DOCSLIB.ORG

Isolation and Characterization of Bioactive Molecules from Endophytic Fungi

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Isolation and Characterization of Bioactive Molecules from Endophytic Fungi Isolation and characterization of bioactive molecules from endophytic fungi A Thesis Submitted in fulfillment of the requirement for the award of the degree of DOCTOR OF PHILOSOPHY IN BIOTECHNOLOGY By M. Vasundhara (Reg. No. 950800006) Department of Biotechnology Thapar University Patiala 147004, India February 2017 -i- -ii- -iii- Acknowledgements Constant inspiration and encouragement given by all concerned was the driving force that enabled me to submit this thesis in the present form. Guidance, direction, cooperation, motivation and support came in my way from many people and it is a moment to acknowledge the same. First of all, I am thankful to the almighty God for blessing me and giving me the strength and perseverance to complete my work successfully. It is a moment of pride to put on record the professional guidance and valuable support that I have received from my guide, Dr. Anil Kumar, Associate Professor, Department of Biotechnology. My sincere thanks to Dr. Dinesh Goyal, Head, Department of Biotechnology for his cooperation and support throughout the research work. Also my special thanks to Dr. Sanjai Saxena, Head, CORE for extending the facilities to carry out my research work. I thank my doctoral committee members Dr. Rajesh Kumar, Dr. S.K. Pandey, Dr. Manoj Baranwal and Dr. Siddharth Sharma for their advice and suggestions. My great appreciation goes to Dr. M. S. Reddy for his valuable and constructive suggestions during the planning and development, execution of research work. I am particularly grateful for the assistance, valuable technical and moral support and encouragement provided by Dr. Manoj Baranwal during by research work. I am also thankful to Dr. N. Tejo Prakash for his continuous help while preparing thesis. I am also thankful to Dr. T.S. Suryanarayanan, Vivekananda Institute of Tropical Mycology, RKM Vidyapith, Chennai for providing the thermotolerant endophytic fungal isolates and Dr. Sunil Kumar Deshmukh, The Energy and Resources Institute, TERI Gram, Gual Pahari, Gurgaon for providing bacterial cultures. My special thanks of gratitude to Dr. N. Sivarasmaiah, Nano Temper Technologies, World Trade Centre, Bangalore, who provided insight and expertise that greatly assisted me in carrying out my research work and I came to know about many new things. -iv- -v- Publications The following publications are the outcome of the present research work 1. M. Vasundhara, Manoj Baranwal, N. Sivaramaiah, Anil Kumar. 2017. Isolation and characterization of trichalasins producing endophytic fungus from Taxus baccata. Annals of Microbiology. DoI: 10.1007/s13213-017-1256-4. 2. M. Vasundhara, Anil Kumar, M.S. Reddy. 2016. Molecular approaches to screen bioactive compounds from endophytic fungi. Front. Microbiol. 7:1774. DoI: 10.3389/fmicb.2016.01774. 3. M. Vasundhara, Manoj Baranwal, Anil Kumar. 2016. Fusarium tricinctum, an endophytic fungus exhibits cell growth inhibition and antioxidant activity. Indian Journal of Microbiology, 56:433-438. -vi- Table of Contents Chapters Page No. I. Introduction 1 II. Review of Literature 5 2.0 Natural products in drug discovery and development 5 2.1 Endophytic fungal diversity 6 2.1.1 Traditional techniques used in endophyte studies 6 2.1.2 Molecular identification of fungi 8 2.1.3 Internal transcribed spacers (ITS) 11 2.2 Antibacterials / Antimycobacterials from endophytic fungi 11 2.3 Antifungals from endophytic fungi 19 2.4 Anticancer, Immunosupressive, Antiinflammatory activities