The Taxonomy and Phylogeny of the Mycangial Fungi from Dendroctonus Brevicomis and D Frontalis
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Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 1996 The at xonomy and phylogeny of the mycangial fungi from Dendroctonus brevicomis and D frontalis (Coleoptera: Scolytidae) Portia Tang-Wung Hsiau Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Botany Commons, and the Plant Pathology Commons Recommended Citation Hsiau, Portia Tang-Wung, "The at xonomy and phylogeny of the mycangial fungi from Dendroctonus brevicomis and D frontalis (Coleoptera: Scolytidae) " (1996). Retrospective Theses and Dissertations. 11374. https://lib.dr.iastate.edu/rtd/11374 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. 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Ml 48106-1346 USA 313/761-4700 800/521-0600 The taxonomy and phylogeny of the mycangial fungi from Dendroctonus brevicomis and D. frontalis (Coleoptera: Scol)rtidae) ty Portia Tang-Wung Hsiau A dissertation submitted to the graduate faculty in partial fulfillment of the requirement for the degree of DOCTOR OF PHILOSOPHY Department: Plant Pathology Major: Plant Pathology Major Professor: Tliomas C. Harrington Iowa State University Ames, Iowa 1996 Copyright © Portia Tang-Wung Hsiau 1996. All rights reserved UNI Number: 9635324 UMIMicrofonn 9635324 Copyright 1996, by UMI Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. UMI 300 North Zeeb Road Ann Arbor, Ml 48103 ii Graduate College Iowa State University This is to certify that the Doctoral dissertation of Portia Tang-Wung Hsiau has met the dissertation requirement of Iowa Sate University Signature was redacted for privacy. Committee Member Signature was redacted for privacy. Committee Member Signature was redacted for privacy. Committee Membe^ Signature was redacted for privacy. Com^fd^tee/Member Signature was redacted for privacy. Major Professor Signature was redacted for privacy. For the Major Department Signature was redacted for privacy. For the Graduate College iii DEDICATION My great appredation is given to my major professor. Dr. Thomas C Harrington for his support, advice and direct contributions to my work. My sincere appreciation is given to my family members, especially my parents for their love and support I am sincerely grateful to Tosak Seelanan for his care and help during my stay at Ames. I would like to thank my friends, Joseph Steimel, Doug McNew, Rich Descenzo and Sharon Parker for their help and cooperation. Also, I thank Diana Six and Scott Kelley for their cooperation in providing materials. I would like to thank my committee members. Dr. Jonathan Wendel, Dr. Edward Braun, Dr. Sandy McNabb and Dr. Lois Tiffany for their assistance. I also appreciate the direct and indirect contributions of E}r. Thomas Bruns, Dr. David Hibbett, Dr. Nils Hallenberg and Dr. Meredith Blackwell. iv TABLE OF CONTENTS USTOFHGURES vi LIST OF TABLES vii ABSTRACT viii CHARTER 1. GENERAL INTRODUCTION 1 Introduction 1 Dissertation Organization 1 Literature Review 2 Literature Cited 5 CHARTER 2. CERATOCYSTIOPSIS BREVICOMISP. NOV., A MYCANGIAL FUNGUS FROM DENDROCTONUS BREVICOMIS (COLEOPTERA:SCOLYTIDAE) 10 Abstract 10 Introduction 10 Materials and Methods 11 Results 13 Discussion 17 Acknowledgment 19 Literature Cited 19 CHAPTERS. PHYLOGENY OF BASIDIOMYCETES ASSOCIATED WITH BARK AND AMBROSIA BEETLES (COLEOPTERA: SCOLYTIDAE) 40 Abstract 40 V Introduction 40 Materials and Methods 43 Results 45 Discussion 48 Acknowledgment 51 Literature Cited 51 CHARTER 4. DERIVATION OF A CLADE OF BARK BEETLE ASSOCIATED FUNGI FROM PENIOPHORA, A GENUS OF WOOD DECAYING BASIDIOMYCETES 60 Abstract 60 Introduction 61 Materials and Methods 62 Results 64 Discussion 68 Acknowledgment 72 Literature Cited 72 CHAPTER 5. GENERAL CONCLUSIONS 81 General Discussion 81 Literatiu-e Cited 82 APPENDDCA. AUGNED MT-SSU-RDNA SEQUENCES 84 APPENDIX B. AUGNED ITS SEQUENCES 92 APPENDIX C. AUGNED IGS SEQUENCES 96 vi USTOFnGURES Figs. 1-6. Anamorph of Ceratocystiopsis brevicomi. 