Journal of Microscopy and Ultrastructure 3 (2015) 29–37

Total Page:16

File Type:pdf, Size:1020Kb

Journal of Microscopy and Ultrastructure 3 (2015) 29–37 Journal of Microscopy and Ultrastructure 3 (2015) 29–37 Contents lists available at ScienceDirect Journal of Microscopy and Ultrastructure jo urnal homepage: www.elsevier.com/locate/jmau Original Article Ultrastructures of silver nanoparticles biosynthesized using endophytic fungi ∗ Lamabam Sophiya Devi, S.R. Joshi Microbiology Laboratory, Department of Biotechnology & Bioinformatics, North-Eastern Hill University, Shillong 793 022, Meghalaya, India a r t i c l e i n f o a b s t r a c t Article history: Three endophytic fungi Aspergillus tamarii PFL2, Aspergillus niger PFR6 and Penicllium Received 12 August 2014 ochrochloron PFR8 isolated from an ethno-medicinal plant Potentilla fulgens L. were used for Received in revised form 6 October 2014 the biosynthesis of silver nanoparticles. Scanning and transmission electron microscopic Accepted 14 October 2014 analysis were performed to study the structural morphology of the biosynthesized silver Available online 28 October 2014 nanoparticles. The electron microscopy study revealed the formation of spherical nano- sized silver particles with different sizes. The nanoparticles synthesized using the fungus A. Keywords: tamarii PFL2 was found to have the smallest average particle size (3.5 ± 3 nm) as compared Electron microscopy to the nanoparticles biosynthesized using other two fungi A. niger PFR6 and P. ochrochloron Endophytic fungi PFR8 which produced average particle sizes of 8.7 ± 6 nm and 7.7 ± 4.3 nm, respectively. Silver nanoparticles Crystalline The energy dispersive X-ray spectroscopy (EDS) technique in conjunction with scanning electron microscopy was used for the elemental analysis of the nanoparticles. The selected area diffraction pattern recorded from single particle in the aggregates of nanoparticles revealed that the silver particles are crystalline in nature. © 2014 Saudi Society of Microscopes. Published by Elsevier Ltd. All rights reserved. 1. Introduction surface defects [1–3]. They have tremendous applications in the area of catalysis, opto-electronics, diagnostic bio- Nanoparticles are the clusters of atoms in the size range logical probes, display devices and photo electrochemical of 1–100 nm. In this size range, materials often develop applications due to their unique size-dependent optical, useful attributes that are distinct from the properties of electrical and magnetic properties [4–9]. the bulk material. Metal particles in the nanometre size There has been a rapid increase in microbes that are exhibit unique physical properties that are different from resistant to conventionally used antibiotics [10]. This has both the ion and the bulk material. Their uniqueness arises resulted in an inevitable and urgent need for develop- specifically from higher surface to volume ratio which ment of novel antimicrobial agents. It has been known results in increased catalytic activity due to morphologies since ancient times that silver and its compounds are with highly active facets; hence, the nanosize materi- effective antimicrobial agents [11–13]. Compared with als are more advantageous than their bulk materials. The other metals, silver exhibits higher toxicity to broad spec- enhanced reactivity of nanomaterials can also be attributed trum of microorganisms while it exhibits lower toxicity to their large number of edges, corners, and high-energy to mammalian cells [14]. Silver ion has been known to be effective against a broad range of microorganisms includ- ing antibiotic-resistant strains [15]. Silver nanoparticles ∗ with higher surface to volume ratio compared to common Corresponding author. Tel.: +91 9436102171; fax: +91 3642550076. E-mail address: [email protected] (S.R. Joshi). metallic silver have shown better antimicrobial activity. http://dx.doi.org/10.1016/j.jmau.2014.10.004 2213-879X/© 2014 Saudi Society of Microscopes. Published by Elsevier Ltd. All rights reserved. 