Biological Species Concepts in Eastern North American Populations of Lentinellus Ursinus Andrew N
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Eastern Illinois University The Keep Masters Theses Student Theses & Publications 1997 Biological Species Concepts in Eastern North American Populations of Lentinellus ursinus Andrew N. Miller Eastern Illinois University This research is a product of the graduate program in Botany at Eastern Illinois University. Find out more about the program. Recommended Citation Miller, Andrew N., "Biological Species Concepts in Eastern North American Populations of Lentinellus ursinus" (1997). Masters Theses. 1784. https://thekeep.eiu.edu/theses/1784 This is brought to you for free and open access by the Student Theses & Publications at The Keep. It has been accepted for inclusion in Masters Theses by an authorized administrator of The Keep. For more information, please contact [email protected]. THESIS REPRODUCTION CERTIFICATE TO: Graduate Degree Candidates {who have written formal theses) SUBJECT: Permission to Reproduce Theses The University Library is receiving a number of requests from other institutions asking permission to reproduce dissertations for inclusion in their library holdings. Although no copyright laws are involved, we feel that professional courtesy demands that permission be obtained from the author before we allow theses to be copied. PLEASE SIGN ONE OF THE FOLLOWING STATEMENTS: Booth Library of Eastern Illinois University has my permission to lend my thesis to a reputable college or university for the purpose of copying it for inclusion in that institution's library or research holdings. Andrew N. Miller u~l.ff~ Author Date 7 I respectfully request Booth Library of Eastern Illinois University not allow my thesis to be reproduced because: Author Date Biological species concepts in eastern North American populations of Lentinellus ursinus (TITLE) BY Andrew N. Miller THESIS SUBMITIED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF Master of Science IN THE GRADUATE SCHOOL, EASTERN ILLINOIS UNIVERSITY CHARLESTON, ILLINOIS 1997 YEAR I HEREBY RECOMMEND THIS THESIS BE ACCEPTED AS FULFILLING THIS PART OF THE GRADUATE DEGREE CITED ABOVE ;.~ - I - <°f7 DATE );;i - / ' '1 '] DATE ABSTRACT As part of a biosystematic study of the Lentinellus ursinus species complex, field and herbarium specimens from North America, Central America, and Europe were examined. Single basidiospore isolates from basidiomata collected in Illinois, Indiana, Iowa, Michigan, Minnesota, Missouri, Tennessee, North Carolina, Wisconsin, Costa Rica, and Austria were crossed in all possible combinations to determine the mating system operating in each population. All populations exhibited bifactorial or tetrapolar mating systems. Tester strains were then crossed to identify intercompatible populations and to determine if morphological species in the L. ursinus species complex are congruent with biological species. Three biological species were recognized and identified as L. ursinus, L. angustifolius, and L. vulpinus. Polyspore isolates were subsequently used in culture mat and phenoloxidase analyses to facilitate identification of somatic cultures. The production of gloeocystidia and/or chlamydospores is taxonomically important for identifying somatic cultures of these species. All three taxa produced laccase, tyrosinase, and peroxidase in culture and were identified as white-rot fungi. This represents the first confirmed report of L. angustifolius from North America. DEDICATION To my mother and father 11 ACKNOWLEDGMENTS For all of her patience, love and encouragement throughout this study, I thank my fiancee, Jennifer. I wish to thank my parents for their endless support, both emotionally and financially. None of this would have been possible if it were not for their love and encouragement. I wish to thank my advisor, Dr. Andrew Methven, for introducing me to the world of mycology. I will always be grateful for all he has done for me and for his patience throughout this project. I also thank Dr. Jim McGaughey and Dr. Bryan Miller for serving on my committee and Dr. Charles Pederson for the use of his microscope. A special thank you is extended to Ors. Irmgard Krisai-Greilhuber, Ronald Petersen, Michael Tansey, and Thomas Volk for providing collections for this study. I thank the Botany Department at Eastern Illinois University for the use of their equipment and supplies throughout this study. I also thank the Lewis Hanford Botany Tiffany Fund at Eastern Illinois University and the Illinois State Academy of Science for providing financial support. Lastly, I thank my friends, Ken Magar, Dan Lloyd, and Dave Gregory, for providing me with the required recreational needs throughout this project. lll TABLE OF CONTENTS PAGE ABSTRACT . DEDICATION ii ACKNOWLEDGEMENTS iii LIST OF TABLES . v LIST OF FIGURES vi &vii I. INTRODUCTION 1 A. Taxonomic and Nomenclatural History . 