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Fungal Structure and Composition in Liverwort-Based Biocrust

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Fungal Structure and Composition in Liverwort-Based Biocrust Fungal structure and composition in liverwort-based biocrust Petra Landmark Guðmundsdóttir Faculty of Biology University of Iceland 2018 Uppbygging og samsetning sveppa í hélumosalífskurn Petra Landmark Guðmundsdóttir 90 eininga ritgerð sem er hluti af Magister Scientiarum gráðu í líffræði Leiðbeinandi Dr. Ólafur Sigmar Andrésson Meistaranefnd Dr. Ólafur Sigmar Andrésson Dr. Snædís Huld Björnsdóttir Prófdómari Dr. Edda Sigurdís Oddsdóttir Líf- og umhverfisvísindadeild Verkfræði- og náttúruvísindasvið Háskóli Íslands Reykjavík, Maí 2018 Fungal structure and composition in liverwort-based biocrust Petra Landmark Guðmundsdóttir 90 ECTS thesis submitted in partial fulfillment of a Magister Scientiarum degree in Biology Advisor Dr. Ólafur Sigmar Andrésson Master's committee Dr. Ólafur Sigmar Andrésson Dr. Snædís Huld Björnsdóttir Examiner Dr. Edda Sigurdís Oddsdóttir Faculty of Biology School of Engineering and Natural Sciences University of Iceland Reykjavik, May 2018 Fungal structure and composition in liverwort-based biocrust 90 ECTS thesis submitted in partial fulfillment of a Magister Scientiarum degree in Biology Copyright © 2018 Petra Landmark Guðmundsdóttir All rights reserved Faculty of Biology School of Engineering and Natural Sciences University of Iceland Sturlugata 7 (Askja) 101, Reykjavik Iceland Telephone: 525 4000 Bibliographic information: Petra Landmark Guðmundsdóttir, 2018, Fungal structure and composition in liverwort- based biocrust, Master’s thesis, Faculty of Biology, University of Iceland, pp. 93. Printing: Háskólaprent Reykjavik, Iceland, May 2018 Abstract Differences in fungal structure between depths (0, 5 and 20 mm) in Icelandic liverwort- based biocrust were determined using light microscopy and by quantification of fungal biomass by direct counting using fluorescence microscopy. Further, the fungal composition in the biocrust was determined and differences between depths (above 5 mm and below 5 mm) were estimated. The biocrust texture and the distribution of fungi differed between depths. The fungal biomass was found to be in a higher amount at the surface where large hyphae, sporangia and fungi within plants were more abundant than at 5 mm and 20 mm depth. Likewise, the composition revealed a difference between depths (above 5 mm and below 5 mm), both for the amount of total fungi, amount within phyla and proportions of phyla. The amount of total fungi was higher in samples above 5 mm. Ascomycota fungi were dominant at both levels while their proportion and amount was higher in samples above 5 mm were they accounted for 93-95%. Basidiomycota were higher in proportion in samples below 5 mm although their amount decreased. Exophiala, Phialocephala and Pseudogymnoascus were the most abundant genera. Útdráttur Sveppir í íslenskri hélumosalífskurn voru greindir með skoðun í ljóssmásjá og talningu í flúrsmásjá. Munur á uppbyggingu sveppa var metinn eftir dýpi (0, 5, 20 mm) og skoðuð samsetning hópa í lífskurninni ofan við 5 mm og neðan við 5 mm. Munur var á áferð lífskurnar og útbreiðslu sveppa eftir dýpi. Lífmassi sveppa var meiri við yfirborð (0 mm) þar sem breiðir sveppþræðir, gróhirslur og sveppir á og í plöntum voru í meira magni en á 5 mm og 20 mm dýpi. Samsetningin var jafnframt mismunandi eftir dýpi hvað varðar heildarmagn sveppa, magn einstakra fylkinga og hlutfall þeirra. Heildarmagn sveppa var hærra í sýnum ofan við 5 mm en neðar. Asksveppir voru ríkjandi í öllum sýnum, bæði ofan við 5 mm og neðan við 5 mm en þeir voru bæði í meira magni og hærra hlutfalli ofan við 5 mm þar sem þeir voru á bilinu 93-95% sveppa. Kólfsveppir voru í meira magni ofan við 5 mm en í hærra hlutfalli neðan við 5 mm. Exophiala, Pialocephala og Pseudogymnoascus voru algengastu ættkvíslirnar. iii Grazie per la pazienza e il supporto, amore mio. v Table of Contents Abstract ............................................................................................................................... iii Útdráttur ............................................................................................................................. iii Table of Contents .................................................................................................................vi List of Figures ................................................................................................................... viii List of Tables ......................................................................................................................... x Acknowledgements .............................................................................................................. xi 1 Introduction ...................................................................................................................... 1 1.1 Fungi......................................................................................................................... 1 1.1.1 Introduction to fungi ...................................................................................... 1 1.1.2 Fungal structure ............................................................................................. 1 1.1.3 Fungal reproduction ....................................................................................... 2 1.1.4 Fungal phyla .................................................................................................. 3 1.1.5 Soil fungi ....................................................................................................... 9 1.1.6 Fungal observation and biomass estimation .................................................. 9 1.2 Biological Soil Crust .............................................................................................. 10 1.2.1 What is biocrust? ......................................................................................... 10 1.2.2 Biocrust worldwide...................................................................................... 14 1.2.3 Biocrust in Iceland ....................................................................................... 15 1.2.4 Ecological roles ........................................................................................... 18 1.2.5 Fungi in biocrust .......................................................................................... 21 1.3 Objectives............................................................................................................... 24 2 Methods .......................................................................................................................... 25 2.1 Sample Collection .................................................................................................. 25 2.1.1 Study sites .................................................................................................... 25 2.1.2 Field setup and sample storage .................................................................... 29 2.2 Structural observation ............................................................................................ 30 2.2.1 Materials and equipment ............................................................................. 30 2.2.2 Preparation of lactophenol cotton blue mount ............................................ 30 2.2.3 70% alcohol ................................................................................................. 31 2.2.4 Slide preparation .......................................................................................... 31 2.2.5 Microscopic observation ............................................................................. 32 2.3 Cross sections stained with calcofluor white ......................................................... 32 2.3.1 Materials and equipment ............................................................................. 32 2.3.2 10% KOH .................................................................................................... 33 2.3.3 Slide preparation .......................................................................................... 33 vi 2.3.4 Observation and image analysis................................................................... 34 2.4 Direct counting by staining with calcofluor white ................................................ 34 2.4.1 Materials and equipment ............................................................................. 34 2.4.2 Slide preparation .......................................................................................... 35 2.4.3 Quantification of fungal biomass................................................................. 35 2.4.4 Estimation of variability in the quantification ............................................. 36 2.5 DAPI ...................................................................................................................... 36 2.5.1 Materials and equipment ............................................................................. 36 2.5.2 DAPI preparation ......................................................................................... 36 2.5.3 Slide preparation .......................................................................................... 37 2.5.4 Observation and image analysis................................................................... 37 2.6 Metagenome analysis ............................................................................................ 37 2.7 Statistical analysis ................................................................................................. 38 3 Results ...........................................................................................................................
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