DISSERTATION ADVANCES IN TORTRICID SYSTEMATICS AND IDENTIFICATION (LEPIDOPTERA: TORTRICIDAE) Submitted by Todd Michael Gilligan Department of Bioagricultural Sciences and Pest Management In partial fulfillment of the requirements For the Degree of Doctor of Philosophy Colorado State University Fort Collins, Colorado Fall 2012 Doctoral Committee: Advisor: Paul Opler Co-Advisor: Boris Kondratieff Terrence Walters John Brown Mark Simmons Copyright by Todd Michael Gilligan 2012 All Rights Reserved ii    ABSTRACT ADVANCES IN TORTRICID SYSTEMATICS AND IDENTIFICATION (LEPIDOPTERA: TORTRICIDAE) The Tortricidae is a large family in the microlepidoptera, consisting of more than 10,300 species in 1,070 genera. Many tortricids are considered to be economically important, and the family contains the highest percentage of pest species in the Lepidoptera. This dissertation provides significant advances in tortricid systematics and identification through the implementation of modern technologies such as matrix-based identification keys and molecular phylogenetics. Chapter 2 focuses on Tortricidae threatening U.S. agriculture. The resulting resource, Tortricids of Agricultural Importance (TortAI – http://www.tortai.org), is designed to aid in the identification of tortricid adults encountered during domestic surveys and tortricid larvae encountered during quarantine inspections at U.S. ports of entry. Chapters 3–5 provide details on the three tortricid pests currently of greatest importance to U.S. agriculture: Epiphyas postvittana (Walker), Lobesia botrana ([Denis and Schiffermüller]), and Thaumatotibia leucotreta (Meyrick). Chapter 6 details the discovery of a new tortricid species with the potential to threaten avocado production in California. Chapter 7 describes the benefits and disadvantages to using DNA barcoding for identification purposes and outlines a novel DNA sequence search tool developed for use in TortAI. Phylogenetic relationships surrounding Eucosma, the largest genus within the Tortricidae, are examined in chapters 8–9. A molecular phylogeny and revised classification for Eucosma and related genera provide the groundwork for future study of these highly diverse taxa. iii    ACKNOWLEDGMENTS This dissertation and all of its associated publications would not have been possible without the assistance of numerous individuals, all of whom I will attempt to list here. The contributions and efforts of everyone are greatly appreciated. I would like to thank my CSU committee for all of their assistance during my time at CSU. Paul Opler and Boris Kondratieff were great advisors and provided constant support, advice, and expertise. Terrence Walters provided funding, supervised cooperative agreements, and the majority of this dissertation would not have been possible without his support. John Brown was a wealth of knowledge for everything related to the Tortricidae and provided access to the excellent collection at the Smithsonian. Mark Simmons provided lab and office space, assistance with all aspects of the phylogenetic analyses presented here, and valuable advice. Special thanks to my home committee, Ella, Tara, and Vixen, for their constant support, patience, and barking. In addition to moving to Colorado and working so that I could play with bugs for the past five years, Ella also helped with web design, databasing, and writing code for the DNA sequence search tool. Thanks to my parents, Mike and Thora, for continued support and encouragement. I would especially like to thank my father for assistance with field work, specimen preparation, and collecting equipment. Marc Epstein, CDFA Sacramento, travelled the world with me collecting tortricids during extended trips to Europe, Australia, and Africa. I would like to thank him for all of his help, despite getting us kidnapped by nuns, eaten by alive by leeches, and almost killed by elephants. Norman Barr, USDA/CPHST Mission Lab, helped with all sequencing projects. I want to thank him for hosting me several times at his lab and for all of his valuable assistance. iv    I would like to thank everyone, past and present, in the Simmons’ lab, including Jennifer Cappa, Luke Tembrock, Jenna McAleer, Salem Eleague, Christine Bacon, Kendra Yakobson, and Jack Munz, for putting up with me in the office, working or helping in the lab, and generally making my time at CSU enjoyable. Thanks to Donald Wright for helping with all aspects of Eucosmini research, providing specimens, and coauthoring numerous publications. Many individuals in BSPM provided valuable assistance though teaching support and help with logistics, paperwork, and account management. Special thanks to Janet Dill, Maggie Hirko, Paula White, Cynthia Brown, Andrew Norton, and Tom Holtzer for all of their help. The following individuals and institutions assisted with, provided logistics for, or provided permits for field work in various countries: Leif Aarvik, Natural History Museum, University of Oslo, Norway; David Agassiz, Weston-Super-Mare, England; Joaquin Baixeras Almela, Institut Cavanilles de Biodiversitat i Biologia Evolutiva, Universitat de Valencia, Spain; Gianfranco Anfora, Claudio Ioriatti, and Valerio Mazzoni, IASMA, S. Michele all'Adige, Italy; Bruno Bagnoli, Consiglio per la Ricerca e la Sperimentazione in Agricoltura, Firenze, Italy; Frans Groenen, Luyksgestel, Netherlands; Bobbie Hitchcock, CSIRO Ecosystem Sciences, Australian National University, Canberra; Marianne Horak, Australian National Insect Collection, CSIRO, Canberra; Margaret Kinnaird, Mpala Research Centre, Kenya; Esther Kioko, Grace Ngugi, and J. Mugambi Ruthiiri, National Museums of Kenya, Nairobi; Andrea Lucchi, University of Pisa, Italy; Max Moulds, David Rentz, and Paul Zborowski, Kuranda, Queensland, Australia; Gionata Pulignani, Mazzei, Castello di Fonterutoli, Castellina in Chianti, Italy; Queensland Parks and Wildlife, Cairns, Queensland; Angus Simpson, Gilgil, Kenya; Marja van der Straten, Plant Protection Service, Wageningen, Netherlands; Daniel Strickman and Rene Sforza, USDA/ARS/European Biological Control Laboratory, Montferrier-sur-Lez, France. The following persons and institutions provided loans of specimens or donated specimens for morphological study and/or DNA sequencing: John Brown, USDA/ARS/SEL, v    Smithsonian, Washington D.C.; Bong-Kyu Byun, Department of Biological Sciences, Hannam University, Korea; Soowon Cho, Department of Plant Medicine, Chungbuk National University, Korea; Jamie Davies, Catherine Young, and Graeme Anderson, Department of Primary Industries, Parks, Water and Environment, Tasmania, Australia; Jason Dombroskie, Department of Entomology, Cornell University, Ithaca; Ted Edwards, Bobbie Hitchcock, and Marianne Horak, Australian National Insect Collection, CSIRO, Canberra; Sabine Gaal-Haszler, Naturhistorisches Museum Wien, Vienna; Roberto Gonzalez, Universidad de Chile, Santiago, Chile; Frans Groenen, Luyksgestel, Netherlands; Mark Hoddle and Doug Yanega, Department of Entomology, University of California, Riverside; Esther Kioko and J. Mugambi Ruthiiri, National Museums of Kenya, Nairobi; Eric LaGasa, Washington State Department of Agriculture, Olympia; Steven Passoa, USDA/APHIS/PPQ, Columbus, Ohio; Jerry Powell, University of California, Essig Museum of Entomology, Berkeley; Obie Sage, California Department of Food and Agriculture, Sacramento; Marja van der Straten, Plant Protection Service, Wageningen, Netherlands; Luis Miguel Torres-Vila, Servicio de Sanidad Vegetal, Spain; Kevin Tuck, The Natural History Museum, London; Richard Worth, Oregon Department of Agriculture, Salem; Doug Yanega, University of California, Riverside. The following individuals and institutions assisted with or provided facilities for DNA sequencing, or provided sequence data used in the DNA sequence search tool: Norman Barr and Lisa Ledezma, USDA/APHIS/PPQ/CPHST, Mission Lab, Edinburg; BOLD Data Systems, Biodiversity Institute of Ontario; Jonathan Brusch, USDA/APHIS/PPQ, Boston; Paul Hebert, University of Guelph, Ontario; Bobbie Hitchcock, CSIRO Ecosystem Sciences, Australian National University, Canberra; Peter Kerr, Scott Kinnee, and Obie Sage, California Department of Food and Agriculture, Sacramento; Richard Newcomb and Leah Tooman, The New Zealand Institute for Plant & Food Research Limited, Auckland, New Zealand; Rodolphe Rougerie, Universite de Rouen, France; Mark Simmons, Colorado State University, Fort Collins; Kendra Yakobson and Jack Munz, formally Colorado State University, Fort Collins. vi    The following persons provided special assistance or access to equipment: Kevin Hoffman and Obie Sage, California Department of Food and Agriculture, Sacramento; Steven Passoa, USDA/APHIS/PPQ, Columbus; Amanda Redford and Christina Southwick, ITP - USDA/APHIS/PPQ/CPHST, Fort Collins. Funding was provided by USDA/APHIS/PPQ/CPHST through the following two cooperative agreements with Colorado State University and California Department of Food and Agriculture (principle investigators: T. M. Gilligan and P. A. Opler): “Diagnostic tools for tortricid moths of immediate and future concern to United States agriculture” ($154,400; January, 2009– January, 2012) and “Diagnostic tools for the identification of western U.S. Tortricidae easily confused with the light brown apple moth” ($42,680; March, 2008–March, 2009). Cooperative agreements were supervised by Terrence Walters, Identification Technology Program, USDA/APHIS/PPQ/CPHST, Fort Collins. vii    TABLE OF CONTENTS ABSTRACT ................................................................................................................................... ii ACKNOWLEDGMENTS ............................................................................................................... iii TABLE OF CONTENTS ............................................................................................................... vii LIST OF TABLES ........................................................................................................................ xiii LIST OF FIGURES ..................................................................................................................... xvi CHAPTER 1: GENERAL INTRODUCTION .................................................................................. 1 Tortricidae ......................................................................................................................... 1 Economic importance ........................................................................................................ 1 Systematics ....................................................................................................................... 3 Summary ........................................................................................................................... 4 References ........................................................................................................................ 5 CHAPTER 2: TORTRICIDS OF AGRICULTURAL IMPORTANCE TO THE UNITED STATES (LEPIDOPTERA: TORTRICIDAE) ................................................................................................ 6 Introduction ....................................................................................................................... 6 Materials and methods .................................................................................................... 10 TortAI adult key ............................................................................................................... 12 Adult key instructions .......................................................................................... 13 TortAI larval key .............................................................................................................. 17 Larval key instructions ......................................................................................... 18 Species accounts ............................................................................................................ 21 Tortricidae: Tortricinae: Tortricini ......................................................................... 