of endophytic 23 fungi 2.5 Antioxidants from endophytic fungi 33 2.6 Analysis of bioactive metabolites 38 2.6.1 Mass Spectrometry 39 2.7 Industrial enzymes from endophytic fungi 40 III. Materials and Methods 43 3.1 Collection of bark samples 43 3.1.1 Isolation of endophytic fungi from Taxus baccata bark 44 samples 3.2 Preparation of fungal extracts for biological assays 44 3.3 Assay for antimicrobial activity 45 3.3.1 Test microorganisms and growth conditions 45 3.3.2 Antimicrobial agent (as control) 46 3.3.3 Turbidity standard for inoculum preparation 46 3.3.4. Prescreen assay: Agar-well diffusion assay 47 3.3.5 Screening: Microplate broth dilution assay 48 3.4 Cell growth inhibition assay 49 3.4.1 Isolation of peripheral blood mononuclear cells 50 3.4.2 Counting of cells 50 -vii- 3.4.3 Lymphocyte proliferation assay 50 3.4.4 Measurement of TNF-α 51 3.4.5 Antioxidant assay 52 3.5 Identification of endophytic fungi 52 3.5.1 Morphological characters 52 3.5.2 Molecular characterization 53 3.5.2.1 Isolation of fungal genomic DNA 53 3.5.2.2 CTAB method for DNA extraction 53 3.5.2.3 Checking of DNA (Agarose Gel Electrophoresis) 54 3.5.2.4 Quantification of DNA using Nano drop 54 3.5.2.5 Amplification of Internal Transcribed Spacer (ITS) 55 region 3.5.2.6 Purification of PCR products and sequencing 55 3.5.2.7 Sequence analysis 56 3.6 Extracellular enzymes production by thermotolerant endophytic fungi 56 3.6.1 Agar Plate Assays 56 3.6.1.1 Amylase activity 56 3.6.1.2 Cellulase activity 56 3.6.1.3 Laccase activity 57 3.6.1.4 Lipase activity 57 3.6.1.5 Pectinase activity 57 3.6.1.6 Proteinase activity 57 3.6.1.7 Tyrosinase activity 57 3.6.1.8 L-asparaginase activity 58 3.7 Quantitative Assay 58 3.7.1 Cellulase Activity 58 3.7.2 Culture conditions for cellulase production 58 3.7.3 Cellulase assay 58 3.7.4 Characterization of enzyme 59 3.7.4.1 Optimization of time for cellulase production 59 3.7.4.2 pH 60 3.7.4.3 Temperature and thermostability 60 3.7.4.4 Lipase Activity 60 -viii- 3.7.4.5 Culture condition for lipase production 61 3.7.4.6 Lipase assay 61 3.7.4.7 Protein assay 62 3.7.4.8 Characterization of enzyme 63 3.7.4.8.1 Optimimization of time for lipase 63 production 3.7.4.8.2 pH 63 3.7.4.8.3 Temperature and thermostability 64 3.8 Analysis of bioactive molecules 64 3.8.1 Sample preparation 64 3.8.2 Mass spectrometry 64 3.8.3 UHPLC-QTOF-MS/MS analysis 65 3.9 Statistical analysis 66 IV. Results and Discussion 67 4.0 Isolation, identification and biological activities of endophytic fungi 67 4.1 Isolation of endophytic fungi from Taxus baccata bark samples 67 4.2 Preliminary screening for bioactivity 68 4.3 Identification of endophytic fungal isolates 68 4.3.1 T1 isolate 69 4.3.2 T2 isolate 74 4.3.3 T5 isolate 78 4.3.4 T6 isolate 82 4.4 Antimicrobial activity 87 4.5 Cytotoxic activity 93 4.6 Antioxidant assay 95 4.7 Cell growth inhibition and antioxidant activity of Diaporthe sp. T1 96 4.7.1 Cytotoxicity 96 4.7.2 Antioxidant activity 68 4.8 Cell growth inhibition and antioxidant activity of Fusarium tricinctum T6 100 4.8.1 Cytotoxicity 100 4.8.2 Anti-proliferative activity of peripheral blood 102 mononuclear cells 4.8.3 TNF- production 103 -ix- 4.8.4 Antioxidant activity 105 5.0 Identification and characterization of putative bioactive molecules 108 5.1 UHPLC-MS analysis of extract of T1 109 5.1.1 Cytosporones C & E 109 5.1.2 Cytochalasins 112 5.2 UHPLC-MS analysis of extract T6 115 5.2.1 Gniditrin 115 5.2.2 7-hydroxyheptaphylline 116 5.2.3 Tirandamycin 118 5.2.4 Fumitremorgin C 120 5.2.5 Paclitaxel (Taxol) 121 5.2.6 10-deacetyl baccatin III (10-DAB) 123 6.0 Extracellular enzymes production by thermotolerant endophytic fungi 126 6.1 Screening for