UNIVERSIDAD DE COSTA RICA SISTEMA DE ESTUDIOS DE POSGRADO TRATAMIENTO SISTEMÁTICO DEL GÉNERO TRICHOSALPINX (ORCHIDACEAE: PLEUROTHALLIDINAE) EN COSTA RICA Tesis sometida a la consideración de la Comisión del Programa de Estudios de Posgrado en Biología para optar al grado y título de Maestría Académica en Biología MELANIA FERNANDEZ CAMPOS Ciudad Universitaria Rodrigo Facio, Costa Rica 2014   ii Dedicatoria Este trabajo está dedicado a Emilia Campos Gaitán, mi mamá, quien me ha acompañado por el hermoso camino de la vida, siempre de la mano. Mi esfuerzo y empeño son inspirados por su ejemplo, y esta es mi manera de reconocerle desde lo más profundo. Deseo agracedecer sinceramente a mis hermanos Leonardo, Alejandro y Manuel, quienes han agregado peso extra a su carga por aligerar la mía durante este proceso de tesis. Finalmente, brindo mi más sincero agradecimiento a Franco Pupulin, mi mentor, por abrirme las puertas a la investigación y haberme entregado su confianza incondicional desde el primer día.   iii Agradecimientos   Quiero brindar un especial agradecimiento a Franco Pupulin por su apoyo y guía durante el desarrollo de este trabajo. A mi profesor Robert L. Dressler, y los miembros de mi comité Diego Bogarín y Mario Blanco por sus observaciones críticas y su instrucción durante la realización de este trabajo. A Franco Pupulin, Diego Bogarín y Adam Karremans por sus consejos y orientación en el trabajo escrito, la elaboración de las ilustraciones y la documentación digital. A todo el personal del Jardín Botánico Lankester, en especial a Jorge Warner por su constante apoyo y consejos, así como al personal del Departamento de Horticultura por el mantenimiento de las colecciones de plantas vivas. A Diego Bogarín, Reinaldo Gómez, Adam Karremans, Luis Sandoval y Rafael Trejos por su compañía y colaboración durante las giras de recolecta. A Maricruz Bonilla por todo el apoyo logístico. A Daniel Jiménez, Hermano Jorge de la Cruz, Ricardo Marín y Gerson Villalobos por facilitar especímenes e información útil para este estudio. A José A. Vargas y Jorge Cortés por sus valiosos comentarios en el anteproyecto. A Hannia Ramírez por la colaboración administrativa. Al Ministerio del Ambiente y Energía (MINAE) y su Sistema Nacional de Áreas de Conservación (SINAC) por emitir los permisos bajo los cuales fueron recolectados los especímenes de este estudio. A los curadores y personal de los herbario CR, INB, JBL y USJ por conceder acceso a las colecciones y brindar información valiosa. A la Vicerrectoría de Investigación de la Universidad de Costa Rica por el apoyo económico y logístico mediante los proyectos institucionales “Flora Costaricensis: Taxonomía y filogenia de la subtribu Pleurothallidinae (Orchidaceae) en Costa Rica” (No. 814-B0-052) e “Inventario y Taxonomía de la flora orquidácea de Costa Rica” (No. 814-   iv A7-015), de los cuales este proyecto es parte. “Esta tesis fue aceptada por la Comisión del Programa de Estudios de Posgrado en Biología de la Universidad de Costa Rica, como requisito parcial para optar al grado y título de Maestría Académica en Biología”.   _______________________________ Ph.D. Óscar Brenes García Representante de la Decana, Sistema de Estudios de Posgrado _______________________________ Dr. Robert L. Dressler Director de Tesis _______________________________ Ph.D. Mario Blanco Coto Asesor _______________________________ M.Sc. Diego Bogarín Cháves Asesor _______________________________ Ph.D. Alfredo Cascante Marín Representante del Director, Programa de Posgrado en Biología _______________________________ Melania Fernández Campos Candidata   v Index Dedicatoria ............................................................................................................................ ii Agradecimientos .................................................................................................................. iii LISTA DE FIGURAS ........................................................................................................... vii PREFACIO ........................................................................................................................ xvii ABSTRACT ........................................................................................................................ xix I. INTRODUCTION ........................................................................................................... 1 II. METHODOLOGY .......................................................................................................... 7 III. TAXONOMIC HISTORY OF TRICHOSALPINX ........................................................ 11 IV. CURRENT PHYLOGENETIC CIRCUMSCRIPTION ................................................. 16 V. MORPHOLOGY ......................................................................................................... 18 Roots ........................................................................................................................... 18 Stems .......................................................................................................................... 18 Ramicaul sheaths ........................................................................................................ 20 Leaves ......................................................................................................................... 21 Inflorescences ............................................................................................................. 23 Flowers ........................................................................................................................ 24 Pollinaria ...................................................................................................................... 25 Fruits ........................................................................................................................... 26 VI. SYSTEMATIC TREATMENT ..................................................................................... 28 1. Key to the Costa Rican subgenera and species of Trichosalpinx ........................... 32 2. Species description and discussion ........................................................................ 37 1. Trichosalpinx arbuscula .......................................................................................... 37 2. Trichosalpinx blaisdellii .......................................................................................... 42 3. Trichosalpinx carinilabia ........................................................................................ 