in the environment. Aspergillus fumigatus is the species most frequently isolated from human infections. In this thesis a novel assay for fingerprinting A. fumigatus is described and

Tracing Conidial Fate and Measuring Host Cell Antifungal Activity Using a Reporter of Microbial Viability in the Lung

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Anupam Jhingran

2012-12-01

Full Text Available Fluorescence can be harnessed to monitor microbial fate and to investigate functional outcomes of individual microbial cell-host cell encounters at portals of entry in native tissue environments. We illustrate this concept by introducing fluorescent Aspergillus reporter (FLARE conidia that simultaneously report phagocytic uptake and fungal viability during cellular interactions with the murine respiratory innate immune system. Our studies using FLARE conidia reveal stepwise and cell-type-specific requirements for CARD9 and Syk, transducers of C-type lectin receptor and integrin signals, in neutrophil recruitment, conidial uptake, and conidial killing in the lung. By achieving single-event resolution in defined leukocyte populations, the FLARE method enables host cell profiling on the basis of pathogen uptake and killing and may be extended to other pathogens in diverse model host organisms to query molecular, cellular, and pharmacologic mechanisms that shape host-microbe interactions.

Changes in the physiological properties and kinetics of citric acid accumulation via carbon ion irradiation mutagenesis of Aspergillus niger *

Science.gov (United States)

Hu, Wei; Chen, Ji-hong; Wang, Shu-yang; Liu, Jing; Song, Yuan; Wu, Qing-feng; Li, Wen-jian

2016-01-01

The objective of this work was to produce citric acid from corn starch using a newly isolated mutant of Aspergillus niger, and to analyze the relationship between changes in the physiological properties of A. niger induced by carbon ion irradiation and citric acid accumulation. Our results showed that the physiological characteristics of conidia in A. niger were closely related to citric acid accumulation and that lower growth rate and viability of conidia may be beneficial to citric acid accumulation. Using corn starch as a raw material, a high-yielding citric acid mutant, named HW2, was obtained. In a 10-L bioreactor, HW2 can accumulate 118.9 g/L citric acid with a residual total sugar concentration of only 14.4 g/L. This represented an 18% increase in citric acid accumulation and a 12.5% decrease in sugar utilization compared with the original strain.

Efeitos de Beauveria bassiana (Bals Vuill e Metarhizium anisopliae (Metsc Sorok sobre fêmeas ingurgitadas de Amblyomma cajennense (Fabricius, 1787 em condições de laboratório Effects of Beauveria bassiana (Bals Vuill and Metarhizium anisopliae (Metsc Sorok on engorged females of Amblyomma cajennense (Fabricius, 1787 in laboratory conditions

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R.C.S. Reis

2004-12-01

Full Text Available The in vitro susceptibility of Amblyomma cajennense engorged females to some isolated of the fungus Beauveria bassiana and Metarhizium anisopliae was verified and lethal concentrations (LC 50 and LC 90 were calculated. The females were dived in conidia suspensions for five minutes, and kept in climatically controlled chambers BOD under 27° C and 80% relative humidity. Each bioassay had four treatments in concentrations of 10(5,10(6,10(7e10(8 conidia/ml. A control group was also used. The following characteristics were evaluated: weight and period of oviposition, indexes of reproductive and nutritional efficiency and percentage of microbiological control. A dose dependent negative effect was observed in ticks treated with the suspension. All isolates tested cause a negative effect on in vitro tests of engorged females of A. cajennense, suggesting its potential for microbiological control of tick's species.

Discrimination of Aspergillus lentulus from Aspergillus fumigatus by Raman spectroscopy and MALDI-TOF MS.

Science.gov (United States)

Verwer, P E B; van Leeuwen, W B; Girard, V; Monnin, V; van Belkum, A; Staab, J F; Verbrugh, H A; Bakker-Woudenberg, I A J M; van de Sande, W W J

2014-02-01

In 2005, a new sibling species of Aspergillus fumigatus was discovered: Aspergillus lentulus. Both species can cause invasive fungal disease in immune-compromised patients. The species are morphologically very similar. Current techniques for identification are PCR-based or morphology-based. These techniques are labour-intense and not sufficiently discriminatory. Since A. lentulus is less susceptible to several antifungal agents, it is important to correctly identify the causative infectious agent in order to optimize antifungal therapy. In this study we determined whether Raman spectroscopy and/or MALDI-TOF MS were able to differentiate between A. lentulus and A. fumigatus. For 16 isolates of A. lentulus and 16 isolates of A. fumigatus, Raman spectra and peptide profiles were obtained using the Spectracell and MALDI-TOF MS (VITEK MS RUO, bioMérieux) respectively. In order to obtain reliable Raman spectra for A. fumigatus and A. lentulus, the culture medium needed to be adjusted to obtain colourless conidia. Only Raman spectra obtained from colourless conidia were reproducible and correctly identified 25 out of 32 (78 %) of the Aspergillus strains. For VITEK MS RUO, no medium adjustments were necessary. Pigmented conidia resulted in reproducible peptide profiles as well in this case. VITEK MS RUO correctly identified 100 % of the Aspergillus isolates, within a timeframe of approximately 54 h including culture. Of the two techniques studied here, VITEK MS RUO was superior to Raman spectroscopy in the discrimination of A. lentulus from A. fumigatus. VITEK MS RUO seems to be a successful technique in the daily identification of Aspergillus spp. within a limited timeframe.

Mummified fruit as a source of inoculum and disease dynamics of olive anthracnose caused by Colletotrichum spp.

Science.gov (United States)

Moral, Juan; Trapero, Antonio

2012-10-01

Anthracnose, caused by Colletotrichum spp., is a destructive disease of olive fruit worldwide. The objective of this study was to investigate the influence of agronomical and weather factors on inoculum production using detached olive fruit and on the development of epidemics in the field. The pathogen produced very large numbers of conidia on rotted (>1.87 × 10(8) conidia/fruit) or mummified (>2.16 × 10(4) conidia/fruit) fruit under optimal conditions. On mummified fruit, conidial production was highest on mummies incubated at 20 to 25°C and 96 h of wetness. Repeated washings of mummies reduced conidial production until it was very low after five washings. When mummies were placed in the tree canopy, conidial production was not reduced after 6 months (May to October); but, when they were held on the soil or buried in the soil, conidial production comparatively decreased up to 10,000 times. Anthracnose epidemics on susceptible 'Hojiblanca' and 'Picudo' during three seasons (2005-08) were influenced by rainfall, temperature, and fruit ripening, and had three main phases: the latent period (May to October); the onset of the epidemic, which coincided with the beginning of fruit ripening (early November); and disease development, which was predicted by the Weibull model (November to March). No epidemics developed on the susceptible cultivars during the driest season (2007-08) or on the resistant 'Picual' olive during any of the three seasons. These results provide the basis for a forecasting system of olive anthracnose which could greatly improve the management of this disease.

Use of Green Fluorescent Protein-Transgenic Strains to Study Pathogenic and Nonpathogenic Lifestyles in Colletotrichum acutatum.

Science.gov (United States)

Horowitz, Sigal; Freeman, Stanley; Sharon, Amir

2002-07-01

ABSTRACT Colletotrichum acutatum, which causes anthracnose disease on strawberry, can also persist on several other plant species without causing disease symptoms. The genetic and molecular bases that determine pathogenic and nonpathogenic lifestyles in C. acutatum are unclear. We developed a transformation system for C. acutatum by electroporation of germinating conidia, and transgenic isolates that express the green fluorescent protein (GFP) were produced. Details of the pathogenic and nonpathogenic lifestyles of C. acutatum were determined by using GFP-transgenic isolates. Major differences between colonization-mediating processes of strawberry and of other plants were observed. On the main host, strawberry, the germinating conidia formed branched, thick hyphae, and large numbers of appressoria were produced that were essential for plant penetration. In strawberry, the fungus developed rapidly, filling the mesophyll with dense mycelium that invaded the cells and caused necrosis of the tissue. In nonpathogenic interactions on pepper, eggplant, and tomato, the conidia germinated, producing thin, straight germ tubes. Appressoria were produced but failed to germinate and penetrate leaf tissue, resulting in epiphytic growth without invasion of the plant. Penetration of the plant occurred only several days after inoculation and was restricted to the intercellular spaces of the first cell layers of infected tissue without causing any visible damage. Much of the new fungal biomass continued to develop on the surface of inoculated organs in the nonpathogenic interaction. The differences in fungal development on strawberry compared with the other plant species suggest that signal molecules, which may be present only in strawberry, trigger appressorial germination and penetration of the primary host.

The Major Chromoblastomycosis Etiologic Agent Fonsecaea pedrosoi Activates the NLRP3 Inflammasome

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Raffael Júnio Araújo de Castro

2017-11-01

Full Text Available Fonsecaea pedrosoi is the main etiologic agent of chromoblastomycosis (CBM, one of the most prevalent subcutaneous mycosis in tropical and subtropical countries. CBM is a poorly characterized chronic infection that commonly starts after transcutaneous inoculation of conidia and saprophytic hyphae of F. pedrosoi. Recently, we have shown that unlike conidia, hyphae and muriform cells (the parasitic morphotype of F. pedrosoi promotes an intense inflammatory response pattern in vivo, which comprises the production of an inflammasome-derived cytokine, IL-1β. Nonetheless, the mechanisms underlying IL-1β production and maturation upon F. pedrosoi infection and its functional output in the course of CBM remains unknown. We show here that F. pedrosoi hyphae, differently from conidia, induce IL-1β secretion in both bone marrow-derived dendritic cells and macrophages. Using inhibitors and knockout cells, we demonstrated that the mechanisms underlying IL-1β production by hyphae-infected macrophages were dependent on dectin-1, -2, and -3 receptors and the Syk-NF-kB signaling pathway. Furthermore, F. pedrosoi promoted a NLRP3-dependent inflammasome activation, which required potassium efflux, reactive oxygen species production, phagolysosomal acidification, and cathepsin B release as triggers. IL-1β processing and release was mediated primarily by caspase-1 and, to a lesser extent, by caspase-8-dependent cleavage. Finally, we showed using a murine CBM model that F. pedrosoi elicits a NLRP3-regulated IL-1β and interleukin-18 release in vivo, but without NLRP3 inflammasome activation interfering in the course of the experimental infection.

The Major Chromoblastomycosis Etiologic Agent Fonsecaea pedrosoi Activates the NLRP3 Inflammasome.

Science.gov (United States)

de Castro, Raffael Júnio Araújo; Siqueira, Isaque Medeiros; Jerônimo, Márcio Sousa; Basso, Angelina Maria Moreschi; Veloso Junior, Paulo Henrique de Holanda; Magalhães, Kelly Grace; Leonhardt, Luiza Chaves; de Oliveira, Stephan Alberto Machado; Bürgel, Pedro Henrique; Tavares, Aldo Henrique; Bocca, Anamélia Lorenzetti

2017-01-01

Fonsecaea pedrosoi is the main etiologic agent of chromoblastomycosis (CBM), one of the most prevalent subcutaneous mycosis in tropical and subtropical countries. CBM is a poorly characterized chronic infection that commonly starts after transcutaneous inoculation of conidia and saprophytic hyphae of F. pedrosoi . Recently, we have shown that unlike conidia, hyphae and muriform cells (the parasitic morphotype) of F. pedrosoi promotes an intense inflammatory response pattern in vivo , which comprises the production of an inflammasome-derived cytokine, IL-1β. Nonetheless, the mechanisms underlying IL-1β production and maturation upon F. pedrosoi infection and its functional output in the course of CBM remains unknown. We show here that F. pedrosoi hyphae, differently from conidia, induce IL-1β secretion in both bone marrow-derived dendritic cells and macrophages. Using inhibitors and knockout cells, we demonstrated that the mechanisms underlying IL-1β production by hyphae-infected macrophages were dependent on dectin-1, -2, and -3 receptors and the Syk-NF-kB signaling pathway. Furthermore, F. pedrosoi promoted a NLRP3-dependent inflammasome activation, which required potassium efflux, reactive oxygen species production, phagolysosomal acidification, and cathepsin B release as triggers. IL-1β processing and release was mediated primarily by caspase-1 and, to a lesser extent, by caspase-8-dependent cleavage. Finally, we showed using a murine CBM model that F. pedrosoi elicits a NLRP3-regulated IL-1β and interleukin-18 release in vivo , but without NLRP3 inflammasome activation interfering in the course of the experimental infection.

Prevalence, persistence, and phenotypic variation of Aspergillus fumigatus in the outdoor environment in Manchester, UK, over a 2-year period.

Science.gov (United States)

Alshareef, Fadwa; Robson, Geoffrey D

2014-05-01

Aspergillus fumigatus, an opportunistic fungal pathogen that causes invasive aspergillosis in immunosuppressed patients, is considered to be the world's most dangerous mould. It is widely distributed in the environment, and airborne asexual conidia serve as the main mode of transport for pulmonary lung infection. It is important to monitor seasonal airborne conidia levels when assessing the risk of acquiring this infection. In this study, air was sampled for total viable fungal spores and viable A. fumigatus conidia monthly over a 2-year period (2009 and 2010) close to Manchester, UK, city center. Total viable airborne fungal counts varied seasonally, peaking in the summer and autumn for both years and reaching levels of approximately 1100-1400 colony-forming units (CFU)/m(3); counts were strongly positively correlated to mean temperature (R(2) = 0.697). By contrast, A. fumigatus viable airborne counts were not seasonally associated; persistent low levels were between 3 and 20 CFU/m(3) and were not correlated with mean temperature (R(2) = 0.018). A total of 220 isolates of A. fumigatus were recovered on potato dextrose agar (PDA) at 45°C, and internal transcribed spacer sequencing and restriction digestion of a partial polymerase chain reaction amplicon of the β-tubulin gene (benA) of 34 randomly selected isolates were used to confirm the isolates as A. fumigatus. When the colony radial growth rates (Kr) were determined, the highest rates were observed on PDA, followed by Vogel's medium supplemented with phosphatidylcholine and Vogel's medium alone. Clinical isolates had a significantly higher mean colony Kr on PDA compared with environmental isolates.

Occurrence of Target Spot on Rosemary Caused by Corynespora cassiicola in Korea

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Wang-Hyu Lee

2013-03-01

Full Text Available The purpose of this experiment was to investigate the development of new spot disease on the leaf and stem of rosemary (Rosmarinus officinalis in commercial greenhouses at Jeonju and Namwon in Korea. Incidence of target spot on rosemary was higher at the end of the rainy season with high humidity. Those symptoms were black ring spots (3−5 mm in diameter and withering on green leaves and stems. Conidiophores and conidia were formed on the infected tissue in moist chamber and conidia were shown as the cylindrical and oval types in chain, ranged from 55 to 275 μm in length, and 7 to 14 μm in width. Conidia with eight to ten pseudosepta were formed on the conidiapore. The optimum growth temperature of isolates was 30oC on the PDA medium under the dark condition. In the pathogenesis test, the target spot and withering symptoms were appeared on the leaves and stems 3 days after inoculation showing similar symptoms compared to those of in nature. The same fungus was re-isolated from infected lesion, indicating that Corynespora cassiicola caused leaf target spot and twig blight on rosemary. The rDNA ITS nucleotide sequences of the pure cultured isolate from the diseased area on rosemary showed 100% similarity to the sequences of C. cassiicola available in the GenBank database (JQ595296, JQ595297, FJ852715 and AY238606. Therefore, we report that the target spot of leaves and stems in rosemary was caused by C. cassiicola.

Morphology and molecular taxonomy of Evlachovaea-like fungi, and the status of this unusual conidial genus.

Science.gov (United States)

Humber, Richard A; Rocha, Luiz F N; Inglis, Peter W; Kipnis, André; Luz, Christian

2013-01-01

The entomopathogenic anamorphic genus Evlachovaea was described to differ from other fungi in forming its conidia obliquely to the axis of the conidiogenous cell and with successive conidia having alternate orientations with a zipper- or chevron-like arrangement resulting in flat, ribbon-like chains. Morphological and molecular studies of six Evlachovaea-like isolates baited from Central Brazilian soils using Triatoma infestans (a vector of Chagas disease) and of other entomopathogens with Evlachovaea-like conidiogenesis led to a re-evaluation of the status of this little known fungal genus. The Brazilian isolates formed two distinct groups based on gene sequences for both the internal transcribed spacer (ITS) and translation elongation factor (EF-1α) genes, morphology, and growth patterns; both groups also differed from the type species, Evlachovaea kintrischica. More detailed studies of these fungi indicated that the alternatingly oblique orientations of forming conidia are neither a stable nor invariant character (even on single phialides). Furthermore, the molecular cladistic analysis unambiguously placed the Evlachovaea isolates firmly within the genus Isaria (Hypocreales: Cordycipitaceae). The ITS sequences of E. kintrischica were very similar or even identical to those of Isaria amoenerosea and Isaria cateniobliqua, thereby suggesting that E. kintrischica is a synonym of one of these species, and that the genus Evlachovaea must be treated as a later synonym of Isaria, which must now be recognized to include several highly divergent modes of conidiogenesis. These taxonomic findings are discussed in the context of dramatic changes recently imposed on the nomenclatural standards used to determine the correct names of all pleomorphic fungi. Published by Elsevier Ltd.

Structural Features of Sugars That Trigger or Support Conidial Germination in the Filamentous Fungus Aspergillus niger

Science.gov (United States)

Hayer, Kimran; Stratford, Malcolm

2013-01-01

The asexual spores (conidia) of Aspergillus niger germinate to produce hyphae under appropriate conditions. Germination is initiated by conidial swelling and mobilization of internal carbon and energy stores, followed by polarization and emergence of a hyphal germ tube. The effects of different pyranose sugars, all analogues of d-glucose, on the germination of A. niger conidia were explored, and we define germination as the transition from a dormant conidium into a germling. Within germination, we distinguish two distinct stages, the initial swelling of the conidium and subsequent polarized growth. The stage of conidial swelling requires a germination trigger, which we define as a compound that is sensed by the conidium and which leads to catabolism of d-trehalose and isotropic growth. Sugars that triggered germination and outgrowth included d-glucose, d-mannose, and d-xylose. Sugars that triggered germination but did not support subsequent outgrowth included d-tagatose, d-lyxose, and 2-deoxy-d-glucose. Nontriggering sugars included d-galactose, l-glucose, and d-arabinose. Certain nontriggering sugars, including d-galactose, supported outgrowth if added in the presence of a complementary triggering sugar. This division of functions indicates that sugars are involved in two separate events in germination, triggering and subsequent outgrowth, and the structural features of sugars that support each, both, or none of these events are discussed. We also present data on the uptake of sugars during the germination process and discuss possible mechanisms of triggering in the absence of apparent sugar uptake during the initial swelling of conidia. PMID:23995938

Method To immobilize the aphid-pathogenic fungus erynia neoaphidis in an alginate matrix for biocontrol

Science.gov (United States)

Shah; Aebi; Tuor

1998-11-01

Erynia neoaphidis is an important fungal pathogen of aphid pests worldwide. There have been few reported attempts to formulate this natural agent for use in biocontrol. In the current study, factors involved in the immobilization of E. neoaphidis hyphae in an alginate matrix were investigated. Hyphae of two isolates cultured in liquid medium were 220 to 620 &mgr;m in length and 7 to 19 &mgr;m in diameter with a 74 to 83% cytoplasmic content. The optimal concentration of low-viscosity sodium alginate for production of conidia from entrapped hyphae was 1.5% (wt/vol), and 0.1 and 0.25 M calcium chloride were equally suitable for use as the gelling solution. Alginate beads were rinsed with 10% sucrose after gelling. However, beads should not be left for longer than 40 min in 0.1 M calcium chloride or 10% sucrose to prevent a 10% loss in conidial production. A 40% (vol/vol) concentration of fungal biomass produced significantly more conidia than either 20% or the standard concentration of 10%. This effect persisted even after beads were dried overnight in a laminar flow hood and stored at 4 degreesC for 4 days. Conidia from freshly produced alginate beads caused 27 to 32% infection in Pea aphids as determined by standardized laboratory bioassays. This finding was not significantly different from infections in aphids inoculated with fresh mycelial mats or plugs from Petri dish cultures. In conclusion, algination appears to be a promising technique for utilizing E. neoaphidis in the biocontrol of aphid pests.

The Proteomic Signature of Aspergillus fumigatus During Early Development*

Science.gov (United States)

Cagas, Steven E.; Jain, Mohit Raja; Li, Hong; Perlin, David S.

2011-01-01

Aspergillus fumigatus is a saprophytic fungus that causes a range of diseases in humans including invasive aspergillosis. All forms of disease begin with the inhalation of conidia, which germinate and develop. Four stages of early development were evaluated using the gel free system of isobaric tagging for relative and absolute quantitation to determine the full proteomic profile of the pathogen. A total of 461 proteins were identified at 0, 4, 8, and 16 h and fold changes for each were established. Ten proteins including the hydrophobin rodlet protein RodA and a protein involved in melanin synthesis Abr2 were found to decrease relative to conidia. To generate a more comprehensive view of early development, a whole genome microarray analysis was performed comparing conidia to 8 and 16 h of growth. A total of 1871 genes were found to change significantly at 8 h with 1001 genes up-regulated and 870 down-regulated. At 16 h, 1235 genes changed significantly with 855 up-regulated and 380 down-regulated. When a comparison between the proteomics and microarray data was performed at 8 h, a total of 22 proteins with significant changes also had corresponding genes that changed significantly. When the same comparison was performed at 16 h, 12 protein and gene combinations were found. This study, the most comprehensive to date, provides insights into early pathways activated during growth and development of A. fumigatus. It reveals a pathogen that is gearing up for rapid growth by building translation machinery, generating ATP, and is very much committed to aerobic metabolism. PMID:21825280

Hessian-based quantitative image analysis of host-pathogen confrontation assays.

Science.gov (United States)

Cseresnyes, Zoltan; Kraibooj, Kaswara; Figge, Marc Thilo

2018-03-01

Host-fungus interactions have gained a lot of interest in the past few decades, mainly due to an increasing number of fungal infections that are often associated with a high mortality rate in the absence of effective therapies. These interactions can be studied at the genetic level or at the functional level via imaging. Here, we introduce a new image processing method that quantifies the interaction between host cells and fungal invaders, for example, alveolar macrophages and the conidia of Aspergillus fumigatus. The new technique relies on the information content of transmitted light bright field microscopy images, utilizing the Hessian matrix eigenvalues to distinguish between unstained macrophages and the background, as well as between macrophages and fungal conidia. The performance of the new algorithm was measured by comparing the results of our method with that of an alternative approach that was based on fluorescence images from the same dataset. The comparison shows that the new algorithm performs very similarly to the fluorescence-based version. Consequently, the new algorithm is able to segment and characterize unlabeled cells, thus reducing the time and expense that would be spent on the fluorescent labeling in preparation for phagocytosis assays. By extending the proposed method to the label-free segmentation of fungal conidia, we will be able to reduce the need for fluorescence-based imaging even further. Our approach should thus help to minimize the possible side effects of fluorescence labeling on biological functions. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

Conidiogenesis-related DNA photolyase gene in Beauveria bassiana.

Science.gov (United States)

Lee, Se Jin; Lee, Mi Rong; Kim, Sihyeon; Kim, Jong Cheol; Park, So Eun; Shin, Tae Young; Kim, Jae Su

2018-03-01

Beauveria bassiana is an entomopathogenic fungi used in environmentally mindful pest management. Its main active ingredient, conidia, is commercially available as a fungal biopesticide. Many studies of conidia production have focused on how to optimize culture conditions for maximum productivity and stability against unfavorable abiotic factors. However, understanding of how conidiogenesis-related genes provide improved conidial production remains unclear. In this study, we focus on identifying conidiogenesis-related genes in B. bassiana ERL1170 using a random mutagenesis technique. Transformation of ERL1170 using restriction enzyme-mediated integration generated one morphologically different transformant, ERL1170-pABeG #163. The transformant was confirmed to represent B. bassiana, and the binary vector was successfully integrated into the genome of ERL1170. Compared to the wild type, transformant #163 showed very slow hyphal growth and within 6 days only produced bassiana exhibits thread-like hyphae and conidiophore structures and circular conidia. To determine the location of the randomly inserted DNA, we conducted thermal asymmetric interlaced (TAIL) PCR and Escherichia coli cloning to clearly sequence the disrupted region. We identified one colony (colony No. 7) with an insertion site identified as DNA photolyase. This was confirmed through a gene knock-out study. It is possible the gene that encodes for DNA photolyase was disrupted during the insertion process and might be involved in fungal conidiogenesis. This work serves as a platform for exploring the function of a variety of B. bassiana genes involved in pest management and their downstream processing. Copyright © 2018 Elsevier Inc. All rights reserved.

Evaluation of a New Entomopathogenic Strain of Beauveria bassiana and a New Field Delivery Method against Solenopsis invicta.