extracellular enzymes 126 6.2 Quantitative studies on extracellular enzymes 134 6.2.1 Cellulase 134 6.2.2 Lipase 139 6.3 Cellulase 146 6.4 Laccase 149 6.5 Amylase 150 6.6 Lipase 151 6.7 Protease 152 6.8 L-asparaginase 153 Summary 155 References 160 Appendices ( I to III) 183 -x- List of Tables Sl. No. Table Description 1 3.1 McFarland Standard 2 4.1 Antimicrobial activity of fungal extracts of T1, T2, T5 and T6 tested with different microorganisms 3 4.2 Effect of the fungal extracts (500 µg/ml) of T1, T2, T5 and T6 isolates on growth inhibition (%) of different microorganisms by MTT assay 4 4.4a Effect of fungal extract of different endophytic fungi on the growth inhibition of MCF cell lines 5 4.4b Effect of different concentration of fungal extract of T1 and T6 isolates on the growth inhibition of MCF cell lines 6 4.5 Effect of different concentrations of fungal extracts of T1, T2, T5 and T6 on the antioxidant activity 7 4.6 Cytotoxic effect of Diaporthe sp. extract against human breast cancer (MCF-7) and human cervical (HeLa) cancer cell lines 8 4.7 Antioxidant effect of Diaporthe sp., extracts based on free radical scavenging activity. Ascorbic acid (AA) (100 µg/ml) was used as positive control 9 4.8 Cytotoxic effect of F. tricinctum T6. extract against human breast cancer (MCF-7) and human cervical (HeLa) cancer cell lines 10 4.9 Immunosuppressive effect of Fusarium tricinctum extract on concanavalin stimulated peripheral blood mononuclear cells 11 4.10 Effect of Fusarium tricinctum extract on TNF-α production in MCF-7 concanavalin A stimulated peripheral blood mononuclear cells (PBMCs) 12 4.11 Antioxidant effect of Fusarium tricinctum extracts based on free radical scavenging activity 13 5.1 LC/ESI-MS analysis data of cytosporone C & cytosporone E from Diaporthe sp. (T1) and their literature reported data 14 5.2 LC/ESI-MS analysis data of trichalasin E, trichalasin F and trichalasin H from Diaporthe sp.
Load more

Recommended publications
  • AR TICLE Recommended Names for Pleomorphic Genera In
    IMA FUNGUS · 6(2): 507–523 (2015) doi:10.5598/imafungus.2015.06.02.14 Recommended names for pleomorphic genera in Dothideomycetes ARTICLE Amy Y. Rossman1, Pedro W. Crous2,3, Kevin D. Hyde4,5, David L. Hawksworth6,7,8, André Aptroot9, Jose L. Bezerra10, Jayarama D. Bhat11, Eric Boehm12, Uwe Braun13, Saranyaphat Boonmee4,5, Erio Camporesi14, Putarak Chomnunti4,5, Dong-Qin Dai4,5, Melvina J. D’souza4,5, Asha Dissanayake4,5,15, E.B. Gareth Jones16, Johannes Z. Groenewald2, Margarita Hernández-Restrepo2,3, Sinang Hongsanan4,5, Walter M. Jaklitsch17, Ruvishika Jayawardena4,5,12, Li Wen Jing4,5, Paul M. Kirk18, James D. Lawrey19, Ausana Mapook4,5, Eric H.C. McKenzie20, Jutamart Monkai4,5, Alan J.L. Phillips21, Rungtiwa Phookamsak4,5, Huzefa A. Raja22, Keith A. Seifert23, Indunil Senanayake4,5, Bernard Slippers3, Satinee Suetrong24, Kazuaki Tanaka25, Joanne E. Taylor26, Kasun M. Thambugala4,5,27, Qing Tian4,5, Saowaluck Tibpromma4,5, Dhanushka N. Wanasinghe4,5,12, Nalin N. Wijayawardene4,5, Saowanee Wikee4,5, Joyce H.C. Woudenberg2, Hai-Xia Wu28,29, Jiye Yan12, Tao Yang2,30, Ying Zhang31 1Department of Botany and Plant Pathology, Oregon State University, Corvallis, Oregon 97331, USA; corresponding author e-mail: amydianer@ yahoo.com 2CBS-KNAW Fungal Biodiversity Institute, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands 3Department of Microbiology and Plant Pathology, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Pretoria 0002, South Africa 4Center of Excellence in Fungal Research, School of Science, Mae Fah
    [Show full text]
  • Occurrence of Pathogenic and Endophytic Fungi and Their Influence on Quality of Medicinal Plants Applied in Management of Neurological Diseases and Mental Disorders