1. Hyphae. 2. Conidia. 3-6. Conidia and conidiophores. 35 Figs. 7-13. Teieomorph of Ceratocystiopsis brevicomi. 7. Perithecial production (arrow) from the pairing of isolates C420 x C436 on pine twig medium in 50-mm-diam Petri dish. 8. Perithecia on twig. Top view (arrow) of a perithecium showing the collar-like structure and the protruding neck. 9-10. Perithecia. 11. Asci from a smashed perithecium. 12-13. Ascospores. 12. Young ascospores; some (arrow) still in asd. 13. Mature ascospores with bulbous swelling toward one end. 37 Fig. 14. Neighbor-joining tree generated from Rogers' genetic distance based on isozyme electromorphs of Ceratocystiopsis species. 39 Fig. 15. One of the most parsimonious trees generated from mt-ssu- rDNA sequences of bark and ambrosia beetle associates and other Holobasidiomycetes. (Tree length = 1025 steps, CI = 0.628, RI = 0.716; base substitutions are shown above branches and decay indices are shown in d values below branches.) Taxa closely associated with bark or ambrosia beetles are indicated by an asterisk. 59 Fig. 16. One of 36 most parsimonious trees from the internal transcribed spacers and 5.8S rDNA sequences of Peniophora and Entomocorticium species. Tree length = 224 steps, CI = 0.665, RI = 0.670; base substitutions are shown above branches and decay indices (d value) and bootstrap values {%) are shown below branches. 79 Fig. 17. One of three most parsimonious trees from the intergenic short spacer sequences of Peniophora and Entomocorticium species. Tree length = 103 steps, CI = 0.922, RI = 0.814; base substitutions are shown above branches and decay indices (d value) and bootstrap values (%) are shown below branches. 80 vii LIST OF TABLES Table L Enzymes, buffer systems and staiiung procedures used in starch gel electrophoresis studies of Ceratocystiopsis species. 23 Table n. Morphological characters of Ceratocystiopsis brevicomi, C. ranaculosus and C. collifera (^m). 25 Table IE. The production of perithecia and ascospores in crosses of MAT-1 and MAT-2 isolates of Ceratocystiopsis brevicomi. 27 Table IV. The production of perithecia and ascospores in crosses between Ceratocystiopsis ranaculosus and C. brevicomi isolates. 29 Table V. Isozyme electromorphs of isolates of Ceratocystiopsis brevicomi, C. ranaculosus, C. collifera and C. minuta. 31 Table VI. Fungal isolations (number) on 1% malt extract agar from surface-sterilized prothoraxes of female Dendroctonus brevicomis collected from the Central Sierra Nevada and Santa Barbara, California. 33 Table Vn. Fungal isolations (colony forming units/beetle) on 1% malt extract agar with cycloheximide and streptomycin sulfate from ground adult Dendroctonus brevicomis females and males from the Central Sierra Nevada of California. 34 Table VIE. Isolate number, beetle associates, substrates and location of origin of isolates used for DNA sequencing. 57 Table IX. Beetle associate, tree host and location of isolates of eight Entompcorticium species used in this study. 75 Table X. Morphological characteristics and growth rate of eight Entomocorticium species and E. dendroctoni. 77 viii ABSTRACT The mycangia of two closely related bark beetles, western pine beetle (Dendroctonus brevicomis, WPB) and southern pine beetle (D. frontalis, SPB), harbor similar fungi. Ceratocystiopsis brevicomi is described as new. It is the mycangial ascomycete of WPB and is distinct from C ranaculosus, a mycangial fungus of SPB and C. collifera, a fungus isolated &om pine tree attacked by D. valens in Mexico, based on isozyme analysis, mating tests and growth rate. Molecular phylogenetic analyses using DNA sequences of the mitochondrial small subunit rRNA (mt-ssu-rRNA) region, the internal transcribed spacers and the integenic short spacer suggested that the mycangial basidiomycetes of WPB and SPB are distinct but closely related