30 L.S. Devi, S.R. Joshi / Journal of Microscopy and Ultrastructure 3 (2015) 29–37 Fig. 1. Reproductive structures of the endophytic fungal isolates as seen under compound microscope: (a) PFL2, (b) PFR6 and (c) PFR8. Most importantly silver nanoparticles are also non-toxic effectively reacting with them leading to the inhibition to the mammalian cells at low concentrations [16]. Sil- of enzyme functions [18,19]. The nanoparticles bind to ver nanoparticles have been known for a long time but proteins and DNA and damage them by inhibiting replica- have not been given due attention [17]. Nanoparticles tion. Thus the silver nanoparticles interrupt the respiratory can disturb functions of cell membranes such as per- chain and cell division leading to cell death [20–22]. In meability and respiration. The silver nanoparticles get addition, complex action mechanisms of metals decrease attached to the cell membrane and also penetrate inside the probability of bacteria developing resistance to them the bacteria. Inside the bacterial cells silver nanoparti- [23]. Thus, one of the promising approaches for overcom- cles can disturb the functions of sulfur-containing proteins ing antibiotic resistance of microorganisms is the use of and phosphorus-containing compounds such as DNA by silver nanoparticles. Fig. 2. Phylogenetic relationships between the three endophytic fungi and the ITS sequences of closely related fungal strains retrieved from NCBI GenBank. L.S. Devi, S.R. Joshi / Journal of Microscopy and Ultrastructure 3 (2015) 29–37 31 Fig. 3. UV–vis absorption spectrum at different time intervals for silver nanoparticles biosynthesized using (a) Aspergillus niger PFR6, (b) Penicillium ochrochloron PFR8 and (c) Aspergillus tamarii PFL2. Owing to the fact that the endophytic fungi provide with 70% alcohol for 1 min. The final rinsing was done a broad variety of bioactive secondary metabolites with with sterile distilled water and blot dried on sterile filter unique structures they could be the explored for their abil- paper. The excess water was dried and the surface sterilized ity to biosynthesis of silver nanoparticles to developed explants were then inoculated onto PDA plates. The plates an efficient environment friendly process. Few studies on were periodically observed for fungal growth. The fungus the biosynthesis of silver nanoparticles using endophytic growing out from the plant explants was then sub-cultured fungi like Amylomyces rouxii, Penicillium sp., P. janthinel- in PDA plates. Three morphologically distinct endophytic lum, Pestalotia sp., Aspergillus clavatus, A. concius, Epicoccum fungi isolated from an ethno-medicinal plant P. fulgens L. nigrum and Phomopsis sp. have shown the proficiency of were used for the biosynthesis of silver nanoparticles (Fig. 1 the endophytic fungi to synthesize silver nanoparticles a–c). possessing antimicrobial activity [24–31]. In the present The fungal isolates were characterized based on the study, an attempt has been made to investigate endophytic analysis of the phylogenetic relationship of the iso- fungi associated with an ethnomedicinal plant Potentilla lates using internal transcribed spacer (ITS) region gene fulgens L. for their ability to synthesize silver nanopar- sequences. Amplification of ITS (ITS1, 5.8S and ITS2) region ticles and the ultrastructure and size distribution of the using primers ITS1 5 -TCCGTAGGTGAACCTGCGG-3 and biosynthesized nanoparticles were studied using electron ITS4 5 -TCCTCCGCTTATTGATATGC-3 [33] were performed microscopy techniques. using a 9700 Gold thermal cycler (Applied Biosystems, UK) under the following conditions: initial denaturation at ◦ ◦ 94 C for 1 min, annealing temperature at 52 C for 1 min 2. Materials and methods ◦ and extension at 72 C for 1 min with an initial dena- turation and final extension for 5 and 10 min at 94 and 2.1. Isolation and characterization of endophytic fungi ◦ 72 C, respectively. For sequencing, the amplified ITS prod- ucts were purified using QIAquick Gel Extraction Spin Kit The endophytic fungi were isolated according to the (QIAGEN, Germany). The purified PCR products were bi- method described by Strobel et al. [32] with minor mod- directionally sequenced using the forward and reverse ifications. The plant material was gently rinsed in running primers. Sequencing of the ITS gene products were per- water to remove any adhering dirt. The samples were then formed using the Big DyeTerminator v3.1 Cycle Sequencing cut into 2 mm blocks and were surface sterilized with 70% Kit (Applied Biosystems, USA). Sequencing of ITS region ethanol for 1 min. Then the samples were soaked in 4% of fungal rDNA resulted in approximately 564 bp long sodium hypochlorite solution for 3 min, and then rinsed 32 L.S. Devi, S.R. Joshi / Journal of Microscopy and Ultrastructure 3 (2015) 29–37 Fig. 4. SEM micrographs of the biosynthesized silver nanoparticles using endophytic fungi (a) Aspergillus