    From Botanical to Medical Research Vol. 63 No. 4 2017 Received: 2017-08-06 DOI: 10.1515/hepo-2017-0025 Accepted: 2017-12-16 Review article Occurrence of pathogenic and endophytic fungi and their influence on quality of medicinal plants applied in management of neurological diseases and mental disorders KATARZYNA WIELGUSZ1, LIDIA IRZYKOWSKA2* 1Institute of Natural Fibers and Medicinal Plants Department of Breeding and Agriculture of Fibrous and Energetic Plants Wojska Polskiego 71b 60-630 Poznań, Poland 2Poznan University of Life Sciences Department of Phytopathology, Seed Science and Technology Dąbrowskiego 159 60-594 Poznań, Poland *corresponding author: phone: 48 61 848 79 27, fax: 48 61 848 79 99, e-mail: [email protected] Summary Due to increasing demand of medicinal plants (MPs), quality and safety more attention to the plant health should be paid. Among herb pathogens, especially fungi cause serious diseases in these plants decreasing yield and quality of herbal raw material. Some species, i.e. Fusarium sp., Alternaria sp., Penicillium sp. are known as mycotoxin producers. Paradoxically, self-treatment with herbal raw material can expose the pa- tient to mycotoxin activity. In tissues of some MPs species, asymptomatically endophytic fungi residue. It is known that they are able to influence a biosynthesis of secondary metabolites in their host plant or produce biologically active compounds. Until recently these microorganisms have been neglected as a component of MPs, the reason why there have unexplored bioactivity and biodiversity. The paper presents an overview of herbal plants that are used in the treatment of nervous system diseases. Pathogenic fungi that infect these plants are described.
    [Show full text]
  • Endophytic Fungi Harbored in Camptotheca Acuminata
    Endophytic fungi harbored in Camptotheca acuminata, Hypericum perforatum and Juniperus communis plants as promising sources of camptothecin, hypericin and deoxypodophyllotoxin D I S S E R T A T I O N Submitted for the degree of Dr. rer. nat. (rerum naturalium) to the Faculty of Chemistry Technische Universität Dortmund by Souvik Kusari 2010 Endophytic fungi harbored in Camptotheca acuminata, Hypericum perforatum and Juniperus communis plants as promising sources of camptothecin, hypericin and deoxypodophyllotoxin APPROVED DISSERTATION Doctoral Committee Chairman: Prof. Dr. Carsten Strohmann Reviewers: 1. Prof. Dr. Dr.h.c. Michael Spiteller 2. Prof. Dr. Oliver Kayser Date of defense examination: October 04, 2010 Chairman of the examination: Prof. Dr. Christof M. Niemeyer “The grand aim of all science is to cover the greatest number of empirical facts by logical deduction from the smallest number of hypotheses or axioms” Albert Einstein (March 14, 1879 – April 18, 1955) THIS THESIS IS DEDICATED TO MY PARENTS … i Declaration Declaration I hereby declare that this thesis is a presentation of my original research work, and is provided independently without any undue assistance. Wherever contributions of others are involved, every effort is made to indicate this clearly, with due reference to the literature(s), and acknowledgement of collaborative research and discussions. This work was done under the guidance and supervision of Professor Dr. Dr.h.c. Michael Spiteller, at the Institute of Environmental Research (INFU) of the Faculty of Chemistry, Chair of Environmental Chemistry and Analytical Chemistry, TU Dortmund, Germany. Dated: August 10, 2010 SOUVIK KUSARI Place: Dortmund, Germany In my capacity as supervisor of the candidate’s thesis, I certify that the above statements are true to the best of my knowledge.