21 Acleris Hübner ..................................................................................................... 21 Acleris comariana (Lienig & Zeller)...................................................................... 22 Acleris flavivittana (Clemens) .............................................................................. 24 Acleris gloverana (Walsingham) .......................................................................... 25 Acleris hastiana (Linnaeus) ................................................................................. 27 Acleris holmiana (Linnaeus) ................................................................................ 29 Acleris minuta (Robinson) ................................................................................... 31 Acleris nivisellana (Walsingham) ......................................................................... 33 Acleris robinsoniana (Forbes) ............................................................................. 35 Acleris senescens (Zeller) ................................................................................... 36 Acleris variana (Fernald) ..................................................................................... 37 Acleris variegana (Denis & Schiffermüller) .......................................................... 39 Tortricidae: Tortricinae: Cochylini ........................................................................ 41 Eupoecilia ambiguella (Hübner) .......................................................................... 43 Henricus umbrabasana (Kearfott) ....................................................................... 45 Tortricidae: Tortricinae: Cnephasiini .................................................................... 47 Cnephasia longana (Haworth) ............................................................................. 48 Decodes basiplagana (Walsingham) ................................................................... 51 viii    Decodes fragariana (Busck) ................................................................................ 52 Tortricidae: Tortricinae: Archipini ......................................................................... 53 Adoxophyes Meyrick ........................................................................................... 54 Adoxophyes furcatana (Walker) .......................................................................... 55 Adoxophyes honmai Yasuda ............................................................................... 56 Adoxophyes negundana (McDunnough) ............................................................. 58 Adoxophyes orana (Fischer von Roslerstamm) .................................................. 59 Archips Hübner .................................................................................................... 62 Archips argyrospila (Walker) ............................................................................... 63 Archips cerasivorana (Fitch) ................................................................................ 67 Archips crataegana (Hübner) .............................................................................. 70 Archips fuscocupreanus Walsingham ................................................................. 72 Archips grisea (Robinson) ................................................................................... 75 Archips mortuana (Kearfott) ................................................................................ 76 Archips podana (Scopoli) .................................................................................... 78 Archips purpurana (Clemens).............................................................................. 81 Archips rosana (Linnaeus) .................................................................................. 83 Archips semiferanus (Walker) ............................................................................. 86 Archips xylosteana (Linnaeus) ............................................................................ 88 Argyrotaenia Stephens ........................................................................................ 90 Argyrotaenia franciscana (Walsingham) .............................................................. 91 Argyrotaenia ljungiana (Thunberg) ...................................................................... 97 Argyrotaenia mariana (Fernald)......................................................................... 100 Argyrotaenia quadrifasciana (Fernald) .............................................................. 102 Argyrotaenia quercifoliana (Fitch)...................................................................... 103 Argyrotaenia velutinana (Walker) ...................................................................... 105 Cacoecimorpha pronubana (Hübner) ................................................................ 108 Choristoneura occidentalis (Walsingham) ......................................................... 111 Choristoneura rosaceana (Harris) ..................................................................... 113 Clepsis Guenée ................................................................................................. 117 Clepsis clemensiana (Fernald) .......................................................................... 118 Clepsis fucana (Walsingham) ............................................................................ 119 Clepsis melaleucanus (Walker) ......................................................................... 122 Clepsis peritana (Clemens) ............................................................................... 123 Clepsis persicana (Fitch) ................................................................................... 125 Clepsis spectrana (Treitschke) .......................................................................... 128 Clepsis virescana (Clemens) ............................................................................. 131 Ctenopseustis obliquana (Walker) ..................................................................... 132 Epichoristodes acerbella (Walker) ..................................................................... 134 Epiphyas postvittana (Walker) (see Chapter 3) ................................................. 136 Isotenes miserana (Walker)............................................................................... 136 Pandemis cerasana (Hübner) ........................................................................... 138 Pandemis heparana (Denis & Schiffermüller) ................................................... 142 Pandemis lamprosana (Robinson) .................................................................... 145 Pandemis limitata (Robinson) ........................................................................... 147 Pandemis pyrusana (Kearfott) ........................................................................... 150 Planotortrix excessana (Walker) [complex] ....................................................... 152 Xenotemna pallorana (Robinson) ...................................................................... 154 Tortricidae: Tortricinae: Sparganothini .............................................................. 156 Amorbia Clemens .............................................................................................. 156 Amorbia cuneana (Walsingham) ....................................................................... 157 ix    Amorbia emigratella Busck ................................................................................ 159 Platynota Clemens ............................................................................................ 162 Platynota flavedana Clemens ............................................................................ 163 Platynota idaeusalis (Walker) ............................................................................ 165 Platynota stultana Walsingham ......................................................................... 167 Sparganothis senecionana (Walsingham) ......................................................... 170 Sparganothis sulfureana (Clemens) .................................................................. 172 Sparganothis tunicana (Walsingham) ............................................................... 175 Tortricidae: Tortricinae: Euliini ........................................................................... 177 Proeulia and related species ............................................................................. 177 Seticosta rubicola Brown & Nishida................................................................... 183 Tortricidae: Olethreutinae: Microcorsini ............................................................. 185 Cryptaspasma Walsingham .............................................................................. 185 Cryptaspasma bipenicilla Brown and Brown ..................................................... 186 Cryptaspasma perseana Gilligan and Brown (see Chapter 6) .......................... 187 Tortricidae: Olethreutinae: Bactrini .................................................................... 187 Endothenia hebesana (Walker) ......................................................................... 187 Tortricidae: Olethreutinae: Olethreutini ............................................................. 190 Lobesia botrana ([Denis & Schiffermüller]) (see Chapter 4) .............................. 190 Paralobesia Obraztsov ...................................................................................... 190 Paralobesia viteana (Clemens) ......................................................................... 191 Tortricidae: Olethreutinae: Eucosmini ............................................................... 193 Crocidosema Zeller ........................................................................................... 193 Crocidosema aporema (Walsingham) ............................................................... 194 Crocidosema plebejana Zeller ........................................................................... 196 Spilonota ocellana ([Denis & Schiffermüller]) .................................................... 199 Tortricidae: Olethreutinae: Enarmoniini ............................................................. 202 Enarmonia formosana (Scopoli) ........................................................................ 202 Tetramoera schistaceana (Snellen)................................................................... 204 Tortricidae: Olethreutinae: Grapholitini .............................................................. 205 Cryptophlebia Walsingham ............................................................................... 205 Cryptophlebia illepida (Butler) ........................................................................... 206 Cryptophlebia ombrodelta (Lower) .................................................................... 209 Cryptophlebia peltastica (Meyrick) .................................................................... 212 Cydia Hübner .................................................................................................... 214 Cydia araucariae (Pastrana) ............................................................................. 215 Cydia caryana (Fitch) ........................................................................................ 216 Cydia coniferana (Saxesen) .............................................................................. 217 Cydia deshaisiana (Lucas) ................................................................................ 219 Cydia fabivora (Meyrick) .................................................................................... 220 Cydia fagiglandana (Zeller) ............................................................................... 222 Cydia garacana (Kearfott) ................................................................................. 224 Cydia glandicolana (Danilevsky) ....................................................................... 