tamarii PFL2, (b) Aspergillus niger PFR6 and (c) Penicillium ochrochloron PFR8. nucleotide sequences. The nucleotide sequences were used flask and agitated again at 120 rpm. After the incubation, for the identification of the fungal isolates. Molecular Evo- the cell filtrate was obtained by filtering it through What- lutionary Genetics Analysis (MEGA version 5) software was man filter paper No. 1. The filtrates were treated with 1 mM used for phylogenetic analyses [34]. The closest homo- silver nitrate (Sigma Aldrich) solution in an Erlenmeyer logues to the sequences were selected and the multiple flask and incubated at room temperature in dark. Control sequence alignments were carried out using the ClustalW containing cell-free
Recommended publications
  • Soil and Litter Invertebrates and Heterotrophic Process: 1997 Revision
    ES/ER/TM-126/R2 Toxicological Benchmarks for Contaminants of Potential Concern for Effects on Soil and Litter Invertebrates and Heterotrophic Process: 1997 Revision This document has been approved by the East Tennessee Technology Park Technical Information Office for release to the public. Date: ES/ER/TM-126/R2 Toxicological Benchmarks for Contaminants of Potential Concern for Effects on Soil and Litter Invertebrates and Heterotrophic Process: 1997 Revision R. A. Efroymson M. E. Will G. W. Suter II Date Issued—November 1997 Prepared for the U.S. Department of Energy Office of Environmental Management under budget and reporting code EW 20 LOCKHEED MARTIN ENERGY SYSTEMS, INC. managing the Environmental Management Activities at the East Tennessee Technology Park Oak Ridge Y-12 Plant Oak Ridge National Laboratory Paducah Gaseous Diffusion Plant Portsmouth Gaseous Diffusion Plant under contract DE-AC05-84OR21400 for the U.S. DEPARTMENT OF ENERGY PREFACE This report presents a standard method for deriving benchmarks for the purpose of “contaminant screening,” performed by comparing measured ambient concentrations of chemicals. The work was performed under Work Breakdown Structure 1.4.12.2.3.04.07.02 (Activity Data Sheet 8304). In addition, this report presents sets of data concerning the effects of chemicals in soil on invertebrates and soil microbial processes, benchmarks for chemicals potentially associated with United States Department of Energy sites, and literature describing the experiments from which data were drawn for benchmark derivation. iii ACKNOWLEDGMENTS The authors would like to thank Carla Gunderson and Art Stewart for their helpful reviews of the document. In addition, the authors would like to thank Christopher Evans and Alexander Wooten for conducting part of the literature review.
    [Show full text]
  • Low-Density Polyethylene Film Biodegradation Potential by Fungal Species from Thailand
    Journal of Fungi Article Low-Density Polyethylene Film Biodegradation Potential by Fungal Species from Thailand Sarunpron Khruengsai 1, Teerapong Sripahco 1 and Patcharee Pripdeevech 1,2,* 1 School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand; [email protected] (S.K.); [email protected] (T.S.) 2 Center of Chemical Innovation for Sustainability (CIS), Mae Fah Luang University, Chiang Rai 57100, Thailand * Correspondence: [email protected] Abstract: Accumulated plastic waste in the environment is a serious problem that poses an ecological threat. Plastic waste has been reduced by initiating and applying different alternative methods from several perspectives, including fungal treatment. Biodegradation of 30 fungi from Thailand were screened in mineral salt medium agar containing low-density polyethylene (LDPE) films. Diaporthe italiana, Thyrostroma jaczewskii, Collectotrichum fructicola, and Stagonosporopsis citrulli were found to grow significantly by culturing with LDPE film as the only sole carbon source compared to those obtained from Aspergillus niger. These fungi were further cultured in mineral salt medium broth containing LDPE film as the sole carbon source for 90 days. The biodegradation ability of these fungi was evaluated from the amount of CO2 and enzyme production. Different amounts of CO2 were released from D. italiana, T. jaczewskii, C. fructicola, S. citrulli, and A. niger culturing with LDPE film, ranging from 0.45 to 1.45, 0.36 to 1.22, 0.45 to 1.45, 0.33 to 1.26, and 0.37 to 1.27 g/L, respectively. These fungi were able to secrete a large amount of laccase enzyme compared to manganese peroxidase, and lignin peroxidase enzymes detected under the same conditions.