Directory of Open Access Journals (Sweden)

Jun Li

Full Text Available Solenopsis invicta Buren is one of the most important pests in China, and control measures are mainly based on the use of synthetic pesticides, which may be inadequate and unsustainable. Hence, there is a growing interest in developing biological control alternatives for managing S. invicta, such as the use of entomopathogenic fungi. To facilitate the commercialization of entomopathogenic fungi against S. invicta, 10 Beauveria bassiana isolates originating from different hosts were tested for virulence in laboratory bioassays, and the most pathogenic strain, ZGNKY-5, was tested in field studies using an improved pathogen delivery system. The cumulative mortality rate reached 93.40% at 1×108 mL-1 conidia after 504 h. The germination and invasion of the spores were observed under a scanning electron microscope, and several conidia adhered to the cuticle of S. invicta after 2 h. Furthermore, the germ tubes of the conidia oriented toward the cuticle after 48 h, and the mycelium colonized the entire body after 96 h. Based on the efficacy observed in the laboratory trials, further experiments were performed with ZGNKY-5 strain to evaluate its utility in an injection control technology against S. invicta in the field. We found that three dosage treatments of ZGNKY-5 strain (500 mL, 750 mL, and 1,000 mL per nest had significant control effects. Our results show that this strain of Beauveria bassiana and our control method were effective against S. invicta in both laboratory and field settings.

Evaluation of a New Entomopathogenic Strain of Beauveria bassiana and a New Field Delivery Method against Solenopsis invicta

Science.gov (United States)

Li, Jun; Guo, Qiang; Lin, Miaofeng; Jiang, Lu; Ye, Jingwen; Chen, Dasong; Li, Zhigang; Dai, Jianqing; Han, Shichou

2016-01-01

Solenopsis invicta Buren is one of the most important pests in China, and control measures are mainly based on the use of synthetic pesticides, which may be inadequate and unsustainable. Hence, there is a growing interest in developing biological control alternatives for managing S. invicta, such as the use of entomopathogenic fungi. To facilitate the commercialization of entomopathogenic fungi against S. invicta, 10 Beauveria bassiana isolates originating from different hosts were tested for virulence in laboratory bioassays, and the most pathogenic strain, ZGNKY-5, was tested in field studies using an improved pathogen delivery system. The cumulative mortality rate reached 93.40% at 1×108 mL-1 conidia after 504 h. The germination and invasion of the spores were observed under a scanning electron microscope, and several conidia adhered to the cuticle of S. invicta after 2 h. Furthermore, the germ tubes of the conidia oriented toward the cuticle after 48 h, and the mycelium colonized the entire body after 96 h. Based on the efficacy observed in the laboratory trials, further experiments were performed with ZGNKY-5 strain to evaluate its utility in an injection control technology against S. invicta in the field. We found that three dosage treatments of ZGNKY-5 strain (500 mL, 750 mL, and 1,000 mL per nest) had significant control effects. Our results show that this strain of Beauveria bassiana and our control method were effective against S. invicta in both laboratory and field settings. PMID:27341441

Structural features of sugars that trigger or support conidial germination in the filamentous fungus Aspergillus niger.

Science.gov (United States)

Hayer, Kimran; Stratford, Malcolm; Archer, David B

2013-11-01

The asexual spores (conidia) of Aspergillus niger germinate to produce hyphae under appropriate conditions. Germination is initiated by conidial swelling and mobilization of internal carbon and energy stores, followed by polarization and emergence of a hyphal germ tube. The effects of different pyranose sugars, all analogues of d-glucose, on the germination of A. niger conidia were explored, and we define germination as the transition from a dormant conidium into a germling. Within germination, we distinguish two distinct stages, the initial swelling of the conidium and subsequent polarized growth. The stage of conidial swelling requires a germination trigger, which we define as a compound that is sensed by the conidium and which leads to catabolism of d-trehalose and isotropic growth. Sugars that triggered germination and outgrowth included d-glucose, d-mannose, and d-xylose. Sugars that triggered germination but did not support subsequent outgrowth included d-tagatose, d-lyxose, and 2-deoxy-d-glucose. Nontriggering sugars included d-galactose, l-glucose, and d-arabinose. Certain nontriggering sugars, including d-galactose, supported outgrowth if added in the presence of a complementary triggering sugar. This division of functions indicates that sugars are involved in two separate events in germination, triggering and subsequent outgrowth, and the structural features of sugars that support each, both, or none of these events are discussed. We also present data on the uptake of sugars during the germination process and discuss possible mechanisms of triggering in the absence of apparent sugar uptake during the initial swelling of conidia.

Large-spored Alternaria pathogens in section Porri disentangled

NARCIS (Netherlands)

Woudenberg, J H C; Truter, M; Groenewald, J Z; Crous, P W

The omnipresent fungal genus Alternaria was recently divided into 24 sections based on molecular and morphological data. Alternaria sect. Porri is the largest section, containing almost all Alternaria species with medium to large conidia and long beaks, some of which are important plant pathogens

Large-spored Alternaria pathogens in section Porri disentangled

NARCIS (Netherlands)

Woudenberg, J.H.C.; Truter, M.; Groenewald, J.Z.; Crous, P.W.

2014-01-01

The omnipresent fungal genus Alternaria was recently divided into 24 sections based on molecular and morphological data. Alternaria sect. Porri is the largest section, containing almost all Alternaria species with medium to large conidia and long beaks, some of which are important plant pathogens

The FlbA-regulated predicted transcription factor Fum21 of Aspergillus niger is involved in fumonisin production

DEFF Research Database (Denmark)

Aerts, David; Hauer, Esther E.; Ohm, Robin A.

2017-01-01

Aspergillus niger secretes proteins throughout the colony except for the zone that forms asexual spores called conidia. Inactivation of flbA that encodes a regulator of G-protein signaling results in colonies that are unable to reproduce asexually and that secrete proteins throughout the mycelium...

Sporulation in Erysiphe necator: signals, differential gene expression and possible implications for disease management

Science.gov (United States)

Abundant production of conidia is a driving factor for epidemics of grape powdery mildew (Erysiphe necator (syn. Uncinula necator). Previous investigations revealed evidence for a signal that coordinates the onset of asexual reproduction. The genetic basis for this signal in powdery mildews had not ...

INFECTIVITY OF TWO MEMBERS OF THE ENTOMOPHTHORA MUSCAE COMPLEX [ZYGOMYCETES: ENTOMOPHTHORALES] FOR MUSCA DOMESTICA [DIPT.: MUSCIDAE

DEFF Research Database (Denmark)

Bellini, R; Mullens, B A; Jespersen, J B

1992-01-01

Dose-mortality studies were conducted with 2 members of the Entomophthora muscae (Cohn) Fresenius complex from southern California (CA) and Denmark (DA) infecting house flies, Musca domestica L., from southern California. Primary conidia of the DA form were significantly more infective (LC50 = 34...

Biodiversity of the genus Cladophialophora

NARCIS (Netherlands)

Badali, H.; Gueidan, C.; Najafzadeh, M.J.; Bonifaz, A.; Gerrits van den Ende, A.H.G.; de Hoog, G.S.

2008-01-01

Cladophialophora is a genus of black yeast-like fungi comprising a number of clinically highly significant species in addition to environmental taxa. The genus has previously been characterized by branched chains of ellipsoidal to fusiform conidia. However, this character was shown to have evolved

The FlbA-regulated predicted transcription factor Fum21 of Aspergillus niger is involved in fumonisin production

NARCIS (Netherlands)

Aerts, David; Hauer, Esther E.; Ohm, Robin A.; Arentshorst, Mark; Teertstra, Wieke R.; Phippen, Christopher; Ram, Arthur F J; Frisvad, Jens C.; Wösten, Han A. B.

2017-01-01

Aspergillus niger secretes proteins throughout the colony except for the zone that forms asexual spores called conidia. Inactivation of flbA that encodes a regulator of G-protein signaling results in colonies that are unable to reproduce asexually and that secrete proteins throughout the mycelium.

High viscosity and anisotropy characterize the cytoplasm of fungal dormant stress resistant spores

NARCIS (Netherlands)

Dijksterhuis, J.; Nijsse, J.; Hoekstra, F.A.; Golovina, E.A.

2007-01-01

Ascospores of the fungus Talaromyces macrosporus are dormant and extremely stress resistant, whereas fungal conidia¿the main airborne vehicles of distribution¿are not. Here, physical parameters of the cytoplasm of these types of spores were compared. Cytoplasmic viscosity and level of anisotropy as

Metacridamides A and B from the biocontrol fungus metarhizium acridum

Science.gov (United States)

Metarhizium acridum, an entomopathogenic fungus, has been commercialized and used successfully for biocontrol of grasshopper pests in Africa and Australia. As part of an effort to catalog the secondary metabolites of this fungus we discovered that its conidia produce two novel 17-membered macrocycl...

Conidia dispersal of Diplodia species  in a French vineyard

Directory of Open Access Journals (Sweden)

P. Kuntzmann

2009-05-01

Full Text Available Diseases caused by species of Botryosphaeriaceae lead to significant losses of grape yield. Species in this family produce foliar symptoms similar to, but distinguishable from esca, and the diseases they cause are generally named black dead arm (BDA. Botryosphaeriaceae species are ascomycetes frequently isolated from grapevine stocks showing decline or dieback symptoms. It is therefore useful to know what is the spore dissemination period of Botryosphaeriaceae in the vineyard. The objective of this study was to determine the peak periods of conidial release by some Diplodia spp. in the Botryosphaeriaceae in grapevines and to ascertain the climatic factors that influence inoculum availability and dispersal. Spore dispersal from Vitis vinifera was studied from 2005 to 2006 in a French vineyard. Spores of Diplodia seriata and D.  mutila were trapped throughout the year. Spore release from D. seriata peaked during the vegetative growth period, while D. mutila  released its spores later.

Evaluation of conidia production and mycelial growth in solid culture ...

African Journals Online (AJOL)

FATIMAYALBERTO

2012-10-02

Oct 2, 2012 ... Yeast peptone dextrose agar (YPDA), Potato dextrose agar (PDA), to name a few (Kamp and Bidochka, 2002). Most of these present a media composition based on source of carbon and one nitrogen which generally vary between them and in the proportions in which they are presented as well as in its ...

Evaluation of conidia production and mycelial growth in solid culture ...

African Journals Online (AJOL)

FATIMAYALBERTO

2012-10-02

Oct 2, 2012 ... liquid culture media of different species of ..... Jackson MA, McGuire MR, Lacey LA, Wraight SP (1997). Liquid culture ... fermentation and the substrate in the production of Trichoderma ... Conidiation induction in Penicillium.

Evaluation of conidia production and mycelial growth in solid culture ...

African Journals Online (AJOL)

It is important to know the ability of native strains to carry out the process of sporulation and growth in different conditions and to determine their possible potential as biological control of pests of agricultural importance, mainly in citrus areas. The objective of this study was to evaluate five different solid culture media for the ...

Systematics of Penicillium simplicissimum based on rDNA sequences, morphology and secondary metabolites

DEFF Research Database (Denmark)

Tuthill, D.E.; Frisvad, Jens Christian; Christensen, M.

2001-01-01

supported by differences in micromorphological characters, particularly of the conidia and phialides, and the production of distinct profiles of secondary metabolites by each species. Group-I introns, located in the SSU rDNA, were identified in six of the 21 isolates; their presence was used to test...

Mummy Berry Fruit Rot and Shoot Blight Incidence in Blueberry: Prediction, Ranking, and Stability in a Long-term Study

Science.gov (United States)

Mummy berry is an important disease of cultivated blueberry. The disease has two distinct phases; a blighting phase initiated by ascospores and a fruit infection stage initiated by conidia. In this study we investigated blueberry cultivar resistance to both phases of the disease and, utilizing ‘stan...

Presence of Fusarium spp. in air and soil associated with sorghum fields

Science.gov (United States)

Sorghum grain, valuable for feed, food and bioenergy, can be colonized by several Fusarium species; therefore, it was of interest to identify possible sources of conidia. Analysis of air and soil samples provided evidence for the presence of propagules from Fusarium genotypes that may cause grain in...

Paecilomyces fumosoroseus blastospore production using liquid ...

African Journals Online (AJOL)

SERVER

2007-09-19

Sep 19, 2007 ... using liquid culture in a bioreactor ... less time required, in comparison to fermentation on solid substratum. ... blastospores in liquid medium, the present study evalua- ... liquid-culture studies, conidia were produced by inoculating Sabou- .... a 250 ml baffled shaker flask containing 50 ml of Sabouraud.

Author Details

African Journals Online (AJOL)

Borisade, OA. Vol 23, No 2 (2015) - Articles Rearing tomato whitefly and field evaluation of modified and unmodified conidia of Beauveria bassiana, Isaria farinosa, Metarhizium anisopliae and low rates of Chlorpyrifos under tropical conditions. Abstract PDF. ISSN: 2072-6589. AJOL African Journals Online. HOW TO USE ...

Standard methods for fungal brood disease research

DEFF Research Database (Denmark)

Jensen, Annette Bruun; Aronstein, Kathrine; Manuel Flores, Jose

2013-01-01

. that are rarely observed, so the impact on colony health is not very well understood. A major concern with the presence of Aspergillus in honey bees is the production of airborne conidia, which can lead to allergic bronchopulmonary aspergillosis, pulmonary aspergilloma, or even invasive aspergillosis in lung...

Protein Kinase A subunits of the ascomycete pathogen Mycosphaerella graminicola regulate asexual fructification, filamentation, melanization and osmosensing

NARCIS (Netherlands)

Mehrabi, R.; Kema, G.H.J.

2006-01-01

As in many fungi, asexual reproduction of Mycosphaerella graminicola in planta is a complex process that requires proper differentiation of the infectious hyphae in the substomatal cavities of foliar tissue before pycnidia with conidia can be formed. In this study, we have investigated the role of

Journal of Genetics | Indian Academy of Sciences

Indian Academy of Sciences (India)

Heterokaryons of Neurospora crassa were generated by transformation of multinucleate conidia of a histidine-3 auxotroph with his-3+ plasmid. In one of the transformants, propagated on a medium with histidine supplementation, a gradual but drastic reduction occurred in the proportion of prototrophic nuclei that contained ...

Summer epidemics of apple scab : The relationship between measurements and their implications for the development of predictive models and threshold levels under different disease control regimes

NARCIS (Netherlands)

Holb, I.J.; Heijne, B.; Jeger, M.J.

2003-01-01

A 2-year study on epidemic progress of apple scab was conducted at Randwijk, the Netherlands, in 1998 and 1999. The summer epidemic caused by conidia was studied instead of the well-described spring season epidemic originating from ascospores. The aim was to investigate relationships between disease

Phylogeny of Hirsutella species (Ophiocordycipitaceae) from the USA: remedying the paucity of Hirsutella sequence data

Science.gov (United States)

Hirsutella (Ophiocordycipitaceae: Hypocreales) is a genus of insect, mite, and nematode pathogens with an asexual morph, which generally produce a mucilaginous cluster of one or several conidia on phialides that are basally subulate and taper to a fine neck. The generic name Hirsutella has been prop...

Comparative efficacy and pathogenicity of keratinophilic soil fungi against Culex quinquefasciatus larvae.

Science.gov (United States)

Mohanty, Suman Sundar; Prakash, Soam

2010-09-01

Out of seven fungal species belonging to four genera isolated from pond and wallow soils using feathers of Pavo cristatus as bait, four species viz., Aspergillus flavus, Aspergillus niger, Chrysosporium pseudomerdarium and Trichophyton ajelloi were most frequent. Chrysosporium and Trichophyton spp. were more pathogenic on Culex quinquefasciatus larvae than Aspergillus and Penicillium. The bioefficacy tests conducted as per the protocol of World Health Organization and the LC(50) values calculated by the Probit analysis showed that 3(rd)-instar C. quinquefasciatus were more susceptible to the conidia of above fungi. Highest mortality was observed in the larvae of C. quinquefasciatus when exposed to T. ajelloi. The density of fungal conidia was greatest on the ventral brush, palmate hair and anal region of the mosquito larvae after exposing for 72 hours. The potentiality of these fungi for use in the control of C. quinquefasciatus is discussed which can be exploited as a suitable biocontrol agent in the tropics.

Survival frequency and spectrum of induced mutations in Penicillium vermiculatum dangeard following

International Nuclear Information System (INIS)

Mitra, Jharna; Chaudhari, K.L.

1974-01-01

A comparative study on the sensitivity of UV, X-rays and gamma rays on conidia of Penicillium vermiculatum has been done. LD50 with UV-rays was shown with less than 60 second treatment at a dose of 6 x 10 3 ergs/mm 2 . Beyond 240 second treatment i.e. 2.4 x 10 4 ergs/mm 2 the survival percent remained almost the same, indicating resistance of surviving spores to the action of UV radiation. With X-rays an increase in the germination percent of conidia was noticed at dosages of 5 and 10 Kr. Abrupt fall in the survival percent was observed with increasing dosage of X-rays. A wide range of biochemical morphological mutants was obtained with UV irradiation as compared to those with X-rays. Nicotinic acid requiring mutants occurred frequently with X-rays and methionine requiring ones with gamma-rays. Compact colonial mutants were induced freely with X-rays and violet pigment producing ones with gamma-rays. (author)

Dispersal of Beauveria bassiana by the activity of nettle insects.

Science.gov (United States)

Meyling, Nicolai V; Pell, Judith K; Eilenberg, Jørgen

2006-10-01

Recent studies have shown that the entomopathogenic fungus Beauveria bassiana occurs naturally on the phylloplanes of several plants, including nettles. Insects could, by their activity, be contributing to this inoculum by dispersing it from other sites. The potential of nettle aphids Microlophium carnosum and their predator Anthocoris nemorum to disperse conidia of B. bassiana from soil to nettles and from sporulating cadavers in the nettle canopy was investigated in laboratory experiments. In petri dish assays, aphids showed potential to distribute B. bassiana from soil to nettle leaves. Predators dispersed inoculum from both soil and cadavers to nettle leaves in petri dishes. In microcosms, aphids did not disperse B. bassiana from the soil or from cadavers confined in the canopy, but A. nemorum were able to transfer inoculum from soil into the nettle canopy and to distribute conidia from cryptic cadavers. In some instances, infections were initiated in aphids and predators as a consequence of dispersal.

Antifungal Activity of a Synthetic Cationic Peptide against the Plant Pathogens Colletotrichum graminicola and Three Fusarium Species

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Eric T. Johnson

2015-09-01

Full Text Available A small cationic peptide (JH8944 was tested for activity against a number of pathogens of agricultural crops. JH8944 inhibited conidium growth in most of the tested plant pathogens with a dose of 50 μg/ml, although one isolate of Fusarium oxysporum was inhibited at 5 μg/ml of JH8944. Most conidia of Fusarium graminearum were killed within 6 hours of treatment with 50 μg/ml of JH8944. Germinating F. graminearum conidia required 238 μg/ml of JH8944 for 90% growth inhibition. The peptide did not cause any damage to tissues surrounding maize leaf punctures when tested at a higher concentration of 250 μg/ml even after 3 days. Liposomes consisting of phosphatidylglycerol were susceptible to leakage after treatment with 25 and 50 μg/ml of JH8944. These experiments suggest this peptide destroys fungal membrane integrity and could be utilized for control of crop fungal pathogens.

Effect of the Combination Hot Water - Calcium Chloride on the In Vitro Growth of Colletotrichum gloeosporioides and the Postharvest Quality of Infected Papaya

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Lidia Elena Ayón-Reyna

2017-12-01

Full Text Available Anthracnose of papaya fruit caused by the fungus Colletotrichum gloeosporioides is one of the most economically important postharvest diseases. Hot water immersion (HW and calcium chloride (Ca treatments have been used to control papaya postharvest diseases; however, the effect of the combination HW-Ca on the pathogen growth and the development of the disease in infected papaya fruit has been scarcely studied. The aim of this study was to evaluate the effect of the HW-Ca treatment on the in vitro growth of C. gloesporioides conidia and the quality of infected papaya. In vitro, the HW-Ca treated conidia showed reduced mycelial growth and germination. In vivo, the HW-Ca treatment of infected papaya delayed for 5 days the onset of the anthracnose symptoms and improved the papaya postharvest quality. The combined treatment HW-Ca was better than any of the individual treatments to inhibit the in vitro development of C. gloeosporioides and to reduce the negative effects of papaya anthracnose.

In vitro antifungal and demelanizing activity of Nepeta rtanjensis essential oil against the human pathogen Bipolaris spicifera

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Ljaljević-Grbić Milica

2011-01-01

Full Text Available The antifungal activity of Nepeta rtanjensis Diklić & Milojević essential oil was tested against the human pathogenic fungus Bipolaris spicifera (Bainier Subramanian via mycelial growth assay and conidia germination assay. The minimally inhibitory concentration (MIC of the oil was determined at 1.0 μg ml-1, while the MIC for the antifungal drug Bifonazole in a positive control was determined at 10.0 μg ml-1. The maximum of conidia germination inhibition was accomplished at 0.6 μg ml-1. In addition, at 0.6 μg ml-1 and 0.8 μg ml-1 the oil was able to cause morphophysiological changes in B. spicifera. The most significant result is the bleaching effect of the melanized conidial apparatus of the test fungi, since the melanin is the virulence factor in human pathogenic fungi. These results showed the strong antifungal properties of N. rtanjensis essential oil, supporting its possible rational use as an alternative source of new antifungal compounds.

Effect of time and temperature of storage on the activity of the etanolic extracts of Lantana camara L. and Heliotropium indicum L. on Colletotrichum gloeosporioides

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Rafael Alejandro Infante

2015-10-01

Full Text Available The effect of time and temperature of storage of ethanolic extracts (EE of Lantana camara and Heliotropium indicum on Colletotrichum gloeosporioides was investigated. The EE were stored at 8±2 and 26±2°C, during one year. Every 2 months they were diluted in potato dextrose agar (PDA at 0; 0.5; 1 y 1.5% (v/v concentrations, fungus disks were placed onto the media, mycelial growth (ICM and sporulation (IE inhibition were measured, as well as conidia germination. The EE stored at 8±2°C maintained their effectiveness on all variables during the 12 months storage, while those stored at 26±2°C started to lose their effectiveness at 6 to 8 months, ending up to 3% ICM, 4% IE and 18% conidia germination at 1.5% concentration. Results indicated that storage temperature and time are determinant for extracts effectiveness.

Parameiosis in the entomopathogenic fungus Metarhizium anisopliae (METSH.) Sorokin

International Nuclear Information System (INIS)

Bagagli, E.; Valadares, M.C.C.; Azevedo, J.L.

1991-01-01

Variations in the parasexual cycle, especially in relation to diploid instability, have been described in several fungal species. The process has been designated parameiosis; it is characterized by the emergence, from heterokaryons, of haploid and diploid recombinants, as well as the typical diploids normally recovered in a parasexual cycle. In the present work the occurrence of a similar process in Metarhizium anisopliae has been investigated. Conidia from heterokaryons formed between well-marked mutant strains, when plated onto appropriate selective media, resulted in the recovery of at least three main groups of colonies. The first group consisted of very unstable diploids or hyperhaploids; these, on plating of their conidia, produced several types of recombinant haploids. The second group consisted of already stable haploid recombinants formed by the breakdown of diploid heterozygous nuclei before conidial formation; and a third group, heterokaryotic colonies, which segregate only parental types. Parameiosis has been found in several Deuteromycetes and may play an important role in increasing genetic variability in these fungi. (author)

Downy mildew of Double Knock Out® rose caused by Peronospora sparsa in Maryland

Science.gov (United States)

Roses are one of the most popular and economically important ornamental plants worldwide. In the last 17 years, Knock Out® roses (Rosa x 'Radtko') have been widely used in public and private gardens across the U.S. due to their disease resistance, self-cleaning, drought tolerance and multiple-bloomi...

