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Stability of Soil Microbial Communities to Applications of the Fungal Biological Control Agent Metarhizium Brunneum

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Stability of Soil Microbial Communities to Applications of the Fungal Biological Control Agent Metarhizium Brunneum Research Collection Doctoral Thesis Stability of soil microbial communities to applications of the fungal biological control agent Metarhizium brunneum Author(s): Mayerhofer, Johanna Publication Date: 2017 Permanent Link: https://doi.org/10.3929/ethz-b-000244602 Rights / License: In Copyright - Non-Commercial Use Permitted This page was generated automatically upon download from the ETH Zurich Research Collection. For more information please consult the Terms of use. ETH Library DISS. ETH Nr. 24581 STABILITY OF SOIL MICROBIAL COMMUNITIES TO APPLICATIONS OF THE FUNGAL BIOLOGICAL CONTROL AGENT METARHIZIUM BRUNNEUM a dissertation submitted to attain the degree of DOCTOR OF SCIENCES of ETH ZURICH (Dr. sc. ETH Zurich) presented by JOHANNA MAYERHOFER MSc Mikrobiologie, Karl-Franzens Universität Innsbruck born on 03.04.1988 citizen of Austria accepted on the recommendation of Prof. Dr. Adrian Leuchtmann Prof. Dr. Bruce McDonald Dr. Jürg Enkerli Prof. Dr. Stefan Vidal 2017 CONTENT Content Content .................................................................................................................................................................... 3 Summary ................................................................................................................................................................. 7 Zusammenfassung ................................................................................................................................................... 9 General introduction ......................................................................................................................................11 1.1 The click beetles Agriotes spp. .............................................................................................................. 11 Distribution and damage potential of Agriotes spp. ........................................................................ 11 Life cycle of Agriotes spp. .............................................................................................................. 12 Control options against Agriotes spp. .............................................................................................. 13 1.2 The Western corn rootworm Diabrotica virgifera virgifera ................................................................. 14 Distribution and damage potential of D. v. virgifera ....................................................................... 14 Life cycle of D. v. virgifera ............................................................................................................. 15 Control options against D. v. virgifera ............................................................................................ 15 1.3 Soil microorganisms .............................................................................................................................. 16 Diversity and function of soil microorganisms ............................................................................... 16 Assessing soil microbial diversity ................................................................................................... 18 Response of soil microbial communities to disturbances ................................................................ 19 1.4 The biocontrol agent and entomopathogenic fungus Metarhizium ....................................................... 20 Biological control using entomopathogenic fungi and its history ................................................... 20 The entomopathogenic fungus Metarhizium ................................................................................... 21 1.4.2.1 Ecology and life cycle of the entomopathogenic fungus Metarhizium ................................... 21 1.4.2.2 Taxonomy and species concept in fungi with focus on Metarhizium ..................................... 22 1.4.2.3 Phylogeny of Metarhizium ..................................................................................................... 23 1.4.2.4 Detection and identification of Metarhizium strains ............................................................... 24 Non-target effects of Metarhizium .................................................................................................. 25 1.5 Objectives .............................................................................................................................................. 27 1.6 Outline of the thesis .............................................................................................................................. 28 Multiplexed microsatellite markers for seven Metarhizium species ..............................................................29 2.1 Abstract ................................................................................................................................................. 29 2.2 Introduction ........................................................................................................................................... 29 2.3 Material and methods ............................................................................................................................ 30 3 CONTENT 2.4 Results and discussion ........................................................................................................................... 30 2.5 Acknowledgements ............................................................................................................................... 33 Assessing effects of the entomopathogenic fungus Metarhizium brunneum on soil microbial communities in Agriotes spp. biological pest control ......................................................................................................................35 3.1 Abstract ................................................................................................................................................. 35 3.2 Introduction ........................................................................................................................................... 35 3.3 Material and methods ............................................................................................................................ 37 Rearing of Agriotes obscurus larvae ............................................................................................... 37 Treatments ....................................................................................................................................... 37 Set-up of the pot experiment ........................................................................................................... 38 Set-up of the field experiment ......................................................................................................... 38 Processing of soil samples, isolation of Metarhizium CFU and identification of applied strain ..... 39 DNA extraction from soil, PCR and Illumina sequencing .............................................................. 40 ITS2 sequence of the applied strain ................................................................................................ 41 Sequence processing and taxonomic classification ......................................................................... 41 Statistical analyses .......................................................................................................................... 41 3.4 Results ................................................................................................................................................... 42 Abundance of the applied Metarhizium strain and efficacy of biocontrol treatments in pots ......... 42 Abundance of the applied Metarhizium strain and efficacy of biocontrol treatments in the field ... 44 Soil microbial communities of the pots ........................................................................................... 45 Abundance of the applied strain and effects of treatments on soil microorganisms in pots ............ 45 Changes of the microbial communities over time in pots ............................................................... 48 Soil microbial communities in the field .......................................................................................... 48 Abundance of the applied strain and effects of treatments on microbial communities in the field . 49 Changes of the microbial communities over time and space in the field ........................................ 49 3.5 Discussion ............................................................................................................................................. 51 3.6 Funding ................................................................................................................................................. 54 3.7 Acknowledgments ................................................................................................................................. 54 Response of soil microbial communities to the application of a formulated Metarhizium brunneum biocontrol strain .......................................................................................................................................................................55 4.1 Abstract ................................................................................................................................................. 55 4.2 Introduction ..........................................................................................................................................
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