    [Show full text]
  • Mycology Proficiency Testing Program
    Mycology Proficiency Testing Program Test Event Critique January 2014 Table of Contents Mycology Laboratory 2 Mycology Proficiency Testing Program 3 Test Specimens & Grading Policy 5 Test Analyte Master Lists 7 Performance Summary 11 Commercial Device Usage Statistics 13 Mold Descriptions 14 M-1 Stachybotrys chartarum 14 M-2 Aspergillus clavatus 18 M-3 Microsporum gypseum 22 M-4 Scopulariopsis species 26 M-5 Sporothrix schenckii species complex 30 Yeast Descriptions 34 Y-1 Cryptococcus uniguttulatus 34 Y-2 Saccharomyces cerevisiae 37 Y-3 Candida dubliniensis 40 Y-4 Candida lipolytica 43 Y-5 Cryptococcus laurentii 46 Direct Detection - Cryptococcal Antigen 49 Antifungal Susceptibility Testing - Yeast 52 Antifungal Susceptibility Testing - Mold (Educational) 54 1 Mycology Laboratory Mycology Laboratory at the Wadsworth Center, New York State Department of Health (NYSDOH) is a reference diagnostic laboratory for the fungal diseases. The laboratory services include testing for the dimorphic pathogenic fungi, unusual molds and yeasts pathogens, antifungal susceptibility testing including tests with research protocols, molecular tests including rapid identification and strain typing, outbreak and pseudo-outbreak investigations, laboratory contamination and accident investigations and related environmental surveys. The Fungal Culture Collection of the Mycology Laboratory is an important resource for high quality cultures used in the proficiency-testing program and for the in-house development and standardization of new diagnostic tests. Mycology Proficiency Testing Program provides technical expertise to NYSDOH Clinical Laboratory Evaluation Program (CLEP). The program is responsible for conducting the Clinical Laboratory Improvement Amendments (CLIA)-compliant Proficiency Testing (Mycology) for clinical laboratories in New York State. All analytes for these test events are prepared and standardized internally.
    [Show full text]
  • Research Article Random Mutagenesis of the Aspergillus Oryzae Genome Results in Fungal Antibacterial Activity
    Hindawi Publishing Corporation International Journal of Microbiology Volume 2013, Article ID 901697, 5 pages http://dx.doi.org/10.1155/2013/901697 Research Article Random Mutagenesis of the Aspergillus oryzae Genome Results in Fungal Antibacterial Activity Cory A. Leonard,1 Stacy D. Brown,2 and J. Russell Hayman1 1 Department of Biomedical Sciences, James H. Quillen College of Medicine, East Tennessee State University, P.O. Box 70577, Johnson City, TN 37614, USA 2 Department of Pharmaceutical Sciences, Bill Gatton College of Pharmacy, East Tennessee State University, P.O. Box 70436, Johnson City, TN 37614, USA Correspondence should be addressed to J. Russell Hayman; [email protected] Received 30 April 2013; Revised 9 July 2013; Accepted 9 July 2013 Academic Editor: Joseph Falkinham Copyright © 2013 Cory A. Leonard et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Multidrug-resistant bacteria cause severe infections in hospitals and communities. Development of new drugs to combat resistant microorganisms is needed. Natural products of microbial origin are the source of most currently available antibiotics. We hypothesized that random mutagenesis of Aspergillus oryzae would result in secretion of antibacterial compounds. To address this hypothesis, we developed a screen to identify individual A. oryzae mutants that inhibit the growth of Methicillin- resistant Staphylococcus aureus (MRSA) in vitro. To randomly generate A. oryzae mutant strains, spores were treated with ethyl methanesulfonate (EMS). Over 3000 EMS-treated A. oryzae cultures were tested in the screen, and one isolate, CAL220, exhibited altered morphology and antibacterial activity.