    [Show full text]
  • Identification and Bioactive Potential of Endophytic Fungi Isolated From
    Qadri et al. SpringerPlus 2013, 2:8 http://www.springerplus.com/content/2/1/8 a SpringerOpen Journal RESEARCH Open Access Identification and bioactive potential of endophytic fungi isolated from selected plants of the Western Himalayas Masroor Qadri1, Sarojini Johri1, Bhahwal A Shah2, Anamika Khajuria3, Tabasum Sidiq3, Surrinder K Lattoo4, Malik Z Abdin5 and Syed Riyaz-Ul-Hassan1* Abstract This study was conducted to characterize and explore the endophytic fungi of selected plants from the Western Himalayas for their bioactive potential. A total of 72 strains of endophytic fungi were isolated and characterized morphologically as well as on the basis of ITS1-5.8S-ITS2 ribosomal gene sequence acquisition and analyses. The fungi represented 27 genera of which two belonged to Basidiomycota, each representing a single isolate, while the rest of the isolates comprised of Ascomycetous fungi. Among the isolated strains, ten isolates could not be assigned to a genus as they displayed a maximum sequence similarity of 95% or less with taxonomically characterized organisms. Among the host plants, the conifers, Cedrus deodara, Pinus roxburgii and Abies pindrow harbored the most diverse fungi, belonging to 13 different genera, which represented almost half of the total genera isolated. Several extracts prepared from the fermented broth of these fungi demonstrated strong bioactivity against E. coli and S. aureus with the lowest IC50 of 18 μg/ml obtained with the extract of Trichophaea abundans inhabiting Pinus sp. In comparison, extracts from only three endophytes were significantly inhibitory to Candida albicans, an important fungal pathogen. Further, 24 endophytes inhibited three or more phytopathogens by at least 50% in co-culture, among a panel of seven test organisms.
    [Show full text]
  • Fungal Endophytes ÂŒ Secret Producers of Bioactive Plant
    REVIEWS Institut für Pharmazeutische Biologie und Biotechnologie, Heinrich-Heine-Universität Düsseldorf, Germany Fungal endophytes – secret producers of bioactive plant metabolites A. H. Aly, A. Debbab, P. Proksch Received November 27, 2012, accepted December 18, 2012 Prof. Dr. Peter Proksch, Institut für Pharmazeutische Biologie und Biotechnologie, Heinrich-Heine-Universität Düsseldorf, Universitätsstrasse 1, Geb. 26.23, 40225 Düsseldorf, Germany [email protected] Dedicated to Professor Theo Dingermann, Frankfurt, on the occasion of his 65th birthday. Pharmazie 68: 499–505 (2013) doi: 10.1691/ph.2013.6517 The potential of endophytic fungi as promising sources of bioactive natural products continues to attract broad attention. Endophytic fungi are defined as fungi that live asymptomatically within the tissues of higher plants. This overview will highlight the uniqueness of endophytic fungi as alternative sources of pharmaceutically valuable compounds originally isolated from higher plants, e.g. paclitaxel, camptothecin and podophyllotoxin. In addition, it will shed light on the fungal biosynthesis of plant associated metabolites as well as new approaches developed to improve the production of commercially important plant derived compounds with the involvement of endophytic fungi. 1. Introduction as potential new sources for therapeutic agents (Aly et al. 2010, 2011; Debbab et al. 2011, 2012) and set the stage for a more Fungal endophytes were first defined by Anton de Bary in 1886 comprehensive examination of the ability of other plants to as microorganisms that colonize internal tissues of stems and yield endophytes producing pharmacologically important nat- leaves (Wilson 1995). More recent definitions denoted that they ural products hitherto only known from plants. are ubiquitous microorganisms present in virtually all plants In this review pharmaceutically valuable plant secondary on earth from the arctic to the tropics (Strobel and Daisy 2003; metabolites which were found to be produced by fungal endo- Huang et al.
