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Fungi Imperfecti) UNIVERSITY OF ALBERTA EOLOMORPH STUDIES OF TEE MICROASCACEAE SEAN P. ABBOTT A thais submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the nquirements for the degree of Doctor of Philosophy DEPARTMENT OF BIOLOGICAL SCIENCES Edmonton, Aiberta, Canada SPRING 2000 National Library Bibliothbque nationale 141 of Canada du Canada Acquisitions and Acquisitions et Bibliographie Services seMces bibliographiques 395 WeUington Street 395,nie Wellington Ottawa ON KIA ON4 OttawaON K1AON4 canada Canada The author has granted a non- L'auteur a accordé une licence non exclusive licence allowing the exclusive permettant à la National Library of Canada to Bibliothèque nationale du Canada de reproduce, loan, distribute or seII reproduire, prêter, distriiuer ou copies of this thesis in microform, vendre des copies de cette thèse sous paper or electronic formats. la forme de microfiche/film, de reproduction sur papier ou sur format électronique. The author retains ownership of the L'auteur conserve la propriété du copyright in this thesis. Neither the droit d'auteur qui protège cette thése. thesis nor substantial extracts fiom it Ni la thèse ni des extraits substantiels may be printed or otherwise de celle-ci ne doivent êeimprimés reproduced without the author's ou autrement reproduits sans son permission. autorisation. For Linda "O know sweet love, I ahvays wrire of you So all my besr is hesing old worh new For as rhe sun is &i& new and old So is my lmaiII telling whis told" (W.Shakespeare, 1609) Morphological, molecular and physiological characters were used to reevaluate the Microasaceae (Microascales, Ascomycota) to include sexudy (teleomorphic) and asexuaiiy re producing (anamo rp hic) species. Anaiysis of sequences fiom small subunit (18s) rDNA ailowed integration of anamorphic taxa into the phylogenetic framework of / the Microascaceae and provided support for monophyly of the family. Life histones were investigated by growing the organisms in axenic culture and revealed that many species exhibited a suite of spore-forming stages that comprised the holomorph. Connections between sexuai and asexual States were made through mating trials. The discovery of the teleomorph of the common mold, Scopuiuriopsis brevicauks was significant because it connected the type species of the anamorphic genus Scopulariopsis with the Microascaceae. Mating trials among isolates of four species in the 'Sco~laripssis brevicaulis Series' demonstrated heterothallism and confirmed connections to teleomorphs in the genus Micrwscus. Single ascospore isolates demonstrated homothallism in Micr04scas nidicola. A reevaluation of definitive characters showed that the genus Pi~hmcuswas synonymous with Micrutzsms. The genus Cephalotrich was redescnbed for synnematous anamorphs of the Microascaceae with annellidic conidiogenesis and conidia in dry chahs, and included synonymy of Doratonryes, Stysamrs and Trichurus. Similarity of banding patterns fkom RFLP anaiysis of the ITS region of rDNA provided support for conspecificity among pleomorphic isolates of Pseu&illescheeria boydii. Tolerance to the antifùngal compound benomyl was consistent among 55 taxa of Microascaceae, and provided a simple test to separate microascaceous species Rom morphologically similar species in other pyrenomycete orders. The Microascaceae were emended to include the teleomorphic genera Microarcu, Pehielh, Pseu&llescherîu, Kemia and tophotrichus, and anarnorphs in the genera Scopdùtiopsis, Cephalotnchum, Ed>inobo+, Wwdomyces, Wardomycopsis, Grqhium and Scedosporim Two telwmorphs were desaibed, Micr- brmèauh and M. soppiii, and six new combinations were proposed, Micromais stoveri, Cephaotrichum coIum~is,C. cylinah'mm, C. dencirocephaum, C. putredimrs, and C. vitdis. Thirty three taxa of Microascaceae were reported frbm Alberta. I ACKNOWLEDGMENTS 1 am pleased to acknowledge Prof Lynne Sigler and Dr. Randy Currah for their supervision of this projed, their input of helpfùl suggestions, and their attention to my training. 1 am sincerely grateful for the guidance and enthsiastic encouragement they have provided throughout the course of this study. 1 wouid also like to thank my supe~sory cornmittee members, Dr. Dale Vitt and Dr. J.P. Tewari for their support and assistance during this program. Appreciation is express& to Arlene Flis and Linda Abbott, University of Alberta Microfungus Collection and Herbarium, for assistance with accessionhg of cultures and laboratory support, Ming Chen for assistance with SEM, GaWi Kemaghan for assistance with DNA extraction and RFLP analysis, Sarah Harnbleton for assistance with DNA sequenchg, Dr. Catherine Larfarge-England and Dr. Sean Graham for their help with sequence analyses, Markus Thorman for translation of German articles, and M. Hertwig-Jaksch for correcting the Latin. Curators and staff of the various culture collections who provided material for this study are sincerely thanked. In addition, Trevor Lumley, Trevor April and Markus Thorman provided cultures of Microascaceae, and their enthusiasm for Iengthy discussions and research collaboration is greatly appreciated. 