    [Show full text]
  • Isolation of Allergic Fungal Microflora from the Aero-Spora of Khammam District, India
    Int.J.Curr.Microbiol.App.Sci (2016) 5(12): 486-492 International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 5 Number 12 (2016) pp. 486-492 Journal homepage: http://www.ijcmas.com Original Research Article http://dx.doi.org/10.20546/ijcmas.2016.512.052 Isolation of Allergic Fungal Microflora from the Aero-Spora of Khammam District, India D. Saikumari* and Neeti Saxena Mycology and Plant Pathology Lab, Department of Botany, University College For women, Koti, Osmania University, Hyderabad, India *Corresponding author ABSTRACT K e yw or ds Allergies like Asthma, contact urticarial, bronchial problems, skin problems, hay fever, and common allergies are caused by fungal spores. Allergic proteins present Fungal spores, in the fungal spores are responsible for the cause of these allergies. Isolation of fungi Allergens, was done by exposed culture plates from three selected areas viz vegetable market Aerospora , area, surrounding area of factories and Government school of Pandurangapuram Human diseases, Khammam district. village, Khammam district, Telangana, India. Isolation was done from three different areas in order to study the variation in the fungal succession. Altogether 76 species32 genera were isolated from these areas. Aspergillus, Alternaria, Pencillium, Article Info Rhizopus, Mucor, Fusarium were commonly isolated. The present study infer/ Accepted: 18 November 2016 showed that maximum fungal spores which are saprophytic, pathogenic and Available Online: toxigenic were isolated from the market area. Wood rotting fungi were isolated from 10 December 2016 the industrial area. Introduction Allergy is one form of human disease which Pasanen et al., 1992; Katz et al., 1999). effects about 20% of human population Fungi are ubiquitous air borne allergens (Sathavahan et al., 2011)).
    [Show full text]
  • Characterization of Indoor Molds with Allergenic Potential After Cryopreservation at -20°C for One Year
    American Journal of Innovative Research and Applied Sciences. ISSN 2429-5396 I www.american-jiras.com ORIGINAL ARTICLE CHARACTERIZATION OF INDOOR MOLDS WITH ALLERGENIC POTENTIAL AFTER CRYOPRESERVATION AT -20°C FOR ONE YEAR | M’boh Epi Reine Elisabeth N'gou 1,2 | Chiaye Claire Antoinette Yapo-Crezoit 2 | Ama Valérie Bonouman 2 | Diane Kouao 2 | Toure offianan 2 | and | Allico Joseph Djaman 1,2 | 1. Félix Houphouët Boigny University | Laboratory of biology and health | 2. Pasteur Institute of Côte d'Ivoire | Unity of Mycology | Unit of Immunology | Unit of biological Ressources | | Received November 22, 2020 | | Accepted December 07, 2020 | | Published December 14, 2020 | | ID Article| M’boh–Ref3-ajira221120| ABSTRACT Background: The cryopreservation for long period of mold strains of medical interest essential for research is complex. Our objective is to evaluate the impact of conservation with 10% of Glycerol and Brain Heart Infusion (BHI Broth) on the survival of molds of medical interest at -20°C. Methodology: This is an analytical study of the cultural and morphological characteristics of 1310 mold strains isolated from environmental samples (air, surface and dust) collected from all azimuths and preserved in 10% Glycerol and BHI Broth at -20°C for one year. Results: Among the 1310 isolated strains and revived, colony growth was observed in 1059 strains (80.84%) against 251 (19.16%) which showed no signs of development and 119 strains (9.08%) were contaminated by other mold species or yeasts. Fungal viable strains observed after cryopreservation are: Aspergillus fumigatus, Aspergillus flavus, Aspergillus versicolor, Aspergillus niger, Aspergillus terreus, Aspergillus ochraceus, Aspergillus clavatus, Aspergillus sp, Penicillium sp, Fusarium solani, Fusarium sp, Curvularia sp, Mucor sp, Rhizomucor sp, and Trichoderma sp.