51 4. Trichosalpinx caudata . ........................................................................................... 55 5. Trichosalpinx cedralensis ...................................................................................... 57   vi 6. Trichosalpinx ciliaris ............................................................................................... 64 7. Trichosalpinx dura .................................................................................................. 67 8. Trichosalpinx fruticosa ........................................................................................... 74 9. Trichosalpinx membraniflora .................................................................................. 77 10. Trichosalpinx memor ........................................................................................... 79 11. Trichosalpinx minutipetala ................................................................................... 91 12. Trichosalpinx nana ............................................................................................... 98 13. Trichosalpinx navarrensis .................................................................................. 101 14. Trichosalpinx orbicularis ..................................................................................... 104 15. Trichosalpinx parsonsii ....................................................................................... 109 16. Trichosalpinx pergrata ....................................................................................... 111 17. Trichosalpinx pusilla ........................................................................................... 115 18. Trichosalpinx reflexa .......................................................................................... 119 19. Trichosalpinx ringens ......................................................................................... 122 20. Trichosalpinx rotundata ..................................................................................... 126 21. Trichosalpinx sanctuarii ..................................................................................... 129 22. Trichosalpinx todziae ......................................................................................... 132 23. Trichosalpinx trachystoma .................................................................................. 135 VII. EXCLUDED SPECIES ........................................................................................... 139 VIII. CONCLUSIONS .................................................................................................... 140 IX. INDEX TO HERBARIUM SPECIMENS EXAMINED ............................................... 143 1. Numerical list of species of Trichosalpinx in Costa Rica ....................................... 143 2. List of Collections .................................................................................................. 144 X. LITERATURE CITED ............................................................................................... 148 XI. FIGURES ................................................................................................................. 158   vii LISTA DE FIGURAS Figure 1. Geographic distribution of the genus Trichosalpinx, with the approximate number of species per region; modified from Pridgeon (2005)......................................................... 3 Figure 2. Figure 2. Elevational range of Trichosalpinx species in Costa Rica, based on herbarium records annotated in this treatment. * indicates species endemic to Costa Rica. -- indicates regional endemisms.......................................................................................... 4 Figure 3. Holotype specimen of Trichosalpinx pusilla (Kunth) Luer (Humboldt & Bonpland s.n.; P-00669678). Credits to the Paris, Muséum National d'Histoire Naturelle, SM......... 11 Figure 4. Holotype specimen and drawing of Trichosalpinx ciliaris (Lindl.) Luer (Loddiges s.n., K-000079852). Reproduced with the kind permission of the Board of Trustees, Royal Botanical Gardens, Kew.................................................................................................... 11 Figure 5. Holotype specimen and drawing of Trichosalpinx orbicularis (Lindl.) Luer (Loddiges s.n.; K-000079889). Reproduced with the kind permission of the Board of Trustees, Royal Botanical Gardens, Kew.......................................................................... 11 Figure 6. Holotype specimen of Trichosalpinx memor (Rchb.f.) Luer (H. Forkel s.n.; W- 0053202). Reproduced with kind permission of the Natural History Museum of Vienna... 11 Figure 7. Holotype specimen of Trichosalpinx blaisdellii (S.Watson) Luer (material framed) mounted in the same sheet (AMES-72460) as Watson s.n. (not a type, 00074089). Reproduced with the kind permission of the Harvard University Herbaria......................... 12   viii Figure 8. Holotype specimen and drawing of Trichosalpinx arbuscula (Lindl.) Luer (Hartweg s.n.; K-000079691). Reproduced with the kind permission of the Board of Trustees, Royal Botanical Gardens, Kew.......................................................................... 12 Figure 9. Holotype specimen and drawing of Trichosalpinx dura (Lindl.) Luer (Jameson s.n.; K-000079890). Reproduced with the kind permission of the Board of Trustees, Royal Botanical Gardens, Kew.................................................................................................... 12   Figure 10. Description of Pleurothallis moschata Rchb.f. by Auguste R. Endrés (W-Rchb.f.- 0020891). Reproduced with kind permission of Natural History Museum of Vienna......... 12 Figure 11. Holotype specimen of Pleurothallis moschata Rchb.f., including color drawing by Reichenbach (Endres s.n.; W-Rchb.f.-0020892). Reproduced with the kind permission of the Natural History Museum of Vienna.......................................................................... 13 Figure 12. Drawing of Pleurothallis moschata Rchb.f. [=Trichosalpinx arbuscula (Lindl.) Luer] by Auguste R. Endrés (not a type drawing; W-Rchb.f.-0020889). Reproduced with the kind permission of the Natural History Museum of Vienna.......................................... 13 Figure 13. Holotype specimen of Trichosalpinx cedralensis (Ames) Luer (Lankester 352; K-000463155). Reproduced with the kind permission of the Board of Trustees, Royal Botanical Gardens, Kew.................................................................................................... 13 Figure 14. Holotype specimen and drawing of Trichosalpinx pergrata (Ames) Luer (Lankester & Sancho 436; AMES-23721). Reproduced with the kind permission of the Harvard University Herbaria............................................................................................... 