Presencia de Phomopsis viticola (Saccardo Saccardo, agente causal de excoriosis, en viñedos de Córdoba, Argentina Presence of Phomopsis viticola (Saccardo Saccardo, causal agent of excoriosis, in vineyards of Córdoba, Argentina

Directory of Open Access Journals (Sweden)

C.I. Cragnolini

2009-12-01

Full Text Available Desde 2001 se observan en viñedos de Córdoba, Argentina, hojas de Vitis vinifera y V. labrusca con manchas cloróticas y centro necrótico. Las plantas presentan nervaduras, pecíolos, raquis y entrenudos con puntos necróticos que al confluir agrietan el tejido cortical. Las bayas pueden marchitarse y desprenderse. En invierno los sarmientos toman color blanquecino con manchas negras, y en ellos y en órganos secos aparecen picnidios con cirros blanco-crema con conidios alfa y beta. El objetivo fue confirmar la presencia de Phomopsis viticola (excoriosis y cuantificar su incidencia. Se realizaron aislamientos a partir de pecíolos, brotes, zarcillos, bayas y sarmientos que produjeron micelio claro con picnidios y conidios alfa (7,5m±1,5 x 2,8m±2 y beta (19m±1 x 1,2m±0,5 o sólo alfa, típicos de P. viticola. La identificación fue confirmada mediante secuenciación molecular. En brotes de las variedades Malbec y Pinot noir inoculados con 3 10(4 conidios/mL, las hojas mostraron lesiones y picnidios 8 días después, y las bayas cirros amarillos a los 15-18 días. P. viticola (Sacc. Sacc. fue reaislado de los cirros y las colonias produjeron picnidios y conidios característicos 10-20 días después. Se detectó excoriosis en 80% de los viñedos evaluados, con mayor incidencia en viñedos añosos de las variedades Pinot noir e Isabella.Leaves with chlorotic spots and necrotic centers have been observed in Vitis vinifera and V. labrusca in vineyards of Córdoba, Argentina, since 2001. Plants exhibit veins, petioles, rachises, and internodes with necrotic spots that coalesce to form cracks in the cortical tissue. Grape berries can shrivel and fall off. Winter canes became whitish with black spots; pycnidia appear in canes and in dry organs, exuding whitish-cream cirri with alpha and beta conidia. The aim of this work was to confirm the presence of Phomopsis viticola (excoriosis and quantify its incidence. Isolates obtained from petioles

Aerobiologia de Cercospora kikuchii Aerobiology of Cercospora kikuchii

Directory of Open Access Journals (Sweden)

Ângela Sathiko Kudo

2011-10-01

Full Text Available Neste estudo, foram quantificados os conídios de Cercospora kikuchii [crestamento foliar em soja (Glycine max] presentes no ar, sua relação com a intensidade da doença e com os fatores ambientais [temperatura, umidade relativa do ar (UR, precipitação e molhamento foliar] no período de novembro a março de 2006/07 e 2007/08 em Brasília, DF. Para tanto, foi instalada uma armadilha volumétrica capta-esporos 'Burkard' de sete dias em campo experimental com soja. Houve maior captura de conídios no período diurno, com mais de 60% dos esporos coletados entre 8h e 15h. Os dados climáticos variaram de acordo com a safra, mas, de modo geral, em ambos os experimentos, o período de maior captura de esporos ocorreu quando houve redução da precipitação e do molhamento foliar. A UR acima de 80% e temperatura entre 20 e 24°C foram mais favoráveis para a captura de conídios e a intensidade do crestamento foliar. Nos dois anos de avaliação, a maior quantidade de esporo capturado ocorreu quando a soja se apresentava no estádio R6-R7, assim como a intensidade da doença foi maior após o início do estágio reprodutivo da cultura. Apesar da quantidade de conídio capturado em 2006/07 ter sido maior que em 2007/08, o padrão da flutuação aérea de conídios foi semelhante.The objective of this study was to evaluate the relation of airborne released conidia of Cercospora kikuchii [cercospora leaf blight of soybean (Glycine max] with the disease intensity and some environmental factors [temperature, relative humidity (RH, precipitation and leaf wetness period under experimental field conditions in Brasília, DF, Brazil. The evaluation was from November to March of 2006/07 and 2007/08. Data for the amount of airborne conidia were collected using a Burkard's seven-day volumetric spore trap set in a field with soybean. Most of the conidia were collected during the day, with over 60% of the spores collected between 8h and 15h. Climatic data varied

Effect of cobalt on the growth of Trichoderma viride and determination of its distribution using 60Co

International Nuclear Information System (INIS)

Frank, V.; Lesny, J.; Babej, L.

1993-01-01

The growth and morphology of filamentous fungus Trichoderma viride grown in the presence of various Co concentrations were studied. Using 60 Co, the cobalt content in mycelium and conidia was determined. No influence of 60 Co-radiation on growth and morphology was observed. (author) 5 refs.; 4 figs

Ranking cultivated blueberry for Mummy Berry Blight and Fruit Infection Incidence

Science.gov (United States)

Mummy berry is an important disease of cultivated blueberry. The disease has two distinct phases; a blighting phase initiated by ascospores and a fruit infection stage initiated by conidia. In this study we investigated the resistance of more than 100 blueberry cultivar to both phases of the disease...

Use of acoustic technology to monitor the time course of Rhynchophorus ferrugineus larval mortality in date palms after treatments with Beauveria bassiana

Science.gov (United States)

Spectral and temporal patterns of insect sound impulses were monitored daily for 23-d periods in 8, 10, or 5 small date palm trees containing larvae dipped in 0 (control), 104 (low), or 108 (high) conidia/ml doses of entomopathogenic fungus, Beauveria bassiana (Bb 203), respectively. Each tree conta...

Maxillary sinusitis from Microascus cinereus and Aspergillus repens.

Science.gov (United States)

Aznar, C; de Bievre, C; Guiguen, C

1989-02-01

Microascus was associated with Aspergillus repens in a left maxillary sinus. Tissue contained septale filaments of two types, conidia, ostiolate perithecia containing ascospores corresponding to Microascus cinereus which was identified by culture. The abundance of sexual fructifications in the tissue indicates that pathogenicity is due to Microascus cinereus.

Soil application of formulated Metarhizium brunneum F52 (Hypocreales: Clavicipitaceae) containing microsclerotia controls eggs of Aedes aegypti (Diptera: Culicidae)

Science.gov (United States)

We evaluated the potential of a granular formulation of Metarhizium brunneum F52 containing microsclerotia (MbMSc granules) for control of Aedes aegypti (L.) by targeting eggs. MbMSc granules produced infective conidia within 14 days after application to moist potting soil, producing 5.9 × 10**5, 2....

Emergomyces africanus in Soil, South Africa.

Science.gov (United States)

Schwartz, Ilan S; Lerm, Barbra; Hoving, J Claire; Kenyon, Chris; Horsnell, William G; Basson, W Joan; Otieno-Odhiambo, Patricia; Govender, Nelesh P; Colebunders, Robert; Botha, Alfred

2018-02-01

We detected Emergomyces africanus, a thermally dimorphic fungus that causes an HIV-associated systemic mycosis, by PCR in 18 (30%) of 60 soil samples from a wide range of habitats in South Africa. Direct and indirect culture techniques were unsuccessful. Experimental intraperitoneal inoculation of conidia induced murine disease.

It’s a Jungle Out There! Abiotic and Biotic Factors That Affect Efficacy and Persistence of the Entomopathogenic Fungi

Science.gov (United States)

One might conclude the soil is a more congenial arena for using entomopathogenic fungi (EPF) than the phylloplane. No ultraviolet light, no rainfall washing conidia from foliage, no rapid attenuation of conidial deposits by rapid plant canopy expansion. The soil is cool, damp and dark – perfect fo...

Histopathologic criteria to confirm white-nose syndrome in bats.

Science.gov (United States)

Meteyer, Carol Uphoff; Buckles, Elizabeth L; Blehert, David S; Hicks, Alan C; Green, D Earl; Shearn-Bochsler, Valerie; Thomas, Nancy J; Gargas, Andrea; Behr, Melissa J

2009-07-01

White-nose syndrome (WNS) is a cutaneous fungal disease of hibernating bats associated with a novel Geomyces sp. fungus. Currently, confirmation of WNS requires histopathologic examination. Invasion of living tissue distinguishes this fungal infection from those caused by conventional transmissible dermatophytes. Although fungal hyphae penetrate the connective tissue of glabrous skin and muzzle, there is typically no cellular inflammatory response in hibernating bats. Preferred tissue samples to diagnose this fungal infection are rostral muzzle with nose and wing membrane fixed in 10% neutral buffered formalin. To optimize detection, the muzzle is trimmed longitudinally, the wing membrane is rolled, and multiple cross-sections are embedded to increase the surface area examined. Periodic acid-Schiff stain is essential to discriminate the nonpigmented fungal hyphae and conidia. Fungal hyphae form cup-like epidermal erosions and ulcers in the wing membrane and pinna with involvement of underlying connective tissue. In addition, fungal hyphae are present in hair follicles and in sebaceous and apocrine glands of the muzzle with invasion of tissue surrounding adnexa. Fungal hyphae in tissues are branching and septate, but the diameter and shape of the hyphae may vary from parallel walls measuring 2 microm in diameter to irregular walls measuring 3-5 microm in diameter. When present on short aerial hyphae, curved conidia are approximately 2.5 microm wide and 7.5 microm in curved length. Conidia have a more deeply basophilic center, and one or both ends are usually blunt. Although WNS is a disease of hibernating bats, severe wing damage due to fungal hyphae may be seen in bats that have recently emerged from hibernation. These recently emerged bats also have a robust suppurative inflammatory response.

Modulation of the immune response by Fonsecaea pedrosoi morphotypes in the course of experimental chromoblastomycosis and their role on inflammatory response chronicity.

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Isaque Medeiros Siqueira

2017-03-01

Full Text Available A common theme across multiple fungal pathogens is their ability to impair the establishment of a protective immune response. Although early inflammation is beneficial in containing the infection, an uncontrolled inflammatory response is detrimental and may eventually oppose disease eradication. Chromoblastomycosis (CBM, a cutaneous and subcutaneous mycosis, caused by dematiaceous fungi, is capable of inducing a chronic inflammatory response. Muriform cells, the parasitic form of Fonsecaea pedrosoi, are highly prevalent in infected tissues, especially in long-standing lesions. In this study we show that hyphae and muriform cells are able to establish a murine CBM with skin lesions and histopathological aspects similar to that found in humans, with muriform cells being the most persistent fungal form, whereas mice infected with conidia do not reach the chronic phase of the disease. Moreover, in injured tissue the presence of hyphae and especially muriform cells, but not conidia, is correlated with intense production of pro-inflammatory cytokines in vivo. High-throughput RNA sequencing analysis (RNA-Seq performed at early time points showed a strong up-regulation of genes related to fungal recognition, cell migration, inflammation, apoptosis and phagocytosis in macrophages exposed in vitro to muriform cells, but not conidia. We also demonstrate that only muriform cells required FcγR and Dectin-1 recognition to be internalized in vitro, and this is the main fungal form responsible for the intense inflammatory pattern observed in CBM, clarifying the chronic inflammatory reaction observed in most patients. Furthermore, our findings reveal two different fungal-host interaction strategies according to fungal morphotype, highlighting fungal dimorphism as an important key in understanding the bipolar nature of inflammatory response in fungal infections.

Muscodor albus Volatiles Control Toxigenic Fungi under Controlled Atmosphere (CA Storage Conditions

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Gordon Braun

2012-11-01

Full Text Available Muscodor albus, a biofumigant fungus, has the potential to control post-harvest pathogens in storage. It has been shown to produce over 20 volatile compounds with fungicidal, bactericidal and insecticidal properties. However, M. albus is a warm climate endophyte, and its biofumigant activity is significantly inhibited at temperatures below 5 °C. Conidia of seven mycotoxin producing fungi, Aspergillus carbonarius, A. flavus, A. niger, A. ochraceus, Penicillium verrucosum, Fusarium culmorum and F. graminearum, were killed or prevented from germinating by exposure to volatiles from 2 g M. albus-colonized rye grain per L of headspace in sealed glass jars for 24 h at 20 °C. Two major volatiles of M. albus, isobutyric acid (IBA and 2-methyl-1-butanol (2MB at 50 µL/L and 100 µL/L, respectively, gave differential control of the seven fungi when applied individually at 20 °C. When the fungi were exposed to both IBA and 2MB together, an average of 94% of the conidia were killed or suppressed. In a factorial experiment with controlled atmosphere storage (CA at 3 °C and 72 h exposure to four concentrations of IBA and 2MB combinations, 50 µL/L IBA plus 100 µL/L 2MB killed or suppressed germination of the conidia of all seven fungi. Controlled atmosphere had no significant effect on conidial viability or volatile efficacy. Major volatiles of M. albus may have significant potential to control plant pathogens in either ambient air or CA storage at temperatures below 5 °C. However, combinations of volatiles may be required to provide a broader spectrum of control than individual volatiles.

Development of a user-friendly delivery method for the fungus Metarhizium anisopliae to control the ectoparasitic mite Varroa destructor in honey bee, Apis mellifera, colonies.

Science.gov (United States)

Kanga, Lambert H B; Adamczyk, John; Patt, Joseph; Gracia, Carlos; Cascino, John

2010-12-01

A user-friendly method to deliver Metarhizium spores to honey bee colonies for control of Varroa mites was developed and tested. Patty blend formulations protected the fungal spores at brood nest temperatures and served as an improved delivery system of the fungus to bee hives. Field trials conducted in 2006 in Texas using freshly harvested spores indicated that patty blend formulations of 10 g of conidia per hive (applied twice) significantly reduced the numbers of mites per adult bee, mites in sealed brood cells, and residual mites at the end of the 47-day experimental period. Colony development in terms of adult bee populations and brood production also improved. Field trials conducted in 2007 in Florida using less virulent spores produced mixed results. Patty blends of 10 g of conidia per hive (applied twice) were less successful in significantly reducing the number of mites per adult bee. However, hive survivorship and colony strength were improved, and the numbers of residual mites were significantly reduced at the end of the 42-day experimental period. The overall results from 2003 to 2008 field trials indicated that it was critical to have fungal spores with good germination, pathogenicity and virulence. We determined that fungal spores (1 × 10(10) viable spores per gram) with 98% germination and high pathogenicity (95% mite mortality at day 7) provided successful control of mite populations in established honey bee colonies at 10 g of conidia per hive (applied twice). Overall, microbial control of Varroa mite with M. anisopliae is feasible and could be a useful component of an integrated pest management program.

Vital role for cyclophilin B (CypB) in asexual development, dimorphic transition and virulence of Beauveria bassiana.

Science.gov (United States)

Chu, Zhen-Jian; Sun, Huan-Huan; Ying, Sheng-Hua; Feng, Ming-Guang

2017-08-01

Cyclophilin B (CypB) was previously revealed as one of many putative secretory proteins in the transcriptome of Beauveria bassiana infection to a lepidopteran pest. Here we show a main localization of CypB in hyphal cell walls and septa and its essential role in the in vitro and in vivo asexual cycles of the fungal insect pathogen. Deletion of cypB reduced colony growth by 16-42% on two rich media and 30 scant media with different carbon or nitrogen sources. The deletion mutant suffered a delayed conidiation on a standard medium and a final 47% reduction in conidial yield, accompanied with drastic transcript depression of several key genes required for conidiation and conidial maturation. The mutant conidia required 10h longer to germinate 50% at optimal 25°C than wild-type conidia. Intriguingly, cultivation of the mutant conidia in a trehalose-peptone broth mimic to insect hemolymph resulted in 83% reduction in blastospore yield but only slight decrease in biomass level, indicating severe defects in transition of hyphae to blastospores. LT 50 for the deletion mutant against Galleria mellonella larvae through normal cuticle infection was prolonged to 7.4d from a wild-type estimate of 4.7d. During colony growth, additionally, the deletion mutant displayed hypersensitivity to Congo red, menadione, H 2 O 2 and heat shock but increased tolerance to cyclosporine A and rapamycin. All of changes were restored by targeted gene complementation. Altogether, CypB takes part in sustaining normal growth, aerial conidiation, conidial germination, dimorphic transition, stress tolerance and pathogenicity in B. bassiana. Copyright © 2017 Elsevier Inc. All rights reserved.

Histopathologic criteria to confirm white-nose syndrome in bats

Science.gov (United States)

Meteyer, Carol U.; Buckles, Elizabeth L.; Blehert, David S.; Hicks, Alan C.; Green, David E.; Shearn-Bochsler, Valerie I.; Thomas, Nancy J.; Gargas, Andrea; Behr, Melissa

2009-01-01

White-nose syndrome (WNS) is a cutaneous fungal disease of hibernating bats associated with a novel Geomyces sp. fungus. Currently, confirmation of WNS requires histopathologic examination. Invasion of living tissue distinguishes this fungal infection from those caused by conventional transmissible dermatophytes. Although fungal hyphae penetrate the connective tissue of glabrous skin and muzzle, there is typically no cellular inflammatory response in hibernating bats. Preferred tissue samples to diagnose this fungal infection are rostral muzzle with nose and wing membrane fixed in 10% neutral buffered formalin. To optimize detection, the muzzle is trimmed longitudinally, the wing membrane is rolled, and multiple cross-sections are embedded to increase the surface area examined. Periodic acid-Schiff stain is essential to discriminate the nonpigmented fungal hyphae and conidia. Fungal hyphae form cup-like epidermal erosions and ulcers in the wing membrane and pinna with involvement of underlying connective tissue. In addition, fungal hyphae are present in hair follicles and in sebaceous and apocrine glands of the muzzle with invasion of tissue surrounding adnexa. Fungal hyphae in tissues are branching and septate, but the diameter and shape of the hyphae may vary from parallel walls measuring 2 ??m in diameter to irregular walls measuring 3-5 ??m in diameter. When present on short aerial hyphae, curved conidia are approximately 2.5 ??m wide and 7.5 ??m in curved length. Conidia have a more deeply basophilic center, and one or both ends are usually blunt. Although WNS is a disease of hibernating bats, severe wing damage due to fungal hyphae may be seen in bats that have recently emerged from hibernation. These recently emerged bats also have a robust suppurative inflammatory response.

AoRim15 is involved in conidial stress tolerance, conidiation and sclerotia formation in the filamentous fungus Aspergillus oryzae.

Science.gov (United States)

Nakamura, Hidetoshi; Kikuma, Takashi; Jin, Feng Jie; Maruyama, Jun-ichi; Kitamoto, Katsuhiko

2016-04-01

The serine-threonine kinase Rim15p is a master regulator of stress signaling and is required for stress tolerance and sexual sporulation in the yeast Saccharomyces cerevisiae. However, in filamentous fungi that reproduce asexually via conidiation, the physiological function of Rim15p homologs has not been extensively analyzed. Here, we functionally characterized the protein homolog of Rim15p in the filamentous fungus Aspergillus oryzae, by deleting and overexpressing the corresponding Aorim15 gene and examining the role of this protein in stress tolerance and development. Deletion of Aorim15 resulted in an increase in the sensitivity of conidia to oxidative and heat stresses, whereas conidia of the Aorim15 overexpressing strain were more resistant to these stresses. These results indicated that AoRim15 functions in stress tolerance, similar to S. cerevisiae Rim15p. Phenotypic analysis revealed that conidiation was markedly reduced by overexpression of Aorim15 in A. oryzae, and was completely abolished in the deletion strain. In addition, the formation of sclerotia, which is another type of developmental structure in filamentous fungi, was decreased by the deletion of Aorim15, whereas Aorim15 overexpression increased the number of sclerotia. These results indicated that AoRim15 is a positive regulator of sclerotia formation and that overexpression of AoRim15 shifts the developmental balance from conidiation towards sclerotia formation. Collectively, we demonstrated that AoRim15 is involved in the stress tolerance of conidia and differentially regulates between the two developmental fates of conidiation and sclerotia formation. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Genetic diversity of Colletotrichum gloeosporioides in Nigeria using ...

African Journals Online (AJOL)

fmodupe

2012-04-24

Apr 24, 2012 ... The genetic variation in C. gloeosporioides is important in choosing strategies to ... conidia size and shape, and presence or absence of setae (Sutton ... E-mail: aduramigbaa@run.edu.ng. ... 8190 Afr. J. Biotechnol. ... different Colletotrichum species (Guthrice et al., 1992). ... 11 blade fitted onto a holder was.

Current status of Colletotrichum capsici strains, causal agents of ...

African Journals Online (AJOL)

Gil

ASIC, 15e Colloque,. Montpellier 7 p. Dabert M (2006). DNA markers in the phylogenetic of the. Acari. Biol. Lett. 43(2):97-107. Damm U, Woudenberg JHC, Cannon PF, Crous PW (2009). Colletotrichum species with curved conidia from herbaceous hosts. Fungal Divers 39:45-87. Emechebe AM (1986). Cowpea pathology.

Phoma negriana, a new invasive pathogen for Moghan's vineyards ...

African Journals Online (AJOL)

SERVER

2008-03-18

Mar 18, 2008 ... Pathogenesity study were carried out on some separating grape- vines located in Moghan Junior College of Agriculture vineyards that were surface sterilized with 0.5% sodium hypochlorite by spraying on aerial parts of them. A spore suspension containing 106 conidia/ml were inoculated using a hand ...

Adulticidal effect of fungal pathogen, Metarhizium anisopliae on ...

African Journals Online (AJOL)

SERVER

2008-03-18

Mar 18, 2008 ... Page 1 ... forms conidiophores or analogous structure and sporu- lates. Alternatively, many species form some type of resting stages capable of forming or releasing a type of spore. ... Mosquitoes landing on the suspension to consume glucose would thus be exposed to conidia through tarsal contact or ...

Author Details

African Journals Online (AJOL)

Sankara, P. Vol 85 (2015) - Articles Assessing the toxicity level of some useful mushrooms of Burkina Faso (West Africa) Abstract PDF · Vol 89 (2015) - Articles In vitro evaluation of the effect of aqueous extracts of Agave sisalana and Cymbopogon citratus on mycelial growth and conidia production of Pyricularia oryzae, ...

Produção de Bipolaris euphorbiae em meios de cultura sólidos e líquidos obtidos de grãos e resíduos agroindustriais Production of Bipolaris euphorbiae in solid and liquid culture media obtained from grains and agricultural industry residues

Directory of Open Access Journals (Sweden)

Mara Cristina Penariol

2008-12-01

Full Text Available A introdução de Bipolaris euphorbiae como bioagente de controle de Euphorbia heterophylla é dependente da produção de conídios em grande quantidade. No presente trabalho, objetivou-se definir meios de cultura sólidos e líquidos, obtidos de grãos ou resíduos da agroindústria, eficazes para a produção de conídios de B. euphorbiae. No preparo dos meios sólidos utilizaram-se grãos de arroz, trigo e sorgo, quirelas de arroz, milho e trigo, sorgo moído, farelos de arroz, trigo e soja, cascas de mandioca e soja, casca de mandioca + farelo de soja, bagaço de cana e bagaço de cana + amido solúvel. No preparo dos meios líquidos, utilizaram-se grãos de arroz, sorgo e trigo, quirela de milho, farelos de trigo, soja e arroz, casca de mandioca e soja, vinhaça de cana e água de prensa da mandioca. Avaliaram-se a produção e a viabilidade dos conídios e a virulência do fungo e, nos meios líquidos, também a biomassa. A produção de conídios é influenciada pelo tipo de meio de cultura, sendo acentuadamente maior nos meios sólidos, destacando-se, como substratos, o sorgo em grão (474 x 10(6 conídios g-1 e a casca de soja (472 x 10(6 conídios g-1. Dentre os meios líquidos obteve-se mais produção usando-se farelo de trigo (1,33 x 10(6 conídios mL-1. A virulência e a viabilidade de B. euphorbiae não são afetadas pelo preparo de meios sólidos ou líquidos e pela composição nutricional dos meios de cultura. Na maioria dos meios sólidos ou líquidos obteve-se viabilidade de conídios maior que 98%; apenas os conídios produzidos nos meios sólidos de quirela de arroz, casca de mandioca + farelo de soja e farelo de soja estavam com viabilidade significativamente menor.The introduction of Bipolaris euphorbiae as a bioagent for Euphorbia heterophylla control depends on fungal conidia production in large amount. This work evaluate to solid and liquid culture media obtained from grains and residues from agriculture industries

Metacridamide B methanol-d4 monosolvate

Science.gov (United States)

The title compound was extracted from conidia of the fungus Metarhizium acridum. Crystals were obtained as a methanol-d4 solvate. The tail part of the 4-methylhexan-2-yl group exhibits disorder over two positions, with an occupancy ratio of 0.682 (9):0.318 (9). The crystal structure confirms the abs...

A Technical and Practical Study of Composting as a Solid Waste Management Alternative for the Air Force

Science.gov (United States)

1992-09-01

52 0C Phvsoderma zeae- Brown spot of corn > 80C maydis Giberella zeae Seedling blight of corn > 651C ascospores Glomeralla gossypii Cotton anthracnose ...Pyrenophora teres Net blotch, barley 45°C conidia 55°C mycelium Septoria lycopersici Leafspot, tomatoes 43°C spores (2:221) 44 The extent of

Primary Infection by Erysiphe graminis f.sp. hordei of Barley Mutants with Resistance Genes in the ml-o Locus

DEFF Research Database (Denmark)

Jørgensen, Jørgen Helms; Mortensen, K.

1977-01-01

by the hosts. An average of about 60% of the germinated conidia formed haustoria, 50% formed hyphae, and 40% formed mildew colonies on the susceptible hosts. In contrast, only about 0.7% formed haustoria, 0.4% formed hyphae, and 0.2% formed colonies on the resistant hosts, on which the great majority...

Differential endophytic colonization of sorghum plant by eight ...

African Journals Online (AJOL)

Virulence of the conidia before and after endophytic growth phases were assessed using Galleria mellonella larvae mortality bioassay in-vitro. All the strains of the fungi colonised the sorghum plant. The strains of I. farinosa and B. bassiana were detected in the roots, the stem and the leaves while M. anisopliae was ...

Potential of a strain of the entomopathogenic fungus Beauveria bassiana (Hypocreales: Cordycipitaceae) as a biological control agent against western flower thrips, Frankliniella occidentalis (Thysanoptera: Thripidae)

Science.gov (United States)

Five Beauveria bassiana strains were evaluated for control of western flower thrips. Strain RSB was the most virulent, causing 69-96% mortality at concentrations of 1×104 – 1×107 conidia mL-1, 10 days after inoculation of first instars. In greenhouse trials, RSB applied to broccoli foliage signifi...