    [Show full text]
  • Taxonomic Revision of Aspergillus Section Clavati Based on Molecular, Morphological and Physiological Data
    available online at www.studiesinmycology.org STUDIE S IN MYCOLOGY 59: 89–106. 2007. doi:10.3114/sim.2007.59.11 Taxonomic revision of Aspergillus section Clavati based on molecular, morphological and physiological data J. Varga1,3, M. Due2, J.C. Frisvad2 and R.A. Samson1 1CBS Fungal Biodiversity Centre, Uppsalalaan 8, NL-3584 CT Utrecht, The Netherlands; 2BioCentrum-DTU, Building 221, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark; 3Department of Microbiology, Faculty of Science and Informatics, University of Szeged, H-6701 Szeged, P.O. Box 533, Hungary *Correspondence: János Varga, [email protected] Abstract: Aspergillus section Clavati has been revised using morphology, secondary metabolites, physiological characters and DNA sequences. Phylogenetic analysis of β-tubulin, ITS and calmodulin sequence data indicated that Aspergillus section Clavati includes 6 species, A. clavatus (synonyms: A. apicalis, A. pallidus), A. giganteus, A. rhizopodus, A. longivesica, Neocarpenteles acanthosporus and A. clavatonanicus. Neocarpenteles acanthosporus is the only known teleomorph of this section. The sister genera to Neocarpenteles are Neosartorya and Dichotomomyces based on sequence data. Species in Neosartorya and Neocarpenteles have anamorphs with green conidia and share the production of tryptoquivalins, while Dichotomomyces was found to be able to produce gliotoxin, which is also produced by some Neosartorya species, and tryptoquivalines and tryptoquivalones produced by members of both section Clavati and Fumigati. All species in section Clavati are alkalitolerant and acidotolerant and they all have clavate conidial heads. Many species are coprophilic and produce the effective antibiotic patulin. Members of section Clavati also produce antafumicin, tryptoquivalines, cytochalasins, sarcins, dehydrocarolic acid and kotanins (orlandin, desmethylkotanin and kotanin) in species specific combinations.
    [Show full text]
  • Antibacterial and Antifungal Activities of Spices
    International Journal of Molecular Sciences Review Antibacterial and Antifungal Activities of Spices Qing Liu 1, Xiao Meng 1, Ya Li 1, Cai-Ning Zhao 1, Guo-Yi Tang 1 and Hua-Bin Li 1,2,* 1 Guangdong Provincial Key Laboratory of Food, Nutrition and Health, Department of Nutrition, School of Public Health, Sun Yat-sen University, Guangzhou 510080, China; [email protected] (Q.L.); [email protected] (X.M.); [email protected] (Y.L.); [email protected] (C.-N.Z.); [email protected] (G.-Y.T.) 2 South China Sea Bioresource Exploitation and Utilization Collaborative Innovation Center, Sun Yat-sen University, Guangzhou 510006, China * Correspondence: [email protected]; Tel.: +86-20-8733-2391 Received: 16 May 2017; Accepted: 11 June 2017; Published: 16 June 2017 Abstract: Infectious diseases caused by pathogens and food poisoning caused by spoilage microorganisms are threatening human health all over the world. The efficacies of some antimicrobial agents, which are currently used to extend shelf-life and increase the safety of food products in food industry and to inhibit disease-causing microorganisms in medicine, have been weakened by microbial resistance. Therefore, new antimicrobial agents that could overcome this resistance need to be discovered. Many spices—such as clove, oregano, thyme, cinnamon, and cumin—possessed significant antibacterial and antifungal activities against food spoilage bacteria like Bacillus subtilis and Pseudomonas fluorescens, pathogens like Staphylococcus aureus and Vibrio parahaemolyticus, harmful fungi like Aspergillus flavus, even antibiotic resistant microorganisms such as methicillin resistant Staphylococcus aureus. Therefore, spices have a great potential to be developed as new and safe antimicrobial agents.