13   ix Figure 15. Lectotype specimen of Trichosalpinx trachystoma (Schltr.) Luer (Brenes 40; AMES-31257). Reproduced with the permission of Harvard University Herbaria.............. 13 Figure 16. Holotype specimen of Trichosalpinx nana (Ames & C.Schweinf.) Luer (Lankester 865; AMES-28677). Reproduced with the kind permission of the Harvard University Herbaria............................................................................................................ 13 Figure 17. Holotype specimen and drawing of Trichosalpinx navarrensis (Ames) Luer (Standley 33888; corresponding material framed) mounted in the same sheet (AMES- 29163) as the paratype specimen of T. navarrensis (Standley 33865). Reproduced with the kind permission of the Harvard University Herbaria..................................................... 14 Figure 18. Holotype specimen of Trichosalpinx rotundata (C.Schweinf.) Luer (Hunter & Allen 561 D; AMES-42016). Reproduced with the kind permission of the Harvard University Herbaria............................................................................................................ 14 Figure 19. Holotype specimen and drawing of Trichosalpinx membraniflora (Ames Luer (Brenes 20.571; AMES-44322). Reproduced with the kind permission of the Harvard University Herbaria............................................................................................................ 14 Figure 20. Holotype specimen of Trichosalpinx carinilabia (Luer) Luer (Smith H 668; AMES-55421). Reproduced with kind permission of Harvard University Herbaria............ 14 Figure 21. Holotype specimen and drawing of Trichosalpinx fruticosa Luer (Luer 12135; MO-5122981). Reproduced with permission of Missouri Botanical Garden Herbarium.... 15 Figure 22. Holotype specimen and drawing of Trichosalpinx lankesteriana Luer (Luer   x 17411; MO-5123145). Reproduced with the kind permission of the Missouri Botanical Garden Herbarium............................................................................................................ 15 Figure 23. Holotype specimen and drawing of Trichosalpinx ringens Luer (Luer 10539; MO-5123191). Reproduced with kind permission of Missouri Botanical Garden............... 15 Figure 24. Isotype specimen of Trichosalpinx parsonsii Luer & Dod (Grantham & Parsons 03 93 90; MO-5123195). Reproduced with the kind permission of the Missouri Botanical Garden Herbarium............................................................................................................. 15 Figure 25. Holotype specimen of Trichosalpinx todziae Luer (Todzia 344; CR-66495). Reproduced with kind permission of the Herbario Nacional de Costa Rica....................... 15 Figure 26. Root morphology in Costa Rican Trichosalpinx. A - T. nana; B - T. cedralensis; C - T. blaisdellii; D - T. minutipetala. A–B: Subgenus Tubella. C–D: Subgenus Trichosalpinx. White arrow points aerial root development. Scale bar = 2 cm.................. 18 Figure 27. Sheath and leaf morphology in Costa Rican Trichosalpinx. A. T. blaisdellii; B. T. reflexa; C. T. memor; D. T. rotundata; E. T. arbuscula; F. T. dura; G. T. pergrata; H. T. cedralensis. A–D Subgenus Trichosalpinx. E–H Subgenus Tubella. Prolific ramicauls are shown in E, F and H; in H, two prolifications were produced at the apex of the proximal ramicaul. Scale bar = 3 cm................................................................................................ 19 Figure 28. Details of sheath morphology in Costa Rican Trichosalpinx. A – Prolification in Trichosalpinx memor. Note perforation of the distalmost sheath for emergence of new tissues. A–B Detail of the sheath morphology in B – subgenusTubella, and C – subgenus   xi Trichosalpinx. Note vernation patterns and marginal ornamentation of the sheath........... 21 Figure 29. Inflorescence morphology in Costa Rican Trichosalpinx. Subgenus Tubella: A - T. arbuscula; B - T. dura; C - T. fruticosa; D - T. todziae. Subgenus Trichosalpinx: E - T. blaisdellii; F - T. caudata; G - T. reflexa; H - T. rotundata; I - T. trachystoma.................... 23 Figure 30. Flower morphology in Costa Rican Trichosalpinx. Subgenus Tubella: A - T. parsonsii; B - T. carinilabia; C - T. pusilla; D - T. pergrata. Subgenus Trichosalpinx: E - T. ringens; F - T. sanctuarii; G - T. ciliaris; H - T. orbicularis; I - T. minutipetala.................... 24 Figure 31. Column morphology in Costa Rican Trichosalpinx. Subgenus Trichosalpinx: A - T. blaisdellii; B - T. memor. Subgenus Tubella: C - T. dura; D - T. arbuscula. A–C: ventral view. D: dorsal view. Note the shape and position of the rostellum, and the shape of the column foot. Scale = 1 mm................................................................................................ 25 Figure 32. Pollinaria morphology in Costa Rican Trichosalpinx. Subgenus Trichosalpinx: A–B T. blaisdellii; C - T. minutipetala; D - T. memor. Subgenus Tubella: E–F T. arbuscula; G - T. cedralensis; H - T. fruticosa. Scale = 0.5 mm.......................................................... 25 Figure 33. Fruit morphology in Costa Rican Trichosalpinx. Subgenus Trichosalpinx: A - T. blaisdellii; B - T. memor; C - T. minutipetala; D - T. reflexa. Subgenus Tubella: E–F T. todziae; G - T. dura. All fruits are shown undehisced. Top row, view from below; bottom row, transverse section. Scale = 1 cm............................................................................... 26 Figure 34. Trichosalpinx arbuscula. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. F – Anther cap and pollinaria. Drawn by M.   xii Fernández from F. Pupulin 4331 (JBL-spirit)..................................................................... 37 Figure 35. Distribution map of Trichosalpinx arbuscula in Costa Rica............................... 41 Figure 36. Trichosalpinx blaisdelli. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. F – Anther cap and pollinaria. Drawn by M. Fernández from D. Bogarín 5022 (JBL-spirit).................................................................... 43 Figure 37. Trichosalpinx blaisdellii (form of T. lankesteriana). A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. F – Anther cap and pollinaria. Drawn by M. Fernández from Dressler s.n. [JBL-00404] (JBL-spirit)......... 