2 B. Generic Description and Disposition of Taxa . 4 C. Lentinellus in Relation to Other Genera. 5 D. Mating Studies . 10 E. Culture Mat Analysis and Phenoloxidase Activity. 12 II. MATERIALS AND METHODS ............. 15 A. Monospore and Polyspore Isolates . 16 B. Self-crosses and Matings . 17 C. Culture Mat Analysis and Phenoloxidase Activity . 18 Ill. RESULTS ....................... .21 A. Description of Taxa . .21 B. Mating Studies. .33 C. Culture Mat Analysis and Phenoloxidase Activity . .34 IV. DISCUSSION . .40 A. Basidiome Morphology .40 B. Mating Studies ..... .41 C. Culture Mat Analysis and Phenoloxidase Activity. .42 V. SUMMARY ..... .47 VI. LITERATURE CITED .48 VII. APPENDICES. .53 A. Tables. 53 B. Figures .58 VIII. VITA .... .81 IV LIST OF TABLES TABLE PAGE I. Phenoloxidase reactions of L. ursinus culture mats at week II . 53 II. Phenoloxidase reactions of L. ursinus culture mats at week VI 53 Ill. Phenoloxidase reactions of L. angustifolius culture mats at week II . 54 IV. Phenoloxidase reactions of L. angustifolius culture mats at week VI. 54 V. Phenoloxidase reactions of L. vulpinus culture mats at week II . 55 VI. Phenoloxidase reactions of L. vulpinus culture mats at week VI . 55 VII. Sources of single basidiospore isolates used in mating studies . 56 VII. lntercompatibility relationships between monokaryotic tester strains representing three biological species within the L. ursinus complex from eastern North America, Austria (ANM 511 ), and Costa Rica (RHP 7880) . 57 v LIST OF FIGURES FIGURE PAGE 1. Self-cross of Lentinellus ursinus ANM 321. .58 2. Self-cross of Lentinellus ursinus ANM 482. .59 3. Self-cross of Lentinellus ursinus ANM 491. .60 4. Self-cross of Lentinellus ursinus ANM 497. 61 5. Additional pairings of Lentinellus ursinus ANM 497 to identify A2B1 mating type . .62 6. Self-cross of Lentinellus ursinus ANM 508. .63 7. Self-cross of Lentinellus ursinus RHP 221 O . .64 8. Self-cross of Lentinellus ursinus RHP 2414. 65 9. Self-cross of Lentinellus ursinus ASM 8027. .66 10. Self-cross of Lentinellus angustifolius ANM 492 .67 11. Additional pairings of Lentinellus ursinus ANM 492 to identify A1B2 mating type .......... .68 12. Self-cross of Lentinel/us angustifolius ANM 495 . .69 13. Additional pairings of Lentinellus ursinus ANM 495 to identify A182 mating type . .70 14. Self-cross of Lentinellus angustifolius RHP 2036 . 71 15. Self-cross of Lentinellus angustifolius RHP 8768 . .72 16. Self-cross of Lentinellus vulpinus RHP 7966 ... .73 17. Mating grid of L. ursinus (ANM 321, ANM 473, ANM 482, ANM 491, ANM 497) and L. vulpinus (RHP 7966) ........... 74 Vl 18. Mating grid of L. ursinus (ANM 508, RHP 2210, RHP 2414, ASM 8027, ASM 8109) and L. vulpinus (RHP 7966) . ..... 75 19. Mating grid of L. ursinus (ANM 321, ANM 473, ANM 482, ANM 491, ANM 497, ANM 508, RHP 2210, RHP 2414, ASM 8027, ASM 8109) . ....... .76 20. Mating grid of L. angustifolius (ANM 492, ANM 495, RHP 2036, RHP 8768) and L. vulpinus (RHP 7966) . .77 21. Mating grid of L. angustifolius (ANM 492, ANM 495, RHP 2036, RHP 8768) and L. ursinus (ANM 321, ANM 473, ANM 482, ANM 491, ANM 497) . 78 22. Mating grid of L. angustifolius (ANM 492, ANM 495, RHP 2036, RHP 8768) and L. ursinus (ANM 508, RHP 2210, RHP 2414, ASM 8027, ASM 8109) ... ....... 79 23. Mating grid of L. angustifolius from Austria (ANM 511 ), Costa Rica (RHP 7880) and eastern North America (ANM 492, ANM 495, RHP 2036, RHP 8768) . 80 Vll I. INTRODUCTION The genus Lentinellus Karsten (Agaricales, Tricholomataceae) includes a group of pleurotoid fungi most commonly coliected in late summer and fall in deciduous and coniferous forests throughout temperate regions. Although most species are found on decaying deciduous and coniferous wood, one species (Lentinellus vulpinus (Fr.) Kuhner & Maire) fruits on the wounds of living elm trees ( Ulmus spp.) and another (Lentinellus semivestitus (Pk.) Singer) has been collected on dead grass roots (Miller and Stewart, 1971 ). Although classical taxonomy has clearly delineated Lentinellus ursinus (Fr.) Kuhner from L. vulpinus, it has not accounted for additional cryptic taxa within the L. ursinus species complex and reveals nothing about reproductive compatibility between populations. Since preliminary data suggested that more than one biological species was represented in the L. ursinus species complex in North America, mating studies were proposed to identify sexually compatible and incompatible populations (i.e., biological species) in the L. ursinus species complex in eastern North America. The hypothesis of this thesis is that more than one biological species occurs in the L. ursinus species complex in eastern North America. The objectives were to: 1) determine the mating systems operating in this species