    [Show full text]
  • Identification and Antifungal Activity of Compounds from the Mangrove
    marine drugs Article Identification and Antifungal Activity of Compounds from the Mangrove Endophytic Fungus Aspergillus clavatus R7 Wensheng Li 1, Ping Xiong 1, Wenxu Zheng 1, Xinwei Zhu 1, Zhigang She 2, Weijia Ding 1,* and Chunyuan Li 1,* 1 College of Materials and Energy, South China Agricultural University, Guangzhou 510642, China; [email protected] (W.L.); [email protected] (P.X.); [email protected] (W.Z.); [email protected] (X.Z.) 2 School of Chemistry and Chemical Engineering, Sun Yat-Sen University, Guangzhou 510275, China; [email protected] * Correspondence: [email protected] (W.D.); [email protected] (C.L.); Tel.: +86-20-85280319 (C.L.) Received: 17 July 2017; Accepted: 17 August 2017; Published: 19 August 2017 Abstract: Two new coumarin derivatives, 4,40-dimethoxy-5,50-dimethyl-7,70-oxydicoumarin (1), 7-(γ,γ-dimethylallyloxy)-5-methoxy-4-methylcoumarin (2), a new chromone derivative, (S)-5-hydroxy-2,6-dimethyl-4H-furo[3,4-g]benzopyran-4,8(6H)-dione (5), and a new sterone derivative, 24-hydroxylergosta-4,6,8(14),22-tetraen-3-one (6), along with two known bicoumarins, kotanin (3) and orlandin (4), were isolated from an endophytic fungus Aspergillus clavatus (collection No. R7), isolated from the root of Myoporum bontioides collected from Leizhou Peninsula, China. Their structures were elucidated using 1D- and 2D- NMR spectroscopy, and HRESIMS. The absolute configuration of compound 5 was determined by comparison of the experimental and calculated electronic circular dichroism (ECD) spectra. Compound 6 significantly inhibited the plant pathogenic fungi Fusarium oxysporum, Colletotrichum musae and Penicillium italicum, compound 5 significantly inhibited Colletotrichum musae, and compounds 1, 3 and 4 greatly inhibited Fusarium oxysporum, showing the antifungal activities higher than those of the positive control, triadimefon.
    [Show full text]
  • Phylogeny, Identification and Nomenclature of the Genus Aspergillus
    available online at www.studiesinmycology.org STUDIES IN MYCOLOGY 78: 141–173. Phylogeny, identification and nomenclature of the genus Aspergillus R.A. Samson1*, C.M. Visagie1, J. Houbraken1, S.-B. Hong2, V. Hubka3, C.H.W. Klaassen4, G. Perrone5, K.A. Seifert6, A. Susca5, J.B. Tanney6, J. Varga7, S. Kocsube7, G. Szigeti7, T. Yaguchi8, and J.C. Frisvad9 1CBS-KNAW Fungal Biodiversity Centre, Uppsalalaan 8, NL-3584 CT Utrecht, The Netherlands; 2Korean Agricultural Culture Collection, National Academy of Agricultural Science, RDA, Suwon, South Korea; 3Department of Botany, Charles University in Prague, Prague, Czech Republic; 4Medical Microbiology & Infectious Diseases, C70 Canisius Wilhelmina Hospital, 532 SZ Nijmegen, The Netherlands; 5Institute of Sciences of Food Production National Research Council, 70126 Bari, Italy; 6Biodiversity (Mycology), Eastern Cereal and Oilseed Research Centre, Agriculture & Agri-Food Canada, Ottawa, ON K1A 0C6, Canada; 7Department of Microbiology, Faculty of Science and Informatics, University of Szeged, H-6726 Szeged, Hungary; 8Medical Mycology Research Center, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8673, Japan; 9Department of Systems Biology, Building 221, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark *Correspondence: R.A. Samson, [email protected] Abstract: Aspergillus comprises a diverse group of species based on morphological, physiological and phylogenetic characters, which significantly impact biotechnology, food production, indoor environments and human health. Aspergillus was traditionally associated with nine teleomorph genera, but phylogenetic data suggest that together with genera such as Polypaecilum, Phialosimplex, Dichotomomyces and Cristaspora, Aspergillus forms a monophyletic clade closely related to Penicillium. Changes in the International Code of Nomenclature for algae, fungi and plants resulted in the move to one name per species, meaning that a decision had to be made whether to keep Aspergillus as one big genus or to split it into several smaller genera.