43 Figure 38. Distribution map of Trichosalpinx blaisdellii in Costa Rica................................ 49 Figure 39. Morphological variability in flowers of Trichosalpinx blaisdellii. A: size, shape and color variability in sepals and petals (Scale = 1 cm). B: Detail of size variations in dorsal sepals of T. blaisdellii (Scale = 5 mm)..................................................................... 50 Figure 40. Trichosalpinx carinilabia. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. F – Anther cap and pollinaria. Drawn by M. Fernández from M. Fernández 631 (JBL-spirit)................................................................. 51 Figure 41. Distribution map of Trichosalpinx carinilabia in Costa Rica.............................. 53 Figure 42. Trichosalpinx caudata. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. F – Anther cap and pollinaria. Drawn by M. Fernández from M. Fernández 546 (JBL-spirit)................................................................. 55   xiii Figure 43. Distribution map of Trichosalpinx caudata in Costa Rica................................. 56 Figure 44. Trichosalpinx cedralensis. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral and lateral view. F – Anther cap and pollinaria. Drawn by M. Fernández from F. Pupulin 7498 (JBL-spirit)............................... 57 Figure 45. Distribution map of Trichosalpinx cedralensis in Costa Rica............................ 62 Figure 46. Trichosalpinx ciliaris. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. F – Anther cap and pollinaria. Drawn by M. Fernández from M. Fernández 328 (JBL-spirit)................................................................. 64 Figure 47. Distribution map of Trichosalpinx ciliaris in Costa Rica.................................... 66 Figure 48. Trichosalpinx dura. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Anther cap and pollinaria. Drawn by M. Fernández from M. Fernández 547 (JBL-spirit)................................................................................................ 67 Figure 49. Distribution map of Trichosalpinx dura in Costa Rica....................................... 71 Figure 50. Morphological variability of flowers of Trichosalpinx dura. Note variations of overall size, and red stains in the lips. Scale = 5 mm........................................................ 73 Figure 51. Trichosalpinx fruticosa. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. F – Anther cap and pollinaria. Drawn by M. Fernández from M. Fernández 617 (JBL-spirit)................................................................. 74   xiv Figure 52. Distribution map of Trichosalpinx fruticosa in Costa Rica................................. 76 Figure 53. Distribution map of Trichosalpinx membraniflora in Costa Rica....................... 78 Figure 54. Trichosalpinx memor, morph 1. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. F – Anther cap and pollinaria. Drawn by M. Fernández from M. Fernández 12 (JBL-spirit)............................................. 79 Figure 55. Trichosalpinx memor, morph 2. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Anther cap and pollinaria. Drawn by M. Fernández from F. Pupulin 5472 (JBL-spirit)....................................................................................... 79 Figure 56. Distribution map of Trichosalpinx memor in Costa Rica................................... 88 Figure 57. Morphological variability of flowers of Trichosalpinx memor. A Morph 1. B Morph 2. Note differences in color of sepals, lip and anther cap. Scale = 3 mm............... 90 Figure 58. Trichosalpinx minutipetala. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. F – Column, ventral view. G – Anther cap and pollinaria. Drawn by M. Fernández from A. Karremans 1333 (JBL-spirit)............ 91 Figure 59. Distribution map of Trichosalpinx minutipetala in Costa Rica........................... 96 Figure 60. Trichosalpinx nana. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Anther cap and pollinaria. Drawn by M. Fernández from A. Karremans 4882 (JBL-spirit).............................................................................................. 98   xv Figure 61. Distribution map of Trichosalpinx nana in Costa Rica...................................... 99 Figure 62. Distribution map of Trichosalpinx navarrensis in Costa Rica.......................... 102 Figure 63. Trichosalpinx orbicularis. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Anther cap and pollinaria. Drawn by M. Fernández from M. Fernández 65 (JBL-spirit)................................................................................................ 104 Figure 64. Distribution map of Trichosalpinx orbicularis in Costa Rica........................... 107 Figure 65. Trichosalpinx parsonsii. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. F – Anther cap and pollinaria. Drawn by M. Fernández from A. Karremans 3305 (JBL-spirit)............................................................. 109 Figure 66. Distribution map of Trichosalpinx parsonsii in Costa Rica. White dot with center black indicates locality of the holotype............................................................................. 110 Figure 67. Trichosalpinx pergrata. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view, not flattened. F – Anther cap and pollinaria. Drawn by M. Fernández from D. Bogarín 6502 (JBL-spirit)............................................. 111 Figure 68. Distribution map of Trichosalpinx pergrata in Costa Rica............................... 114 Figure 69. Trichosalpinx pusilla. A – Habit. B – Flower. C – Dissected perianth. D – Lip, ventral view. E – Anther cap and pollinaria. Drawn by M. Fernández from A. Karremans 5642 (JBL-spirit)............................................................................................................... 115   xvi Figure 70. Distribution map of Trichosalpinx pusilla in Costa Rica.................................. 117 Figure 71. Trichosalpinx reflexa. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Column, ventral view. F – Anther cap and pollinaria. Drawn by D. Bogarín from D. Bogarín 1674 (JBL-spirit)............................................................. 119 Figure 72. Distribution map of Trichosalpinx reflexa in Costa Rica................................. 121 Figure 73. Trichosalpinx ringens. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. F – Anther cap and pollinaria. Drawn by M. Fernández from D. Bogarín 7187 (JBL-spirit).................................................................. 