225 Cydia kurokoi (Amsel) ....................................................................................... 226 Cydia latiferreana (Walsingham) ....................................................................... 228 Cydia pomonella (Linnaeus) .............................................................................. 229 Cydia splendana (Hübner) ................................................................................ 232 Cydia strobilella (Linnaeus) ............................................................................... 234 Cydia toreuta (Grote) [complex] ........................................................................ 235 Ecdytolopha Zeller ............................................................................................. 237 Ecdytolopha insiticiana Zeller ............................................................................ 238 x    Grapholita Treitschke ........................................................................................ 239 Grapholita delineana (Walker) ........................................................................... 240 Grapholita funebrana (Treitschke) ..................................................................... 242 Grapholita interstinctana (Clemens) .................................................................. 244 Grapholita molesta (Busck) ............................................................................... 245 Grapholita packardi (Zeller) ............................................................................... 248 Grapholita prunivora (Walsingham) ................................................................... 250 Gymnandrosoma Dyar ...................................................................................... 252 Gymnandrosoma aurantianum Lima ................................................................. 253 Gymnandrosoma punctidiscanum Dyar ............................................................ 255 Pammene fasciana (Linnaeus) .......................................................................... 256 Talponia batesi (Heinrich) ................................................................................. 258 Thaumatotibia leucotreta (Meyrick) (see Chapter 5) ......................................... 259 Plates .......................................................................................................................... 260 Adults ................................................................................................................ 261 Male genitalia .................................................................................................... 290 Female genitalia ................................................................................................ 304 References .................................................................................................................... 321 CHAPTER 3: THE DISCOVERY OF LIGHT BROWN APPLE MOTH, EPIPHYAS POSVITTANA (WALKER) IN CALIFORNIA (LEPIDOPTERA: TORTRICIDAE) .............................................. 334 Introduction ................................................................................................................... 334 Nomenclature ................................................................................................................ 335 Geographic distribution ................................................................................................. 335 Larval hosts and damage .............................................................................................. 336 Life cycle and biology .................................................................................................... 336 Morphology ................................................................................................................... 341 Discovery of LBAM in California .................................................................................... 347 Conclusions ................................................................................................................... 349 References .................................................................................................................... 350 CHAPTER 4: DISCOVERY OF LOBESIA BOTRANA ([DENIS & SCHIFFERMÜLLER]) IN CALIFORNIA: AN INVASIVE SPECIES NEW TO NORTH AMERICA (LEPIDOPTERA: TORTRICIDAE) ........................................................................................................................ 353 Introduction ................................................................................................................... 353 Materials and methods .................................................................................................. 354 Results and discussion ................................................................................................. 356 Nomenclature .................................................................................................... 356 Geographic distribution ..................................................................................... 356 Larval hosts and damage .................................................................................. 357 Life cycle and biology ........................................................................................ 359 Morphology ........................................................................................................ 361 Discovery of Lobesia botrana in California ........................................................ 370 Conclusions ................................................................................................................... 371 References .................................................................................................................... 373 xi    CHAPTER 5: DISCOVERY OF FALSE CODLING MOTH, THAUMATOTIBIA LEUCOTRETA (MEYRICK), IN CALIFORNIA (LEPIDOPTERA: TORTRICIDAE) ............................................ 377 Introduction ................................................................................................................... 377 Materials and methods .................................................................................................. 378 Results and discussion ................................................................................................. 379 Nomenclature and distribution ........................................................................... 379 Larval hosts and biology .................................................................................... 380 Morphology ........................................................................................................ 382 Discovery of Thaumatotibia leucotreta in California .......................................... 386 Conclusions ................................................................................................................... 387 References .................................................................................................................... 389 CHAPTER 6: A NEW AVOCADO PEST IN CENTRAL AMERICA (LEPIDOPTERA: TORTRICIDAE) WITH A KEY TO LEPIDOPTERA LARVAE THREATENING AVOCADOS IN CALIFORNIA .......................................................................................................................... 392 Introduction ................................................................................................................... 392 Materials and methods .................................................................................................. 394 Description .................................................................................................................... 395 Cryptaspasma (Cryptaspasma) perseana Gilligan and Brown .......................... 395 Discussion ..................................................................................................................... 404 Amorbia spp. (Tortricidae: Tortricinae) .............................................................. 406 Argyrotaenia franciscana (Walsingham) (Tortricidae: Tortricinae) .................... 407 Cacoecimorpha pronubana (Hübner) (Tortricidae: Tortricinae) ......................... 407 Epiphyas postvittana (Walker) (Tortricidae: Tortricinae) ................................... 408 Caloptilia, Marmara, and Phyllocnistis spp. (Gracillariidae: Gracillariinae, Phyllocnistinae) ................................................................................................. 408 Platynota stultana Walsingham (Tortricidae: Tortricinae) .................................. 408 Sabulodes aegrotata (Guenée) (Geometridae: Ennominae) ............................. 409 Stenoma catenifer Walsingham (Elachistidae: Stenomatinae) .......................... 409 Thaumatotibia leucotreta (Meyrick) (Tortricidae: Olethreutinae) ....................... 410 Other Lepidoptera pests .................................................................................... 410 Key to Lepidoptera larvae threatening avocado in California ........................................ 410 References .................................................................................................................... 413 CHAPTER 7: A SEQUENCE SEARCH TOOL FOR THE IDENTIFICATION OF LEPIDOPTERA THROUGH DNA BARCODING ................................................................................................ 418 Introduction ................................................................................................................... 418 Public barcode databases ............................................................................................. 421 NCBI-GenBank .................................................................................................. 421 BOLD ................................................................................................................. 422 Species-specific tools .................................................................................................... 425 DNA barcoding for regulatory decisions ........................................................................ 426 Tortricid DNA sequence search tool ............................................................................. 428 Materials and methods .................................................................................................. 428 Results .......................................................................................................................... 430 Instructions .................................................................................................................... 434 Conclusions ................................................................................................................... 438 References .................................................................................................................... 439 xii    CHAPTER 8: THE TYPE SPECIES OF EUCOSMA HÜBNER (LEPIDOPTERA: TORTRICIDAE: EUCOSMINI) .......................................................................................................................... 443 Introduction ................................................................................................................... 443 Materials and methods .................................................................................................. 447 Discussion and results .................................................................................................. 448 Descriptions and redescriptions .................................................................................... 451 Eucosma circulana Hübner ............................................................................... 451 Eucosma gemellana Heinrich, new status ......................................................... 455 Eucosma paragemellana Gilligan & Wright, new species ................................. 459 Eucosma fraudabilis Heinrich ............................................................................ 