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Distribution of Vegetative Compatibility Groups in the Wood Decay Fungus, Phellopilus Nigrolimitatus at a Local Scale

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Distribution of Vegetative Compatibility Groups in the Wood Decay Fungus, Phellopilus Nigrolimitatus at a Local Scale Distribution of vegetative compatibility groups in the wood decay fungus, Phellopilus nigrolimitatus at a local scale Andreas Langeland Jenssen Master of Science Thesis Department of Biosciences Faculty of Mathematics and Natural Sciences University of Oslo 01.12.2017 © Andreas Langeland Jenssen 2017 Distribution of vegetative compatibility groups in the wood decay fungus, Phellopilus nigrolimitatus at a local scale. Andreas Langeland Jenssen http://www.duo.uio.no/ Trykk: Reprosentralen, Universitetet i Oslo II Abstract The boreal forests of Fennoscandia have gone through extensive structural changes and fragmentation due to forest management practices. One direct consequence is a significant reduction in dead wood availabilities and occurrence of wood dependent fungi. In Norway, there has been a considerable reduction in fungal diversity, with 29% of the evaluated fungal species lining up on the IUCN Red List. Fungi are important ecological contributors in nutrient recycling, symbiotic relationships and other biological processes in ecosystem functioning. There is still limited knowledge about the local population structure of wood-inhabiting fungi, including how they distribute and organize in single substrate units. Within substrates, fungi use self-nonself recogniction (allorecognition), also known as vegetative incompatibility, to separate own mycelia from others. Hence, this is a central mechanism for organization of mycelia at a local scale. In this thesis, I investigate the local population structure of the rare and red-listed fungus Phellopilus nigrolimitatus by in vitro vegetative incompatibility experiments. Among 321 samples collected from six late-stage decay Picea abies logs, I identify, delimit, and estimate size of 53 P. nigrolimitatus vegetative compatibility groups (VCGs). I demonstrate that the size of the poroid surface area of fruit bodies is positively correlated with the estimated size of VCGs. I find high phenotypic variation among the P. nigrolimitatus cultures in growth morphology and coloration, where cultural morphology and colouration is consistent within, but varied between, the VCGs. Hence, growth morphology and colouration can, to some extent, be used to determine VCG belonging in P. nigrolimitatus. Vegetative compatibility tests of cultures isolated from different logs reveals that VCGs were not distributed across logs, but that dispersal of VCGs is facilitated with substrate continuity. I suggest that vegetative dispersal of VCGs is short ranged and slow, due to probable reliance on substrate continuity and establishment success. III IV Acknowledgements Thanks to my supervisor Sandy Maurice for her tremendous help in pretty much every aspect of this journey. You have helped me gain knowledge and new skills, as well as providing me with valuable critique and guidance, for which I am very grateful. I am also thankful to my co- supervisor Håvard Kauserud for sharing his vast knowledge and experience with me, and his many helpful insights during discussions. Thanks to Jérôme Guerrand for his fantastic help in sampling in the field, and during the laboratory work. Also thanks to Anders Wollan for his great field work, as well as his pleasant company. I would also like to thank Einar Heegaard and Tom Andresen for their input on statistical procedures. I am also thankful to all my university friends and the rest of the pleasant people from the Oslo mycology group, who helped me with their input and helpful comments. V Contents 1 Introduction ....................................................................................................................1 2 Materials and methods ....................................................................................................6 2.1 Sampling ..................................................................................................................6 2.2 Laboratory work .......................................................................................................9 2.2.1 Fruit bodies measurement and image analysis ...................................................9 2.2.2 Media preparation .............................................................................................9 2.2.3 Culture isolation .............................................................................................. 10 2.2.4 Phenotypic characterization ............................................................................. 11 2.2.5 Vegetative compatibility tests .......................................................................... 11 2.3 Statistical analyses.................................................................................................. 15 3 Results .......................................................................................................................... 17 3.1 Wood sample and fruit body isolation success ........................................................ 17 3.2 Vegetative compatibility group location and size .................................................... 18 3.3 Consistency of fruit body measurements................................................................. 22 3.4 Fruit body size, abundance and their respective VCG correlation ........................... 23 3.4.1 Effect of subsampling the surface area dataset ................................................. 23 3.4.2 Fruit body surface area and VCG size.............................................................. 25 3.4.3 Effect of subsampling the fruit body abundance dataset ................................... 27 3.4.4 Fruit body abundance and VCG size ................................................................ 28 3.5 Phenotypic variation and VCG delimitation ............................................................ 29 3.6 Common VCGs across separated wood logs ........................................................... 33 4 Discussion .................................................................................................................... 34 4.1 Isolation success of Phellopilus nigrolimitatus ....................................................... 34 4.2 Vegetative compatibility groups in Phellopilus nigrolimitatus ................................ 34 4.2.1 How are VCGs distributed within logs? ........................................................... 34 4.2.2 Are differences in size of VCGs reflecting individual fitness? ......................... 36 4.2.3 Variation in nonself-recognition ...................................................................... 36 4.3 Fruit body size, abundance and their respective VCG correlation ........................... 37 4.4 Are distinct phenotypes predicting individuals? ...................................................... 38 4.5 Small scale dispersal .............................................................................................. 40 5 Concluding remarks and future work ............................................................................ 41 VI References ............................................................................................................................ 43 Appendix .............................................................................................................................. 48 Appendix 1: Defined phenotypic groups ........................................................................... 48 Appendix 2: Measurements correlation ............................................................................. 54 Appendix 3 Fruit body surface area subset plot ................................................................. 56 Appendix 4: Fruit body abundance model ......................................................................... 57 Table of figures and tables Figure 1. Map of Norway and Skotjernfjell. .......................................................................6 Figure 2 An example of the sampling design. ......................................................................7 Table 1. Overview of number of wood samples and fruit bodies collected from six Norway spruce logs. ..............................................................................................................8 Equation 1. Binomial coefficient. ....................................................................................... 11 Figure 3. Diagram of vegetative compatibility test dishes................................................. 12 Figure 4. Vegetative compatibility tests. ............................................................................ 13 Figure 5. Compatibility testing procedure. ........................................................................ 14 Figure 6. Isolation success. ................................................................................................. 17 Figure 7. Vegetative incompatibility groups on log 5. ....................................................... 18 Table 2 summary of vegetative compatibility groups in all logs. ...................................... 19 Figure 8. Cumulative richness of VCGs in Phellopilus nigrolimitatus. ............................. 21 Figure 9. Linear regression models for fruit body surface area and diagnostic plot. ...... 22 Figure 10. Comparison of surface area from ImageJ. ...................................................... 23 Table 3 Analysis of deviance output from (fruit body surface area) linear mixed effect models (LMM). ..................................................................................................................
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