122 Figure 74. Distribution map of Trichosalpinx ringens in Costa Rica................................ 124 Figure 75. Trichosalpinx rotundata. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. Drawn by M. Fernández from A. Karremans 4120 (JBL-spirit)............................................................................................................... 126 Figure 76. Distribution map of Trichosalpinx rotundata in Costa Rica............................. 128 Figure 77. Trichosalpinx sanctuarii. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. F – Column, ventral view. G – Anther cap and pollinaria. Drawn by M. Fernández from M. Fernández 529 (JBL-spirit).................. 129 Figure 78. Distribution map of Trichosalpinx sanctuarii in Costa Rica............................. 130 Figure 79. Trichosalpinx todziae. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. F – Pollinaria. Drawn by M. Fernández from   xvii D. Bogarín 49 (JBL-spirit)................................................................................................ 132 Figure 80. Distribution map of Trichosalpinx todziae in Costa Rica................................. 134 Figure 81. Trichosalpinx trachystoma from type locality. A – Habit. B – Flower. C – Dissected perianth. D – Pollinaria and anther cap. Drawn by F. Pupulin from F. Pupulin 101 (USJ, possibly lost)................................................................................................... 135 Figure 82. Trichosalpinx trachystoma from Tilarán, Guanacaste. A – Habit. B – Flower. C – Dissected perianth. D – Lip and column, lateral view. E – Lip, ventral view. F – Column, ventral view. G – Anther cap and pollinaria. Drawn by M. Fernández from M. Fernández 536 (JBL-spirit)................................................................................................................. 135 Figure 83. Distribution map of Trichosalpinx trachystoma in Costa Rica......................... 137   xviii PREFACIO La revisión taxonómica de Trichosalpinx (Orchidaceae) en Costa Rica es presentada. La historia taxonómica del género, su posición filogenética y problemas nomenclaturiales son discutidos. El género es tratado como un grupo de 23 especies en el país, y claves dicotómicas a nivel de subgénero y especies son proveídas. Los taxa son descritos e ilustrados con base en material costarricense, y su distribución en el país es evaluada. Se proveen mapas de distribución para todos los taxa. Se presenta la distribución general, la derivación del nombre, notas sobre su ecología y caracteres diagnósticos para cada taxon.   xix ABSTRACT A taxonomic revision of Trichosalpinx (Orchidaceae) in Costa Rica is presented. The taxonomic history of the genus, its phylogenetic position and nomenclatural issues are discussed. Characters of vegetative and floral morphology are addressed and illustrated, and their taxonomic significance is discussed. The genus is treated as comprising 23 species in the country, and a key to subgenera and species is provided. Taxa are described and illustrated on the basis of Costa Rican material, and their distribution in the country is assessed. Distribution maps for all the taxa are given. Overall distribution, derivation of name, notes on species ecology, and diagnostic features are presented for each taxon.   1   I. INTRODUCTION The Neotropical subtribe Pleurothallidinae Lindl. comprises about 60–80 genera and approximately 4100 species (Pridgeon et al. 2005). The group represents about 15–16% of all known orchid species worldwide. It is distributed from south Florida to northern Argentina, with peaks of diversity in the montane and sub-montane rain and cloud forests of Central America and the Andes (Jørgensen & León-Yánez 1999, Vásquez & Ibisch 2000, Dodson 2004, Stenzel 2004). In Mesoamerica, Costa Rica holds the highest number of genera within the Pleurothallidinae (Ossenbach et al. 2007). The hyperdiversity of the subtribe, which represents one of the most diverse monophyletic groups of plants on Earth, poses extremely interesting questions from the evolutionary point of view. Despite the several efforts, none of the morphological, anatomical or molecular approaches (Pridgeon 1982, Luer 1986, 2002, Neyland et al. 1995, Pridgeon et al. 2001, Pridgeon & Chase 2001, 2005, van den Berg et al. 2005) have been able to achieve an estable generic and subgeneric classification in this monophyletic subtribe (sensu Pridgeon et al. 2001, 2005). Members of this subtribe are distinguished by the sympodial growth, unifoliate (except for Dilomilis Raf., Frondaria Luer and Tomzanonia Nir.), nonpseudobulbous ramicauls (the term used to designate the part of the stem above of the last reiteration bud in Pleurothallids; see Stern & Pridgeon 1984), conduplicate leaves, velamentous roots, and an articulation between the pedicel and the ovary (except for Dilomilis, Neocogniauxia Schltr. and Tomzanonia) instead of at the base of the pedicel as in most the other orchids (Luer 1986, van den Berg et al. 2005, Pridgeon et al. 2005). Genera in the Pleurothallidinae have been traditionally defined on the basis of number of pollinia, number of stigma lobes, degree of sepal connation, resupination, shape of the various perianth   2   parts, and presence or absence of lip mobility (Garay 1979, Luer 1986, 1987, 2000, Stenzel 2004). Plants in the Pleurothallidinae are epiphytic, terrestrial or lithophytic, and inhabit dry, humid, and cloud forests, mangroves and paramo. In Costa Rica, they are mainly epiphytic, and inhabit forests between 0 to more than 3000 m elevation, although many have restricted distributions in humid forests between 1000 and 2000 m. Plants can be as long as 1 m, but most of them are less than 30 cm long. The pollination system of most species remains unknown. The vast majority of the documented cases represent pollination by dipterans, some by deceit (Borba & Semir 2001, Borba et al. 2001, Blanco & Barboza 2005, Pupulin et al. 2012). Pollination studies in Acianthera Scheidw. (Singer & Cocucci 1999, Borba & Semir 2001, Borba et al. 2001), Dracula Luer (Endara et al. 2010), Lepanthes Sw. (Blanco & Barboza 2005, Blanco & Vieira 2011), Masdevallia Ruiz & Pav. (Cuervo 2012, Dodson 1962), Pleurothallis R.Br. (Duque-Buitrago et al. 2014) and Specklinia Lindl. (Pupulin et al. 2012) have mostly revealed myophily, or pollination by flies. Instead of a single strategy, Pleurothallids use a variety of different ways of attracting pollinators, including odor attractants and structural mimicry (e.g. fungus or flesh-like structures and insect sexual