461 References .................................................................................................................... 465 CHAPTER 9: MOLECULAR PHYLOGENY OF EUCOSMA HÜBNER AND RELATED GENERA (LEPIDOPTERA: TORTRICIDAE: EUCOSMINI): A REVISED CLASSIFICATION AND UPDATED WORLD CHECKLIST ............................................................................................. 468 Introduction ................................................................................................................... 468 Methods ........................................................................................................................ 473 Taxon sampling ................................................................................................. 473 Morphological characters .................................................................................. 473 Molecular methods ............................................................................................ 474 Data analysis ..................................................................................................... 475 Results .......................................................................................................................... 477 Process partitions .............................................................................................. 478 Systematic implications ..................................................................................... 480 Discussion ..................................................................................................................... 481 Generic descriptions and redescriptions ....................................................................... 487 Eucosma ........................................................................................................... 487 Pelochrista ......................................................................................................... 493 Eucopina ........................................................................................................... 498 Phaneta ............................................................................................................. 502 Epiblema ........................................................................................................... 505 References .................................................................................................................... 508 APPENDICES Appendix I: Morphological character states .................................................................. 513 Appendix II: List of sampled specimens ........................................................................ 548 Appendix III: Morphological characters and character states........................................ 552 Appendix IV: Supplementary trees ................................................................................ 556 Appendix V: Revised World Catalogue of Eucopina, Eucosma, Pelochrista, and Phaneta .................................................................................................................. 562 Eucopina ........................................................................................................... 563 Eucosma ........................................................................................................... 564 Pelochrista ......................................................................................................... 583 Phaneta ............................................................................................................. 600 Sonia ................................................................................................................. 601 Zeiraphera ......................................................................................................... 601 Eucosmini unplaced .......................................................................................... 601 xiii    LIST OF TABLES CHAPTER 2 Table 1: Primary target species ........................................................................................ 8 Table 2: Secondary target species .................................................................................... 8 Table 3: Acleris comariana host plants ........................................................................... 24 Table 4: Acleris flavivittana host plants ........................................................................... 25 Table 5: Acleris gloverana host plants ............................................................................ 27 Table 6: Acleris hastiana host plants .............................................................................. 29 Table 7: Acleris holmiana host plants ............................................................................. 31 Table 8: Acleris minuta host plants ................................................................................. 33 Table 9: Acleris nivisellana host plants ........................................................................... 34 Table 10: Acleris robinsoniana host plants ..................................................................... 36 Table 11: Acleris senescens host plants ......................................................................... 37 Table 12: Acleris variana host plants .............................................................................. 38 Table 13: Acleris variegana host plants .......................................................................... 41 Table 14: Eupoecilia ambiguella host plants ................................................................... 45 Table 15: Henricus umbrabasana host plants ................................................................. 46 Table 16: Cnephasia longana host plants ....................................................................... 49 Table 17: Decodes basiplagana host plants ................................................................... 51 Table 18: Decodes fragariana host plants ...................................................................... 53 Table 19: Adoxophyes furcatana host plants .................................................................. 55 Table 20: Adoxophyes honmai host plants ..................................................................... 57 Table 21: Adoxophyes negundana host plants ............................................................... 59 Table 22: Adoxophyes orana host plants ........................................................................ 61 Table 23: Archips argyrospila host plants ....................................................................... 65 Table 24: Archips cerasivorana host plants .................................................................... 69 Table 25: Archips crataegana host plants ....................................................................... 71 Table 26: Archips fuscocupreanus host plants ............................................................... 73 Table 27: Archips grisea host plants ............................................................................... 76 Table 28: Archips mortuana host plants .......................................................................... 78 Table 29: Archips podana host plants ............................................................................. 80 Table 30: Archips purpurana host plants ........................................................................ 82 Table 31: Archips rosana host plants .............................................................................. 84 Table 32: Archips semiferanus host plants ..................................................................... 87 Table 33: Archips xylosteana host plants ........................................................................ 89 Table 34: Argyrotaenia franciscana host plants .............................................................. 93 Table 35: Argyrotaenia ljungiana host plants .................................................................. 99 Table 36: Argyrotaenia mariana host plants ................................................................. 101 Table 37: Argyrotaenia quadrifasciana host plants ....................................................... 103 Table 38: Argyrotaenia quercifoliana host plants .......................................................... 104 Table 39: Argyrotaenia velutinana host plants .............................................................. 107 Table 40: Cacoecimorpha pronubana host plants ........................................................ 110 Table 41: Choristoneura occidentalis host plants ......................................................... 113 Table 42: Choristoneura rosaceana host plants ........................................................... 115 Table 43: Clepsis clemensiana host plants ................................................................... 119 Table 44: Comparison of Clepsis species. .................................................................... 120 Table 45: Clepsis fucana host plants ............................................................................ 121 Table 46: Clepsis melaleucanus host plants ................................................................. 123 xiv    Table 47: Clepsis peritana host plants .......................................................................... 125 Table 48: Clepsis persicana host plants ....................................................................... 126 Table 49: Clepsis spectrana host plants ....................................................................... 130 Table 50: Clepsis virescana host plants ........................................................................ 132 Table 51: Ctenopseustis obliquana host plants ............................................................ 133 Table 52: Epichoristodes acerbella host plants ............................................................. 135 Table 53: Isotenes miserana host plants ...................................................................... 137 Table 54: Comparison of Pandemis species. ................................................................ 140 Table 55: Pandemis cerasana host plants .................................................................... 141 Table 56: Pandemis heparana host plants .................................................................... 143 Table 57: Pandemis lamprosana host plants ................................................................ 147 Table 58: Pandemis limitata host plants ....................................................................... 149 Table 59: Pandemis pyrusana host plants .................................................................... 151 Table 60: Planotortrix excessana host plants ............................................................... 153 Table 61: Xenotemna pallorana host plants .................................................................. 155 Table 62: Amorbia cuneana host plants ........................................................................ 158 Table 63: Amorbia emigratella host plants .................................................................... 160 Table 64: Platynota flavedana host plants .................................................................... 164 Table 65: Platynota idaeusalis host plants .................................................................... 166 Table 66: Platynota stultana host plants ....................................................................... 168 Table 67: Sparganothis senecionana host plants ......................................................... 171 Table 68: Sparganothis sulfureana host plants ............................................................. 174 Table 69: Sparganothis tunicana host plants ................................................................ 176 Table 70: Proeulia and related species host plants ...................................................... 181 Table 71: Seticosta rubicola host plants ....................................................................... 