    [Show full text]
  • Fungal Endophytes As Efficient Sources of Plant-Derived Bioactive
    microorganisms Review Fungal Endophytes as Efficient Sources of Plant-Derived Bioactive Compounds and Their Prospective Applications in Natural Product Drug Discovery: Insights, Avenues, and Challenges Archana Singh 1,2, Dheeraj K. Singh 3,* , Ravindra N. Kharwar 2,* , James F. White 4,* and Surendra K. Gond 1,* 1 Department of Botany, MMV, Banaras Hindu University, Varanasi 221005, India; [email protected] 2 Department of Botany, Institute of Science, Banaras Hindu University, Varanasi 221005, India 3 Department of Botany, Harish Chandra Post Graduate College, Varanasi 221001, India 4 Department of Plant Biology, Rutgers University, New Brunswick, NJ 08901, USA * Correspondence: [email protected] (D.K.S.); [email protected] (R.N.K.); [email protected] (J.F.W.); [email protected] (S.K.G.) Abstract: Fungal endophytes are well-established sources of biologically active natural compounds with many producing pharmacologically valuable specific plant-derived products. This review details typical plant-derived medicinal compounds of several classes, including alkaloids, coumarins, flavonoids, glycosides, lignans, phenylpropanoids, quinones, saponins, terpenoids, and xanthones that are produced by endophytic fungi. This review covers the studies carried out since the first report of taxol biosynthesis by endophytic Taxomyces andreanae in 1993 up to mid-2020. The article also highlights the prospects of endophyte-dependent biosynthesis of such plant-derived pharma- cologically active compounds and the bottlenecks in the commercialization of this novel approach Citation: Singh, A.; Singh, D.K.; Kharwar, R.N.; White, J.F.; Gond, S.K. in the area of drug discovery. After recent updates in the field of ‘omics’ and ‘one strain many Fungal Endophytes as Efficient compounds’ (OSMAC) approach, fungal endophytes have emerged as strong unconventional source Sources of Plant-Derived Bioactive of such prized products.
    [Show full text]
  • Why Plants Harbor Complex Endophytic Fungal Communities: Insights from Perennial Bunchgrass Stipagrostis Sabulicola in the Namib Sand Sea
    fmicb-12-691584 June 2, 2021 Time: 17:58 # 1 ORIGINAL RESEARCH published: 08 June 2021 doi: 10.3389/fmicb.2021.691584 Why Plants Harbor Complex Endophytic Fungal Communities: Insights From Perennial Bunchgrass Stipagrostis sabulicola in the Namib Sand Sea Anthony J. Wenndt1, Sarah E. Evans2,3, Anne D. van Diepeningen4, J. Robert Logan2,3, Peter J. Jacobson5, Mary K. Seely6 and Kathryn M. Jacobson5* 1 Plant Pathology and Plant-Microbe Biology, School of Integrative Plant Sciences, Cornell University, Ithaca, NY, Edited by: United States, 2 Department of Integrative Biology, Michigan State University, East Lansing, MI, United States, 3 W.K. Kellogg Raffaella Balestrini, Biological Station, Michigan State University, Hickory Corners, MI, United States, 4 B.U. Biointeractions and Plant Health, Institute for Sustainable Plant Wageningen Plant Research, Wageningen University and Research, Wageningen, Netherlands, 5 Department of Biology, Protection, National Research Council Grinnell College, Grinnell, IA, United States, 6 Desert Research Foundation of Namibia, Windhoek, Namibia (CNR), Italy Reviewed by: All perennial plants harbor diverse endophytic fungal communities, but why they tolerate Iñigo Zabalgogeazcoa, Institute of Natural Resources these complex asymptomatic symbioses is unknown. Using a multi-pronged approach, and Agrobiology of Salamanca we conclusively found that a dryland grass supports endophyte communities comprised (IRNASA), Spain George Newcombe, predominantly of latent saprophytes that can enhance localized nutrient recycling after University of Idaho, United States senescence. A perennial bunchgrass, Stipagrostis sabulicola, which persists along a *Correspondence: gradient of extreme abiotic stress in the hyper-arid Namib Sand Sea, was the focal Kathryn M. Jacobson point of our study. Living tillers yielded 20 fungal endophyte taxa, 80% of which [email protected] decomposed host litter during a 28-day laboratory decomposition assay.