Bio-Sustainable Control of the Blue Stain Fungi Aureobasidium pullulans on Exterior Wood Coatings

DEFF Research Database (Denmark)

Stenbæk, Jonas

on the coating has been studied. In this study, manipulation of the micro structure on the coating surface has been performed and revealed that small scratches-structure (between 4-20 μm) had an inhibitory effect on the attachment on fungal conidia from moulds while bigger structure scratches (>50 μm) did...

Effect of seasonal abiotic conditions and field margin habitat on the activity of Pandora neoaphidis inoculum on soil.

Science.gov (United States)

Baverstock, J; Clark, S J; Pell, J K

2008-03-01

The ability of the aphid pathogenic fungus Pandora neoaphidis to remain active in the absence of a resting stage through a combination of continuous infection and as conidia deposited on soil was assessed alongside the potential for planted field margins to act as a refuge for the fungus. P. neoaphidis was able to infect the pea aphid, Acyrthosiphon pisum, when maintained under controlled conditions that simulated those that occur seasonally in the UK. Although there was a significant inverse relationship between temperature and time-to-kill, with death occurring after 4.2, 6.9 and 13.6 days when maintained under fluctuating summer, autumn and winter temperatures, respectively, there were no additional statistically significant effects of photoperiod. The activity of inoculum on soil was indirectly assessed by baiting with A. pisum. Under controlled conditions P. neoaphidis remained active on soil and was able to infect aphids for up to 80 days. However, the percentage of aphids that became infected decreased from 76% on day 1 to 11% on day 80. Whereas there was little difference in the activity of conidia that had been maintained at 4 degrees C and 10 degrees C, activity at 18 degrees C was considerably reduced. Under field conditions the activity of inoculum was strongly influenced by season. On day 49 there was little or no activity during spring, summer or winter. However, during autumn a mean proportion of 0.08 aphids still became infected with P. neoaphidis. Margin type did not affect the activity of conidia nor was there a difference in activity between blocks that had regenerated naturally and those that had been planted. These results suggest that P. neoaphidis can infect aphids and remain active on soil under the abiotic conditions that occur seasonally in the UK and that this fungus may be able to persist annually without a resting stage.

Studies on the infection process by Erysiphe polygoni in resistant and susceptible peas

International Nuclear Information System (INIS)

Cirulli, M.; Montemurro, G.; Ciccarese, F.; Smilari, F.

1976-01-01

The infection of pea by Erysiphe polygoni was investigated. The susceptible cultivar ''Sprinter'' and the resistant varieties ''Stratagem Resistant'' (er 1 er 1 Er 2 Er 2 ) and ''Mexique-4'' (er 1 er 1 er 2 er 2 ) were used. Observations of germination of conidia, formation of primary appressoria, differentiation of secondary hyphae, number of secondary hyphae per conidium, and number of branches on the longest hyphae per conidium were made at different times from inoculation. The rate of conidial germination was not affected by the host genotypes. The formation of short germ tubes with primary appressoria were found to be similar on the susceptible ''Sprinter'' as well as on the ''Stratagem Resistant'' and ''Mexique-4''. No statistical difference in the growth of germ tubes with appressorium between susceptible and resistant peas was observed at 2, 4, 6, 10 and 12 hrs after inoculation. In resistant peas formation of primary appressoria was not followed by further mycelial growth. Conversely, in the susceptible pea, germinating conidia produced multiple germ tubes and branching hyphae. The establishment of a compatible relationship between host and pathogen appears to occur at or near the stage of formation of the primary appressorium. Most conidia on the leaves formed primary appressoria as early as 2 hrs after inoculation. The Course of the host/pathogen relationship is apparently decided at a very early stage after pathogen contact with the host, such as the length of the longest hyphae, number of germ tubes per conidium and branching of the longest, and the difference in the macroscopic fungal fructification. E. polygoni activity in susceptible pea is evidently influenced by temperature whereas the gene action of the genetic factors remains unaffected in resistant pea

Identification and evaluation of a new entomopathogenic fungal strain against Riptortus pedestris (Hemiptera: Alydidae) and its two egg parasitoids

Science.gov (United States)

Lim, Un Taek

2018-01-01

A strain (ARP14) of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin was isolated from field-collected Riptortus pedestris (Fabricius) (Hemiptera: Alydidae). The lethal median concentration of the ARP14 strain was compared with that of a commercialized strain (GHA) of the same fungus against R. pedestris and its two egg parasitoids, Ooencyrtus nezarae Ishii (Hymenoptera: Encyrtidae) and Gryon japonicum (Ashmead) (Hymenoptera: Platygastridae). Mortality and mycosis rates were evaluated after exposure to five concentrations of the fungus, i.e., 1×109, 1×108, 1×107, 1×106, and 1×105 conidia/mL, using a glass scintillation vial as an exposure arena in 25.0 ± 0.5°C and 93.7 ± 2.9% RH. The lethal median concentrations (LC50) for 2nd and 4th instar nymphs, and adults of R. pedestris were not significantly different between the two strains of B. bassiana. However, the mycosis rate of ARP14 was 1.3 and 1.8 times higher than that of the GHA strain in 4th instar nymphs and adult females of R. pedestris, respectively, at the 1×108 conidia/mL concentration. More interestingly, the mycosis rates at 1×108 conidia/mL concentration in the parasitoids G. japonicum and O. nezarae were much lower in the ARP14 strain (15.0 and 0%) than in the GHA strain (73.3 and 66.0%), respectively, suggesting that the B. bassiana strain ARP14 is less virulent to these parasitoids than the commercially available strain. Our results suggest that B. bassiana ARP14 may be a potential new biopesticide against R. pedestris with fewer negative effects on beneficial parasitoids than currently available options. PMID:29664929

Phacidiopycnis washingtonensis--a new species associated with pome fruits from Washington State.

Science.gov (United States)

Xiao, C L; Rogers, J D; Kim, Y K; Liu, Q

2005-01-01

A new species of Phacidiopycnis associated with pome fruits is described. The fungus causes fruit rot on apples during storage and is associated with a twig dieback and canker disease of crabapple trees and dead twigs of pear trees. To characterize the biology of the fungus and compare it with Ph. piri, the type species of the genus, effects of nine media and light on mycelial growth and pycnidial production, mycelial growth in response to temperature and mode of conidial germination in response to nutrient were determined. Apple-juice agar, pear-juice agar, prune-juice agar, potato-dextrose agar (PDA) and malt-extract agar, Czapek-Dox agar and oatmeal agar (OMA) favored mycelial growth. Cornmeal agar (CMA) did not favor mycelial growth. Light effect on pycnidial formation was medium dependent. Abundant pycnidia with mature conidia formed in 14 d old PDA and OMA cultures at 20 C, regardless of light, whereas none or very few pycnidia formed on other media in the dark. Fluorescent light stimulated formation of pycnidia except on CMA. The fungus grew at -3-25 C, with optimum growth at 15-20 C. Conidia germinated either by forming germ tubes or less often by budding. Budding of conidia occurred in 1 and 10% pear-juice solutions but not in 100% pear-juice solution. Six isolates of Ph. washingtonensis from different species of pome fruits had identical ITS sequences. The sizes of the ITS region were the same for both Ph. washingtonensis and Ph. piri, and four polymorphic nucleotide sites were found in the ITS region between Ph. washingtonensis and Ph. piri. The similarity in ITS sequences between these two taxa is confirmatory evidence for the erection of the new species of Phacidiopycnis associated with pome fruits we describe here.

Surface ultrastructural studies on the germination, penetration and conidial development of Aspergillus flavus Link:Fries infecting silkworm, Bombyx mori Linn.

Science.gov (United States)

Kumar, Vineet; Singh, G P; Babu, A M

2004-01-01

Aspergillosis is a common disease of the silkworm Bombyx mori Linn., caused by an insect mycopathogen Aspergillus flavus Link:Fries. The present study reveals the germination, penetration and conidial development of A. flavus on the larval integument of B. mori under SEM. Four different strains (NB18, KA, NB4D2 and NB7) of B. mori was surface inoculated with ca. of 1 x 10(6) conidia/ml. Each conidium germinated on the cuticle approximately 6 h after inoculation, forming a humpy or suctorial appressoria within 24 h. The hyphae which entered into haemocoel 2 day post-inoculation, grew and multiplied extensively, forming a mycelial complex, causing death of the host larva in about 4-5 days. This occurred with minimal breakdown of the internal tissues. Death of the host was followed by ramification of the fungus through the mesodermal and epidermal tissues, leading to larval mummification about 5-6 days after inoculation. Extensive fungal growths on the entire larval body followed, consisting of aerial hyphae, which developed branched conidiophores. The aerial hyphae with abundant conidiophores formed a confluent yellowish green fungal mat over the entire larval body in 6-7 days of post-inoculation. The tip of each emerging conidiophores gradually dilated and developed to become a bulbous head known as the vesicle. A large number of conidiogenous cells were produced over the entire surface of vesicle, which later developed into finger-like projections termed as sterigmata or phialides. The phialides matured within 2 days after the aerial hyphae emerged as evidenced by chains of conidia at their tips. The conidia were globose with externally roughened walls. The life cycle of the fungus on B. mori was completed in six to seven days.

Selection of entomopathogenic fungi against the red spider mite Tetranychus kanzawai (Kishida (Tetranychidae: Acarina

Directory of Open Access Journals (Sweden)

Yayan Sanjaya

2013-12-01

Full Text Available The pathogenicity of three entomopathogenic fungal species to Tetranychus kanzawai was investigated. Seven isolates of Metarhizium anisopliae, six isolates of Beauveria bassiana, and an isolate of Paecilomyces lilacinus from the Philippines and Indonesia were evaluated. The following studies were undertaken: (1 screening of M. anisoplae, B. bassiana and P. lilicanus pathogenic to T. kanzawai, and (2 bioefficacy studies of the selected entomopathogenic fungi under greenhouse conditions. Conidia of each isolate were mass-produced on potato dextrose agar (PDA at 26+-1 oC and a 12-hour photophase for a maximum of 21 days. Preliminary screening for the most pathogenic isolate within the same species was determined using suspension with 104 to 108 conidia ml-1. At 4 days after treatment (DAT, the pathogenicity within M. anisopliae isolates in decreasing order was Ma5>Ma6>Ma4>Ma2>Ma1>Ma3>Ma7 while for B. bassiana, was Bb6>Bb5>Bb4>Bb3>Bb1>Bb2. The top three most pathogenic isolates within the two species were subjected to further studies to determine the most virulent isolate against T. kanzawai. At 5 DAT, the LC50 values of M. anisopliae isolates ranged from 5.0 x102 to 1.4x103 while for B. bassiana ranged from 1.2 x 103 to 2.4x 103 conidia ml-1. Based on LC50, the virulence of the fungal isolates within the species in decreasing order was Ma6>Ma5>Ma4 and Bb6>Bb5>Bb4. However, the LC50 values are not significantly different from each other. Green house trials showed that the epizootic of entomopathogenic fungus can regulate the population of mites. The fungal isolates used in the study, although not originally isolated from mites were virulent to T. kanzawai, indicating their wide host range.

In vitro evaluation of the effect of aqueous extracts of Agave sisalana ...

African Journals Online (AJOL)

Objectives: To evaluate the effects of aqueous extracts of Agave sisalana (sisal) and Cymbopogon citratus (lemon grass) on mycelial growth and conidia production of Pyricularia oryzae, causal agent of Rice Blast. Methodology and Results: The plants aqueous extracts were used at concentrations 0.1; 0.2; 0.3; 0.4; 0.5;1; 2; ...

Infection of adult Aedes aegypti and Ae. albopictus mosquitoes with the entomopathogenic fungus Metarhizium anisopliae

NARCIS (Netherlands)

Scholte, E.J.; Takken, W.; Knols, B.G.J.

2007-01-01

This study describes a laboratory investigation on the use of the insect-pathogenic fungus Metarhizium anisopliae against adult Aedes aegypti and Ae. albopictus mosquitoes. At a dosage of 1.6 × 1010 conidia/m2, applied on material that served as a mosquito resting site, an average of 87.1 ± 2.65% of

Three new species of Trichoderma with hyaline ascospores from China.

Science.gov (United States)

Zhu, Z X; Zhuang, W Y

2015-01-01

Collections of Trichoderma having hyaline ascospores from different areas of China were examined. Using combined analyses of morphological data, culture characters and phylogenetic information based on rDNA sequences of partial nuc translation elongation factor 1-α encoding gene (TEF1-α) and the gene encoding the second largest nuc RNA polymerase subunit (RPB2), three new species, Trichoderma applanatum, T. oligosporum and T. sinoluteum, were discovered and are described. Trichoderma applanatum produces continuous flat to pulvinate, white to cream stromata with dense orange or pale brown ostioles, and simple acremonium-like to verticillium-like conidiophores, belongs to the Hypocreanum clade and is closely related to T. decipiens. Trichoderma oligosporum forms reddish brown stromata with a downy surface, hyaline conidia and gliocladium-like conidiophores, and is closely related to but distinct from T. crystalligenum in the Psychrophila clade. Trichoderma sinoluteum, as a member of the Polysporum clade, is characterized by pale yellow stromata, white pustulate conidiomata, pachybasium-like conidiophores, and hyaline conidia. Differences between the new species and their close relatives are discussed. © 2015 by The Mycological Society of America.

Two new hyaline-ascospored species of Trichoderma and their phylogenetic positions.

Science.gov (United States)

Qin, W T; Zhuang, W Y

2016-01-01

Collections of hypocrealean fungi found on decaying wood in subtropical regions of China were examined. Two new species, Trichoderma confluens and T. hubeiense, were discovered and are described. Trichoderma confluens is characterized by its widely effuse to rarely pulvinate, yellow stromata with densely disposed yellowish brown ostioles, simple acremonium- to verticillium-like conidiophores, hyaline conidia and multiform chlamydospores. Trichoderma hubeiense has pulvinate, grayish yellow stromata with brownish ostioles, trichoderma- to verticillium-like conidiophores and hyaline conidia. The phylogenetic positions of the two fungi were investigated based on sequence analyses of RNA polymerase II subunit b and translation elongation factor 1-α genes. The results indicate that T. confluens belongs to the Hypocreanum clade and is associated with but clearly separated from T. applanatum and T. decipiens. Trichoderma hubeiense belongs to the Polysporum clade and related to T. bavaricum but obviously differs from other members of the clade in sequence data. Morphological distinctions between the new species and their close relatives are noted and discussed. © 2016 by The Mycological Society of America.

Seven wood-inhabiting new species of the genus Trichoderma (Fungi, Ascomycota) in Viride clade.

Science.gov (United States)

Qin, Wen-Tao; Zhuang, Wen-Ying

2016-06-01

More than 200 recent collections of Trichoderma from China were examined and 16 species belonging to the Viride clade were identified based on integrated studies of phenotypic and molecular data. Among them, seven wood-inhabiting new species, T. albofulvopsis, T. densum, T. laevisporum, T. sinokoningii, T. sparsum, T. sphaerosporum and T. subviride, are found. They form trichoderma- to verticillium-like conidiophores, lageniform to subulate phialides and globose to ellipsoidal conidia, but vary greatly in colony features, growth rates, and sizes of phialides and conidia. To explore their taxonomic positions, the phylogenetic tree including all the known species of the Viride clade is constructed based on sequence analyses of the combined RNA polymerase II subunit b and translation elongation factor 1 alpha exon genes. Our results indicated that the seven new species were well-located in the Koningii, Rogersonii and Neorufum subclades as well as a few independent terminal branches. They are clearly distinguishable from any existing species. Morphological distinctions and sequence divergences between the new species and their close relatives were discussed.

Cobweb, a serious pathology in mushroom crops: A review

Directory of Open Access Journals (Sweden)

Jaime Carrasco

2017-07-01

Full Text Available Cobweb is a fungal disease of commercially cultivated mushrooms. Several members of the ascomycete genus Cladobotryum sp. have been reported as causal agents. White button mushroom is the most frequently cited host, but a wide range of cultivated edible mushrooms suffer cobweb. The pathology causes production losses and reduces the crop surface available. The parasite produces a great number of harmful conidia that can be released easily and distributed throughout the mushroom farm to generate secondary points of infection. To prevent initial outbreaks, hygiene is of primary importance within the facilities dedicated to mushroom cultivation, while additional measures must be implemented to control and reduce cobweb if there is an outbreak, including chemical and biological methods. This review summarizes and discusses the knowledge available on the historic occurrence of cobweb and its impact on commercial mushroom crops worldwide. Causal agents, disease ecology, including the primary source of infection and the dispersal of harmful conidia are also reviewed. Finally, control treatments to prevent the disease from breaking out are discussed.

Histopathology of Colletotrichum gloeosporioides on guava fruits (Psidium guajava L.

Directory of Open Access Journals (Sweden)

Sylvia Raquel Gomes Moraes

2013-06-01

Full Text Available Anthracnose, caused by Colletotrichum gloeosporioides, produces brown lesions on guava fruits, causing severe losses on postharvest. In this study, the infection and colonization of guava fruits by C. gloeosporioides has been examined using scanning and transmission electron microscopy. Fruits at the physiologically mature stage were inoculated with a 10(5 conidia/mL spore suspension. Afterward, fruits were incubated at 25 °C in a wet chamber for periods of 6, 12, 24, 48, 96 and 120 h to allow examination of the infection and colonization process. Conidia germination and appressoria formation occurred six hours after inoculation (h.a.i. Penetration occurred directly via penetration pegs from appressoria, which penetrated the host cuticle 48 h.a.i. Notably, the appressoria did not produce an appressorial cone surrounding the penetration pore. Infection vesicles were found in epidermal cells 96 h.a.i. The same fungal structures were found in epidermal and parenchymal cells of the host 120 h.a.i. Colonization strategy of C. gloeosporioides on guava fruit was intracellular hemibiotrophic.

Cobweb, a serious pathology in mushroom crops: A review

International Nuclear Information System (INIS)

Carrasco, J.; Navarro, M.J.; Gea, F.

2017-01-01

Cobweb is a fungal disease of commercially cultivated mushrooms. Several members of the ascomycete genus Cladobotryum sp. have been reported as causal agents. White button mushroom is the most frequently cited host, but a wide range of cultivated edible mushrooms suffer cobweb. The pathology causes production losses and reduces the crop surface available. The parasite produces a great number of harmful conidia that can be released easily and distributed throughout the mushroom farm to generate secondary points of infection. To prevent initial outbreaks, hygiene is of primary importance within the facilities dedicated to mushroom cultivation, while additional measures must be implemented to control and reduce cobweb if there is an outbreak, including chemical and biological methods. This review summarizes and discusses the knowledge available on the historic occurrence of cobweb and its impact on commercial mushroom crops worldwide. Causal agents, disease ecology, including the primary source of infection and the dispersal of harmful conidia are also reviewed. Finally, control treatments to prevent the disease from breaking out are discussed.

Cobweb, a serious pathology in mushroom crops: A review

Energy Technology Data Exchange (ETDEWEB)

Carrasco, J.; Navarro, M.J.; Gea, F.

2017-07-01

Cobweb is a fungal disease of commercially cultivated mushrooms. Several members of the ascomycete genus Cladobotryum sp. have been reported as causal agents. White button mushroom is the most frequently cited host, but a wide range of cultivated edible mushrooms suffer cobweb. The pathology causes production losses and reduces the crop surface available. The parasite produces a great number of harmful conidia that can be released easily and distributed throughout the mushroom farm to generate secondary points of infection. To prevent initial outbreaks, hygiene is of primary importance within the facilities dedicated to mushroom cultivation, while additional measures must be implemented to control and reduce cobweb if there is an outbreak, including chemical and biological methods. This review summarizes and discusses the knowledge available on the historic occurrence of cobweb and its impact on commercial mushroom crops worldwide. Causal agents, disease ecology, including the primary source of infection and the dispersal of harmful conidia are also reviewed. Finally, control treatments to prevent the disease from breaking out are discussed.

Growth and glucoamylase production by the thermophilic fungus Thermomyces lanuginosus in a synthetic medium

Energy Technology Data Exchange (ETDEWEB)

Haasum, I; Eriksen, S H; Jensen, B; Olsen, J [Copenhagen Univ. (Denmark). Dept. of General Microbiology

1991-02-01

The production of glucogenic amylase from the thermophilic fungus Thermomyces lanuginosus was studied in shake flasks and laboratory fermentors. As conidia were not able to germinate in media without yeast extract, pregerminated conidia were applied as inoculum. By this procedure it was possible to use different NH{sub 4}{sup +} salts as the sole source of nitrogen for growth and amylase formation in a synthetic medium. In pH-controlled fermentors a fourfold increase in the extracellular glucogenic amylase activity was obtained with (NH{sub 4})H{sub 2}PO{sub 4} as the nitrogen source as compared with yeast extract. However, by fractionation of these activities, comparable yields of partially purified glucoamylases were obtained. The glucoamylase preparation from fermentations with either of the nitrogen sources had a temperature optimum at 70deg C and showed similar thermal stability. By incubation without substrate at 60deg C, 90% of the activity was still present after 5 h. At 70deg C, 50% of the activity was retained after 30 min incubation. (orig.).

Fungitoxic effect of concentrations of essential oils on Colletotrichum gloeosporioides, isolated from the passion fruit

Directory of Open Access Journals (Sweden)

Ismail Teodoro Souza Júnior

2009-09-01

Full Text Available This study aimed to evaluate the effect of different concentrations of essential oils on the fungus Colletotrichum gloeosporioides. The essential oils were obtained from shoots of the plants Lippia sidoides, Ocimum gratissimum, Lippia citriodora, Cymbopogon citratus and Psidium guayava var. pomifera. After the incorporation of oil in water-agar and PDA medium, in concentrations of 1μL/mL, 3μL/mL 5μL/mL and 10μL/mL and witness at 0μL/mL (oil/the culture medium, the inhibitions of conidia germination and mycelial growth, respectively, were evaluates. The results showed that from the concentration of 1μL/mL, essential oils of all plant species had an effect on conidia germination, with an inhibition of 100%. The species Lippia sidoides, Ocimum gratissimum, Lippia citriodora and Cymbopogon citratus inhibited the growth of the fungus mycelium by 100%. The oil of Psidium guayava var. pomifera had progressive control over the mycelium of C. gloeosporioides, in accordance with the increase in concentrations of essential oil.

Discovery, Development, and Evaluation of a Horn Fly-Isolated (Diptera: Muscidae) Beauveria bassiana (Hypocreales: Cordyciptaceae) Strain From Florida, USA

Science.gov (United States)

Holderman, Christopher J.; Wood, Lois A.; Geden, Christopher J.

2017-01-01

The horn fly, Haematobia irritans (L.) is an important cattle pest and traditionally has been managed using insecticides; however, many horn fly populations are insecticide-resistant in United States. Use of alternative control techniques has been limited because of the challenges of managing a fly pest on pastured cattle. After the discovery of a wild horn fly infected with Beauveria bassiana in Florida, the fungus was cultured and evaluated for efficacy against laboratory-reared horn flies. This fungal strain was selected for increased virulence by passage through laboratory-reared horn fly hosts to shorten interval from infection to fly death and subsequent conidia formation, properties important to future use of the fungus as a biological control agent against horn flies. After seven passages through horn fly hosts, fly mortality was not significantly accelerated as evaluated through LT50 values, but conidia were readily produced from these killed flies. Although further development is needed to improve fungal efficacy, this fungal strain holds promise as a biological control agent for inclusion in horn fly integrated pest management programs. PMID:28423414

Nannizziopsis guarroi infection in 2 Inland Bearded Dragons (Pogona vitticeps): clinical, cytologic, histologic, and ultrastructural aspects.

Science.gov (United States)

Le Donne, Viviana; Crossland, Nicholas; Brandão, João; Sokolova, Yuliya; Fowlkes, Natalie; Nevarez, Javier G; Langohr, Ingeborg M; Gaunt, Stephen D

2016-06-01

Chrysosporium-related infections have been increasingly reported in reptiles over the last 2 decades. In this report, we describe clinical, cytologic, histopathologic, and ultrastructural aspects of Chrysosporium-related infection in 2 Inland Bearded Dragons (Pogona vitticeps). Case 1 was presented for an enlarging raised lesion over the left eye and multiple additional masses over the dorsum. Case 2 was submitted to necropsy by the referring veterinarian for suspected yellow fungus disease. Impression smears of the nodules in case 1 revealed granulomatous to pyogranulomatous inflammation and many septate, variably long, 4-10 μm wide, often undulated hyphae, and very rare conidia. Postmortem impression smears of the superficial lesions of case 2 contained large numbers of solitary conidia and arthroconidia and low numbers of hyphae with similar morphology to case 1. Histopathology of the 2 cases revealed severe, multifocal, chronic, ulcerative, nodular pyogranulomatous dermatitis, with myriad intralesional septate hyphae, and arthroconidia. Fungal culture and molecular sequencing in both cases indicated infection with Nannizziopsis guarroi. © 2016 American Society for Veterinary Clinical Pathology.