184 Table 72: Cryptaspasma bipenicilla host plants ............................................................ 187 Table 73: Endothenia hebesana host plants ................................................................. 188 Table 74: Paralobesia viteana host plants .................................................................... 193 Table 75: Crocidosema aporema host plants ............................................................... 195 Table 76: Crocidosema plebejana host plants .............................................................. 198 Table 77: Spilonota ocellana host plants ...................................................................... 200 Table 78: Enarmonia formosana host plants ................................................................ 203 Table 79: Tetramoera schistaceana host plants ........................................................... 205 Table 80: Comparison of Cryptophlebia species. ......................................................... 207 Table 81: Cryptophlebia illepida host plants ................................................................. 208 Table 82: Cryptophlebia ombrodelta host plants ........................................................... 211 Table 83: Cryptophlebia peltastica host plants ............................................................. 214 Table 84: Cydia araucariae host plants ......................................................................... 216 Table 85: Cydia caryana host plants ............................................................................. 217 Table 86: Cydia coniferana host plants ......................................................................... 218 Table 87: Cydia deshaisiana host plants ...................................................................... 220 Table 88: Cydia fabivora host plants ............................................................................. 222 Table 89: Cydia fagiglandana host plants ..................................................................... 223 Table 90: Cydia garacana host plants ........................................................................... 224 Table 91: Cydia glandicolana host plants ..................................................................... 226 Table 92: Cydia kurokoi host plants .............................................................................. 227 Table 93: Cydia latiferreana host plants ........................................................................ 229 Table 94: Cydia pomonella host plants ......................................................................... 231 Table 95: Cydia splendana host plants ......................................................................... 233 Table 96: Cydia strobilella host plants .......................................................................... 235 Table 97: Cydia toreuta host plants .............................................................................. 236 xv    Table 98: Ecdytolopha insiticiana host plants ............................................................... 239 Table 99: Grapholita delineana host plants ................................................................... 241 Table 100: Grapholita funebrana host plants ................................................................ 244 Table 101: Grapholita interstinctana host plants ........................................................... 245 Table 102: Grapholita molesta host plants .................................................................... 247 Table 103: Grapholita packardi host plants ................................................................... 250 Table 104: Grapholita prunivora host plants ................................................................. 251 Table 105: Gymnandrosoma aurantianum host plants ................................................. 254 Table 106: Gymnandrosoma punctidiscanum host plants ............................................ 256 Table 107: Pammene fasciana host plants ................................................................... 258 Table 108: Talponia batesi host plants ......................................................................... 259 CHAPTER 3 Table 109: Partial list of E. postvittana host plants ....................................................... 337 CHAPTER 4 Table 110: Documented host plants for L. botrana ....................................................... 357 CHAPTER 5 Table 111: Documented host plants for T. leucotreta ................................................... 380 CHAPTER 6 Table 112: Tortricidae reported to feed on avocado ..................................................... 405 CHAPTER 7 Table 113: COI primers used in this study .................................................................... 429 CHAPTER 8 Table 114: Pattern elements present in Hübner’s E. circulana illustration scored for candidate species (Pe. = Pelochrista; Ph. = Phaneta) .................................................. 450 CHAPTER 9 Table 115: Data matrix and tree statistics. CI = ensemble consistency index (Kluge and Farris, 1969) on the most parsimonious trees for the parsimony-informative characters. RI = ensemble retention index (Farris 1989). *Bootstrap statistics calculated from the combined molecular likelihood tree. .............................................................................. 477 xvi    LIST OF FIGURES CHAPTER 2 Figure 1. Tortricinae; a–e: Acleris comariana; f–i: Acleris flavivittana; j–l: Acleris gloverana. ........................................................................................ 261 Figure 2. Tortricinae; a: Acleris gloverana; b–i: Acleris hastiana; j–k: Acleris holmiana; l: Acleris minuta. ................................................................................................ 262 Figure 3. Tortricinae; a: Acleris minuta; b–c: Acleris nivisellana; d–g: Acleris robinsoniana; h–k: Acleris senescens; l: Acleris variana. .................................. 263 Figure 4. Tortricinae; a–b: Acleris variana; c–f: Acleris variegana; g–h: Tortrix viridana; i–j: Eupoecilia ambiguella; k–l: Henricus umbrabasana. ................................... 264 Figure 5. Tortricinae; a: Henricus umbrabasana; b–e: Cnephasia longana; f–g: Decodes basiplagana; h–j: Decodes fragariana; k–l: Adoxophyes furcatana. .................. 265 Figure 6. Tortricinae; a: Adoxophyes furcatana; b–d: Adoxophyes negundana; e–j: Adoxophyes orana; k–l: Archips argyrospila. .............................................. 266 Figure 7. Tortricinae; a–b: Archips argyrospila; c–e: Archips cerasivorana; f–i: Archips crataegana; j–l: Archips fuscocupreanus........................................................... 267 Figure 8. Tortricinae; a–c: Archips grisea; d–e: Archips mortuana; f–j: Archips podana; k–l: Archips purpurana. ..................................................................................... 268 Figure 9. Tortricinae; a–c: Archips purpurana; d–g: Archips rosana; h–k: Archips semiferanus; l: Archips xylosteana. ................................................................... 269 Figure 10. Tortricinae; a–b: Archips xylosteana; c–f: Argyrotaenia franciscana; g–i: Argyrotaenia ljungiana; j–l: Argyrotaenia mariana. ............................................ 270 Figure 11. Tortricinae; a–c: Argyrotaenia quadrifasciana; d–e: Argyrotaenia quercifoliana; f–h: Argyrotaenia velutinana; i–l: Cacoecimorpha pronubana. ... 271 Figure 12. Tortricinae; a: Cacoecimorpha pronubana; b–d: Choristoneura occidentalis; e–h: Choristoneura rosaceana; i–j: Clepsis clemensiana; k–l: Clepsis fucana. ........................................................................................... 272 Figure 13. Tortricinae; a: Clepsis fucana; b–d: Clepsis melaleucanus; e–h: Clepsis peritana; i–l: Clepsis spectrana. ........................................................................ 273 Figure 14. Tortricinae; a–c: Clepsis virescana; d–g: Ctenopseustis obliquana; h–j: Epichoristodes acerbella; k–l: Epiphyas postvittana. .................................. 274 Figure 15. Tortricinae; a–b: Epiphyas postvittana; c–d: Isotenes miserana; e–h: Pandemis cerasana; i–l: Pandemis heparana. .................................................. 275 Figure 16. Tortricinae; a–d: Pandemis lamprosana; e–g: Pandemis limitata; h–j: Pandemis pyrusana; k–l: Planotortrix excessana. ............................................. 276 Figure 17. Tortricinae; a: Planotortrix excessana; b–c: Xenotemna pallorana; d–g: Amorbia cuneana; h–k: Amorbia emigratella; l: Platynota flavedana. ............... 277 Figure 18. Tortricinae; a–c: Platynota flavedana; d–g: Platynota idaeusalis; h–j: Platynota stultana; k–l: Sparganothis senecionana. .......................................... 278 Figure 19. Tortricinae; a–b: Sparganothis senecionana; c–e: Sparganothis sulfureana; f–h: Sparganothis tunicana; i–j: Accuminulia buscki; k–l: Chileulia stalactitis. .. 279 Figure 20. Tortricinae; a–b: Proeulia auraria; c–e: Proeulia chrysopteris; f–g: Proeulia triquetra; h: Proeulia apospasta; i–k: Seticosta rubicola. ................................... 280 Figure 21. Olethreutinae; a–b: Cryptaspasma bipenicilla; c–d: Cryptaspasma perseana; e: Endothenia hebesana; f–h: Lobesia botrana; i–j: Paralobesia viteana; k: Paralobesia lireodendrana; l: Paralobesia monotropana. .............................. 281 xvii    Figure 22. Olethreutinae; a: Paralobesia palliolana; b: Paralobesia rhoifructana; c: Paralobesia yaracana; d: Crocidosema aporema (photo credit Joaquín Baixeras Almela); e–h: Crocidosema plebejana; i–k: Spilonota ocellana; l: Enarmonia formosana. ........................................................................................................ 282 Figure 23. Olethreutinae; a–b: Enarmonia formosana; c–d: Tetramoera schistaceana; e–g: Cryptophlebia illepida; h–j: Cryptophlebia ombrodelta; k–l: Cryptophlebia peltastica. .......................................................................................................... 283 Figure 24. Olethreutinae; a: Cryptophlebia peltastica; b–c: Cydia araucariae; d–e: Cydia caryana; f–h: Cydia coniferana; i–j: Cydia deshaisiana; k–l: Cydia fabivora. .... 284 Figure 25. Olethreutinae; a–c: Cydia fagiglandana; d: Cydia garacana; e–g: Cydia glandicolana; h–i: Cydia kurokoi; j: Cydia latiferreana; k–l: Cydia pomonella. .. 285 Figure 26. Olethreutinae; a: Cydia pomonella; b–f: Cydia splendana; g–i: Cydia strobilella; j: Cydia toreuta; k–l: Ecdytolopha insiticiana. ................................... 286 Figure 27. Olethreutinae; a: Ecdytolopha insiticiana; b: Ecdytolopha mana; c: Ecdytolopha occidentana; d: Grapholita delineana; e–f: Grapholita funebrana; g: Grapholita interstinctana; h–i: Grapholita molesta; j–l: Grapholita packardi. ..... 287 Figure 28. Olethreutinae; a–b: Grapholita prunivora; c–e: Gymnandrosoma aurantianum; f–h: Gymnandrosoma punctidiscanum; i–k: Pammene fasciana; l: Talponia batesi. .............................................................................................. 288 Figure 29. Olethreutinae; a: Talponia batesi; b–f: Thaumatotibia leucotreta................. 289 Figure 30. Male genitalia; a: A. comariana; b: A. flavivittana; c: A. gloverana; d: A. hastiana; e: A. holmiana; f: A. minuta; g: A. nivisellana; h: A. robinsoniana. ............................................................................................ 290 Figure 31. Male genitalia; a: A. senescens; b: A. variana; c: A. variegana; d: T. viridana; e: E. ambiguella; f: H. umbrabasana; g: C. longana; h: D. basiplagana. ........... 291 Figure 32. Male genitalia; a: D. fragariana; b: A. furcatana; c: A. negundana; d: A. orana; e: A. argyrospila; f: A. cerasivorana; g: A. crataegana; h: A. fuscocupreanus. ........................................................................................ 292 Figure 33. Male genitalia; a: A. grisea; b: A. mortuana; c: A. podana; d: A. purpurana; e: A. rosana; f: A. semiferanus; g: A. xylosteana; h: A. franciscana. ................. 