AN ABSTRACT OF THE THESIS OF Abdulmagid M. Alsheikh for the degree of Doctor of Philosophy in Botany and Plant Pathology presented on July 8. 1994. Title: Taxonomy and Mycorrhizal Ecology of the Desert Truffles in the Genus Terfezia. Redacted for Privacy Abstract approved: James M. Trappe Mycorrhizal fungi in arid and semiarid rangelands have received little study in southwest Asia and north Africa, although they are important in food production and rangeland productivity. The desert truffles of the genera Terfezia and Tirmania form fruiting-bodies and mycorrhizae with rangeland plants. This thesis treats the taxonomy and the ecology of the genus Terfezia. A second part describes experimental studies of the mycorrhizal associations between two Tirmania and two Terfezia spp. and six annual and perennial spp. of Cistaceae. Terfezia has been found in all continents except Australia and Antarctica. Most species and specimens have been collected from north Africa to west and southwest Asia and southern Europe. Terfezia is characterized by smooth, brown to dark brown ascomata with a gleba of brown, fertile, globose to elongated pockets separated by paler, sterile but otherwise undifferentiated tramal veins; globose to subglobose, reniform or saccate asci with (3-) 5-8 (14) spores; and mostly globose, hyaline to golden brown spores prominently ornamented with spines, rods, cones, warts, papillae or reticula. Before this study, names had been proposed for 57 species, 9 varieties and 1 form within the genus Terfezia. This work retains 18 species and no varietal names. No collections were located of one species and five synonyms. Three new species are provisionally proposed. Most Terfezia species are widespread in desert habitats and rangelands except T. gigantea and T. terfezioides which grow in northern temperate, mesic forests. Spores of species growing in deserts are apparently dispersed by the abrasion of exposed ascomata, dried in-situ, by wind-blown sand or other storm systems. Desert animals may also participate in spore dispersal. Some Terfezia spp. have been demonstrated to form mycorrhizae with annual and perennial Helianthemum and other Cistaceae spp., and the mycorrhizal habit may be assumed for all. Mycorrhizae of 4 species of desert truffles and 6 annual and perennial species of Cistaceae were synthesized. Pure cultures originating from in-vitro spore germination of Terfezia boudieri, T. claveryi, Tirmania nivea and T. pinoyi served as inoculum. Hosts were Helianthemum ledifolium, H. salicifolium, H. kahiricum, H. lippii, Cistus albidus and Fumana procumbens. Not all fungus-host combinations formed the distinctive "helianthemum" type of mycorrhizae. Some ecological factors prevalent in desert habitats, but not included in the experimental design, proved to be crucial in the mycorrhiza synthesis. Considerations for future research on desert truffle mycorrhizae are proposed. Taxonomy and Mycorrhizal Ecology of the Desert Truffles in the Genus Terfezia by Abdulmagid M. Alsheikh A THESIS submitted to Oregon State University in partial fulfillment of the requirements for the degree of Doctor of Philosophy Completed July 8, 1994 Commencement June, 1995 APPROVED: Redacted for Privacy Professor of Botany and Plant Pathology in charge of major Redacted for Privacy Head of Department of Botany and Plant Pathology Redacted for Privacy C4­ Dean of Graduat hool Redacted for Privacy Date thesis is presented JULY 8th, 1994 Typed by: Ginny Bissel, Sheila Till, Sarah Berry, and Kim Osieczanek for Abdulmagid M. Alsheikh. DEDICATION In memory of my brothers, Abdulmonem and Zein-Al-Abedeen, and my grandmother, Latifah Jassem Almureikhi, who wanted to see this work completed ACKNOWLEDGMENTS My everlasting appreciation and gratitude to Dr. James M. Trappe, my mentor and friend, for placing his confidence in my ability, for his inspiration, infinite patience, guidance and encouragement over the years. Without his generous support and optimism this work would not have been possible. Dr. Trappe taught me to be a student of all truffles, not only those of the desert. I thank my advisors, Drs. William Denison, William Chilcote and Robert Linderman for their guidance and support, and for serving on my Ph.D. Committee. Dr. Randy Molina, leader of the Soil Microbial Ecology Team of the U.S. Department of Agriculture, Forest Service, Pacific Northwest Research Station, Forestry Sciences Laboratory, Corvallis, Oregon, graciously furnished space and facilities. His encouragement and support to finish this work is greatly appreciated. I thank Dr. Michael Castellano for his help with the final preparation of this thesis. Dr. Dan Luoma made logistical contributions and Chris Maser helped in editing of Chapter 3. I thank the Forest Service