    [Show full text]
  • Multimycotoxin Analysis of South African Aspergillus Clavatus Isolates
    Short research communication Multimycotoxin analysis of South African Aspergillus clavatus isolates Authors: C. J. Botha1 M. Truter2 M. Sulyok3 Affiliations: 1 Department of Paraclinical Sciences, Faculty of Veterinary Science, University of Pretoria, Private bag X04, Onderstepoort, 0110, South Africa 2 Biosystematics Division, Agricultural Research Council-Vegetable and Ornamental Plants, Private Bag X293, Pretoria, 0001, South Africa 3 Centre for Analytical Chemistry, Department of Agrobiotechnology (IFA-Tulln), University of Natural Resources and Life Sciences, Konrad Lorenzstr. 20, A-3430, Tulln, Austria CJ Botha [email protected] Tel.: +27 125298023; fax +27 125298304. ORCID CJ Botha 0000 0003 1535 9270 1 Abstract Aspergillus clavatus poisoning is a neuromycotoxicosis of ruminants that occurs sporadically across the world after ingestion of infected feedstuffs. Although various toxic metabolites are synthesized by the fungus it is not clear which specific or group of mycotoxins induces the syndrome. A. clavatus isolates were deposited in the culture collection of the Biosystematics Division, Plant Protection Research Institute, Agricultural Research Council during incidences of livestock poisoning (1988 – 2016). Six isolates were still viable and these plus three other South African isolates that were also previously deposited in the collection were positively identified as A. clavatus based on morphology and ß-tubulin sequence data. The cultures were screened for multiple mycotoxins using a liquid chromatography-tandem mass spectrometric (LC-MS/MS) method. Twelve A. clavatus metabolites were detected. The concentrations of the tremorgenic mycotoxins (i.e. tryptoquivaline A and its related metabolites deoxytryptoquivaline A and deoxynortryptoquivaline) were higher than patulin and cytochalasin E. Livestock owners should not feed A.
    [Show full text]
  • Isolation of Ascomycetous Fungi from a Tertiary Institution Campus Soil
    JASEM ISSN 1119-8362 Full-text Available Online at J. Appl. Sci. Environ. Manage. December, 2008 All rights reserved www.bioline.org.br/ja Vol. 12(4) 57 - 61 Isolation of Ascomycetous Fungi from a Tertiary Institution Campus Soil DUROWADE, K. A1; *KOLAWOLE, O. M2; UDDIN II, R. O3; ENONBUN, K.I2 1Department of Epidemiology and Community Medicine, College of Medicine, University of Ilorin. Kwara State. 2Department of Microbiology, Faculty of Science. University of Ilorin. P.M.B 1515, Ilorin, Kwara State. Tel: 08060088495. Email: [email protected] 3 Department of Crop Protection, Faculty of Agriculture, University of Ilorin. Kwara State ABSTRACT: Studies were carried out on the ascomycetous fungi present in six different but carefully selected sites on the University of Ilorin permanent site soil. Fungi isolation was done by the soil dilution method incubated at 27oC for 72 hours. The predominant Ascomycetous fungi isolated include among others; Aspergillus niger, Fusarium solani, Fusarium oxysporum, Penicillium italicum, Fusarium acuminatum, Fusarium culmorum, Candida albicans, Botrytis cinerea, Geotrichum candidum, Trichoderma viride, Verticillium lateritum, Curvularia palescens, Penicillium griseofulvum, Penicillium janthinellum, Penicillium chrysogenum, Aspergillus terreus, Aspergillus flavus, Aspergillus fumigatus, Aspergillus glaucus, Aspergillus clavatus, Cladosporium resinae, Alternaria alternate, Trichothecium roseum, Phialophora fastigiata, Aspergillus nidulans, Aspergillus wentii, Humicola grisea, Trichophyton rubrum, Helminthosporium cynodontis, Penicillium funiculosum, Penicillium purpurogenum, Saccharomyces cerevisiae, Trichoderma harzianum, Scopulariopsis candida.The physico- chemical characteristics of soil samples was found to affect the distribution and population of fungi. The colony count in the study are ranged between 5.8 x 10″ per gram of soil to 1.63 x 10″ per gram of soil.
    [Show full text]