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A Survey of Ballistosporic Phylloplane Yeasts in Baton Rouge, Louisiana Louisiana State University LSU Digital Commons LSU Master's Theses Graduate School 2012 A survey of ballistosporic phylloplane yeasts in Baton Rouge, Louisiana Sebastian Albu Louisiana State University and Agricultural and Mechanical College, [email protected] Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_theses Part of the Plant Sciences Commons Recommended Citation Albu, Sebastian, "A survey of ballistosporic phylloplane yeasts in Baton Rouge, Louisiana" (2012). LSU Master's Theses. 3017. https://digitalcommons.lsu.edu/gradschool_theses/3017 This Thesis is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Master's Theses by an authorized graduate school editor of LSU Digital Commons. For more information, please contact [email protected]. A SURVEY OF BALLISTOSPORIC PHYLLOPLANE YEASTS IN BATON ROUGE, LOUISIANA A Thesis Submitted to the Graduate Faculty of the Louisiana Sate University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Master of Science in The Department of Plant Pathology by Sebastian Albu B.A., University of Pittsburgh, 2001 B.S., Metropolitan University of Denver, 2005 December 2012 Acknowledgments It would not have been possible to write this thesis without the guidance and support of many people. I would like to thank my major professor Dr. M. Catherine Aime for her incredible generosity and for imparting to me some of her vast knowledge and expertise of mycology and phylogenetics. Her unflagging dedication to the field has been an inspiration and continues to motivate me to do my best work. I am also very fortunate to have three other wonderful committee members-Dr. Meredith Blackwell, Dr. Ken Damann and Dr. Lowell Urbatsch- who have given me invaluable advice and have always been available when I have needed their assistance. Additionally, Dominik Begerow was extremely kind to provide several of his Kordyana sequences that were used in the analysis of the Exobasidiomycetes. I have been priveledged to work with a number of people in my lab who are not only great at what they do, but have taught me valuable tools which I have used in my research. I am grateful to Alistair McTaggart for his patience in teaching me about maximum likelihood and RAxML, Merje Toome and Geg Heller for their help with various molecular techniques and Ramandeep Kaur for making the lab run smoothly. Hector Urbina and Rebecca Melanson’s help with formatting and assembling this thesis is very much appreciated. My fellow students Tomas Rush, Andrew Rodriguez, Jorge Diaz, Youwen Gong, Don Nelsen, Rachel Koch and I have not only worked together, but our friendship extends outside of the lab and I know we will always feel a sense of deep camaraderie for one another. I would also like to extend many thanks to Paige Muse, Shiquita Brooks and Catherine Perino who have provided a great of help with culture work. I am also grateful to the Louisiana Board of Regents and for an NSF AFTOL grant for providing the funding for this research. Finally, I want to acknowledge my family for years of unconditional support, love and understanding. Some of my wildest speculations have resulted from late night conversations with my brother, Alex who also appreciates a wild edible mushroom on any hiking trip. Finally, I would like to thank my parents, Valeriu and Magdalena Albu. Mere words will never express the gratitude that I feel towards them and everything they have given me. ii Table of Contents Acknowledgments........................................................................................................................... ii Abstract ........................................................................................................................................... v Chapter 1. Literature Review .......................................................................................................... 1 1.1 Yeasts .................................................................................................................................... 1 1.2 The Phylloplane .................................................................................................................... 1 1.3 Ballistosporic Discharge ....................................................................................................... 2 1.4 Seasonal Studies.................................................................................................................... 3 1.5 Physiological Adaptations .................................................................................................... 4 1.6 Biodiversity ........................................................................................................................... 5 1.7 Yeast Systematics ................................................................................................................. 6 1.8 Objectives ............................................................................................................................. 7 Chapter 2. A Phylogenetic Analysis of Basidiomycete Yeasts Isolated from Seven Ferns ........... 8 2.1 Introduction ........................................................................................................................... 8 2.2 Methods and Materials .......................................................................................................... 9 2.2.1 Sample Collection and Yeast Isolation .......................................................................... 9 2.2.2 Polymerase Chain Reaction (PCR) and Sequencing ................................................... 10 2.2.3 Phylogenetic Analysis .................................................................................................. 10 2.2.4 Climate Data Collection ............................................................................................... 11 2.2.5 Data Analysis ............................................................................................................... 11 2.3 Results ................................................................................................................................. 11 2.3.1 Statistics ....................................................................................................................... 12 2.3.1.1 Chi Squares Goodness of Fit Test ......................................................................... 12 2.3.1.2 Analysis of Variance ............................................................................................. 13 2.3.2 Diversity of Yeasts by Class ........................................................................................ 13 2.3.3 Phylogenetic Analyses ................................................................................................. 25 2.4 Discussion ........................................................................................................................... 85 Chapter 3. Description of Two Yeasts in Ustilaginales ................................................................ 90 3.1 Introduction ......................................................................................................................... 90 3.2 Materials and Methods ........................................................................................................ 90 3.2.1 Sample Collection and Yeast Isolation ........................................................................ 90 3.2.2 Measurements and Light Microscopy .......................................................................... 91 3.2.3 Assimilation ................................................................................................................. 91 3.2.4 PCR Amplification and Sequencing ............................................................................ 91 3.2.5 Phylogenetic Analysis .................................................................................................. 92 3.3 Results ................................................................................................................................. 92 3.4 Taxonomy ........................................................................................................................... 96 3.4.1 Description of Farysia sp. nov. Albu, Rush and Aime ................................................ 96 3.4.2 Description of Pseudozyma sp. nov. Albu, Rush and Aime ........................................ 97 3.5 Discussion ........................................................................................................................... 98 iii Chapter 4. Conclusions ............................................................................................................... 101 References ................................................................................................................................... 103 Appendix A. Yeast Strain and Host Information ........................................................................ 116 Appendix B. Statistical Tables and Climate Information ........................................................... 128 Appendix C. Reference Information for Genbank Taxa ............................................................. 132 Vita .............................................................................................................................................. 151 iv Abstract A study documenting basidiomycete yeast biodiversity
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