parts). Although myophily and self-incompatibility have been reported to be basal characteristics in the former delimitation of the Pleurothallidinae and possibly have been kept in the derived lineages (Barbosa et al. 2009), alternative pollinators, including pollination by microhymenoptera and Coleoptera, have also been reported to occur (Duque 1993). Pleurothallidinae include several of the largest genera within Orchidaceae, such as Pleurothallis, Stelis Sw. and Lepanthes. The latter is one of the genera bearing lepanthiform sheaths on the ramicauls, along with Lepanthopsis (Cogn.) Ames,   3   Draconanthes (Luer) Luer and Trichosalpinx Luer. Lepanthiform sheaths are tubular sheaths that cover the ramicaul, more or less imbricate, with longitudinal ribs and dilated ostia with thickened, ciliate margins. Trichosalpinx was described in 1983 and typified by the widely distributed Specklinia ciliaris Lindl. (Luer 1983). The name comes from the Greek trichos, “hair”, and salpinx, “a trumpet”, referring to the ciliate ribs and margins of the trumpet-like sheaths of the ramicaul (Luer 1983). At present, around 160 species have been described in Trichosalpinx after Luer (1997) relegated 20 names to the synonymy of other species, and transferred 10 species to other genera such as Lepanthopsis, Pabstiella Brieger & Senghas and Pleurothallis. Populations of Trichosalpinx are found throughout the Neotropics, with evident species concentration in the Andes and the high mountain ranges of Costa Rica and western Panama. The moist and wet forests of the Andes between 1600 m and 2700 m harbor the majority of the Trichosalpinx species described to date (Luer 1997; Fig. 1). Species distribution in Trichosalpinx is variable. Some species are found all throughout the Neotropics, as in the cases of T. memor (Rchb.f.) Luer and T. dura (Lindl.) Luer. Other species are restricted to a regional scale (e.g. T. pergrata [Ames] Luer, found only in Costa Rica and Panama). Furthermore, regional endemisms occur on a regular basis, as in the cases with T. reflexa Mel.Fernández & Bogarín, restricted to the Pacific lowlands of Costa Rica, and T. systremmata (Luer) Luer, only located in the rich, cloud forests of Loja in Ecuador.   4   Before the beginning of this study, 20 species were recognized to be represented in Costa Rica (Luer 1997, Ossenbach et al. 2007). After that, two new species, Trichosalpinx reflexa (Fernández & Bogarín 2011), and T. sanctuarii Mel.Fernández & Bogarín (Fernández & Bogarín 2013) were added, while T. minutipetala (Ames & C.Schweinf.) Luer (previously considered a synonym of T. memor) was recognized as a distinct species (Fernández 2011). Therefore, the present study recognizes a total of 23 species, twelve of them belong to the subgenus Trichosalpinx, and 11 belong to the subgenus Tubella Luer. No species of the other two subtribes, Pseudolepanthes Luer and Xenia Luer, which are escentially South American, are known in Costa Rica. In Costa Rica, the elevational distribution of Trichosalpinx ranges from 100 m in the northwestern Pacific lowlands to 2500 m in the cloud forests of the Cordillera de Tilarán and the Cordillera Volcánica Central, with a maximum concentration of species between 800 and 1500 m (Fig. 2). Individuals are commonly found growing epiphytically in scattered trees in open areas and riparian forests, as well as in secondary and primary premontane and lower montane forests (life zones according to Holdridge 1967, 1987). Pollination of Trichosalpinx species is unknown (Blanco & Barboza 2005, Pridgeon et al. 2005). Nevertheless, floral morphology suggests myophily, especially because of the brownish or dull reddish floral colors and mobile lip, similar to those of other fly-pollinated orchids such as Bulbophylum Thouars (Teixeira et al. 2004, Jersáková et al. 2006) and Acianthera (Borba & Semir 2001). Further studies on scent and reward recognition are still needed. Plants of Trichosalpinx are epiphytic and caespitose or repent; some species are prolific (with ramicauls borne on top of each other), and all of them bear lepanthiform sheaths.   5   The leaves are apical on the ramicauls, and shape varies from broadly to narrowly elliptical, with acute to obtuse apexes; the inflorescence is apical, racemose and rarely single-flowered. The flowers are resupinate, with the lateral sepals separate or partially or completely connate, while the petals are membranous. The lip is entire or basally lobed, and is attached to the column foot by a flexible articulation. The column is semi-cylindrical, with a short foot. The anther is incumbent, operculate with two pyriform pollinia (Luer 2003). Luer proposed an infrageneric classification that has been used for more than 30 years now (Luer 1983, 1997). He organized Trichosalpinx in four subgenera: Tubella, Trichosalpinx, Xenia and Pseudolepanthes. Differences are based on the presence of prolific ramicauls, the relationship between the length of the leaf and that of the inflorescence, the shape and margins of the petals and the presence of a column-foot. Morphological and anatomical data was used by Archila (2000) to support the elevation of Tubella (to which he also added species from subgenus Xenia without providing evidence) and Pseudolepanthes to generic status. Trichosalpinx was included in recent phylogenetic analyses. Based on molecular data, Pridgeon and colleagues (2001) revealed that Trichosalpinx (sensu Luer) is a polyphyletic genus. Moreover, their results in the consensus tree show that subgenus Trichosalpinx is also polyphyletic, with two member of the subgenus strongly supported in one clade but separated from a third member, T. berlineri (Luer) Luer, which was found to be close to Zootrophion Luer with low support. The only species of subgenus Tubella evaluated, the widely distributed T. arbuscula (Lindl.) Luer, failed to provide phylogenetic information in the consensus tree. In a more recent study, Karremans (2014) evaluated the phylogenetic   6   relationships within Anathallis Barb.Rodr. and related genera, which confirmed the polyphyletic nature of genus and subgenus Trichosalpinx, and the need for a recircumscription to attain monophyly. His study also included a sample of T. arbuscula sister to the other Trichosalpinx species, with low bootstrap support (0.44). A phylogenetic analysis of genus Trichosalpinx (Fernández et al. in prep.), including several species of subgenera Trichosalpinx and Tubella, further evidences the need for a complete re- circumscription of this highly polyphyletic genus, which is diversely