Trichoderma evansii and T. lieckfeldtiae: two new T. hamatum-like species.

Science.gov (United States)

Samuels, Gary J; Ismaiel, Adnan

2009-01-01

The new species, Trichoderma evansii and T. lieckfeldtiae, resemble the closely related T. hamatum and T. pubescens in forming discrete, setose conidial pustules within which arise smooth, green conidia from pachybasium-like conidiophores. The phylogenetic position of these species was determined with combined partial sequences of ITS, translationelongation factor 1-alpha, RNA polymerase II subunit and actin genes. All are members of the Viride clade. Trichoderma evansii forms a sister group relationship with a clade that includes T. hamatum and T. pubescens. It differs from the latter two species in having subglobose conidia; it was isolated as an endophyte from sapwood of Lophira alata (Ochnaceae) and Cola verticillata (Malvaceae) in Cameroon and Theobroma gileri (Malvaceae) in Peru. Trichoderma lieckfeldtiae occupies an unresolved position in the Viride clade despite being virtually morphologically indistinguishable from T. hamatum; it was isolated from fruit of cacao infected with Moniliophthora roreri in Colombia, pseudostroma of Moniliophthora roreri on pods of Theobroma cacao in Peru and from soil in a cacao farm in Cameroon (central Africa).

Disease and colony foundation in the dampwood termite Zootermopsis angusticollis: The survival advantage of nestmate pairs

Science.gov (United States)

Calleri, Daniel V.; Rosengaus, Rebeca B.; Traniello, James F. A.

2005-06-01

To determine the impact of inbreeding and outbreeding on disease resistance and survival during colony foundation, nestmate (NM) and non-nestmate (NON) primary reproductives of the dampwood termite Zootermopsis angusticollis were exposed to a single or double dose of conidia of the entomopathogenic fungus Metarhizium anisopliae. Male and female primary reproductive pairs originating from the same parent colony had higher survivorship than NON pairs in control and conidia-exposure treatments. The survival advantage of NM primary reproductives increased with the intensity of pathogen challenge and was significantly greater in the single- and double-dose treatments than in the controls. Although NM pairs had significantly lower mortality than NON pairs, the survivorship of colonies stabilized as they matured and inbred and outbred colonies did not differ in offspring production. These results demonstrate that colony foundation by NON male and female reproductives may have a disease-related survival cost during this critical phase of their life cycle. There may also be a cost associated with lower offspring heterozygosity, but in the first generation this does not appear to significantly impact colony growth.

Repetophragma calongeii sp. nov. and other interesting dematiaceous hyphomycetes from the North of Spain

Directory of Open Access Journals (Sweden)

Guarro, Josep

2009-12-01

Full Text Available Repetophragma calongeii sp. nov., a dematiaceous hyphomycete found on unidentified dead wood collected in the Pagoeta Natural Park, Spain, is described and illustrated. The fungus is recognized by numerous percurrent proliferations in the conidiophores and its 12-27 euseptate, subcylindrical or subfusiform conidia. A key to species of Repetophragma without rostrate conidia is given and 18 additional dematiaceous hyphomycetes are firtly reported for Spain.

Se describe e ilustra un hifomicete dematiáceo, Repetophragma calongeii sp. nov., encontrado sobre madera muerta colectada en el Parque Natural de Pagoeta, España. Este hongo se caracteriza por presentar conidióforos con numerosas proliferaciones percurrentes y conidios subcilíndricos o subfusiformes, 12-17 euseptados. Se propone una clave dicotómica para las especies de Repetophragma que no presentan conidios rostrados o apendiculados. Además, se describen otras 18 especies de hifomicetes dematiáceos que son nuevas citas para el catálogo micológico español.

New ochratoxin A producing species of Aspergillus section Circumdati

DEFF Research Database (Denmark)

Frisvad, Jens Christian; Frank, J.M.; Houbraken, J.A.M.P.

2004-01-01

Aspergillus section Circumdati contains species with yellow to ochre conidia and non-black sclerotia that produce at least one of the following extrolites: ochratoxins, penicillic acids, xanthomegnins or melleins. The exception to this is A. robustus, which produces black sclerotia, phototropic...... ochratoxin A production in coffee, rice, beverages and other foodstuffs are A. ochraceus, A. westerdijkiae and A. steynii....

Occurrence of toxicity among protease, amylase, and color mutants of a nontoxic soy sauce koji mold

International Nuclear Information System (INIS)

Kalayanamitr, A.; Bhumiratana, A.; Flegel, T.W.; Glinsukon, T.; Shinmyo, A.

1987-01-01

A soy sauce koji mold, Aspergillus flavus var. columnaris Raper and Fennel (ATCC 44310), was treated with UV irradiation to obtain mutant strains possessing high protease activities, high amylase activities, and light-colored conidia. Selected mutant strains were tested for toxicity, and some were found acutely toxic to weanling rats, although all were negative for aflatoxin production

The weak acid preservative sorbic acid inhibits conidial germination and mycelial growth of Aspergillus niger through intracellular acidification

NARCIS (Netherlands)

Plumridge, A.; Hesse, S.J.A.; Watson, A.J.; Lowe, K.C.; Stratford, M.; Archer, D.B.

2004-01-01

The growth of the filamentous fungus Aspergillus niger, a common food spoilage organism, is inhibited by the weak acid preservative sorbic acid (trans-trans-2,4-hexadienoic acid). Conidia inoculated at 105/ml of medium showed a sorbic acid MIC of 4.5 mM at pH 4.0, whereas the MIC for the amount of

Variability of Germinative Potential among Pathogenic Species of Aspergillus

OpenAIRE

Araujo, Ricardo; Rodrigues, Acacio Gonçalves

2004-01-01

The objective of our study was to evaluate parameters influencing the germination of Aspergillus conidia. Inoculum concentration and age significantly influenced germination. Different incubation temperatures revealed significant differences among Aspergillus species. The internal human milieu provides the ideal conditions for the development of invasive disease by Aspergillus fumigatus but restricts invasion by Aspergillus flavus and Aspergillus niger.

THE CHARACTERISATION OF FUNGI Passalora fulva (Cooke U.Braun&Crous PATHOTYPES, CAUSER OF TOMATO LEAF MOLD IN CROATIA

Directory of Open Access Journals (Sweden)

Adrijana Novak

2012-12-01

Full Text Available Tomato is considered one of the most widely grown vegetable crops in the world and production of it is rising every year in Croatia as well as in the world. This research paper shows results on tomato leaf mould caused by Passalora fulva (Cooke U.Braun & Crous (syn. Cladosporium fulvum (Cooke, which can cause significant reduction of yield and quality of tomato. Other than economic loss, tomato leaf mould can be the cause of allergies because it produces lots of spores. During research, the following materials and methods have been used: collecting samples of plant material infected with tomato leaf mould and assessments of disease intensity, isolation and determination of the pathogen, detecting pathotypes using biassay, susceptibility testing of different tomato genotypes infected with P. fulva, molecular identification and testing the effects of essential oils in vitro on conidia germination. Visual controls of greenhouse and outdoor growing tomato were conducted during the period from 2006 to 2011. Samples (17 from 14 localities in 10 counties (Istra County, Koprivnica-Križevci County, KrapinaZagorje County, Međimurje County, Osijek-BaranjaCounty, Primorje-Gorski Kotar County, Varaždin County, Virovitica-Podravina County, Zadar County and Zagreb County were collected. Symptoms were detected on 6 different tomato genotypes: Belle, Jeremy, Minaret, Amaneta, Gravitet and test genotype. Outdoors, P. fulva was not found. For isolation and determination of P. fulva 4 different growing media were used: PDA (potato – dextrose agar, tomato – agar, carrot – agar and V8 – agar. By measuring the size of conidia on carrot – agar it was determined that they were the closest in size to the conidia from infected tomato leaf. Bioassay for the first time in Croatia confirmed presence of two different physiological pathotypes of P. fulva, Cf – 0 and Cf – 5. Other pathotypes obtained by bioassay were not confirmed by sequencing because the

Resistance Against Basil Downy Mildew in Ocimum Species.

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Ben-Naim, Yariv; Falach, Lidan; Cohen, Yigal

2015-06-01

Downy mildew, caused by the oomycete Peronospora belbahrii, is a devastating disease of sweet basil. In this study, 113 accessions of Ocimum species (83 Plant Introduction entries and 30 commercial entries) were tested for resistance against downy mildew at the seedling stage in growth chambers, and during three seasons, in the field. Most entries belonging to O. basilicum were highly susceptible whereas most entries belonging to O. americanum, O. kilimanadascharicum, O. gratissimum, O. campechianum, or O. tenuiflorum were highly resistant at both the seedling stage and the field. Twenty-seven highly resistant individual plants were each crossed with the susceptible sweet basil 'Peri', and the F1 progeny plants were examined for disease resistance. The F1 plants of two crosses were highly resistant, F1 plants of 24 crosses were moderately resistant, and F1 plants of one cross were susceptible, suggesting full, partial, or no dominance of the resistance gene(s), respectively. These data confirm the feasibility of producing downy mildew-resistant cultivars of sweet basil by crossing with wild Ocimum species.

Temperature effect on rose downy mildew development under environmental controlled conditions

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Juan José Filgueira D.

2014-04-01

Full Text Available The rose downy mildew disease, caused by Peronospora sparsa Berkeley, is one of the most important that affect rose crops in Colombia. To manage this disease, flower growers must deal with high-costs due to the excessive application of fungicides, but without good results. Studies on P. sparsa behavior have shown its narrow relationship with environmental conditions. In this study, the temperature effect was evaluated during the infection and sporulation of P. sparsa in Charlotte leaflets, a susceptible commercial variety, through an environmental controlled conditions system. Infection and sporulation were observed at different temperatures in a range of from 4 to 40°C. Infection with the absence of or very low sporulation was observed at 4°C. The most favorable pathogen responses were between 15 and 18°C in terms of inoculum concentration and sporulation percentage. There was no infection or leaflet change above 35°C. According to the results, sporulation can occur from 4 to 33°C, confirming the fact that P. sparsa is able to reproduce throughout a wide temperature range.

POTENTIAL IMPACT OF METARHIZIUM ANISOPLIAE ON THE DIAMONDBACK MOTH (LEPIDOPTERA: PLUTELLIDAE AND ITS PARASITOID DIADEGMA SEMICLAUSUM (HYMENOPTERA: ICHNEUMONIDAE

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Rosma Hasibuan, Nilly Christalia, F.X. Susilo1, and Nur Yasin .

2011-11-01

Full Text Available Potential Impact of Metarhizium anisopliae on the Diamondback Moth (Lepidoptera: Plutellidae and Its Parasitoid Diadegma semiclausum (Hymenoptera: Ichneumonidae.  Laboratory studies were conducted to evaluate the effect of the Metarhizium anisopliae against the diamondback moth, Plutella xylostella and its parasitoid, Diadegma  semiclausum. A completely randomized design consisted of 5 treatments (4 concentrations of conidial suspension: 5 x 104, 3.5 x 105, 2.5 x 106, 1.2 x 107 conidia/ml and control was used.  The results indicated that the mortality of P. xylostella larvae were significantly induced by the fungal treatments.  A significant reduction in pupation and adult emergence of P. xylostella was also detected in all treatments when compared with that in the control. The fungus might also result in a male-biased sex ratio of the surviving P. xylostella. When applied at a concentration of 1.2 x 107 conidia/ml, M.  anisopliae might significantly reduce the survival of the parasitoid, D. semiclausum. Thus, despite its potential as a biological control agent against P. xylostella, the entomomogenous fungus M. anisopliae was also detrimental to the larvae parasitoid D. semiclausum.

Effect of nutrition and environmental factors on the endoparasitic fungus Esteya vermicola, a biocontrol agent against pine wilt disease.

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Xue, Jianjie; Zhang, Yongan; Wang, Chunyan; Wang, Yuzhu; Hou, Jingang; Wang, Zhen; Wang, Yunbo; Gu, Lijuan; Sung, Changkeun

2013-09-01

The nematophagous fungus Esteya vermicola has tremendous potential for biological control. This species exhibits strong infectious activity against pinewood nematodes, whereas the study on the effect of nutrition and environmental factors is still of paucity. Carbon (C), nitrogen (N), pH value, temperature, and water activity have great impact on the fungal growth, sporulation, and germination. In nutrition study, the greatest number of conidia (2.36 × 10(9) per colony) was obtained at the C:N ratio of 100:1 with a carbon concentration 32 g l(-1). In addition, the germination rate and radial growth of E. vermicola were used to evaluate the effects of environmental conditions and they were optimized as following: pH 5.5, 26 °C and water activity of 0.98. Our results also confirmed that variation of environmental factors has a detrimental influence on the efficacy of active conidia and growth of fungus. Moreover, under above optimal condition, the biocontrol efficacy was significantly improved in regard to the increase of adhesive and mortality rate, which highlight the study on the application of E. vermicola as pine wilt disease biocontrol agent.

Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis.

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Chen, Chun; Xie, Tingna; Ye, Sudan; Jensen, Annette Bruun; Eilenberg, Jørgen

2016-01-01

The selection of suitable reference genes is crucial for accurate quantification of gene expression and can add to our understanding of host-pathogen interactions. To identify suitable reference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expression of three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongation factor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reaction at different developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae), and under different nutritional conditions. We calculated the expression stability of candidate reference genes using four algorithms including geNorm, NormFinder, BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking of candidate reference genes from the most stable to the least stable was 18S (1.189), 28S (1.414) and EF1 (3). The 18S was, therefore, the most suitable reference gene for real-time RT-PCR analysis of gene expression under all conditions. These results will support further studies on gene expression in P. neoaphidis. Copyright © 2015 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis

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Chun Chen

2016-03-01

Full Text Available Abstract The selection of suitable reference genes is crucial for accurate quantification of gene expression and can add to our understanding of host–pathogen interactions. To identify suitable reference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expression of three traditional candidate genes including 18S rRNA(18S, 28S rRNA(28S and elongation factor 1 alpha-like protein (EF1, were measured by quantitative polymerase chain reaction at different developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae, and under different nutritional conditions. We calculated the expression stability of candidate reference genes using four algorithms including geNorm, NormFinder, BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking of candidate reference genes from the most stable to the least stable was 18S (1.189, 28S (1.414 and EF1 (3. The 18S was, therefore, the most suitable reference gene for real-time RT-PCR analysis of gene expression under all conditions. These results will support further studies on gene expression in P. neoaphidis.

Identification and validation of reference genes for qRT-PCR studies of the obligate aphid pathogenic fungus Pandora neoaphidis during different developmental stages.

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Zhang, Shutao; Chen, Chun; Xie, Tingna; Ye, Sudan

2017-01-01

The selection of stable reference genes is a critical step for the accurate quantification of gene expression. To identify and validate the reference genes in Pandora neoaphidis-an obligate aphid pathogenic fungus-the expression of 13classical candidate reference genes were evaluated by quantitative real-time reverse transcriptase polymerase chain reaction(qPCR) at four developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae). Four statistical algorithms, including geNorm, NormFinder, BestKeeper and Delta Ct method were used to rank putative reference genes according to their expression stability and indicate the best reference gene or combination of reference genes for accurate normalization. The analysis of comprehensive ranking revealed that ACT1and 18Swas the most stably expressed genes throughout the developmental stages. To further validate the suitability of the reference genes identified in this study, the expression of cell division control protein 25 (CDC25) and Chitinase 1(CHI1) genes were used to further confirm the validated candidate reference genes. Our study presented the first systematic study of reference gene(s) selection for P. neoaphidis study and provided guidelines to obtain more accurate qPCR results for future developmental efforts.

Aspergillus fumigatus melanins: Interference with the host endocytosis pathway and impact on virulence

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Thorsten eHeinekamp

2013-01-01

Full Text Available The opportunistic human pathogenic fungus Aspergillus fumigatus produces at least two types of melanin, namely pyomelanin and dihydroxynaphthalene (DHN melanin. Pyomelanin is produced during tyrosine catabolism via accumulation of homogentisic acid. Although pyomelanin protects the fungus against reactive oxygen species and acts as a defense compound in response to cell wall stress, mutants deficient for pyomelanin biosynthesis do not differ in virulence when tested in a murine infection model for invasive pulmonary aspergillosis. DHN melanin is responsible for the characteristic grey-greenish color of A. fumigatus conidia. Mutants lacking a functional polyketide synthase PksP, the enzyme responsible for the initial step in DHN-melanin formation, i.e., the synthesis of naphthopyrone, produce white spores and are attenuated in virulence. The activity of PksP was found to be essential not only for inhibition of apoptosis of phagocytes by interfering with the host PI3K/Akt signaling cascade but also for effective inhibition of acidification of conidia-containing phagolysosomes. These features allow A. fumigatus to survive in phagocytes and thereby to escape from human immune effector cells and to become a successful pathogen.

Aspergillus atacamensis and A. salisburgensis: two new halophilic species from hypersaline/arid habitats with a phialosimplex-like morphology.

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Martinelli, Livia; Zalar, Polona; Gunde-Cimerman, Nina; Azua-Bustos, Armando; Sterflinger, Katja; Piñar, Guadalupe

2017-07-01

Halophilic fungal strains isolated from historical wooden staircase in a salt mine in Austria, and from wall biofilm and soil of a cave in the Coastal Range of the hyperarid Atacama Desert in Chile were characterised and described newly as Aspergillus salisburgensis and Aspergillus atacamensis. Morphological characters including solitary phialides producing solitary conidia and conidia in chains and/or heads suggested affinity to Aspergillus subgenus Polypaecilum. Strains required salt for growth, grew optimally on media with 10-25% NaCl and at 15-28 °C. These values are similar to those observed for Aspergillus salinarus comb. nov. (Phialosimplex salinarum), while the ex-type strains of Aspergillus sclerotialis, Aspergillus chlamydosporus and Aspergillus caninus (all belonging to Aspergillus subgen. Polypaecilum) grew optimally at 0-5% NaCl and showed fastest growth at 28-37 °C. Phylogenetic analyses on the basis of rDNA sequences, RAPD-PCR fingerprint patterns, and cellobiohydrolase gene (cbh-I) polymorphism clustered the strains into three groups and supported their taxonomic recognition as A. salinarus, A. atacamensis and A. salisburgensis. On the basis of phylogenetic inferences, also Sagenomella keratitidis is newly combined as Aspergillus keratitidis and inferred as a species of Aspergillus subgenus Polypaecilum.

Keragaman Morfologi, Genetika, dan Patogenisitas Colletotrichum acutatum PenyebabAntraknosa Cabai di Jawa dan Sumatera

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Roy Ibrahim

2017-05-01

Full Text Available Colletotrichum acutatum is known as the major causal agents of anthracnose disease of chilli pepper in Indonesia. This research was aimed to study genetic variability of C. acutatum in Java and Sumatera based on morphological and molecular characteristics and to evaluate pathogenicity of several isolates. Observation on morphological characteristics involved colour and growth of each colony as well as shape and size of conidia. Molecular identification was performed by DNA amplification using specific primer for C. acutatum, i.e. CaInt2/ITS4 followed by sequencing and nucleotide sequence analysis. Pathogenicity test for each isolate on Capsicum annuum and C. frutescens was conducted in the laboratory using detached chilli. Morphology variability was observed from 40 isolates of C. acutatum. Most of the isolates have white/beige colonies with fusiform conidia of 6.11–9.73 µm in length and 2.24­–2.73 µm in width. Pathogenicity of 3 C. acutatum isolates varies from moderate to high based on lesions size on infected chilli. Sequence analysis of 8 C. acutatum isolates indicated high homology among isolates from Java and Sumatera

Inducible immune proteins in the dampwood termite Zootermopsis angusticollis

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Rosengaus, Rebeca B.; Cornelisse, Tara; Guschanski, Katerina; Traniello, James F. A.

2007-01-01

Dampwood termites, Zootermopsis angusticollis (Isoptera: Termopsidae), mount an immune response to resist microbial infection. Here we report on results of a novel analysis that allowed us to electrophoretically assess changes in hemolymph proteins in the same individual before and after exposure to a pathogen. We demonstrate that contact with a sublethal concentration of the entomopathogenic fungus Metarhizium anisopliae (Deuteromycotina:Hypomycetes) induces the production of protective proteins in nymphs, pseudergates (false workers), and soldiers. Termites exposed to an immunizing dosage of fungal conidia consistently showed an enhancement of constitutive proteins (62-85 kDa) in the hemolymph as well as an induction of novel proteins (28-48 kDa) relative to preimmunization levels. No significant differences in protein banding patterns relative to baseline levels in control and naïve termites were observed. Incubating excised and eluted induced proteins produced by immunized pseudergates or immunized soldiers with conidia significantly reduced the germination of the fungus. The fungistatic effect of eluted proteins differed significantly among five colonies examined. Our results show that the upregulation of protective proteins in the hemolymph underscores the in vivo immune response we previously recorded in Z. angusticollis.

Bioregulation of aflatoxin biosynthesis by unirradiated and irradiated conidia of Aspergillus flavus

International Nuclear Information System (INIS)

Aziz, N.H.; Abu-Shady, M.R.; El-Fouly, M.Z.; Moussa, L.A.

1996-01-01

A sequential technique involving the transfer of mycelia from peptone-based, aflatoxin-non-supporting medium to glucose based, aflatoxin-supporting medium was used to study the effect of γ-irradiation on the regulation of aflatoxin biosynthesis by Aspergillus flavus. Analysis indicated that irradiation at a dose of 1.00 kGy produced enhancement of aflatoxin biosynthesis in peptone-glucose mineral salt cultures with an increase of adenine nucleotide levels and fatty acid patterns of microsomes and mitochondria. The results suggest that aflatoxin synthesis is not regulated by the overall energy status of the fungal cell but that lipoperoxidation by γ-irradiation plays a role in aflatoxin biosynthesis

Proteomic analysis of the signaling pathway mediated by the heterotrimeric G? protein Pga1 of Penicillium chrysogenum

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Carrasco-Navarro, Ulises; Vera-Estrella, Rosario; Barkla, Bronwyn J.; Z??iga-Le?n, Eduardo; Reyes-Vivas, Horacio; Fern?ndez, Francisco J.; Fierro, Francisco

2016-01-01

Background The heterotrimeric G? protein Pga1-mediated signaling pathway regulates the entire developmental program in Penicillium chrysogenum, from spore germination to the formation of conidia. In addition it participates in the regulation of penicillin biosynthesis. We aimed to advance the understanding of this key signaling pathway using a proteomics approach, a powerful tool to identify effectors participating in signal transduction pathways. Results Penicillium chrysogenum mutants with ...

Apple scab control with grapefruit seed extract: no alternative to chemical fungicides

OpenAIRE

Spitaler, Renate; Marschall, Klaus; Zidorn, Christian; Kelderer, Markus; Zelger, Roland; Stuppner, Hermann

2004-01-01

The growth inhibiting effect of four commercially available grapefruit seed extracts on the causal organism of apple scab Venturia inaequalis was tested. Germination of the conidia of Venturia inaequalis was pronouncedly inhibited by all tested extracts. The commercial products were analyzed by high pressure liquid chromatography and thin layer chromatography. All samples contained at least one preserving agent. These substances were identified as either benzethonium chloride,...

Transcriptome Analyses Shed New Insights into Primary Metabolism and Regulation of Blumeria graminis f. sp. tritici during Conidiation

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Fan-Song Zeng

2017-06-01

Full Text Available Conidia of the obligate biotrophic fungal pathogen Blumeria graminis f. sp. tritici (Bgt play a vital role in its survival and rapid dispersal. However, little is known about the genetic basis for its asexual reproduction. To uncover the primary metabolic and regulatory events during conidiation, we sequenced the transcriptome of Bgt epiphytic structures at 3 (vegetative hyphae growth, 4 (foot cells initiation, and 5 (conidiophore erection days post-inoculation (dpi. RNA-seq analyses identified 556 and 404 (combined 685 differentially expressed genes (DEGs at 4 and 5 dpi compared with their expression levels at 3 dpi, respectively. We found that several genes involved in the conversion from a variety of sugars to glucose, glycolysis, the tricarboxylic acid cycle (TAC, the electron transport chain (ETC, and unsaturated fatty acid oxidation were activated during conidiation, suggesting that more energy supply is required during this process. Moreover, we found that glucose was converted into glycogen, which was accumulated in developing conidiophores, indicating that it could be the primary energy storage molecule in Bgt conidia. Clustering for the expression profiles of 91 regulatory genes showed that calcium (Ca2+, H2O2, and phosphoinositide (PIP signaling were involved in Bgt conidiation. Furthermore, a strong accumulation of H2O2 in developing conidiophores was detected. Application of EGTA, a Ca2+ chelator, and trifluoperazine dihydrochloride (TFP, a calmodulin (CaM antagonist, markedly suppressed the generation of H2O2, affected foot cell and conidiophore development and reduced conidia production significantly. These results suggest that Ca2+ and H2O2 signaling play important roles in conidiogenesis and a crosslink between them is present. In addition to some conidiation-related orthologs known in other fungi, such as the velvet complex components, we identified several other novel B. graminis-specific genes that have not been previously

Production of extracellular traps against Aspergillus fumigatus in vitro and in infected lung tissue is dependent on invading neutrophils and influenced by hydrophobin RodA.