1 would like to acknowledge the various fùnding agencies that supported this research Phary support was provided by a Natural Sciences and Engineering Research Council of Canada (NSERC) Post Graduate Scholarship and a Izaak Walton Killarn Memoial Scholarship. Field and laboratory work was supported by an award from the Challenge Grants in Biodiversity Program, jointly sponsored by the Alberta Conservation Association and the University of Alberta Department of Biological Sciences. I am pleased to acknowledge, with sincere thanks, the recognition and support received from other agencies during the course of my doctoral research including the Mywlogical Society of America (Myron Bakkus Award), the Canadian Botanical Association (Luella K. Weresub Award, John Macoun Travel Bursary), and the University of Alberta (Andrew Stewart Mernorial Graduate Prize, Walter H. Johns Graduate FeIIowship). Finaily, 1 must express my deepest gratitude to my de, Linda, who once again has provided the tremendous assistance needed to get me through a graduate research project, and without whose unfailhg support through the final stages, this project could not have been completed. Thank you for your love and companionship on our voyage 'into the grrat wide open'. TABLE OF CONTENTS CHAPTER 1. INTRODUCTION Introduction ................................ ......................................................................... 1 Research Rationale and Objectives ...................................................................... -3 Literature Cited .................................................................................................... 5 CHAPTER 2. MICROASCUS BREYICIUZIS SP. NOV., THE TELEOMORPH OF SCOPULAMOPSLT BREYIC4CIWS. SUPPORTS PLACEMENT OF SCOflULARlOPPSLT WITH THE MICROASCACEAE Introduction ...................................................................................................... 8 Materials and Methods ......................................................................................... 8 Results ............................. .. ................................................................................ 9 Discussion ................. .. ....................................................................................... 11 Literature Cited .................................................................................................. 17 CHAPTER 3. @TEROTHALLISM IN THE MICROASCACEAE DEMONSTRATED BY TEIREE SPECIES IN THE SCOPULARIOPSIS BREWC4 ULIS SERIES Introduction ......................... ............................................................................. 19 Materials and Methods ....................................................................................... 20 Taxonomie Part ........................... ,., .................................................................... 21 Holomorph Concepts ......................................................................................... 27 Literature Cited .................................................................................................. 35 CHAPTER 4. USE OF HOLOMORPH CEARACTERS TO DISTINGUISH M?CRO!CUSNIDICOLA AND MICROASCUS SOPPII SP. NOV., WITH NOTES ON TEE GENUS PmOASCUS Introduction ....................................~...~~.~..~........~.....................~.............~....~.....37 Materials and Methods ....................................................................................... 38 Taxonomic Part .................,.......................................................................... 39 Discussion. ...................................................................................................... 41 ...,... Literature Cited .................... ,, ...... .. ...,... .50 CHAPTER 5. RECOGNlTION OF CONSPECIFICITY AMONG PLEOMORPHIC ISOLATES OF PSEUDWC7HERL4 BODI' (MICROASCACEAE) CAPABLE OF DEGRADMG HYDROCARBONS IN CRUDE OIL Introduction .................... ,,, .............................................................................. 52 Materials and Methods ..................... ,,, ....~~...~.~.............................................53 Resuits ................................. ...,, ........................................................................ 54 Discussion ............ ,. ..,.. .., .............................................................................. 56 Literature Cited .. ..............,,,...... ,,,
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