interrelated with other genera as Anathallis, Lepanthes, Zootrophion and the recently proposed Lankesteriana Karremans. Since the systematic relationships of Trichosalpinx sensu lato (i.e., including all the taxa that in some moment were assigned to it) are still unclear, I prefer to maintain the traditional circumscription of Trichosalpinx in the present contribution. Although Costa Rica has a well-established tradition in orchidology, the local knowledge on genus Trichosalpinx has been based on a relatively low number of herbarium collections, which naturally limits the understanding of their natural variation and the geographical and ecological patterns associated to their distribution. Additionally, many herbarium specimens are erroneously identified due to the difficulty ib recognizing characters in their small, dried flowers. Likewise, major difficulties in delimiting taxa in the several species complexes of the genus have not yet been resolved, so a taxonomic revision of the genus in Costa Rica is needed. As part of a major project aimed toward the completion of the treatment of Pleurothallidinae for the Flora Costaricensis, the present work aims to produce the first taxonomic revision of Trichosalpinx in Costa Rica, based on an extensive survey of living and preserved specimens and their correponding morphological variation and distributional and ecological data.   7   II. METHODOLOGY The taxonomic work was mainly conducted at Lankester Botanical Garden, University of Costa Rica (located in Dulce Nombre, Cartago province, 9°50’25.9” N 83°53’19.4” W at 1370 m), from 2011 to 2014. Collection of samples In this study live plants from fieldtrips were acquired. Different localities were visited according to the following cri teria: 1) their type locality (for species described from Costa Rican plants), 2) other Costa Rican localities reported in the protologue, 3) other Costa Rican localities recorded in herbarium samples and the literature. Botanical exploration in several traditionally under- collected areas of Costa Rica was also conducted. Whenever possible, at least 5 plants per locality and/or population were collected. For the purposes of the present work, a population was defined as all the individuals occupying the same phorophyte (following Tremblay 1997). Flowerless plants were cultivated at the Lankester Botanical Garden facilities until floral material was obtained for study. The herbarium specimens saved in the dried and spirit-preserved collections of the Costa Rican National Herbarium (CR), the Luis Fournier Origgi Herbarium (USJ), the National Biodiversity Institute Herbarium (INB) and the Lankester Botanical Garden Herbarium (JBL) were studied. Digital images of type specimens, when deposited outside Costa Rica, were also studied.   8   A database associated with the geographical information from living plants and herbarium specimens was created, which includes the following information: a) name of the species; b) locality (province, county, district, other locality data); c) geographic coordinates; d) vegetation type and life zone; e) habit; f) collecting date; g) collector name and number; h) herbarium of deposition. For spirit-preserved specimens at JBL lacking collector, the term “JBL-” followed by the Living Plants Collection identification number, all in italics, was used. For plants collected in the field during the course of this study, geographical reference data were obtained with a Garmin eTrex Vista GPS unit. Life zone classification was based on Holdridge´s Life Zone System (Holdridge 1967, 1987). Collecting data are partially available through the Lankester Botanical Garden Herbarium web site (http://www.jbl.ucr.ac.cr/php/investigacion/herbConsultEspecif.php). Description and documentation of specimens Unless indicated otherwise, botanical descriptions were prepared from living plants and spirit-preserved specimens at JBL, supplemented with information from preserved material at the other national herbaria. All herbarium material studied was annotated. Each plant collected from the field or otherwise flowered in cultivation at JBL was photographed, illustrated, preserved in spirit, and deposited in the Spirit Collection at JBL. The date indicated in all specimens deposited at JBL cited in the section “Additional Material Studied” refer to the collecting dates, not to flowering dates as in the material from other herbaria. When duplicates were available, they were dried and deposited at CR or USJ.   9   Recording of morphological characters, including size and shape, as well as drawings and photographs, were executed using Leica MZ 9.5 and M80 stereomicroscopes, provided with a drawing tube and Leica IC80 and Leica DFC295 digital cameras. Images of stems, flowers and fruits were obtained using Epson Perfection 2400 and Epson Perfection V600 scanners, at resolutions between 1000 and 3000 dpi. When live plants were available, taxa were illustrated with a composite line drawing based on living material. Composite plates were diagrammed by using Adobe Photoshop®. Ink drawings were prepared in smooth Fabriano® paper of 240 g/m2 with a Rotring® Rapidograph 0.1mm using black capillary cartridges and traced in Artograph LightPad® A920. Character variations of morphologically variable species were also drawn. Illustrations include at least the plant habit, flower, dissected perianth, column, anther and pollinaria. When possible, uniformity in floral views and plate composition was maintained to facilitate species comparisons. When the flower structure of a herbarium specimen was not easily observed, or if it did not seem to correspond to any of the species of Trichosalpinx studied from living plants, one or (preferably) two flowers were rehydrated by placing them in concentrated ammonia (NH3) in a closed vial for 6 to 24 hours, and then transferred to tap water. These flowers were examined, dissected and drawn using a stereomicroscope as specified above. Once completed, the flower parts were put in a lignin-free paper envelope, dried, and returned to the herbarium specimen along with a copy of the drawing and identification label where necessary. Phenological data (specifically flowering dates) were acquired from field collections and exsiccata. Phenological data obtained from cultivated plants were summarized separately   10   from that of plants in the field, as their phenology can be influenced by the conditions in cultivation. Phenology of populations from the Caribbean and Pacific versants was summarized separately to detect any differences for those species that occur in both sides of the continental divide. After documentation, individuals were classified based on their morphological similarities. Descriptions were made based on Stearn (1987). For the application of species names, type specimens or the corresponding images of species of Trichosalpinx found in Costa Rica were studied. The documented material and descriptions were compared with protologue descriptions and available type material of Trichosalpinx species known. Interpretation of size, shape, texture, arrangement and position characters were based on Stearn (1987) and Harris & Harris (2001). Reference material, illustrations, descriptions and images, as well as the articles generated from this project, are available online through the web site www.epidendra.org (Pupulin 2007, 2009).   