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Sandra Bruns

2010-04-01

Full Text Available Aspergillus fumigatus is the most important airborne fungal pathogen causing life-threatening infections in immunocompromised patients. Macrophages and neutrophils are known to kill conidia, whereas hyphae are killed mainly by neutrophils. Since hyphae are too large to be engulfed, neutrophils possess an array of extracellular killing mechanisms including the formation of neutrophil extracellular traps (NETs consisting of nuclear DNA decorated with fungicidal proteins. However, until now NET formation in response to A. fumigatus has only been demonstrated in vitro, the importance of neutrophils for their production in vivo is unclear and the molecular mechanisms of the fungus to defend against NET formation are unknown. Here, we show that human neutrophils produce NETs in vitro when encountering A. fumigatus. In time-lapse movies NET production was a highly dynamic process which, however, was only exhibited by a sub-population of cells. NETosis was maximal against hyphae, but reduced against resting and swollen conidia. In a newly developed mouse model we could then demonstrate the existence and measure the kinetics of NET formation in vivo by 2-photon microscopy of Aspergillus-infected lungs. We also observed the enormous dynamics of neutrophils within the lung and their ability to interact with and phagocytose fungal elements in situ. Furthermore, systemic neutrophil depletion in mice almost completely inhibited NET formation in lungs, thus directly linking the immigration of neutrophils with NET formation in vivo. By using fungal mutants and purified proteins we demonstrate that hydrophobin RodA, a surface protein making conidia immunologically inert, led to reduced NET formation of neutrophils encountering Aspergillus fungal elements. NET-dependent killing of Aspergillus-hyphae could be demonstrated at later time-points, but was only moderate. Thus, these data establish that NET formation occurs in vivo during host defence against A

Powdery mildew of Ruta graveolens in Brazil caused by Oidiopsis haplophylli Oídio de Ruta graveolens no Brasil causado por Oidiopsis haplophylli

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José Ricardo Liberato

2006-03-01

Full Text Available Oidiopsis haplophylli is described causing powdery mildew on Ruta graveolens for the first time in Brazil (Viçosa, MG. The fungus causes yellowing of infected leaves accompanied by the presence of a whitish mycelial colony abaxially. The following combination of characters typical of this species was observed on the specimens collected in Viçosa: mycelium hypophyllous, hemiendophytic (partly external and partly internal, entering the leaves through the stomata; conidiophores hypophyllous, produced from the internal mycelium emerging through the stomata, cylindrical, hyaline, smooth; conidia, isolate, dimorphic - primary conidia lanceolate, 66.5-91.5 x 11.0-20.0 mm l/w ratio 3.5-6.2, secondary conidia cylindrical with rounded ends, 57.0-81.5 x 13.5-20.0 mm, l/w ratio 3.1 - 5.3, aseptate, hyaline, smooth.Oidiopsis haplophylli é relatado causando oídio em arruda (Ruta graveolens pela primeira vez no Brasil (Viçosa, MG. O fungo causa um amarelecimento de folhas de arruda sem bordos definidos acompanhado de cobertura fúngica branca na superfície abaxial. O seguinte conjunto de características, típico desta espécie foi observado nos espécimes coletados em Viçosa: presença de micélio hemiendofítico (parcialmente interno e parcialmente externo, tênue a denso com penetração das folhas através dos estômatos; conidióforos originando do micélio interno, emergindo através dos estômatos, cilíndricos, hialinos, lisos; conídios unicelulares, hialinos, produzidos isoladamente, dimórficos - conídios primários lanceolados, 66,5-91,5 x 11,0-20,0 mm, razão c/l 3,5-6,2, conídios secondários cilíndricos com extremidades arredondadas, 57,0-81,5 x 13,5-20,0 mm, razão c/l 3,1-5,3.

Towards the development of an autocontamination trap system to manage populations of emerald ash borer (Coleoptera: Buprestidae) with the native entomopathogenic fungus, Beauveria bassiana.

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Lyons, D Barry; Iavallée, Robert; Kyei-Poku, George; Van Frankenhuyzen, Kees; Johny, Shajahan; Guertin, Claude; Francese, Joseph A; Jones, Gene C; Blais, Martine

2012-12-01

Emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae) is an invasive species from Asia that was discovered in North America Canada, in 2002. Herein, we describe studies to develop an autocontamination trapping system to disseminate Beauveria bassiana to control beetle populations. The standard trap for emerald ash borer in Canada is a light green prism trap covered in an insect adhesive and baited with (Z)-3-hexenol. We compared of green multifunnel traps, green intercept panel traps (both with and without fluon coating) and green prism traps for capturing emerald ash borer in a green ash plantation. The coated green multifunnel traps captured significantly more males and more females than any other trap design. We examined the efficacy of two native B. bassiana isolates, INRS-CFL and L49-1AA. In a field experiment the INRS-CFL isolate attached to multifunnel traps in autocontamination chambers retained its pathogenicity to emerald ash borer adults for up to 43 d of outdoor exposure. Conidia germination of the INRS-CFL isolate was >69% after outdoor exposure in the traps for up to 57 d. The L49-1AA isolate was not pathogenic in simulated trap exposures and the germination rate was extremely low (<5.3%). Mean (+/- SEM) conidia loads on ash borer adults after being autocontaminated in the laboratory using pouches that had been exposed in traps out of doors for 29 d were 579,200 (+/- 86,181) and 2,400 (+/- 681) for the INRS-CFL and the LA9-1AA isolates, respectively. We also examined the fungal dissemination process under field conditions using the L49-1AA isolate in a green ash plantation. Beetles were lured to baited green multifunnel traps with attached autocontamination chambers. Beetles acquired fungal conidia from cultures growing on pouches in the chambers and were recaptured on Pestick-coated traps. In total, 2,532 beetles were captured of which 165 (6.5%) had fungal growth that resembled B. bassiana. Of these 25 beetles were positive for

Survival and mutant production induced by mutagenic agents in Metarhizium anisopliae Sobrevivência e obtenção de mutantes induzidos por agentes mutagênicos em Metarhizium anisopliae

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V. Kava - Cordeiro

1995-12-01

Full Text Available A wild strain of Metarhizium anisopliae, an entomopathogenic fungus, was submitted to three mutagenic agents: gamma radiation, ultraviolet light and nitrous acid. Survival curves were obtained and mutants were selected using different mutagenic doses which gave 1 to 5% survival. Morphological and auxotrophic mutants were isolated. Morphological mutants were grouped in a class with yellow conidia and other with pale vinaceous conidia as opposed to the green wild type conidia. Auxotrophic mutants had requirements for vitamin and aminoacid biosynthesis. More than 58% of the total auxotrophk mutants required proline/aipnine. Gamma radiation showed to be the most efficient mutagenic agent giving 0.2% of auxotrophk mutants followed by ultraviolet light (0.12% and nitrous acid (0.06%.The conidial colour and auxotrophk mutants isolated until now from M. anisopliae were reviewed.Uma linhagem selvagem do fungo entomopatogênico Metarhizium anisopliae foi submetida à ação de três agentes mutagênicos: radiação gama, luz ultravioleta e ácido nitroso. Curvas de sobrevivência foram obtidas para cada mutagênicos utilizado e mutantes foram selecionados a partir de doses dos mutagênicos que proporcionassem de 1 a 5% de sobrevivência. Mutantes morfológicos para a coloração de conídios e mutantes auxotróficos foram isolados. Mutantes para coloração de conidios foram agrupados em duas classes, uma com conídios amarelos e outra com conídios vinho pálido. Os mutantes auxotróficos obtidos foram deficientes para aminoácidos e vitaminas e mais de 58% deles eram auxotróficos para prolina/argmina. Radiação gama foi o mutagênico mais eficiente com uma porcentagem de obtenção de mulantes auxotróficos de aproximadamente 0,2%, seguido pela luz ultravioleta (0.12% e pelo ácido nitroso (0.06%.Os mulantes morfológicos e auxotróficos obtidos até o momento em Metarhizium anisopliae foram revistos.

Selección de Cepas de Hongos Entomopatógenos para el Manejo de Anastrepha obliqua (Macquart, 1835 (Diptera: Tephritidae en Colombia Selection of Strains of Entomopathogenic Fungi for Management of Anastrepha obliqua (Macquart, 1835 (Diptera: Tephritidae in Colombia

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Armando Osorio-Fajardo

2011-12-01

Full Text Available Se evaluaron 15 cepas de los hongos entomopatógenos Beauveria bassiana y Metarhizium anisopliae sobre adultos de un día de edad de la mosca de la fruta Anastrepha obliqua. El trabajo se realizó con el fin de seleccionar las cepas más virulentas al insecto y estudiar el efecto sobre los adultos jóvenes cuando el hongo era aplicado antes de la emergencia. Mediante un screening con una concentración de 1x10(7 conidias/mL se seleccionaron las tres cepas más virulentas, siendo dos de ellas de Beauveria y una de Metarhizium, las cuales causaron mortalidades del 77%, 71% y 66%. Valores de CL50 de 2,38x10(6, 1,81x10(6 y 9,94x10(6 conidias/mL, respectivamente, fueron determinados para cada una de estas cepas y un TL50 respectivo de 48,12; 56 y 42,75 horas. No se encontraron diferencias significativas en la mortalidad entre hembras y machos. La aspersión de la CL90 de las cepas seleccionadas sobre el medio de pupación de la mosca de la fruta produjo 34-48% de mortalidad durante las 120 horas de evaluación. Los hongos entomopatógenos pueden ser utilizados fácilmente para el control biológico de A. obliqua aplicándolos de manera dirigida a los adultos jóvenes bajo la copa de los árboles, en programas de manejo integrado de plagas.Fifteen strains from entomopathogenic Beauveria bassiana and Metarhizium anisopliae fungi were evaluated on one day-old adults of Anastrepha obliqua fruit fly. Tested were carried out for selecting the most virulent strains and the effectiveness of their use on young adult when the entomopathogen were applied before emergence were studied too. A screening with a 1x10(7 conidia/mL concentration was used for selecting the three most pathogenic isolates, two from Beauveria and one from Metarhizium, having 77, 71 and 66% mortality. The LC50 for these isolates were 2.38x10(6, 1.81x10(6 and 9.94x10(6 conidia/mL, respectively, and a respective LT50 were 48.12, 56 and 42.75 hours. No significant differences were found

Ethylene not responsible for inhibition of conidium germination by soils volatiles

Energy Technology Data Exchange (ETDEWEB)

Schippers, B.; Boerwinkel, D.J.; Konings, H.

1978-01-01

Volatile factors inhibiting the germination of Botrytis cinerea and Cochlibolus sativus conidia on membrane filters were emanated from six alkaline and two acid soils of different organic content and origin. Addition of lime or chitin increased the inhibitory action. Evidence is presented that ethylene is not involved either directly or indirectly in the inhibition of conidium germination above the different soils tested. 16 references, 4 figures, 2 tables.

Morphological and Pathogenic Characteristics of the Fungus Cladobotryum dendroides, the Causal Agent of Cobweb Disease of the Cultivated Mushroom Agaricus bisporus in Serbia

OpenAIRE

Ivana Potočnik; Emil Rekanović; Svetlana Milijašević; Biljana Todorović; Miloš Stepanović

2008-01-01

Twenty isolates were isolated from diseased fruiting bodies of Agaricus bisporus collected from Serbian mushroom farms during 2003-2007. The isolates formed white, cottony, aerial colonies on agar media. With age, conidia and colonies turned yellow and redish. Pathogenicity of these isolates was confirmed by inoculation of harvested basidiomes of A. bisporus and by casing inoculation. Symptoms similar to natural infection were recorded. Based on pathogenicity tests and morphological character...

Detection of Fusarium verticillioides by PCR-ELISA based on FUM21 gene.

Science.gov (United States)

Omori, Aline Myuki; Ono, Elisabete Yurie Sataque; Bordini, Jaqueline Gozzi; Hirozawa, Melissa Tiemi; Fungaro, Maria Helena Pelegrinelli; Ono, Mario Augusto

2018-08-01

Fusarium verticillioides is a primary corn pathogen and fumonisin producer which is associated with toxic effects in humans and animals. The traditional methods for detection of fungal contamination based on morphological characteristics are time-consuming and show low sensitivity and specificity. Therefore, the objective of this study was to develop a PCR-ELISA based on the FUM21 gene for F. verticillioides detection. The DNA of the F. verticillioides, Fusarium sp., Aspergillus sp. and Penicillium sp. isolates was analyzed by conventional PCR and PCR-ELISA to determine the specificity. The PCR-ELISA was specific to F. verticillioides isolates, showed a 2.5 pg detection limit and was 100-fold more sensitive than conventional PCR. In corn samples inoculated with F. verticillioides conidia, the detection limit of the PCR-ELISA was 1 × 10 4 conidia/g and was also 100-fold more sensitive than conventional PCR. Naturally contaminated corn samples were analyzed by PCR-ELISA based on the FUM21 gene and PCR-ELISA absorbance values correlated positively (p PCR-ELISA developed in this study can be useful for F. verticillioides detection in corn samples. Copyright © 2018 Elsevier Ltd. All rights reserved.

Induction of mutation in Aspergillus niger for conversion of cellulose into fuel precursor

International Nuclear Information System (INIS)

Helmi, S.; Khalil, A.; Tahoun, M.K.; Khairy, A.H.

1990-01-01

Induction of mutation in Aspergillus niger ATCC 1004 was achieved by exposure of conidia suspension to uv-light 2,400 erg/m 2 (240 J/m 2 ) for 120 s to obtain 5% survivors. The mutants were tested for the activity of different cellulases and were selected according to (1) rate and percentage of germination, (2) mycelial dry weight, (3) colony morphology, and (4) diameter of conidia. The efficiency of the mutants to convert celllose into reducing sugars was detected by the assay of the activity of different cellulases. Three of the selected mutants indicated higher Avicel activity, 0.0183, 0.0205, and 0.0295 μ mol reducing sugars/ml/min compared to 0.0153 for the wild type. The uv-irradiation also induced mutants that gave elevated CMCase activity, 3.06, 3.44, 5.30, 6.47, and 7.20 μmol reducing sugars/ml/min as compared with the wild type, 2.94 μmol reducing sugars/ml/min. Highly significant (F-test) increases in the activity of β-glucosidase were obtained for some of the selected mutants, 196.50, 255.00, 255.70, and 267.5 μmol P-nitrophenol/ml/min compared to the parent strain, 167.20 μmol P-nitrophenol/ml/min

Identification and validation of reference genes for qRT-PCR studies of the obligate aphid pathogenic fungus Pandora neoaphidis during different developmental stages.

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Shutao Zhang

Full Text Available The selection of stable reference genes is a critical step for the accurate quantification of gene expression. To identify and validate the reference genes in Pandora neoaphidis-an obligate aphid pathogenic fungus-the expression of 13classical candidate reference genes were evaluated by quantitative real-time reverse transcriptase polymerase chain reaction(qPCR at four developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae. Four statistical algorithms, including geNorm, NormFinder, BestKeeper and Delta Ct method were used to rank putative reference genes according to their expression stability and indicate the best reference gene or combination of reference genes for accurate normalization. The analysis of comprehensive ranking revealed that ACT1and 18Swas the most stably expressed genes throughout the developmental stages. To further validate the suitability of the reference genes identified in this study, the expression of cell division control protein 25 (CDC25 and Chitinase 1(CHI1 genes were used to further confirm the validated candidate reference genes. Our study presented the first systematic study of reference gene(s selection for P. neoaphidis study and provided guidelines to obtain more accurate qPCR results for future developmental efforts.

Evidence for Anti-Pseudogymnoascus destructans (Pd Activity of Propolis

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Soumya Ghosh

2017-12-01

Full Text Available White-nose syndrome (WNS in bats, caused by Pseudogymnoascus destructans (Pd, is a cutaneous infection that has devastated North American bat populations since 2007. At present, there is no effective method for controlling this disease. Here, we evaluated the effect of propolis against Pd in vitro. Using Sabouraud dextrose agar (SDA medium, approximately 1.7 × 107 conidia spores of the Pd strain M3906-2/mL were spread on each plate and grown to form a consistent lawn. A Kirby–Bauer disk diffusion assay was employed using different concentrations of propolis (1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, in plates incubated at 8 °C and 15 °C. At 8 °C and 15 °C, as the concentration of propolis increased, there was an increasing zone of inhibition (ZOI, reaching the highest degree at 10% and 25% concentrations, respectively. A germule suppression assay showed a similar effect on Pd conidia germination. A MALDI-TOF-MS analysis of propolis revealed multiple constituents with a potential anti-Pd activity, including cinnamic acid, p-coumaric acid, and dihydrochalcones, which could be further tested for their individual effects. Our study suggests that propolis or its individual constituents might be suitable products against Pd.

Evidence for Anti-Pseudogymnoascus destructans (Pd) Activity of Propolis.

Science.gov (United States)

Ghosh, Soumya; McArthur, Robyn; Guo, Zhi Chao; McKerchar, Rory; Donkor, Kingsley; Xu, Jianping; Cheeptham, Naowarat

2017-12-21

White-nose syndrome (WNS) in bats, caused by Pseudogymnoascus destructans ( Pd ), is a cutaneous infection that has devastated North American bat populations since 2007. At present, there is no effective method for controlling this disease. Here, we evaluated the effect of propolis against Pd in vitro. Using Sabouraud dextrose agar (SDA) medium, approximately 1.7 × 10⁷ conidia spores of the Pd strain M3906-2/mL were spread on each plate and grown to form a consistent lawn. A Kirby-Bauer disk diffusion assay was employed using different concentrations of propolis (1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%), in plates incubated at 8 °C and 15 °C. At 8 °C and 15 °C, as the concentration of propolis increased, there was an increasing zone of inhibition (ZOI), reaching the highest degree at 10% and 25% concentrations, respectively. A germule suppression assay showed a similar effect on Pd conidia germination. A MALDI-TOF-MS analysis of propolis revealed multiple constituents with a potential anti- Pd activity, including cinnamic acid, p-coumaric acid, and dihydrochalcones, which could be further tested for their individual effects. Our study suggests that propolis or its individual constituents might be suitable products against Pd .

Reactive oxygen species production, induced by atmospheric modification, alter conidial quality of Beauveria bassiana.

Science.gov (United States)

Pérez-Guzmán, D; Montesinos-Matías, R; Arce-Cervantes, O; Gómez-Quiroz, L E; Loera, O; Garza-López, P M

2016-08-01

The aim of this study was to determine the relationship between reactive oxygen species (ROS) production and conidial infectivity in Beauveria bassiana. Beauveria bassiana Bb 882.5 was cultured in solid-state culture (SSC) using rice under three oxygen conditions (21%, or pulses at 16 and 26%). Hydrophobicity was determined using exclusion phase assay. Bioassays with larvae or adults of Tenebrio molitor allowed the measurements of infectivity parameters. A fluorometric method was used for ROS quantification (superoxide and total peroxides). NADPH oxidase (NOX) activity was determined by specific inhibition. Conidial hydrophobicity decreased by O2 pulses. Mortality of larvae was only achieved with conidia harvested from cultures under 21% O2 ; whereas for adult insects, the infectivity parameters deteriorated in conidia obtained after pulses at 16 and 26% O2 . At day 7, ROS production increased after 16 and 26% O2 treatments. NOX activity induced ROS production at early stages of the culture. Modification of atmospheric oxygen increases ROS production, reducing conidial quality and infectivity. This is the first study in which conidial infectivity and ROS production in B. bassiana has been related, enhancing the knowledge of the effect of O2 pulses in B. bassiana. © 2016 The Society for Applied Microbiology.

Transcription factor Afmac1 controls copper import machinery in Aspergillus fumigatus.

Science.gov (United States)

Kusuya, Yoko; Hagiwara, Daisuke; Sakai, Kanae; Yaguchi, Takashi; Gonoi, Tohru; Takahashi, Hiroki

2017-08-01

Copper (Cu) is an essential metal for all living organisms, although it is toxic in excess. Filamentous fungus must acquire copper from its environment for growth. Despite its essentiality for growth, the mechanisms that maintain copper homeostasis are not fully understood in filamentous fungus. To gain insights into copper homeostasis, we investigated the roles of a copper transcription factor Afmac1 in the life-threatening fungus Aspergillus fumigatus, a homolog of the yeast MAC1. We observed that the Afmac1 deletion mutant exhibited not only significantly slower growth, but also incomplete conidiation including a short chain of conidia and defective melanin. Moreover, the expressions of the copper transporters, ctrA1, ctrA2, and ctrC, and metalloreductases, Afu8g01310 and fre7, were repressed in ∆Afmac1 cells, while those expressions were induced under copper depletion conditions in wild-type. The expressions of pksP and wetA, which are, respectively, involved in biosynthesis of conidia-specific melanin and the late stage of conidiogenesis, were decreased in the ∆Afmac1 strain under minimal media condition. Taken together, these results indicate that copper acquisition through AfMac1 functions in growth as well as conidiation.

Fungal Volatiles Can Act as Carbon Sources and Semiochemicals to Mediate Interspecific Interactions Among Bark Beetle-Associated Fungal Symbionts.

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Jonathan A Cale

Full Text Available Mountain pine beetle (Dendroctonus ponderosae has killed millions of hectares of pine forests in western North America. Beetle success is dependent upon a community of symbiotic fungi comprised of Grosmannia clavigera, Ophiostoma montium, and Leptographium longiclavatum. Factors regulating the dynamics of this community during pine infection are largely unknown. However, fungal volatile organic compounds (FVOCs help shape fungal interactions in model and agricultural systems and thus may be important drivers of interactions among bark beetle-associated fungi. We investigated whether FVOCs can mediate interspecific interactions among mountain pine beetle's fungal symbionts by affecting fungal growth and reproduction. Headspace volatiles were collected and identified to determine species-specific volatile profiles. Interspecific effects of volatiles on fungal growth and conidia production were assessed by pairing physically-separated fungal cultures grown either on a carbon-poor or -rich substrate, inside a shared-headspace environment. Fungal VOC profiles differed by species and influenced the growth and/or conidia production of the other species. Further, our results showed that FVOCs can be used as carbon sources for fungi developing on carbon-poor substrates. This is the first report demonstrating that FVOCs can drive interactions among bark beetle fungal symbionts, and thus are important factors in beetle attack success.

Passive vectoring of entomopathogenic fungus Beauveria bassiana among the wax moth Galleria mellonella larvae by the ectoparasitoid Habrobracon hebetor females.

Science.gov (United States)

Kryukov, Vadim Yu; Kryukova, Natalia A; Tyurin, Maksim V; Yaroslavtseva, Olga N; Glupov, Viktor V

2017-03-15

Females of the ectoparasitoid Habrobracon hebetor attack and envenomate numerous host individuals during oviposition. The vectoring of the entomopathogenic fungus Beauveria bassiana during the adhesion stage by ectoparasitoid females among the wax moth larvae Galleria mellonella was explored under laboratory conditions. Vectoring occurred both from infected parasitoids to wax moth larvae and from infected to healthy wax moth larvae by parasitoids. The efficacy of vectoring in both cases was dose dependent. Parasitoid females were unable to recognize infected larvae in a labyrinth test. In addition, the presence of H. hebetor females significantly (1.5-13 fold) increased the mycoses level in clusters of G. mellonella, with 40% of the larvae infected with fungal conidia. Envenomation by H. hebetor increased conidia germination on the cuticles of the wax moth larvae by 4.4 fold. An enhanced germination rate (2 fold) was registered in the n-hexane epicuticular extract of envenomated larvae compared to that of healthy larvae. Both envenomation and mycoses enhanced the phenoloxidase (PO) activity in the integument of G. mellonella and, in contrast, decreased the encapsulation rate in hemolymphs. We hypothesize that changes in the integument property and inhibition of cellular immunity provide the highest infection efficacy of entomopathogenic fungi with H. hebetor. © 2017 Institute of Zoology, Chinese Academy of Sciences.

