SYSTEMATICS AND SPECIES DELIMITATION IN PESTALOTIA AND PESTALOTIOPSIS S.L. (AMPHISPHAERIALES, ASCOMYCOTA) Dissertation zur Erlangung des Doktorgrades der Naturwissenschaften vorgelegt beim Fachbereich 15 Biowissenschaften der Goethe -Universität in Frankfurt am Main von Caroline Judith-Hertz aus Darmstadt Frankfurt am Main, Oktober 2016 (D30) vom Fachbereich Biowissenschaften der Johann Wolfgang Goethe - Universität als Dissertation angenommen. Dekanin: Prof. Dr. Meike Piepenbring Gutachter: Prof. Dr. Meike Piepenbring Prof. Dr. Imke Schmitt Datum der Disputation: Contents 1 Abstract ....................................................................................................................... 1 2 Zusammenfassung ....................................................................................................... 3 3 List of abbreviations and symbols ............................................................................... 8 3.1 Abbreviations of herbaria and institutions ........................................................... 8 3.2 General abbreviations ........................................................................................... 8 3.3 Symbols .............................................................................................................. 10 4 Introduction ............................................................................................................... 11 4.1 Preface ................................................................................................................ 11 4.2 History of Pestalotia, Pestalotiopsis, and closely related genera ....................... 12 4.3 Pestalotiopsis s.l. ................................................................................................ 14 4.3.1 Systematic position ...................................................................................... 15 4.3.2 Morphology ................................................................................................. 16 4.3.3 The sexual morph ......................................................................................... 20 4.3.4 Ecology ........................................................................................................ 20 4.4 Morphological characters for species recognition of Pestalotiopsis s.l. ............ 21 4.5 Molecular phylogenetic studies of Pestalotiopsis s.l. ........................................ 23 4.6 Generic concepts in this study ............................................................................ 24 4.7 Aims of the study ................................................................................................ 25 5 Material and Methods ............................................................................................... 26 5.1 Specimen collection ............................................................................................ 26 5.2 Selection of herbarium material ......................................................................... 26 5.3 Identification of host plants ................................................................................ 27 5.4 Morphological methods ...................................................................................... 27 5.4.1 Light microscopy and morphometrics ......................................................... 27 5.4.2 Microscopic drawings and image capture .................................................... 28 5.4.3 Statistical analysis of morphological data .................................................... 28 I 5.5 Isolation and cultivation ..................................................................................... 29 5.5.1 Moist incubation chambers .......................................................................... 30 5.5.2 Induce sporulation ........................................................................................ 30 5.5.3 Culture media ............................................................................................... 30 5.6 Molecular methods ............................................................................................. 31 5.6.1 Marker selection .......................................................................................... 31 5.6.2 DNA Extraction, Amplification and Sequencing ........................................ 32 5.6.3 Phylogenetic analyses of Pestalotiopsis ...................................................... 36 5.6.4 Species delimitation in Pestalotiopsis ......................................................... 37 5.6.5 Phylogenetic analyses of Neopestalotiopsis ................................................ 37 5.6.6 Clade naming ............................................................................................... 38 5.6.7 Phylogenetic analyses of the proposed epitype of Pestalotia pezizoides .... 40 6 Results ....................................................................................................................... 43 6.1 Taxonomic results .............................................................................................. 43 6.1.1 Taxonomic results in the genus Neopestalotiopsis ...................................... 43 6.1.2 Taxonomic results in the genus Pestalotia .................................................. 45 6.1.3 Taxonomic results in the genus Pestalotiopsis ............................................ 53 6.1.4 Taxonomic results in the genus Pseudopestalotiopsis ................................. 68 6.1.5 Taxonomic results in the genus Truncatella ................................................ 69 6.2 Further species accounts from examined herbarium material ............................ 69 6.3 Phylogenetic analyses ......................................................................................... 78 6.3.1 DNA sequences ............................................................................................ 78 6.3.2 Model test and alignment properties ............................................................ 78 6.3.3 Phylogenetic analyses of Pestalotiopsis ...................................................... 78 6.3.4 Species delimitation analysis of Pestalotiopsis ........................................... 85 6.3.5 Clade naming ............................................................................................... 85 6.3.6 Pestalotiopsis spp. host association ............................................................. 87 6.3.7 Morphological species discrimination ......................................................... 87 II 6.3.8 Phylogenetic analyses of Neopestalotiopsis ................................................ 96 7 Discussion ................................................................................................................. 99 7.1 The importance of systematics ........................................................................... 99 7.2 Species concepts and delimitation in fungi ...................................................... 100 7.3 Species delimitation in Pestalotiopsis s.l. ........................................................ 102 7.4 Phylogenetic marker performance .................................................................... 106 7.5 Taxonomic species relationship within the Pestalotia-Pestalotiopsis complex 109 7.6 Naming species in Pestalotiopsis s.l. ............................................................... 110 Acknowledgements ....................................................................................................... 112 References ..................................................................................................................... 115 8 Appendices .............................................................................................................. 132 8.1 Eidesstattliche Versicherung ............................................................................ 150 8.2 Curriculum Vitae .............................................................................................. 151 8.3 Supplementary files on CD-ROM .................................................................... 155 List of Figures Figure 1: Morphological characters of Pestalotiopsis s.l. species .................................. 19 Figure 2: Map showing origin of samples used in this study. ......................................... 26 Figure 3: ML phylogenetic trees of members of Pestalotiopsidaceae showing the placements of Pestalotiopsis simitheae and P. coffeae-arabicae within the Pseudopestalotiopsis and Neopestalotiopsis clades, respectively ........................... 44 Figure 4: ML TEF1 single locus phylogenetic tree of members of Pestalotiopsidaceae showing the placements of Pestalotiopsis simitheae and P. coffeae-arabicae within the Pseudopestalotiopsis and Neopestalotiopsis clades, respectively ......... 45 Figure 5: Morphology of Pestalotia pezizoides (epitype) transferred to Seiridium ........ 47 Figure 6: ML phylogenetic tree for members of the Amphisphaeriales of a concatenated dataset of ITS and LSU sequences. ......................................................................... 52 III Figure 7: Morphology of Pestalotiopsis adusta (BPI 910046) ....................................... 57 Figure 8: Morphology of the lectotype of Pestalotiopsis guepinii (BPI 405955). .......... 65 Figure 9: Morphology of the proposed epitype of Pestalotiopsis maculans .................. 66 Figure 10: Morphology of Pestalotiopsis neglecta (isotype, BPI 406415) ...................