293 Figure 34. Male genitalia; a: A. ljungiana; b: A. mariana; c: A. quadrifasciana; d: A. quercifoliana; e: A. velutinana; f: C. pronubana; g: C. occidentalis; h: C. rosaceana. ................................................................................................ 294 Figure 35. Male genitalia; a: C. clemensiana; b: C. fucana; c: C. melaleucanus; d: C. peritana; e: C. persicana; f: C. spectrana; g: C. virscana; h: C. obliquana. ....... 295 Figure 36. Male genitalia; a: E. acerbella; b: E. postvittana; c: I. miserana; d: P. cerasana; e: P. heparana; f: P. lamprosana; g: P. limitata; h: P. pyrusana. .................................................................................................. 296 Figure 37. Male genitalia; a: P. excessana; b: X. pallorana; c: A. cuneana; d: A. emigratella; e: P. flavedana; f: P. idaeusalis; g: P. stultana; h: S. senecionana. ............................................................................................. 297 Figure 38. Male genitalia; a: S. sulfureana; b: S. tunicana; c: S. rubicola; d: C. bipenicilla; e: C. perseana; f: E. hebesana; g: L. botrana; h: P. viteana. ............................. 298 Figure 39. Male genitalia; a: C. aporema (photo credit Joaquín Baixeras Almela); b: C. plebejana; c: S. ocellana; d: E. formosana; e: T. schistaceana; f: C. illepida; g: C. ombrodelta; h: C. peltastica. ..................................................................... 299 Figure 40. Male genitalia; a: C. araucariae; b: C. caryana; c: C. coniferana; d: C. deshaisiana; e: C. fabivora; f: C. fagiglandana; g: C. garacana; h: C. glandicolana. ............................................................................................. 300 Figure 41. Male genitalia; a: C. kurokoi; b: C. latiferreana; c: C. pomonella; d: C. splendana; e: C. strobilella; f: C. toreuta; g: E. insiticiana; h: G. delineana. ...... 301 xviii    Figure 42. Male genitalia; a: G. funebrana; b: G. interstinctana; c: G. molesta; d: G. packardi; e: G. prunivora; f: G. aurantianum; g: G. punctidiscanum; h: P. fasciana. .................................................................................................... 302 Figure 43. Male genitalia; a: T. batesi; b: T. leucotreta. ................................................ 303 Figure 44. Female genitalia; a: A. comariana; b: A. flavivittana; c: A. gloverana; d: A. hastiana; e: A. holmiana; f: A. minuta. ...................................................... 304 Figure 45. Female genitalia; a: A. nivisellana; b: A. robinsoniana; c: A. senescens; d: A. variana; e: A. variegana; f: T. viridana. ..................................................... 305 Figure 46. Female genitalia; a: E. ambiguella; b: H. umbrabasana; c: C. longana; d: D. basiplagana; e: D. fragariana; f: A. furcatana. .......................................... 306 Figure 47. Female genitalia; a: A. orana; b: A. argyrospila; c: A. cerasivorana; d: A. crataegana; e: A. fuscocupreanus; f: A. grisea. ........................................ 307 Figure 48. Female genitalia; a: A. mortuana; b: A. podana; c: A. purpurana; d: A. rosana; e: A. semiferanus; f: A. xylosteana. ................................................................... 308 Figure 49. Female genitalia; a: A. franciscana; b: A. ljungiana; c: A. mariana; d: A. quadrifasciana; e: A. quercifoliana; f: A. velutinana. ................................. 309 Figure 50. Female genitalia; a: C. pronubana; b: C. occidentalis; c: C. rosaceana; d: C. clemensiana; e: C. fucana; f: C. melaleucanus. ........................................ 310 Figure 51. Female genitalia; a: C. peritana; b: C. persicana; c: C. spectrana; d: C. virescana; e: C. obliquana; f: E. acerbella. ............................................... 311 Figure 52. Female genitalia; a: E. postvittana; b: P. cerasana; c: P. heparana; d: P. lamprosana; e: P. limitata; f: P. pyrusana. ......................................................... 312 Figure 53. Female genitalia; a: P. excessana; b: X. pallorana; c: A. cuneana; d: A. emigratella; e: P. flavedana; f: P. idaeusalis...................................................... 313 Figure 54. Female genitalia; a: P. stultana; b: S. senecionana; c: S. sulfureana; d: S. tunicana; e: S. rubicola; f: C. bipenicilla. ........................................................... 314 Figure 55. Female genitalia; a: C. perseana; b: E. hebesana; c: L. botrana; d: P. viteana; e: C. aporema (photo credit Joaquín Baixeras Almela); f: C. plebejana. ........... 315 Figure 56. Female genitalia; a: S. ocellana; b: E. formosana; c: T. schistaceana; d: C. illepida; e: C. ombrodelta; f: C. peltastica. ......................................................... 316 Figure 57. Female genitalia; a: C. araucariae; b: C. caryana; c: C. coniferana; d: C. deshaisiana; e: C. fabivora; f: C. fagiglandana. ........................................ 317 Figure 58. Female genitalia; a: C. garacana; b: C. glandicolana; c: C. kurokoi; d: C. latiferreana; e: C. pomonella; f: C. splendana. .................................................. 318 Figure 59. Female genitalia; a: C. toreuta; b: E. insiticiana; c: G. funebrana; d: G. interstinctana; e: G. packardi; f: G. delineana; g: G. molesta. ........................... 319 Figure 60. Female genitalia; a: G. prunivora; b: G. aurantianum; c: G. punctidiscanum; d: P. fasciana; e: T. batesi; f: T. leucotreta. ....................................................... 320 CHAPTER 3 Figure 61. Epiphyas postvittana immature stages; 1: egg mass; 2: pupa; 3–6: larvae. ........................................................................................................ 342 Figure 62. Epiphyas postvittana larval morphology; 7: setal map; 8: head; 9: mandible. ....................................................................................................... 343 Figure 63. Epiphyas postvittana adults; 10–16: males; 17–18: females. ...................... 345 Figure 64. Epiphyas postvittana genitalia; 19: male, aedeagus removed; 20: aedeagus; 21: female. ......................................................................................................... 347 xix    CHAPTER 4 Figure 65. Adult wing patterns; 1–2: Lobesia botrana; 3–4: Paralobesia viteana. ........ 355 Figure 66. Male and female genitalia; 5: L. botrana female; 6: L. botrana male; 7: L. botrana male, arrow denotes gap in spines on valva; 8: P. viteana male; 9: P. viteana female. .......................................................................................... 360 Figure 67. Lobesia botrana larvae; 10: larva feeding on Botrytis cinerea; 11: larva on grape; 12: typical damage to grapes (courtesy Roberto H. González, Universidad de Chile); 13: details of head and prothoracic shield; 14: dorsal aspect of segment A9 showing D2s on “saddle” pinaculum, and D1 and SD1 on same pinaculum; 15: ventral aspect of segment A8 showing relative spacing of V1 setae; 16: ventral aspect of segment A9 showing relative spacing of V1 setae. ........................................................................................................... 362 Figure 68. Lobesia botrana eggs and pupae; 17: pupa and cocoon; 18: pupa; 19: pupa, dorsal aspect of A10 with patch of spines; 20: egg on grape; 21: egg on grape; 22: embryo inside of translucent egg. ............................................................... 364 Figure 69. L. botrana larval details; 23: complete setal map; 24: head with setal map; 25: mandible. ..................................................................................................... 366 Figure 70. SEMs of L. botrana larva; 26: head; 27: spinneret; 28: dorsal aspect of segments A9-10 with anal shield; 29: ventral aspect of segments A9-10 showing spacing of V1 setae, 4 setae on the anal proleg, and anal comb with 5 teeth. ....................................................................................................... 368 CHAPTER 5 Figure 71. Adult T. leucotreta wing patterns; 1–3: males; 4: female; 5: female holotype. ............................................................................................................ 383 Figure 72. Morphological details of T. leucotreta adults; 6: male genitalia; 7: female genitalia; 8: pocket of opalescent scales on hind wing; 9–10: hind tibia. .......... 385 CHAPTER 6 Figure 73. C. perseana adult characters; 1: male holotype; 2: female; 3: male genitalia; 4: female genitalia; 5: male eighth abdominal sternite; 6: male coremata, slide mounted; 7: head; 8: male coremata, posterior aspect; 9: male coremata, lateral aspect. ............................................................................................................... 396 Figure 74. SEM of male coremata scales. .................................................................... 398 Figure 75. Cryptaspasma perseana larval morphology; 11: complete setal map; 12: setal map of head; 13: mandible; 14: SD pinaculum on A1–8; 15: D1 and D2 setae on A8; 16: anal shield; 17: D1, D2, and SD1 setae on A9. ..................................... 400 Figure 76. Cryptaspasma perseana immature stages; 18–19: egg masses; 20: larval damage to avocado fruit; 21–22: larvae; 23–25: pupae; 26: pupa exuvium extruding from avocado; 27: adult resting on avocado. ..................................... 402 CHAPTER 7 Figure 77: An obvious error in the BOLD database. The specimen illustrated is not a tortricid, but rather a noctuid. This mistake (screenshot on August 1, 2009) has since been corrected. ........................................................................................ 424 xx    Figure 78: Example of the XML file format used to house DNA data. A complete record, including species and DNA information, is contained between the tags. ................................................................................................................... 431 Figure 79: The HTML front-end for the DNA sequence search tool incorporated into TortAI. ................................................................................................................ 431 Figure 80: Javascript code used to query the XML database file. ................................. 432 Figure 81: Results of a successful query using the DNA sequence search tool. Exact sequence matches are showing in red. ............................................................. 435 CHAPTER 8 Figure 82. Original description of E. circulana from Hübner (1823). ............................. 444 Figure 83. Original illustrations of E. circulana by Hübner (Figs. 363, 364; Plate [63]). ......................................................................................................... 445 Figure 84. Pattern elements in Hübner’s figure 363. ..................................................... 449 Figure 85. Eucosma circulana and closely related species; 4: Hübner’s illustration; 5–7: E. circulana sensu Heinrich; 8–11: E. gemellana; 12–13: E. paragemellana; 14–15: E. fraudabilis. ......................................................................................... 453 Figure 86. Species easily confused with E. circulana; 16–18: Pelochrista scintillana; 19: P. fratruelis; 20–21: Phaneta verna; 22–24: Phaneta autumnana. .............. 456 Figure 87. Male genitalia; 26: E. circulana; 27: E. gemellana; 28: E. fraudabilis; 29: E. paragemellana. ....................................................................................... 460 Figure 88. Female genitalia; 30: E. circulana; 31: E. gemellana; 32: E. fraudabilis; 34: E. paragemellana. ....................................................................................... 462 Figure 89. Distribution map for E. circulana, E. gemellana, E. fraudabilis, and E. paragemellana. ............................................................................................. 