    [Show full text]
  • Endophytic Fungi of Olive Tree and Their Exploitation in the Biological
    ESCUELA SUPERIOR Y TÉCNICA DE INGENIERÍA AGRARIA INGENIERÍA DE BIOSISTEMAS Endophytic fungi of olive tree and their exploitation in the biological control of olive anthracnose Doctoral Thesis Maria de Fátima Tomé Martins Director: Professora Doutora Paula Cristina Santos Baptista León 2020 ESCUELA SUPERIOR Y TÉCNICA DE INGENIERÍA AGRARIA INGENIERÍA DE BIOSISTEMAS Hongos endofíticos del olivo y su aprovechamiento para el control biológico de la antracnosis del olivo Tesis Doctoral Maria de Fátima Tomé Martins Director: Profesora Doctora Paula Cristina Santos Baptista León 2020 This research was supported by FEDER funds through the COMPETE (Operational Programme for Competitiveness Factors) and by the Foundation for Science and Technology (FCT, Portugal) within the POCI-01-0145-FEDER-031133 project and FCT/MCTES to CIMO (UIDB/00690/2020), as well as the Horizon 2020, the European Union’s Framework Programme for Research and Innovation, for financial support the project PRIMA/0002/2018. Fátima Martins also thanks the individual research grant ref. SFRH / BD / 112234/2015 award by FCT. The studies presented in this thesis were performed at the AgroBioTechnology Laboratory, at the Mountain Research Centre (CIMO), School of Agriculture Polytechnic Institute of Bragança. Às minhas filhas Acknowledgements Neste momento, é com enorme gosto e satisfação que agradeço a todos aqueles que direta ou indiretamente contribuíram para a realização e conclusão deste trabalho. Em primeiro lugar gostaria de agradecer em especial à minha orientadora. À Professora Doutora Paula Cristina dos Santos Baptista, da Escola Superior Agrária, por toda a orientação prestada durante a realização do trabalho ao nível laboratorial e escrito, bem como, por todo o conhecimento transmitido, paciência, disponibilidade e acima de tudo pela amizade e carinho demonstrados ao longo destes anos.
    [Show full text]
  • Non-Rainfall Moisture Activates Fungal Decomposition of Surface Litter in the Namib Sand Sea
    RESEARCH ARTICLE Non-Rainfall Moisture Activates Fungal Decomposition of Surface Litter in the Namib Sand Sea Kathryn Jacobson1☯*, Anne van Diepeningen2, Sarah Evans3, Rachel Fritts1, Philipp Gemmel1, Chris Marsho1, Mary Seely4, Anthony Wenndt1, Xiaoxuan Yang1, Peter Jacobson1☯ 1 Biology Department, Grinnell College, Grinnell, Iowa, United States of America, 2 CBS-KNAW Fungal Biodiversity Centre, Utrecht, The Netherlands, 3 Kellogg Biological Station, Michigan State University, Hickory Corners, Michigan, United States of America, 4 Gobabeb Research and Training Centre, Gobabeb, Namibia ☯ These authors contributed equally to this work. * [email protected] Abstract The hyper-arid western Namib Sand Sea (mean annual rainfall 0–17 mm) is a detritus- based ecosystem in which primary production is driven by large, but infrequent rainfall OPEN ACCESS events. A diverse Namib detritivore community is sustained by minimal moisture inputs Citation: Jacobson K, van Diepeningen A, Evans S, from rain and fog. The decomposition of plant material in the Namib Sand Sea (NSS) has Fritts R, Gemmel P, Marsho C, et al. (2015) Non- long been assumed to be the province of these detritivores, with beetles and termites alone Rainfall Moisture Activates Fungal Decomposition of accounting for the majority of litter losses. We have found that a mesophilic Ascomycete Surface Litter in the Namib Sand Sea. PLoS ONE 10 (5): e0126977. doi:10.1371/journal.pone.0126977 community, which responds within minutes to moisture availability, is present on litter of the perennial
    [Show full text]
  • A Worldwide List of Endophytic Fungi with Notes on Ecology and Diversity
    Mycosphere 10(1): 798–1079 (2019) www.mycosphere.org ISSN 2077 7019 Article Doi 10.5943/mycosphere/10/1/19 A worldwide list of endophytic fungi with notes on ecology and diversity Rashmi M, Kushveer JS and Sarma VV* Fungal Biotechnology Lab, Department of Biotechnology, School of Life Sciences, Pondicherry University, Kalapet, Pondicherry 605014, Puducherry, India Rashmi M, Kushveer JS, Sarma VV 2019 – A worldwide list of endophytic fungi with notes on ecology and diversity. Mycosphere 10(1), 798–1079, Doi 10.5943/mycosphere/10/1/19 Abstract Endophytic fungi are symptomless internal inhabits of plant tissues. They are implicated in the production of antibiotic and other compounds of therapeutic importance. Ecologically they provide several benefits to plants, including protection from plant pathogens. There have been numerous studies on the biodiversity and ecology of endophytic fungi. Some taxa dominate and occur frequently when compared to others due to adaptations or capabilities to produce different primary and secondary metabolites. It is therefore of interest to examine different fungal species and major taxonomic groups to which these fungi belong for bioactive compound production. In the present paper a list of endophytes based on the available literature is reported. More than 800 genera have been reported worldwide. Dominant genera are Alternaria, Aspergillus, Colletotrichum, Fusarium, Penicillium, and Phoma. Most endophyte studies have been on angiosperms followed by gymnosperms. Among the different substrates, leaf endophytes have been studied and analyzed in more detail when compared to other parts. Most investigations are from Asian countries such as China, India, European countries such as Germany, Spain and the UK in addition to major contributions from Brazil and the USA.