Participation of the phosphoinositide metabolism in the hypersensitive response of Citrus limon against Alternaria alternata

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XIMENA ORTEGA

2001-01-01

Full Text Available Lemon seedlings inoculated with Alternaria alternata develop a hypersensitive response (HR that includes the induction of Phenylalanine ammonia-lyase (PAL, E. C. 4.3.1.5 and the synthesis of scoparone. The signal transduction pathway involved in the development of this response is unknown. We used several inhibitors of the Phosphoinositide (PI animal system to study a possible role of Inositol-1,4,5-triphosphate (IP3 in the transduction of the fungal conidia signal in Citrus limon. The HR was only partially inhibited by EGTA, suggesting that not only external but internal calcium as well are necessary for a complete development of the HR. In this plant system, Alternaria alternata induced an early accumulation of the second messenger IP3. When lemon seedlings were watered long term with LiCl, an inhibitor of the phosphoinositide cycle, the IP3 production was reduced, and the LiCl-watered plants could neither induce PAL nor synthesize scoparone in response to fungal conidia. Furthermore, neomycin, a Phospholipase C (PLC, E. C. 3.1.4.3 inhibitor, also inhibited PAL induction and scoparone synthesis in response to A. alternata. These results suggest that IP3 could be involved in the signal transduction pathway for the development of the HR of Citrus limon against A. alternata

Effects of Simultaneous Radiofrequency Radiation and Chemical Exposure of Mammalian Cells. Volume 2

Science.gov (United States)

1988-07-01

chromosome - - - - - - -I aberrations and sister chromatid exchanges (SCE). Yao (1982) exposed rat kangaroo RH5 and RH1l6 cells to 2.45 GHz radiation, and...control was reported in chromosome aberrations. Yac (1982) investigated the cytogenetic consequences of chronic microwave exposure on rat kangaroo RH5...was said to be 280C. The cells were exposed both as conidia, which are "rather inactive metabolically ," and also after DNA replication had been

Control of Root-Knot Nematodes on Tomato by the Endoparasitic Fungus Meria coniospora

OpenAIRE

Jansson, Hans-Börje; Jeyaprakash, A.; Zuckerman, Bert M.

1985-01-01

The endoparasitic nematophagous fungus Meria coniospora reduced root-knot nematode galling on tomatoes in greenhouse pot trials. The fungus was introduced to pots by addition of conidia at several inoculum levels directly to the soil or addition of nematodes infected with M. coniospora to the soil; both methods reduced root galling by root-knot nematodes. These studies represent a part of a recently initiated effort to evaluate the potential of endoparasitic nematophagous fungi for biocontrol...

Ekofyziologie mikroskopické houby Pseudogymnoascus destructans

OpenAIRE

Homutová, Karolína

2014-01-01

A microscopic fungus Pseudogymnoascus destructans (Ascomycota: Pseudeurotiaceae) causes illness known as white-nose syndrome (WNS) causing death of bats during hibernation. The illness occurs in the North America and in Europe. The fungus is characteristic by asymmetrically curved conidia, by slow growth and growth at low temperatures (below 20 řC). The aim of this study is to clarify properties responsible for unique ecelogy of Pseudogymnoascus destructans by comparison with ecological relat...

Resource capture and competitive ability of non-pathogenic Pseudogymnoascus spp. and P. destructans, the cause of white-nose syndrome in bats

OpenAIRE

Wilson, Michael B.; Held, Benjamin W.; Freiborg, Amanda H.; Blanchette, Robert A.; Salomon, Christine E.

2017-01-01

White-nose syndrome (WNS) is a devastating fungal disease that has been causing the mass mortality of hibernating bats in North America since 2006 and is caused by the psychrophilic dermatophyte Pseudogymnoascus destructans. Infected bats shed conidia into hibernaculum sediments and surfaces, but it is unknown if P. destructans can form stable, reproductive populations outside its bat hosts. Previous studies have found non-pathogenic Pseudogymnoascus in bat hibernacula, and these fungi may pr...

Modern methods in cereal grain mycology

OpenAIRE

Olsson, Johan

2000-01-01

A simple rapid DNA extraction method, equally suitable for spores and mycelia is proposed. Heating samples in NaOH and SDS provides DNA of high purity, suitable for Polymerase Chain Reaction (PCR) analysis. For Penicillium roqueforti the detection limit was 6 x lo3 conidia and 1 mg (fresh weight) mycelium in the extraction liquid. The method proved efficient with Aspergillus jlavus, Fusarium graminearum, Rhizopus stolonifer, Eurotium herbariorum, and Cladosporium herbarum, as well. An optimis...

Optimization of Factors Affecting Beauveria bassiana Fungus Ability in Control of Greater Wax Moth (Galleria mellonella L. by Response Surface Method

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Ali Heidari

2017-03-01

Full Text Available Introduction: Stored product pests are a major problem in the storage of agricultural products that cause damage from harvest until consumption. Greater Wax Moth (Galleria mellonella L. is one of the most important pests of stored products and beehives. The most common method to control this pest in many countries is use of chemical compounds. However, these compounds have disadvantages such as pesticide residues in wax, the development of resistance in pest and irreversible effects on the environment and humans. The use of insect pathogenic fungi due to its low risk on mammals and natural enemies can be a good alternative to conventional chemical pesticides. Response surface methodology (RSM is a statistical technique that is employed to optimize processes that are affected by several variables. This technique uses regression analysis to obtain optimal equations to estimate the values. Using this method, while maintaining the quality in the experiments, the number of those could be reduced. Therefore, this study was aimed to evaluate response surface methodology to determine the effect of optimum lethal level of concentration of B. bassiana conidia, temperature as well as humidity variables on the mortality of fifth instar larvae of greater wax moth. Materials and Methods: Wax moth-eating insects were raised in plastic containers containing artificial food and old black wax at 30 ± 1 ° C and a relative humidity of 85 ± 1 % and photoperiod of 14:10 h (L: D. Isolation of insect pathogenic fungus B. bassiana was done by using Galleria Bait Method (GBM. For this purpose, after preparation of the fungus suspension from the infected larvae, 1 ml volume of the suspension was transferred to the water-agar 1.2% and then sealed petri dishes incubated at 30 ° C for three days. After identifying the single colony and formation of pure isolates, microscopic slides were prepared and eventually recovered isolates were recognized as B. bassiana. The

Modelo climático para previsão do míldio da cebola

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Leandro Luiz Marcuzzo

2016-03-01

Full Text Available RESUMO A partir de dados publicados na literatura, elaborou-se um modelo de previsão do míldio da cebola, causado por Peronospora destructor. O modelo desenvolvido baseia-se nas condições climáticas favoráveis para o processo infeccioso, baseado na temperatura e na duração do molhamento foliar. Obteve-se a interação que representa os dois fatores sobre a probabilidade de infecção. Com auxílio da planilha eletrônica Excell, calculou-se a duração do período de molhamento foliar para obter diferentes probabilidades de infecção do patógenos em diferentes temperaturas. Com os dados foi elaborada uma tabela que poderá ser usada na programação computacional que registra a duração do período de molhamento foliar e a temperatura média e automaticamente o valor diário de probabilidade de ocorrência de infecção. O modelo será validado em experimentos conduzidos no campo e com infecção natural, em Rio do Sul, SC.

First report of Peronospora sp. causing downy mildew disease on Geum sp. in the northeastern United States

Science.gov (United States)

Plants in the genus Geum are clump-forming perennials in the family Rosaceae. Known for long bloom times and the ability to attract pollinators, Geum hybrids are popular ornamental plants commonly grown in borders and rock gardens (Brickell & Cathey, 2004). In early April 2017, the entire crop of Ge...

Metodologia para produção de Metarhizium anisopliae (METSCH. sorokin em cultivo submerso: esporulação da biomassa, efeito da concentração de açúcar e custo do inoculant Methodology for production of Metarhizium anisopliae (METSCH. sorokin in submerged cultivation: biomass sporulation, sugar concentration effect and inoculant cost

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Sonia Regina de Mello Pereira

1999-09-01

Full Text Available Desenvolveram-se um método de cultivo e um meio de cultura para produção massal do fungo Metarhizium anisopliae (Metsch. Sorokin, 1883, com maior pureza e concentração de conídios. Este método envolveu o cultivo submerso da linhagem M-61 do entomopatógeno em meio líquido de arroz parboilizado, extrato de levedura, extrato do percevejo da soja (Nezara viridula (L., 1758 Hemiptera, Pentatomidae, sob seis diferentes níveis de concentração de açúcar (0, 2, 4, 6, 8, 10g l-1, além do meio convencional sólido de arroz em grão. As biomassas obtidas foram separadas através de tela de nylon (63 mesh e dispostas em estufa para a esporulação. Os efeitos dos tratamentos foram avaliados pelos parâmetros pesos fresco e seco do micélio, número de conídios por grama de substrato, viabilidade e patogenicidade dos conídios sobre o percevejo. Observou-se que 2.0g l-1 de açúcar em meio de cultura de extrato de N. viridula produziu o dobro do número de conídios por grama de substrato em relação à concentração de 10.0g l-1, a um custo 51 vezes inferior ao obtido no processo convencional de produção do fungo. A viabilidade não foi afetada nos diferentes meios utilizados. Não ocorreram diferenças significativas na patogenicidade em função dos meios de cultura e métodos de cultivo.A method of cultivation and a culture medium were developed aiming at the mass production of fungus Metarhizium anisopliae (Metsch. Sorokin, 1883, with great concentration and purity of conidia. This method involved the M-61 strain of entomopathogenic fungus in liquid medium of rice, yeast extract, soybean bug extract (Nezara viridula (L., 1758 Hemiptera, Pentatomidae, under six differents concentrations of sugar (0, 2, 4, 6, 8, 10g l-1, and the solid conventional medium of rice grains. The biomasses obtained were filtered and put in an incubator to promote sporulation. The treatments were evaluated through the parameters wet and dry-weight of micelium

Controle de nematóides parasitos gastrintestinais por Arthrobotrys musiformis em ovinos naturalmente infestados mantidos em pastagens Control of gastrintestinal parasitic nematodes by Arthrobotrys musiformis in naturally infested sheep maintained on pastures

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Érika Barbosa Neves Graminha

2005-09-01

Full Text Available Este trabalho objetivou avaliar, in vitro e in vivo, o fungo predador Arthrobotrys musiformis, administrado via oral na forma de conídios microencapsulados em alginato ou in natura em meio líquido, após passagem pelo trato gastrintestinal de ovinos. No teste in vitro, avaliou-se a manutenção da atividade predatória, e no teste in vivo, a capacidade de controlar nematóides parasitos de ovinos naturalmente infectados, mantidos em pastagens. O resultado do teste in vitro confirmou a viabilidade de A. musiformis (95,5%, mesmo após a passagem pelo trato gastrintestinal. Na avaliação in vivo, não houve diferença estatística entre o número de ovos por grama de fezes dos grupos tratados e controle, provavelmente em razão da baixa lotação dos piquetes, da quantidade e periodicidade insuficientes de oferecimento de conídios aos ovinos, o que teria proporcionado dispersão e migração das larvas na pastagem. Contudo, os helmintos recuperados na necropsia dos animais traçadores mostraram que A. musiformis reduziu em 50,9% e 57,6% o número de Trichostrongylus colubriformis dos grupos tratados com conídios microencapsulados e in natura, respectivamente, e em 95% o número de L4 de Haemonchus do grupo tratado com conídios in natura. Embora os resultados não sejam conclusivos, A. musiformis mostrou-se promissor agente no biocontrole de nematóides parasitos gastrintestinais.The objective of the present study was to assess in vitro and in vivo the predatory fungus Arthrobotrys musiformis administered orally, in the form of alginate-microencapsulated conidia, or in natura in liquid medium after passage through the gastrointestinal tract of sheep. Maintenance of the predatory activity of the fungus was evaluated in the in vitro test, and its ability to control parasitic nematodes in naturally infested sheep maintained on pastures was analyzed in the in vivo test. The result of the in vitro test confirmed the viability of A. musiformis (95

Mechanisms and Characterization of Trichoderma longibrachiatum T6 in Suppressing Nematodes (Heterodera avenae in Wheat

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Shuwu Zhang

2017-09-01

Full Text Available Heterodera avenae is an important soil-borne pathogen that affects field crops worldwide. Chemical nematicides can be used to control the nematode, but they bring toxicity to the environment and human. Trichoderma longibrachiatum has been shown to have the ability to control H. avenae cysts, but detailed microscopic observations and bioassays are lacking. In this study, we used microscopic observations and bioassays to study the effect of T. longibrachiatum T6 (TL6 on the eggs and second stage juveniles (J2s of H. avenae, and investigate the role of TL6 in inducing the resistance to H. avenae in wheat seedling at physiological and biochemical levels. Microscopic observations recorded that TL6 parasitized on the H. avenae eggs, germinated, and produced a large number of hyphae on the eggs surface at the initial stage, thereafter, the eggs were completely surrounded by dense mycelia and the contents of eggs were lysed at the late stage. Meanwhile, the conidia suspension of TL6 parasitized on the surface of J2s, produced a large number of hyphae that penetrated the cuticle and caused deformation of the nematodes. TL6 at the concentration of 1.5 × 107 conidia ml−1 had the highest rates of parasitism on eggs and J2s, reflected by the highest hatching-inhibition of eggs and the mortality of J2s. In the greenhouse experiments, wheat seedlings treated with TL6 at 1.5 × 107 conidia ml−1 had reduced H. avenae infection, and increased plant growth significantly compared to the control. The cysts and juveniles in soil were reduced by 89.8 and 92.7%, the juveniles and females in roots were reduced by 88.3 and 91.3%, whereas the activity of chitinase and β-1, 3-glucanase, total flavonoids and lignin contents in wheat roots were increased significantly at different stage after inoculation with the eggs and TL6 conidia in comparison to the control. Maximum activity of chitinase and β-1, 3-glucanase were recorded at the 20th and 15th Days after

Unintended consequences of invasive predator control in an Australian forest: overabundant wallabies and vegetation change.

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Nick Dexter

Full Text Available Over-abundance of native herbivores is a problem in many forests worldwide. The abundance of native macropod wallabies is extremely high at Booderee National Park (BNP in south-eastern Australia. This has occurred because of the reduction of exotic predators through an intensive baiting program, coupled with the absence of other predators. The high density of wallabies at BNP may be inhibiting the recruitment of many plant species following fire-induced recruitment events. We experimentally examined the post-fire response of a range of plant species to browsing by wallabies in a forest heavily infested with the invasive species, bitou bush Chrysanthemoides monilifera. We recorded the abundance and size of a range of plant species in 18 unfenced (browsed and 16 fenced (unbrowsed plots. We found the abundance and size of bitou bush was suppressed in browsed plots compared to unbrowsed plots. Regenerating seedlings of the canopy or middle storey tree species Eucalyptus pilularis, Acacia implexa, Allocasuarina littoralis, Breynia oblongifolia and Banksia integrifolia were either smaller or fewer in number in grazed plots than treatment plots as were the vines Kennedia rubicunda, Glycine tabacina and Glycine clandestina. In contrast, the understorey fern, Pteridium esculentum increased in abundance in the browsed plots relative to unbrowsed plots probably because of reduced competition with more palatable angiosperms. Twelve months after plots were installed the community structure of the browsed and unbrowsed plots was significantly different (P = 0.023, Global R = 0.091. The relative abundance of C. monilifera and P. esculentum contributed most to the differences. We discuss the possible development of a low diversity bracken fern parkland in Booderee National Park through a trophic cascade, similar to that caused by overabundant deer in the northern hemisphere. We also discuss its implications for broad scale fox control in southern

Laboratory and field evaluation of Metarhizium anisopliae var. anisopliae for controlling subterranean termites

OpenAIRE

Hussain, A; Ahmed, S; Shahid, M

2011-01-01

The efficacy of the Metarhizium anisopliae strain ARSEF 6911 was determined in the laboratory and field against two sugarcane pests, Microtermes obesi Holmgren and Odontotermes obesus Rambur (Termitidae: Isoptera). The susceptibility of both termite species to different conidial suspensions (1 × 10(10), 1 × 10(8), 1 × 10(6) and 1 × 10(4) conidia/ml) was determined in laboratory. All conidial suspensions were able to induce mortality. Termite mortality caused by the fungal suspensions was dose...

Antifungal Metabolites (Monorden, Monocillin IV, and Cerebrosides) from Humicola fuscoatra Traaen NRRL 22980, a Mycoparasite of Aspergillus flavus Sclerotia

OpenAIRE

Wicklow, Donald T.; Joshi, Biren K.; Gamble, William R.; Gloer, James B.; Dowd, Patrick F.

1998-01-01

The mycoparasite Humicola fuscoatra NRRL 22980 was isolated from a sclerotium of Aspergillus flavus that had been buried in a cornfield near Tifton, Ga. When grown on autoclaved rice, this fungus produced the antifungal metabolites monorden, monocillin IV, and a new monorden analog. Each metabolite produced a clear zone of inhibition surrounding paper assay disks on agar plates seeded with conidia of A. flavus. Monorden was twice as inhibitory to A. flavus mycelium extension (MIC > 28 μg/ml) ...

Simple sequence repeat markers useful for sorghum downy mildew (Peronosclerospora sorghi and related species

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Odvody Gary N

2008-11-01

Full Text Available Abstract Background A recent outbreak of sorghum downy mildew in Texas has led to the discovery of both metalaxyl resistance and a new pathotype in the causal organism, Peronosclerospora sorghi. These observations and the difficulty in resolving among phylogenetically related downy mildew pathogens dramatically point out the need for simply scored markers in order to differentiate among isolates and species, and to study the population structure within these obligate oomycetes. Here we present the initial results from the use of a biotin capture method to discover, clone and develop PCR primers that permit the use of simple sequence repeats (microsatellites to detect differences at the DNA level. Results Among the 55 primers pairs designed from clones from pathotype 3 of P. sorghi, 36 flanked microsatellite loci containing simple repeats, including 28 (55% with dinucleotide repeats and 6 (11% with trinucleotide repeats. A total of 22 microsatellites with CA/AC or GT/TG repeats were the most abundant (40% and GA/AG or CT/TC types contribute 15% in our collection. When used to amplify DNA from 19 isolates from P. sorghi, as well as from 5 related species that cause downy mildew on other hosts, the number of different bands detected for each SSR primer pair using a LI-COR- DNA Analyzer ranged from two to eight. Successful cross-amplification for 12 primer pairs studied in detail using DNA from downy mildews that attack maize (P. maydis & P. philippinensis, sugar cane (P. sacchari, pearl millet (Sclerospora graminicola and rose (Peronospora sparsa indicate that the flanking regions are conserved in all these species. A total of 15 SSR amplicons unique to P. philippinensis (one of the potential threats to US maize production were detected, and these have potential for development of diagnostic tests. A total of 260 alleles were obtained using 54 microsatellites primer combinations, with an average of 4.8 polymorphic markers per SSR across 34

Simple sequence repeat markers useful for sorghum downy mildew (Peronosclerospora sorghi) and related species.

Science.gov (United States)

Perumal, Ramasamy; Nimmakayala, Padmavathi; Erattaimuthu, Saradha R; No, Eun-Gyu; Reddy, Umesh K; Prom, Louis K; Odvody, Gary N; Luster, Douglas G; Magill, Clint W

2008-11-29

A recent outbreak of sorghum downy mildew in Texas has led to the discovery of both metalaxyl resistance and a new pathotype in the causal organism, Peronosclerospora sorghi. These observations and the difficulty in resolving among phylogenetically related downy mildew pathogens dramatically point out the need for simply scored markers in order to differentiate among isolates and species, and to study the population structure within these obligate oomycetes. Here we present the initial results from the use of a biotin capture method to discover, clone and develop PCR primers that permit the use of simple sequence repeats (microsatellites) to detect differences at the DNA level. Among the 55 primers pairs designed from clones from pathotype 3 of P. sorghi, 36 flanked microsatellite loci containing simple repeats, including 28 (55%) with dinucleotide repeats and 6 (11%) with trinucleotide repeats. A total of 22 microsatellites with CA/AC or GT/TG repeats were the most abundant (40%) and GA/AG or CT/TC types contribute 15% in our collection. When used to amplify DNA from 19 isolates from P. sorghi, as well as from 5 related species that cause downy mildew on other hosts, the number of different bands detected for each SSR primer pair using a LI-COR- DNA Analyzer ranged from two to eight. Successful cross-amplification for 12 primer pairs studied in detail using DNA from downy mildews that attack maize (P. maydis & P. philippinensis), sugar cane (P. sacchari), pearl millet (Sclerospora graminicola) and rose (Peronospora sparsa) indicate that the flanking regions are conserved in all these species. A total of 15 SSR amplicons unique to P. philippinensis (one of the potential threats to US maize production) were detected, and these have potential for development of diagnostic tests. A total of 260 alleles were obtained using 54 microsatellites primer combinations, with an average of 4.8 polymorphic markers per SSR across 34 Peronosclerospora, Peronospora and Sclerospora

Influence of entomopathogenic fungus, Metarhizium anisopliae, alone and in combination with diatomaceous earth and thiamethoxam on mortality, progeny production, mycosis, and sporulation of the stored grain insect pests.

Science.gov (United States)

Ashraf, Misbah; Farooq, Muhammad; Shakeel, Muhammad; Din, Naima; Hussain, Shahbaz; Saeed, Nadia; Shakeel, Qaiser; Rajput, Nasir Ahmed

2017-12-01

The stored grain insects cause great damage to grains under storage conditions. Synthetic insecticides and fumigants are considered as key measures to control these stored grain insect pests. However, the major issue with these chemicals is grain contamination with chemical residues and development of resistance by insect pests to these chemicals. Biological control is considered as a potential alternative to chemical control especially with the use of pathogens, alone or in combination with selective insecticides. The present study was conducted to evaluate the synergism of Metarhizium anisopliae with diatomaceous earth (DE) and thiamethoxam against four insect pests on the stored wheat grains. In the first bioassay, the M. anisopliae was applied at 1.4 × 10 4 and 1.4 × 10 6 conidia/ml alone and in integration with two concentrations (250 and 500 ppm) of tested DE. The tested fungus when combined with DE and thiamethoxam possessed synergistic impact as compared to their individual efficacy. Adult mortality increased with respect to increased exposure interval and doses. In the second bioassay, M. anisopliae was applied at 1.4 × 10 4 conidia/ml individually and in combination with three concentrations (0.50, 0.75, and 1.00 ppm) of thiamethoxam. Results concluded that M. anisopliae integrated with DE and thiamethoxam provides more effective control of stored grain insect pests.

Asp f6, an Aspergillus allergen specifically recognized by IgE from patients with allergic bronchopulmonary aspergillosis, is differentially expressed during germination.

Science.gov (United States)

Schwienbacher, M; Israel, L; Heesemann, J; Ebel, F

2005-11-01

Aspergillus fumigatus is a pathogenic mould causing allergic and invasive respiratory diseases. Allergic bronchopulmonary Aspergillosis (ABPA) is a severe pulmonary complication resulting from hypersensitivity to A. fumigatus proteins. Aspergillus allergen Asp f6 is recognized by IgE from ABPA patients, but not from sensitized individuals, a fact that can be used to differentiate between these two groups of allergic patients. Proteins from hyphae, resting and germinating conidia of A. fumigatus were compared by SDS-PAGE. Protein identification was performed using MALDI-TOF mass spectrometry. Recombinant A. fumigatus allergens were used to isolate specific monoclonal antibodies (mab) from a hybridoma bank generated against Aspergillus proteins. A hyphae-specific 23 kDa A. fumigatus protein was identified as the allergen Asp f6/manganese-dependent superoxide dismutase (MnSOD). Differential expression of MnSOD was confirmed by immunoblot using a specific mab. In contrast, Asp f8 another intracellular, but not ABPA-specific allergen, was detected in hyphae and conidia. Aspergillus fumigatus is able to colonize its environment by the formation of hyphae. Hyphae are found in the lung of ABPA patients, but not in patients suffering from atopic asthma. Our finding that Asp f6 is specifically expressed in hyphae might explain why an IgE response to Asp f6 is specific for ABPA patients.

Zombie soldier beetles: Epizootics in the goldenrod soldier beetle, Chauliognathus pensylvanicus (Coleoptera: Cantharidae) caused by Eryniopsis lampyridarum (Entomophthoromycotina: Entomophthoraceae).

Science.gov (United States)

Steinkraus, Donald C; Hajek, Ann E; Liebherr, Jim K

2017-09-01

Adult goldenrod soldier beetles, Chauliognathus pensylvanicus, were found infected by the fungus Eryniopsis lampyridarum (Entomophthoromycotina) in Arkansas during September - October (1996, 2001, 2015 and 2016). Living and dead infected beetles were found on flowering frost aster, Symphyotrichum pilosum, common boneset, Eupatorium perfoliatum, and Canada goldenrod, Solidago canadensis. Live and dead beetles (n=446) were collected in 1996 from S. pilosum flowers and held individually in the laboratory for determination of fungal prevalence. Of the beetles collected, 281 (63%) were males and 165 (37%) were females. A total of 90 beetles were infected with E. lampyridarum, an overall prevalence of 20.2%. Prevalence in males was 19.6% (n=55 infected/281 males total) and prevalence in females was 21.2% (n=35 infected /165 females total). Conidia were produced from 57% of the infected beetles, 23% of the infected beetles produced resting spores, and 20% contained the hyphal body stage. Infected beetles produced either conidia or resting spores but never both in the same host. Post-mortem morphological changes in the hosts due to E. lampyridarum were observed periodically for 24h. Shortly before death, by unknown mechanisms, dying infected beetles tightly clamped their mandibles into flower heads and ca. 15-22h later (between 2400 and 0700h) the fungus caused dead beetles to raise their elytra and expand their metathoracic wings. Copyright © 2017. Published by Elsevier Inc.