464 CHAPTER 9 Figure 90. Simultaneous analysis parsimony strict consensus tree. Parsimony JK values are above each branch and likelihood BS are below each branch. The single clade that was contradicted by ≥ 50% BS support is indicated by *XX* with BS support for the contradictory clade. The base of the Eucosmini is indicated with a star. Ingroup taxa are labeled with generic names proposed in this revision and color-coded as follows: Eucopina green; Epiblema purple; Pelochrista blue; Eucosma red. Female sterigmata types are mapped onto the tree for the Epiblema-Eucosma-Pelochrista clade and indicated with the following symbols: type 1, circle; type 2, square; type 3, triangle. Moth photos represent typical wing patterns found in each ingroup clade. ............................................................... 479 Figure 91. Examples of Eucosma adults; a: E. circulana; b: E. giganteana; c: E. sombreana; d: E. autumnana; e: E. radiatana; f: E. ochrocephala; g: E. argutipunctana; h: E. convergana; i: E. canusana; j: E. parmatana; k: E. influana; l: E. argenticostana. .................................................................... 489 Figure 92. Examples of Eucosma male genitalia; a: E. circulana; b: E. giganteana; c: E. sombreana; d: E. radiatana; e: E. convergana; f: E. montanana; g: E. rusticana; h: E. ochrocephala; i: E. autumnana; j: E. awemeana; k: E. canusana; l: E. argutipunctana. ................................................................. 490 Figure 93. Examples of Eucosma sterigmata; a: E. giganteana; b: E. sombreana; c: E. radiatana; d: E. montanana; e: E. rusticana; f: E. ochrocephala; g: E. autumnana; h: E. awemeana; i: E. canusana; j: E. argutipunctana. ......... 491 xxi    Figure 94. Comparison of Eucosma and Pelochrista females; a: E. circulana; b: P. scintillana. ................................................................................................. 492 Figure 95. Examples of Pelochrista adults; a: P. mancipiana; b: P. corosana; c: P. fuscosparsa; d: P. gilligani; e: P. scintillana; f: P. kingi; g: P. derelicta; h: P. agricolana; i: P. ridingsana; j: P. pulveratana; k: P. ragonoti; l: P. canariana. .................................................................................................. 495 Figure 96. Examples of Pelochrista male genitalia; a: P. scintillana; b: P. rorana; c: P. argenteana; d: P. corosana; e: P. milleri; f: P. pallidipalpana; g: P. agricolana; h: P. matutina; i: P. ridingsana; j: P. robinsonana; k: P. derelicta; l: P. similiana. .................................................................................................... 496 Figure 97. Examples of Pelochrista sterigmata; a: P. scintillana; b: P. rorana; c: P. argenteana; d: P. pallidipalpana; e: P. milleri; f: P. agricolana; g: P. ridingsana; h: P. robinsonana; i: P. derelicta; j: P. similiana. .................... 497 Figure 98. Examples of Eucopina (a–h) and Epiblema (i–l) adults; a: Eucopina bobana; b: E. cocana; c: E. ponderosa; d: E. monitorana; e: E. tocullionana; f: E. siskiyouana; g: E. gloriola; h: E. sonomana; i: Epiblema strenuana; j: E. carolinana; k: E. otiosana; l: E. abruptana.................................................. 500 Figure 99. Examples of Eucopina male (a–c) and female (d–f) genitalia; a, d: E. cocana; b, e: E. tocullionana; c, f: E. gloriola. ................................................................. 501 Figure 100. Phaneta pauperana; a: adult male; b: male genitalia; c: female genitalia. ............................................................................................ 504 Figure 101. Examples of Epiblema genitalia; a: E. walsinghami female; b: E. strenuana male; c: E. obfuscana male. .............................................................................. 507 Figure 102. Parsimony strict consensus gene tree for 28S rDNA. Parsimony JK values are above each branch and likelihood BS values are below each branch. ....... 557 Figure 103. Parsimony strict consensus gene tree for COI. Parsimony JK values are above each branch and likelihood BS values are below each branch. ............. 558 Figure 104. Parsimony strict consensus gene tree for EF-1a. Parsimony JK values are above each branch and likelihood BS values are below each branch. ............. 559 Figure 105. Parsimony strict consensus gene tree for CAD. Parsimony JK values are above each branch and likelihood BS values are below each branch. ............. 560 Figure 106. Combined molecular maximum likelihood tree showing relative branch lengths. .............................................................................................................. 561 1 Chapter 1 General Introduction Tortricidae The Tortricoidea is one of the most diverse superfamilies in the microlepidoptera, second only to the Gelechioidea in number of described species (Horak 1998). It contains a single family, Tortricidae, which is composed of more than 10,300 species in approximately 1,070 genera (Gilligan et al. 2012). Tortricid adults are characterized by a combination of the following character states: head rough-scaled above; scaling of lower frons short, appressed and upwardly directed; proboscis well developed and unscaled; labial palpi three-segmented and generally held horizontally or porrect, with apical segment short and blunt; maxillary palpi reduced; ocelli and chaetosema present; and ovipositor lobes flat (Horak 2006). The family is divided into three subfamilies: Tortricinae, Olethreutinae, and Chlidanotinae (Horak 1998). The entire family has been recently catalogued (Brown 2005), and an updated online taxonomic database is available (Gilligan et al. 2012). Economic importance Agriculture in the United States is constantly threatened by invasive species. Crop losses in the U.S. because of introduced insect pests are estimated to exceed $14 billion USD annually (Pimentel et al. 2005), and many of these pests are Lepidoptera. Worldwide, Tortricidae contains close to 700 economically important species, a number that is third only to the Noctuoidea and Pyraloidea (Zhang 1994). Identification of tortricid pests can be difficult, and the availability of diagnostic resources prior to an invasion is essential for responding in a timely and appropriate manner to new introductions. This dissertation significantly contributes towards 2 the protection of U.S. agriculture from invasive Lepidoptera pests by providing adult and larval identification resources, descriptions of new pest species, and novel molecular diagnostic tools. Chapter 2 focuses on Tortricidae threatening U.S. agriculture. The resulting resource, Tortricids of Agricultural Importance (TortAI – http://www.tortai.org), is designed to aid in the identification of tortricid adults encountered during domestic surveys and tortricid larvae encountered during quarantine inspections at U.S. ports of entry. Identification is facilitated through interactive identification keys that include both adult and larval characters along with detailed fact sheets for each tribe, genus, and species, and numerous photographs of adults, larvae, and important morphological characters. This is the first study to use interactive keys for the identification of tortricid pests. Chapters 3–5 provide details on the three tortricid pests currently of greatest importance to U.S. agriculture: Epiphyas postvittana (Walker), Lobesia botrana ([Denis and Schiffermüller]), and Thaumatotibia leucotreta (Meyrick). Epiphyas postvittana, the light brown apple moth, was discovered in California in 2006 (Brown 2010). It rapidly spread throughout the state, resulting in quarantine restrictions on agricultural products that made identification of both adults and larvae a top priority. Chapter 3 details the adult and larval morphology of this pest, and discusses characters that can be used to distinguish it from similar species. Lobesia botrana, the European grapevine moth, was discovered in Napa Valley, California in 2009. It is one of the most destructive pests of grape in the Palearctic, and its arrival in California seriously threatens that state’s wine industry. Chapter 4 provides an account of the discovery of L. botrana, details diagnostic character states, and discusses closely related species present in the eastern U.S. Thaumatotibia leucotreta, the false codling moth, is one of the most destructive pests of avocado, citrus, and cotton in Africa. A single male was discovered in California in 2008. Chapter 5 provides a detailed description of this species that can be used by identifiers during domestic surveys for this important pest. 3 The list of species threatening U.S. agriculture includes undescribed species as well as well-documented pests. Chapter 6 details the discovery of a new tortricid species with the potential to threaten avocado production in California. Cryptaspasma perseana Gilligan and Brown is described as new. Adults and larvae are described in detail and its life history is reviewed. A worldwide list of avocado-feeding tortricids is provided along with a key to identifying Lepidoptera larvae threatening avocados in California. DNA barcoding, or the use of a standardized segment of DNA for species identification and discrimination, is a widely accepted procedure that has been used to identify economically important insects. Chapter 7 describes the benefits and disadvantages to using DNA barcoding for identification purposes, and assesses the utility of the two largest public DNA barcode databases, NCBI-GenBank and BOLD. The criteria for implementing a successful DNA barcoding system for quarantine and regulatory functions are discussed. Details of a DNA barcode identification tool developed to identify tortricid pests and implemented in TortAI (Chapter 2) are provided. Systematics Only recently has molecular data been used to infer phylogenetic relationships within the Tortricidae and between the Tortricoidea and other superfamilies. Tortricids are included in the non-obtectomeran Apoditrysia, and have been hypothesized to be related to the Cossoidea or the Cossoidea + Sesioidea by various authors (Common 1990; Horak and Brown 1991, Horak 1998). Recent molecular studies using molecular data to determine relationships within the Ditrysia failed to define a sister group for the Tortricoidea, although the superfamily was consistently recovered as monophyletic (Regier et al. 2009, 2012). Regier et al. (2012) were the first to publish a comprehensive molecular phylogeny of the Tortricidae. They included all three subfamilies and 19 out of 22 tribes in their analysis, and used the results to examine life history evolution within the family. Dombroskie (2011) constructed a molecular phylogeny for the 4 Archipini using both nuclear and mitochondrial data. He used the results to examine the evolution of secondary sexual characters within the tribe. No published molecular studies have examined relationships below the tribal level within the Olethreutinae. In chapters 8–9, I examine phylogenetic relationships within Eucosma, the largest genus within the Tortricidae, and related genera. Chapter 8 investigates the type species of Eucosma, E. circulana, for which the type specimen has been lost or destroyed. Candidate taxa are evaluated to match the original description, and a neotype is proposed for E. circulana. Chapter 9 presents the first study to infer phylogenetic relationships within the Eucosmini using a combination of molecular and morphological data. The monophyly of Eucosma, Epiblema, Pelochrista, and Phaneta is tested in a simultaneous parsimony analysis consisting of 2692 bp of DNA data, 27 morphological characters, and five gap characters. Morphological characters are identified to separate and better define the four genera. The pine-feeding Eucosma species are resolved as a monophyletic lineage and placed in a new genus. A new world catalogue of Eucosma-Pelochrista-Phaneta is produced that aligns the generic definitions of these genera in the Nearctic and Palearctic. The world catalogue contains 504 species and 320 new combinations. Summary This dissertation provides significant advances in tortricid systematics and identification through the implementation of modern technologies such as matrix-based identification keys and molecular phylogenetics. It incorporates studies of worldwide pest species that threaten U.S. agriculture as well as non-pest species that comprise the largest genus in the family. The identification resources presented here are already in use by identifiers at the local, state, and federal levels. A molecular phylogeny and revised classification for genera in the Eucosmini provides the groundwork for future study of taxa in this highly diverse tribe. 