    [Show full text]
  • Fungal Biodiversity in Extreme Environments and Wood Degradation Potential
    http://waikato.researchgateway.ac.nz/ Research Commons at the University of Waikato Copyright Statement: The digital copy of this thesis is protected by the Copyright Act 1994 (New Zealand). The thesis may be consulted by you, provided you comply with the provisions of the Act and the following conditions of use: Any use you make of these documents or images must be for research or private study purposes only, and you may not make them available to any other person. Authors control the copyright of their thesis. You will recognise the author’s right to be identified as the author of the thesis, and due acknowledgement will be made to the author where appropriate. You will obtain the author’s permission before publishing any material from the thesis. Fungal biodiversity in extreme environments and wood degradation potential A thesis submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biological Sciences at The University of Waikato by Joel Allan Jurgens 2010 Abstract This doctoral thesis reports results from a multidisciplinary investigation of fungi from extreme locations, focusing on one of the driest and thermally broad regions of the world, the Taklimakan Desert, with comparisons to polar region deserts. Additionally, the capability of select fungal isolates to decay lignocellulosic substrates and produce degradative related enzymes at various temperatures was demonstrated. The Taklimakan Desert is located in the western portion of the People’s Republic of China, a region of extremes dominated by both limited precipitation, less than 25 mm of rain annually and tremendous temperature variation.
    [Show full text]
  • Fungal Pathogens of Proteaceae
    Persoonia 27, 2011: 20–45 www.ingentaconnect.com/content/nhn/pimj RESEARCH ARTICLE http://dx.doi.org/10.3767/003158511X606239 Fungal pathogens of Proteaceae P.W. Crous 1,3,8, B.A. Summerell 2, L. Swart 3, S. Denman 4, J.E. Taylor 5, C.M. Bezuidenhout 6, M.E. Palm7, S. Marincowitz 8, J.Z. Groenewald1 Key words Abstract Species of Leucadendron, Leucospermum and Protea (Proteaceae) are in high demand for the interna- tional floriculture market due to their brightly coloured and textured flowers or bracts. Fungal pathogens, however, biodiversity create a serious problem in cultivating flawless blooms. The aim of the present study was to characterise several cut-flower industry of these pathogens using morphology, culture characteristics, and DNA sequence data of the rRNA-ITS and LSU fungal pathogens genes. In some cases additional genes such as TEF 1- and CHS were also sequenced. Based on the results of ITS α this study, several novel species and genera are described. Brunneosphaerella leaf blight is shown to be caused by LSU three species, namely B. jonkershoekensis on Protea repens, B. nitidae sp. nov. on Protea nitida and B. protearum phylogeny on a wide host range of Protea spp. (South Africa). Coniothyrium-like species associated with Coniothyrium leaf systematics spot are allocated to other genera, namely Curreya grandicipis on Protea grandiceps, and Microsphaeropsis proteae on P. nitida (South Africa). Diaporthe leucospermi is described on Leucospermum sp. (Australia), and Diplodina microsperma newly reported on Protea sp. (New Zealand). Pyrenophora blight is caused by a novel species, Pyrenophora leucospermi, and not Drechslera biseptata or D.
    [Show full text]