The Aspergillus flavus Homeobox Gene, hbx1, Is Required for Development and Aflatoxin Production

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Jeffrey W. Cary

2017-10-01

Full Text Available Homeobox proteins, a class of well conserved transcription factors, regulate the expression of targeted genes, especially those involved in development. In filamentous fungi, homeobox genes are required for normal conidiogenesis and fruiting body formation. In the present study, we identified eight homeobox (hbx genes in the aflatoxin-producing ascomycete, Aspergillus flavus, and determined their respective role in growth, conidiation and sclerotial production. Disruption of seven of the eight genes had little to no effect on fungal growth and development. However, disruption of the homeobox gene AFLA_069100, designated as hbx1, in two morphologically different A. flavus strains, CA14 and AF70, resulted in complete loss of production of conidia and sclerotia as well as aflatoxins B1 and B2, cyclopiazonic acid and aflatrem. Microscopic examination showed that the Δhbx1 mutants did not produce conidiophores. The inability of Δhbx1 mutants to produce conidia was related to downregulation of brlA (bristle and abaA (abacus, regulatory genes for conidiophore development. These mutants also had significant downregulation of the aflatoxin pathway biosynthetic genes aflC, aflD, aflM and the cluster-specific regulatory gene, aflR. Our results demonstrate that hbx1 not only plays a significant role in controlling A. flavus development but is also critical for the production of secondary metabolites, such as aflatoxins.

Effects of Two Salts Compounds on Mycelial Growth, Sporulation, and Spore Germination of Six Isolates of Botrytis cinerea in the Western North of Algeria

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Boualem Boumaaza

2015-01-01

Full Text Available Six isolates of Botrytis cinerea were isolated from leaves and stems of different tomato varieties taken from four areas in the northwest of Algeria where tomato is mostly grown in greenhouses and high tunnels. The purpose of this research was to determine the effect of two salts, NaCl and CaCl2, on three stages of Botrytis cinerea’s life cycle. All isolates tested were stimulated in 50 to 150 ppm; NaCl was the most effective treatment to increase mycelial growth at two tested concentrations. However, at 300 ppm concentration, CaCl2 completely inhibited the growth of mycelium; they reach 34.78% for the isolate TR46 and 26.72% for isolate F27. The sodium and calcium salts stimulated conidia production in liquid culture. We noticed that the effect of calcium chloride on sporulation was average while sodium chloride. In the medium containing 50 ppm, calcium chloride and sodium chloride increased the germination capacity of most isolates compared with the control. Other calcium salts, at 100 or 300 ppm, decreased the germination percentage of the conidia. With the exception of sodium salts, the inhibitions of germination reduce at 150 or 300 compared with the control. Conidial germination was slightly inhibited by sodium chloride only when the concentration was over 300 ppm.

Development of races of Passalora sojina on different substrates and light regimes

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Juliane Nicolodi Camera

2014-09-01

Full Text Available The leaf “frog-eye” spot is a disease that often occurs in Argentina and Brazil is becoming important in soy producing regions. The difficulty in achieving inoculum for studies with P. sojina led to the development of this work was to determine the sporulation of races P. sojina in different culture media and light regimes and check difference in sporulation of races of this fungus. The plates with the culture media BDA, STT , AI , V8 and FAA containing the three races of P. sojina were subjected to a photoperiod of 12/12 h (light and dark and continuous dark. The experimental design was a factorial 5x2x3 (culture medium, light regimes, races of P. sojina with four replications. The greatest number of conidia cm- 2 of the fungus was found for race 23 in the middle of STT culture and races 24 e 25 in the middle of V8 culture. The fungus race 25, with the largest number of conidia cm-2, regardless of the medium and the light regimen. With respect to light regime for all races of the fungus, the largest esporulation were checked at a photoperiod of 12/12 h. When the fungus was grown in culture medium AI, this had the largest area of the colony, regardless of breed fungus and light regime

A prospective survey of air and surface fungal contamination in a medical mycology laboratory at a tertiary care university hospital.

Science.gov (United States)

Sautour, Marc; Dalle, Frédéric; Olivieri, Claire; L'ollivier, Coralie; Enderlin, Emilie; Salome, Elsa; Chovelon, Isabelle; Vagner, Odile; Sixt, Nathalie; Fricker-Pap, Véronique; Aho, Serge; Fontaneau, Olivier; Cachia, Claire; Bonnin, Alain

2009-04-01

Invasive filamentous fungi infections resulting from inhalation of mold conidia pose a major threat in immunocompromised patients. The diagnosis is based on direct smears, cultural symptoms, and culturing fungi. Airborne conidia present in the laboratory environment may cause contamination of cultures, resulting in false-positive diagnosis. Baseline values of fungal contamination in a clinical mycology laboratory have not been determined to date. A 1-year prospective survey of air and surface contamination was conducted in a clinical mycology laboratory during a period when large construction projects were being conducted in the hospital. Air was sampled with a portable air system impactor, and surfaces were sampled with contact Sabouraud agar plates. The collected data allowed the elaboration of Shewhart graphic charts. Mean fungal loads ranged from 2.27 to 4.36 colony forming units (cfu)/m(3) in air and from 0.61 to 1.69 cfu/plate on surfaces. Strict control procedures may limit the level of fungal contamination in a clinical mycology laboratory even in the context of large construction projects at the hospital site. Our data and the resulting Shewhart graphic charts provide baseline values to use when monitoring for inappropriate variations of the fungal contamination in a mycology laboratory as part of a quality assurance program. This is critical to the appropriate management of the fungal risk in hematology, cancer and transplantation patients.

Interactions involving ozone, Botrytis cinerea, and B. squamosa on onion leaves

Energy Technology Data Exchange (ETDEWEB)

Rist, D.L.

1983-01-01

Interactions involving Botrytis cinerea Pers., B. squamosa Walker, and ozone on onion (alium cepae L.) were investigated. Initially, threshold dosages of ozone required to predispose onion leaves to greater infection by B. cinerea and B. squamosa were determined under controlled conditions in an ozone-exposure chamber. Subsequent experiments supported the hypothesis that nutrients leaking out of ozone-injured cells stimulated lesion production by B. cinerea. The electrical conductivity of, and carbohydrate concentration in, dew collected from leaves of onion plants which had been exposed to ozone were greater than the electrical conductivity of, and carbohydrate concentration in, dew collected from leaves of other, non-exposed onion plants. When conidia of B. cinerea were suspended in dew collected from leaves of plants which had been exposed to ozone and the resulting suspension atomized onto leaves of non-exposed plants, more lesions were induced than on leaves of other non-exposed plants inoculated with conidia suspended in dew collected from plants which had not been exposed to ozone. EDU protected onion leaves from ozone-induced predisposition to these fungi under controlled conditions. Experiments designed to detect interaction between B. cinerea and B. squamosa in onion leaf blighting indicated that such interaction did not occur. Leaves were blighted rapidly when inoculated with B. squamosa whether B. cinerea was present or absent.

Invasion of Solanum tuberosum L. by Aspergillus terreus: a microscopic and proteomics insight on pathogenicity.

Science.gov (United States)

Louis, Bengyella; Waikhom, Sayanika Devi; Roy, Pranab; Bhardwaj, Pardeep Kumar; Singh, Mohendro Wakambam; Chandradev, Sharma K; Talukdar, Narayan Chandra

2014-06-10

Aspergillus terreus is one of the most harmful filamentous fungal pathogen of humans, animals and plants. Recently, researchers have discovered that A. terreus can cause foliar blight disease in potato (Solanum tuberosum L.). We used light and scanning electron microscopy, and performed proteomics analysis in an attempt to dissect the invasion process of A. terreus in this important crop. Microscopic study revealed that invasion of leaf tissue is marked by rapid germination of A. terreus phialidic conidia (PC) by 4 h after inoculation. By 8 h after inoculation, primary germ tubes from PC differentiated into irregular protuberance, often displayed stomata atropism, and failed to penetrate via the epidermal cells. Colonization of leaf tissues was associated with high rate of production of accessory conidia (AC). These analyses showed the occurrence of a unique opposing pattern of AC, tissue-specific and produced on melanized colonizing hyphae during the infection of leaf tissue. A significant proteome change hallmarked by differential expression of class I patatin, lipoxygenase, catalase-peroxidase complex, and cysteine proteinase inhibitor were observed during tuber colonization. These proteins are often involved in signal transduction pathways and crosstalk in pathogenic responses. A. terreus abundantly produced AC and multipolar germinating PC to invade potato leaf tissue. Additionally, A. terreus differentially induced enzymes in potato tuber during colonization which facilitates rapid disease development.

Molecular Insights Into Development and Virulence Determinants of Aspergilli: A Proteomic Perspective

Science.gov (United States)

Shankar, Jata; Tiwari, Shraddha; Shishodia, Sonia K.; Gangwar, Manali; Hoda, Shanu; Thakur, Raman; Vijayaraghavan, Pooja

2018-01-01

Aspergillus species are the major cause of health concern worldwide in immunocompromised individuals. Opportunistic Aspergilli cause invasive to allergic aspergillosis, whereas non-infectious Aspergilli have contributed to understand the biology of eukaryotic organisms and serve as a model organism. Morphotypes of Aspergilli such as conidia or mycelia/hyphae helped them to survive in favorable or unfavorable environmental conditions. These morphotypes contribute to virulence, pathogenicity and invasion into hosts by excreting proteins, enzymes or toxins. Morphological transition of Aspergillus species has been a critical step to infect host or to colonize on food products. Thus, we reviewed proteins from Aspergilli to understand the biological processes, biochemical, and cellular pathways that are involved in transition and morphogenesis. We majorly analyzed proteomic studies on A. fumigatus, A. flavus, A. terreus, and A. niger to gain insight into mechanisms involved in the transition from conidia to mycelia along with the role of secondary metabolites. Proteome analysis of morphotypes of Aspergilli provided information on key biological pathways required to exit conidial dormancy, consortia of virulent factors and mycotoxins during the transition. The application of proteomic approaches has uncovered the biological processes during development as well as intermediates of secondary metabolite biosynthesis pathway. We listed key proteins/ enzymes or toxins at different morphological types of Aspergillus that could be applicable in discovery of novel therapeutic targets or metabolite based diagnostic markers. PMID:29896454

Neurospora ribosomal DNA sequences are indistinguishable within cell types but distinguishable among heterothallic species

International Nuclear Information System (INIS)

Chambers, C.; Dutta, S.K.

1983-01-01

High molecular nuclear DNAs were isolated from three developmental cell types of N. crassa: conidia, mycelia and germinated conidia, and from mycelial cells of two other heterothallic species, N. intermedia and N. sitophila. These nuclear DNAs were treated with several restriction enzymes: EcoR1, Bam H1, Hind III, Hinc II, Bgl II, Sma I and Pst 1. All seven restriction enzymes were tested on 0.7% agarose gels. EcoR1, Hind III, Pst 1, and Hinc II showed band differences among the species, but not among the cell types. Southern blot transfers of restricted DNA gels were then hybridized with 32 P-labelled pMF2 rDNAs (probe). This later DNA was prepared from N. crassa rDNA cloned into pBR322 plasmid, obtained from Dr. Robert Metzenberg of the University of Wisconsin. Autoradiograms of these hybrids between southern blots and probe DNA revealed similar rDNA band patterns confirming the observations on restriction gels. In the case of EcoR1 restriction analysis there were differences in fragments on 0.7% agarose gel, but after hybridization of southern blots no differences in band patterns were seen in autoradiograms. This raises the question whether the background bands were all of rDNA sequences. These studies are being continued using ITS (internal transcribed spacer) sequences of N. crassa rDNAs cloned in pBR322 plasmid

Scanning electron microscopy and histopathological observations of Beauveria bassiana infection of Colorado potato beetle larvae.

Science.gov (United States)

Duan, Yulin; Wu, Hui; Ma, Zhiyan; Yang, Liu; Ma, Deying

2017-10-01

Beauveria bassiana is a potential candidate for use as an environmentally friendly bio-pesticide. We studied the infection process and histopathology of B. bassiana strain NDBJJ-BFG infection of the Colorado potato beetle (Leptinotarsa decemlineata) using scanning electron microscopy and hematoxylin-eosin staining of tissue sections. The results show that the fungus penetrated the insect epidermis through germ tubes and appressoria after spraying the larvae with conidial suspensions. The conidia began to germinate after 24 h and invade the epidermis. After 48 h, the conidia invaded the larvae with germ tubes and began to enter the haemocoel. By 72 h, hyphae had covered the host surface and had colonized the body cavity. The dermal layer was dissolved, muscle tissues were ruptured and adipose tissue was removed. The mycelium had damaged the intestinal wall muscles, and invaded into intestinal wall and midfield cells resulting in cell separation and tracheal deformation. After 96 h of inoculation, the internal structure of the larvae was destroyed. The research shows that B. bassiana NDBJJ-BFG surface inoculation resulted in a series of histopathological changes to the potato beetle larvae that proved lethal within 72 h. This indicated that this fungus has a high pathogenicity to Colorado potato beetle larvae. Copyright © 2017. Published by Elsevier Ltd.

Alkane-grown Beauveria bassiana produce mycelial pellets displaying peroxisome proliferation, oxidative stress, and cell surface alterations.

Science.gov (United States)

Huarte-Bonnet, Carla; Paixão, Flávia R S; Ponce, Juan C; Santana, Marianela; Prieto, Eduardo D; Pedrini, Nicolás

2018-06-01

The entomopathogenic fungus Beauveria bassiana is able to grow on insect cuticle hydrocarbons, inducing alkane assimilation pathways and concomitantly increasing virulence against insect hosts. In this study, we describe some physiological and molecular processes implicated in growth, nutritional stress response, and cellular alterations found in alkane-grown fungi. The fungal cytology was investigated using light and transmission electron microscopy while the surface topography was examined using atomic force microscopy. Additionally, the expression pattern of several genes associated with oxidative stress, peroxisome biogenesis, and hydrophobicity were analysed by qPCR. We found a novel type of growth in alkane-cultured B. bassiana similar to mycelial pellets described in other alkane-free fungi, which were able to produce viable conidia and to be pathogenic against larvae of the beetles Tenebrio molitor and Tribolium castaneum. Mycelial pellets were formed by hyphae cumulates with high peroxidase activity, exhibiting peroxisome proliferation and an apparent surface thickening. Alkane-grown conidia appeared to be more hydrophobic and cell surfaces displayed different topography than glucose-grown cells. We also found a significant induction in several genes encoding for peroxins, catalases, superoxide dismutases, and hydrophobins. These results show that both morphological and metabolic changes are triggered in mycelial pellets derived from alkane-grown B. bassiana. Copyright © 2017 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Molecular Insights Into Development and Virulence Determinants of Aspergilli: A Proteomic Perspective

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Jata Shankar

2018-05-01

Full Text Available Aspergillus species are the major cause of health concern worldwide in immunocompromised individuals. Opportunistic Aspergilli cause invasive to allergic aspergillosis, whereas non-infectious Aspergilli have contributed to understand the biology of eukaryotic organisms and serve as a model organism. Morphotypes of Aspergilli such as conidia or mycelia/hyphae helped them to survive in favorable or unfavorable environmental conditions. These morphotypes contribute to virulence, pathogenicity and invasion into hosts by excreting proteins, enzymes or toxins. Morphological transition of Aspergillus species has been a critical step to infect host or to colonize on food products. Thus, we reviewed proteins from Aspergilli to understand the biological processes, biochemical, and cellular pathways that are involved in transition and morphogenesis. We majorly analyzed proteomic studies on A. fumigatus, A. flavus, A. terreus, and A. niger to gain insight into mechanisms involved in the transition from conidia to mycelia along with the role of secondary metabolites. Proteome analysis of morphotypes of Aspergilli provided information on key biological pathways required to exit conidial dormancy, consortia of virulent factors and mycotoxins during the transition. The application of proteomic approaches has uncovered the biological processes during development as well as intermediates of secondary metabolite biosynthesis pathway. We listed key proteins/ enzymes or toxins at different morphological types of Aspergillus that could be applicable in discovery of novel therapeutic targets or metabolite based diagnostic markers.

Molecular Insights Into Development and Virulence Determinants of Aspergilli: A Proteomic Perspective.

Science.gov (United States)

Shankar, Jata; Tiwari, Shraddha; Shishodia, Sonia K; Gangwar, Manali; Hoda, Shanu; Thakur, Raman; Vijayaraghavan, Pooja

2018-01-01

Aspergillus species are the major cause of health concern worldwide in immunocompromised individuals. Opportunistic Aspergilli cause invasive to allergic aspergillosis, whereas non-infectious Aspergilli have contributed to understand the biology of eukaryotic organisms and serve as a model organism. Morphotypes of Aspergilli such as conidia or mycelia/hyphae helped them to survive in favorable or unfavorable environmental conditions. These morphotypes contribute to virulence, pathogenicity and invasion into hosts by excreting proteins, enzymes or toxins. Morphological transition of Aspergillus species has been a critical step to infect host or to colonize on food products. Thus, we reviewed proteins from Aspergilli to understand the biological processes, biochemical, and cellular pathways that are involved in transition and morphogenesis. We majorly analyzed proteomic studies on A. fumigatus, A. flavus, A. terreus , and A. niger to gain insight into mechanisms involved in the transition from conidia to mycelia along with the role of secondary metabolites. Proteome analysis of morphotypes of Aspergilli provided information on key biological pathways required to exit conidial dormancy, consortia of virulent factors and mycotoxins during the transition. The application of proteomic approaches has uncovered the biological processes during development as well as intermediates of secondary metabolite biosynthesis pathway. We listed key proteins/ enzymes or toxins at different morphological types of Aspergillus that could be applicable in discovery of novel therapeutic targets or metabolite based diagnostic markers.

European apple canker: morphophysiological variability and pathogenicity in isolates of Neonectria ditissima in southern Brazil

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Jonatas da Silva Campos

Full Text Available ABSTRACT: European apple canker (EC is caused by Neonectria ditissima, a pathogen officially registered as a quarantine pest in 2012. Thirty-five isolates of N. ditissima of different geographical regions of southern Brazil from apple branches showing symptoms of EC were identified by the specific pair primers Ch1 and Ch2 and analyzed concerning the virulence on Gala apple cultivar and morphophysiological characteristics. The disease symptoms were characterized and the isolates compared based on average mycelium growth (AMG, mycelium growth index (MGI, colony color, conidia type, dimensions and growth on potato dextrose agar (PDA, malt agar (AM, and synthetic SNAY (SN culture media. Nineteen isolates showed the greatest AMG on PDA, forming three growth groups of 35.56 (GI, 52.71 (GII, and 62.67mm (GIII. Seven isolates showed MGI greater than 4.0mm diameter on PDA compared with that on AM and SN. The highest conidia production was on SN, and the predominant colony color in all media was white to beige with central pigmentation of brown and borders colored in shades of beige. There were significant differences among the average dimensions of micro- and macroconidia on PDA, AM, and SN. The pathogenicity was confirmed for all isolates despite of different morphophysiological characteristics. There was no correlation among isolates morphophysiological variability, virulence, and geographical origin.

Central Role of IL-23 and IL-17 Producing Eosinophils as Immunomodulatory Effector Cells in Acute Pulmonary Aspergillosis and Allergic Asthma.

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Guerra, Evelyn Santos; Lee, Chrono K; Specht, Charles A; Yadav, Bhawna; Huang, Haibin; Akalin, Ali; Huh, Jun R; Mueller, Christian; Levitz, Stuart M

2017-01-01

Aspergillus fumigatus causes invasive pulmonary disease in immunocompromised hosts and allergic asthma in atopic individuals. We studied the contribution of lung eosinophils to these fungal diseases. By in vivo intracellular cytokine staining and confocal microscopy, we observed that eosinophils act as local sources of IL-23 and IL-17. Remarkably, mice lacking eosinophils had a >95% reduction in the percentage of lung IL-23p19+ cells as well as markedly reduced IL-23 heterodimer in lung lavage fluid. Eosinophils killed A. fumigatus conidia in vivo. Eosinopenic mice had higher mortality rates, decreased recruitment of inflammatory monocytes, and decreased expansion of lung macrophages after challenge with conidia. All of these functions underscore a potential protective role for eosinophils in acute aspergillosis. Given the postulated role for IL-17 in asthma pathogenesis, we assessed whether eosinophils could act as sources of IL-23 and IL-17 in models where mice were sensitized to either A. fumigatus antigens or ovalbumin (OVA). We found IL-23p19+ IL-17AF+ eosinophils in both allergic models. Moreover, close to 95% of IL-23p19+ cells and >90% of IL-17AF+ cells were identified as eosinophils. These data establish a new paradigm in acute and allergic aspergillosis whereby eosinophils act not only as effector cells but also as immunomodulatory cells driving the IL-23/IL-17 axis and contributing to inflammatory cell recruitment.

Genetic analysis of conidiation regulatory pathways in koji-mold Aspergillus oryzae.

Science.gov (United States)

Ogawa, Masahiro; Tokuoka, Masafumi; Jin, Feng Jie; Takahashi, Tadashi; Koyama, Yasuji

2010-01-01

Conidia of koji-mold Aspergillus oryzae are often used as starters in the fermented food industry. However, little is known about conidiation regulation in A. oryzae. To improve the productivity of conidia in A. oryzae, it is necessary to understand conidiation regulation in the strain. Therefore, we analyzed the conidiation regulatory system in A. oryzae using 10 kinds of conidiation regulatory gene disruptants. The phenotypes of AorfluG, AorflbA, AorflbB, AorflbC, AorflbD, AorflbE, AorbrlA, AorabaA, AorwetA, and AorfadA mutants are almost identical to those of the corresponding mutants in Aspergillus nidulans. The results indicated that the functions of conidiation regulatory genes are almost conserved between A. oryzae and A. nidulans. However, the severely reduced conidiation phenotype of the AorfluG disruptant in A. oryzae differs from the phenotype of the corresponding mutant in Aspergillus fumigatus in air-exposed culture conditions. These results suggest that A. oryzae, A. nidulans, and A. fumigatus have a G-protein signaling pathway and brlA orthologs in common, and only A. fumigatus has particular brlA activation pathways that are independent of the fluG ortholog. Furthermore, the analyses of AorflbA disruptant and AorfadA dominant-active mutants implicated that AorFadA-mediated G-protein signaling suppresses vegetative growth of A. oryzae.

Large-spored Alternaria pathogens in section Porri disentangled

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Woudenberg, J.H.C.; Truter, M.; Groenewald, J.Z.; Crous, P.W.

2014-01-01

The omnipresent fungal genus Alternaria was recently divided into 24 sections based on molecular and morphological data. Alternaria sect. Porri is the largest section, containing almost all Alternaria species with medium to large conidia and long beaks, some of which are important plant pathogens (e.g. Alternaria porri, A. solani and A. tomatophila). We constructed a multi-gene phylogeny on parts of the ITS, GAPDH, RPB2, TEF1 and Alt a 1 gene regions, which, supplemented with morphological an...

Polyphasic taxonomy of Aspergillus section Aspergillus (formerly Eurotium), and its occurrence in indoor environments and food

DEFF Research Database (Denmark)

Chen, A.J.; Hubka, Vit; Frisvad, Jens Christian

2017-01-01

, epiheveadrides, auroglaucins and anthraquinone bisanthrons, and to be consistent in strains of nearly all species. Other extrolites are species-specific, and thus valuable for identification. Several extrolites show antioxidant effects, which may be nutritionally beneficial in food and beverages. Important...... not distinguish species. All species can be differentiated using CaM or RPB2 sequences. For BenA, Aspergillus brunneus and A. niveoglaucus share identical sequences. Ascospores and conidia morphologyw, growth rates at different temperatures are most useful characters for phenotypic species identification....

Caracterización de la respuesta de tres variedades de rosa a la infección de Peronospora sparsa Berkeley, bajo condiciones de invernadero

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Gómez Sandra

2005-12-01

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Plantas sanas de rosa de las variedades ‘Charlotte’, ‘Classy’ y ‘First Red’ injertadas en los portainjertos ‘Natal Briar’ y ‘Manetti’ y como estacas se inocularon con una suspensión de esporangios de Peronospora sparsa. Posterior a la inoculación, las plantas se mantuvieron por un período de 12 h bajo

Efecto de la temperatura en el periodo de latencia y producción de esporangios de Peronospora sparsa Berkeley en tres variedades de rosa

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Gómez Sandra

2005-12-01

="Subtle Emphasis" />

El alto costo y la limitada eficacia de las medidas de manejo para el mildeo velloso ocasionado por Peronospora sparsa, determinan su importancia en el cultivo de rosas de exportación. Esta investigación tuvo como objetivo ampliar el conocimiento sobre la biología del patógeno y evaluar su relación con la temperatura y la

Influencia de la luz y la temperatura en la germinacion de esporangios y en la esporulacion de peronospora sparsa berkeley, en rosa cultivar charlotte

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Giraldo Buitrago Sandra Liliana

2002-12-01

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Se evaluó la influencia de la luz y la temperatura en la germinación de esporangios en agar agua, y en la esporulación en foliolos de rosa con mildeo velloso Peronospora sparsa