5 References Brown, J. W. 2005. World catalogue of insects. Volume 5: Tortricidae (Lepidoptera). Apollo Books, Stenstrup, Denmark. 741 pp. Brown, J. W., M. E. Epstein, T. M. Gilligan, S. Passoa and J. A. Powell. 2010. Biology, identification, and history of the light brown apple moth, Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae: Archipini) in California: an example of the importance of local faunal surveys to document the establishment of exotic insects. American Entomologist. 56: 34–43. Common, I. F. B. 1990. Moths of Australia. Melbourne University Publishing. 535 pp. Dombroskie, J. J. 2011. Aspects of archipine evolution (Lepidoptera: Tortricidae). University of Alberta, Department of Biological Sciences. Ph.D. dissertation. 488 pp. Gilligan, T. M., J. Baixeras, J. W. Brown and K. R. Tuck. 2012. T@RTS: Online World Catalogue of the Tortricidae (Ver. 2.0). http://www.tortricid.net/catalogue.asp. Horak, M. 1991. Morphology, pp. 1–22. In: L. P. S. van der Geest, H. H. Evenhuis (eds.), Tortricid Pests: Their Biology, Natural Enemies and Control. Elsevier, Amsterdam, The Netherlands. Horak, M. 1998. The Tortricoidea, pp. 199–215. In: N. P. Kristensen (ed.), Handbook of Zoology, Lepidoptera, Moths and Butterflies, Vol 1: Evolution, Systematics, and Biogeography. Walter de Gruyter, Berlin, New York. Horak, M. 2006. Olethreutine moths of Australia (Lepidoptera: Tortricidae). Monographs on Australian Lepidoptera, Vol. 10. 522 pp. Horak, M. and R. L. Brown. 1991. Taxonomy and phylogeny, pp. 23–50. In: L. P. S. van der Geest, H. H. Evenhuis (eds.), Tortricid Pests: Their Biology, Natural Enemies and Control. Elsevier, Amsterdam, The Netherlands. Pimentel, D., R. Zuniga and D. Morrison. 2005. Update on the environmental and economic costs associated with alien-invasive species in the United States. Ecological Economics. 52: 273–288. Regier, J.C., A. Zwick, M. P. Cummings, A. Y. Kawahara, S. Cho, S. J. Weller, A. D. Roe, J. Baixeras-Almela, J. W. Brown, C. S. Parr, D. R. Davis, M. E. Epstein, W. Hallwachs, A. Hausmann, D. H. Janzen, I. J. Kitching, M. A. Solis, S.-H. Yen, A. Bazinet, C. Mitter. 2009. Toward reconstructing the evolution of advanced moths and butterflies (Lepidoptera: Ditrysia): an initial molecular study. BMC Evolutionary Biology 9: 280. Regier, J. C., J. W. Brown, C. Mitter, J. Baixeras, S. Cho, M. P. Cummings and A. Zwick. 2012. A molecular phylogeny for the leaf-roller moths (Lepidoptera: Tortricidae) and its implications for classification and life history evolution. PLoS ONE. 7: e35574. doi:10.1371/journal.pone.0035574. Zhang, B.-C. 1994. Index of economically important Lepidoptera. CAB International, Wallingford, UK. 599 pp. 6 Chapter 2 Tortricids of Agricultural Importance to the United States (Lepidoptera: Tortricidae)* Introduction Many tortricids are important agricultural pests and the family contains a large number of pest species. Zhang (1994) lists 687 tortricids as economically important, a number third only to the highly diverse Noctuidae, with 1034 pest species, and Pyralidae (+ Crambidae) with 748 recognized pest species. The economic importance of the family is demonstrated by the amount of publications dedicated to the biology, physiology, identification, and control of tortricid moths, and entire volumes have been published on these subjects (e.g., van der Geest and Evenhuis 1991). Perhaps there is no better example of a ubiquitous tortricid pest than the “worm” that is found after biting into an apple, which is actually the larva of the codling moth, Cydia pomonella (L.). In today’s world of global trade and travel, the potential for invasive pests to be transported outside of their native range, either accidentally or intentionally, is higher than ever before. The *Portions of this chapter are published in: Gilligan, T. M. & M. E. Epstein. 2009. LBAM ID: Tools for diagnosing light brown apple moth and related western U.S. leafrollers (Tortricidae: Archipini). CD-ROM. Center for Plant Health Science and Technology, USDA/APHIS/PPQ, Raleigh, NC. *A full version of this chapter is published as: Gilligan, T. M. & M. E. Epstein. 2012. TortAI, Tortricids of Agricultural Importance to the United States (Lepidoptera: Tortricidae). CD-ROM. Identification Technology Program (ITP), USDA/APHIS/PPQ/CPHST, Fort Collins, CO. 7 introduction of pest species to new locations has the potential to greatly affect local agricultural production, and Pimentel et al. (2000) estimate that crop losses in the United States due to damage by introduced insect pests exceeded $14 billion USD in 1998. In addition to supplying food to its 300,000,000+ residents, U.S. agriculture accounts for over $96 billion USD in annual exports (ERS/USDA 2010). The most important tortricid pests that threaten U.S. agriculture 1) are generally not established in North America, and 2) have the greatest potential to contribute to the decline of domestic food production and/or agricultural exports. The USDA has compiled a list of the top ten tortricid pests threatening U.S. agriculture. These species have been or are currently being surveyed under the USDA’s Cooperative Agriculture Pest Survey Program (CAPS 2010) and are listed as primary target species (Table 1). We have compiled a list of 15 additional tortricids using a combination of prior CAPS targets, recent introductions into North America, and introduction potentials, which are listed as secondary target species (Table 2). The majority (60%) of the target species listed here are native to the Palearctic; others are native to Australia, South America, Africa, or Asia. Two species of Cryptophlebia, although native to Hawaii, could significantly affect fruit production if imported into the continental U.S. Taxonomic resources for tortricid adult identification are numerous. The Palearctic fauna has been well documented, and large monographs have been published that illustrate nearly all of the tortricids described from that region (e.g., Bradley et al., 1973, 1979, Kuznetsov 1978, Razowski 2002, 2003, 2008, etc.). The Oriental fauna has been treated by Yasuda (1969, 1972, 1975), Diakonoff (1968, 1973, 1975), and others. The Australian Olethreutine genera have been recently revised by Horak (2006). Relatively few publications exist on the Neotropical and African faunas, although treatments for genera and tribes have been published by various authors (e.g., Diakonoff 1963, Obraztsov 1966, Powell 1986, Aarvik 2004, etc.). The Nearctic fauna is well documented, especially for the Olethreutinae. Heinrich (1923, 1926), Miller (1987), and Gilligan et al. (2008) provide coverage of most of the North American olethreutine species. 8 The Nearctic Tortricinae has not been treated in its entirety other than by Freeman (1958) for the Archipini and Powell (1964) for the California tortricines. Most recently, Powell and Opler (2009) treat approximately 25% of the western North American Tortricidae. Table 1: Primary target species Genus Species Author Common name Target status Region Adoxophyes orana (Fi scher von Rös lers tamm) summer frui t tortrix primary Pa learctic Archips podana (Scopol i ) fruit tree  tortrix primary Pa learctic Archips xylosteanus (Linnaeus) golden variegated leafrol ler primary Pa learctic Crocidosema aporema (Wals ingham) bean shoot moth primary S. America Epiphyas postvittana (Walker) l ight brown apple  moth primary Austra l ia Eupoecilia ambiguella (Hübner) vine  moth/grape  berry moth primary Pa learctic Grapholita funebrana (Trei tschke) plum frui t moth primary Pa learctic Lobesia botrana ([Denis  & Schi ffermül ler]) grape  berry moth primary Pa learctic Thaumatotibia leucotreta (Meyrick) fa l se  codl ing moth primary Africa Tortrix viridana Linnaeus European oak leafrol ler primary Pa learctic Table 2: Secondary target species Genus Species Author Common name Target status Region Acleris comariana (Lienig & Zel ler) s trawberry tortrix secondary Holarctic Archips crataegana (Hübner) brown oak tortrix secondary Pa learctic Archips fuscocupreanus Wals ingham apple  tortrix secondary Orienta l Argyrotaenia ljungiana (Thunberg) grape  tortrix secondary Pa learctic Cacoecimorpha  pronubana (Hübner) carnation tortrix secondary Pa learctic Clepsis spectrana (Trei tschke) cyclamen tortrix secondary Pa learctic Cryptaspasma perseana Gi l l igan & Brown [avocado moth] secondary C. America Cryptophlebia illepida (Butler) koa  seedworm secondary Hawai i Cryptophlebia ombrodelta (Lower) l i tchi  frui t moth secondary Aust./Hawai i Cydia coniferana (Saxesen) pine  res in moth secondary Pa learctic Cydia fabiovra (Meyrick) pod moth secondary S. America Cydia splendana (Hübner) chestnut tortrix/acorn moth secondary Pa learctic Enarmonia formosana (Scopol i ) cherry  bark tortrix secondary Pa learctic Pandemis cerasana (Hübner) barred frui t tree  tortrix secondary Pa learctic Pandemis heparana ([Denis  & Schi ffermül ler]) dark frui t tree  tortrix secondary Pa learctic Although detailed larval descriptions have been published for many economically important tortricids, very few comparative taxonomic works have been produced on tortricid larvae. Swatschek (1958) published the first extensive treatment of the Palearctic Tortricidae, 9 where he described the larvae of more than 350 species. MacKay (1959, 1962) published larval descriptions for more than 280 species of Nearctic Tortricidae in two volumes, providing detailed illustrations and keys to species, species groups, and genera. Yasuda (1969) published descriptions and illustrations of the Japanese tortricid fauna. Other important works including tortricid larval systematics are Brown’s (1987) treatment of the family and Dugdale et al.’s (2005) guide to quarantine pests, which includes molecular as well as morphological characters. Despite the relative abundance of literature, identification of tortricid adults and larvae remains problematic. Adult identification often requires genitalic dissection and prior knowledge of intra- versus interspecific variation in wing patterns, sibling species, and other information that may only be available to a specialist with many years of experience. Larval identification is much more difficult and, in many aspects, more important, since it is the larva that causes economic damage and this is the life stage often intercepted during quarantine inspections. Tortricid larvae are surprisingly homogeneous, with very few distinguishing characters available to separate species, or even genera. Further complicating the problem of larval identification is the paucity of material in collections. Published or informal keys to tortricid larvae often fail in practice due to the low sample size examined during construction of the key. Tortricids of Agricultural Importance (TortAI) is designed to aid in the identification of tortricid adults encountered during domestic surveys and tortricid larvae encountered during quarantine inspections at U.S. ports of entry. This resource [published as a CD-ROM and also available on the Internet at http://www.tortai.org] includes interactive identification keys, detailed fact sheets, an illustrated glossary, information on dissecting and preparing specimens, identification thumbnail galleries, and a database of DNA barcode sequences. The interactive identification keys run in a web browser as a Lucid3 Java Applet. Because the world tortricid fauna is too large to treat as a whole, this resource is not designed to identify every tortricid encountered, but rather to reliably eliminate or confirm target taxa if or when they are encountered. It may not be possible to identify many tortricid larvae using only morphological 10 characters. A molecular sequence search tool is provided to allow identifiers with sequencing capabilities the ability to confirm DNA barcode sequences for the larvae