Soil Microbial Ecology Team and the other Pacific Northwest Research Station personnel for their support and encouragement, especially Darr M. Duff for preparation of excellent microscopic sections, and Drs. Mauro Martignoni, Earl Nelson, C. Y. Li, Iwan Ho, Charlie Sartwell, and Gody Spycher. Ginny Bissell, Sheila Till and Sarah Berry typed parts of this thesis. Wes Colgan and Ari Jumponnen helped me on illustration preparation. Laura D. de Toledo helped in preparation of plates, while Jane Smith assisted in preparing the final draft on the computer. I am grateful to the many herbaria which lent specimens. Mr. Louis Riousset sent French and Italian Terfezia specimens and Dr. Efren Cazares provided specimens from Mexico. Mr. Gerard Chevalier provided Terfezia and Tirmania cultures and some seeds. Dr. Ali Alraawi from Kuwait University sent me Helianthemum seeds from the Kuwait desert. Special thanks are due to the scholarship committee at the Kuwait Institute for Scientific Research (KISR), which provided financial support for my graduate studies. The active support and goodwill of many people in KISR is much appreciated, especially Drs. Homood Ergobah, Adnan Shihab-el-Din, Kazem Behbehani, Messrs. Nizar Hussein, Mohammed Alasfoor, Abdullah Alfadhel, Drs. Mohammed Al attaar, Dr. Najla Alnassaar, Dr. Samirah Omar, Dr. Adel Salman, Messrs. Mohammed Awameh, Mahmood Saeed, Miss Fayizah Alawadhi. Miss Hibah Alhosseini from the Kuwait University Office in the Embassy of Kuwait in Washington, D.C. was very supportive and understanding during the work on my thesis. My studies were also supported in part by the Kuwait Foundation for the Advancement of Science, Ministries of Education in the United Arab Emirates and the State of Bahrain, and the Embassy of the Kingdom of Saudi Arabia, Washington, D.C. Study of the western North American species was supported in part by National Science Foundation Grant BSR 9007186 to Drs. Nancy S. Weber, James M. Trappe and William C. Denison. I greatly appreciate the support and encouragement of my friends in Corvallis and at home who never let me give up over the years, especially Jalaal Alrumoh, Salman Alzayani, Mohammed A. Almishari, Abdul la J. Alnajadah, Abdullah Althuweikh, Mohammed Almijrin, Ali Alshehry, Ali A. Bamoudah, Eid A. Abdulaal, Mohammed F. Dosari, Fatma A. Douiri, Kareem Hamdy, Mohammed Alayan, Maysoon Salamah, Irene Hazou, Mona Matar, Ayman Asadi, Faten Sak-allah, Farha Elkindy and Kim Osieczanek. They and others made the years spent in Corvallis an experience to remember and cherish always. My former instructor in the University of Kuwait, Dr. Abdul aal H. Moubasher inspired me to study mycology and showed me what a fascinating field it is. This work could not have been finished without the moral and financial support of my family. My mother realized the importance of education in our better future and always encouraged me and my brothers and sisters to learn more. She stayed with me during the last part of my studies. My brother Taarik helped in typing and correcting the final draft and knew what it meant to be alone working to put a thesis together. All of my family suffered greatly during my absence from home. My extended family provided always their encouragement and support over the years. I thank them all, because since my early childhood I always enjoyed eating our desert truffles and wanted to know more about them. TABLE OF CONTENTS Page CHAPTER 1 : Introduction 1 Historical Background 1 The Names of Desert Truffles, Scientific and Otherwise 3 Considerations in Truffle Taxonomy 4 Use of Mycorrhizal Fungi in Desert Habitats 4 Effect of War on Truffle Habitat in Kuwait 5 Future Directions 8 Literature Cited 9 CHAPTER 2 : Desert Truffles: The Genus Terfezia 13 Abstract 14 Introduction 15 Taxonomic History 16 Ecology 18 Phylogeny 29 The Orders Tuberales and Pezizales 29 Families of the Order Pezizales 30 Phylogeny of the Genera of the family Terfeziaceae 31 Phylogeny of the Species of Terfezia 32 Considerations for a Suggested Hypothesis 34 Recent Chemical Methods in Studying Phylogeny 35 Utilization and Ethnobotany 39 Culture Trials and Spore Germination 43 Physiological Literature 44 Taxonomic Materials and Methods 46 Herbaria 48 Key to Genera and Species of Desert Truffles in Kuwait 50 Synopsis of the Genera of Terfeziaceae 51 The Genus Terfezia - Generic Diagnosis 52 Key to Species of Terfezia 53 Species Description 55 Terfezia arenaria 55 Terfezia arizonica 63 Terfezia austroafricana 68 Terfezia boudieri 72 Terfezia claveryi 80 Terfezia decaryi 90 Terfezia eremita 96 Terfezia gigantea 100 Terfezia indica 105 Terfezia lahorensis 110 Terfezia leptoderma 115 Terfezia longii 121 Terfezia metaxasi 125 Terfezia mexicana 130 Terfezia olbiensis 135 Terfezia pfeilii 142