11   III. TAXONOMIC HISTORY OF TRICHOSALPINX 1. The first discoveries The first species that would be eventually assigned to Trichosalpinx was described in 1816 as Dendrobium pusillum by Kunth, from a collection made by Humboldt, Bonpland & Kunth in Loja, Ecuador (Humboldt & Bonpland 1815; Fig. 3). Lindley (1835) transferred the species to Specklinia on the basis of its partially connate sepals that form a hump. Shortly later the same author transferred it to Pleurothallis (Lindley 1842: 82–83). Almost fifty years later Kuntze made a new combination of the species in Humboldtia Ruiz & Pav. (Kuntze 1891). Nonetheless, this genus is illegitimate in the Orchidaceae for being a later homonym (for a few months) of another genus proposed in 1794 in the Fabaceae (Vahl 1794). Finally, Luer (1983) made it the type species of his new genus Trichosalpinx Luer (Luer 1983). The type material is preserved at the National Museum of Natural History in Paris, where Humboldt and Bonpland deposited much of the material gathered during their voyage through South and North America. Lindley described Pleurothalis arbuscula Lindl. in 1842, and P. dura Lindl. in 1859. They were found in Loja (Ecuador) and Peru respectively (Lindley 1842, 1859; Figs. 8, 9). In 1838, Lindley added Specklinia ciliaris and S. orbicularis Lindl. (now Trichosalpinx ciliaris and T. orbicularis, respectively; Figs. 4, 5), two species described from plants cultivated in the greenhouses of the Loddiges family (Lindley 1838). Apparently, S. ciliaris was imported from somewhere in Mexico, while S. orbicularis was brought to the greenhouses from Demerara in the Guyanese coast. Shortly after, Pleurothallis memor Rchb.f. (now Trichosalpinx memor) was described by Reichenbach (1856). It was based on a plant with doubtful locality, cultivated by Forkel in Hamburg (Fig. 6). The species is rather common   12   from Mexico to the Andes and the Antilles. Its high morphological variability promoted the proliferation of synonyms, which according to Luer (1997) includes P. brevis Schltr., P. gnomonifera Ames, Trichosalpinx greenwoodiana Soto Arenas and T. nageliana Soto Arenas), as well as the entangling with other similar species in herbaria and literature (e.g., T. ciliaris [Lindl.] Luer and T. blaisdellii [S.Watson] Luer). Pleurothallis blaisdellii S.Watson (now Trichosalpinx blaisdellii [S.Watson] Luer) was described by Watson (1888) from a plant collected in the Chocón forests of Guatemala (Fig. 7). It is distributed from Mexico to Panama and grows in sites located between 300 and 1800 m of elevation (Luer 1997). At least five species names have been considered synonyms of T. blaisdellii afterwards (Luer 1997). 2. History of Costa Rican Trichosalpinx species Auguste R. Endrés documented the first records of Trichosalpinx in Costa Rica. He botanized in the country between the end of 1866 and the beginning of 1874. His specimens, descriptions and illustrations were sent to his principal correspondent, Professor Reichenbach in Hamburg. Today, the materials of Endrés are preserved at the Museum of Natural History of Wien, where Reichenbach's orchid herbarium was deposited after his death (Ossenbach et al. 2013). Amongst those materials the type of Pleurothallis moschata Rchb.f., the first species of Trichosalpinx described from Costa Rica, is found. Although the type locality is unknown, Endrés collected this plant during one of his many expeditions to the interior of Costa Rica. He described it as having “[…] prolific stems […] with obtuse, oblong leaves […] and inflorescences of white-yellowish flowers […]” (Fig. 10) (Pupulin et al. 2011). As usual, Endrés elaborated a set of fine sketches of the plant and   13   flower parts, which he sent to Reichenbach, along with the type material (Ossenbach et al. 2013) (Figs. 11 & 12). Reichenbach published the species in 1881 in his Xenia Orchidacea. After transfering it to Trichosalpinx, Luer (1997) reduced T. moschata (Rchb.f.) Luer to the synonymy of T. arbuscula, considering that “the collections from Central America known as T. moschata […] are not specifically different from those from the Andes, although the habit is smaller”. In 1923 Ames and Schlechter described Pleurothallis cedralensis Ames (1923), P. pergrata Ames (1923) and P. trachystoma Schltr. (Schlechter 1923) (Figs. 13, 14 & 15, respectively). The first two were collected around 1919 by Charles Lankester at El Guarco, south of Cartago in the Cordillera de Talamanca. Pleurothallis cedralensis is a commonly found species in Costa Rica, Panama, Ecuador, Venezuela and Bolivia with multiple inflorescence axes and plain yellow flowers (Luer 1997). Trichosalpinx pergrata is perhaps the most attractive species of the genus in Costa Rica; Ames called it “pergrata” in reference to the “very pleasant” appearance of the plant, characterized by the flowers with short peduncles and long, pink to red sepals. Trichosalpinx trachystoma (Schltr.) Luer was collected in 1921 in the vicinity of San Pedro de San Ramón, in the Cordillera de Tilarán, by Alberto M. Brenes, and sent to Schlechter for description. The type specimen at Berlin was destroyed during World War II, forcing it to be neotypified based on a drawing of the species (Barringer 1986). Two years later Ames and Schweinfurth described Pleurothallis nana Ames & C.Schweinf. naming it from the Latin nanus, or dwarf, in allusion to the tiny size of the plant (Ames & Schweinfurth 1925; Fig. 16). Lankester collected it at “Pejibaye de Turrialba” [now Jiménez], along the Caribbean watershed of the Cordillera de Talamanca. For several decades this was the only sample known, until he found it again in 1963 in Heredia,   14   somewhere between Poás and Barva volcanoes in the Cordillera Volcánica Central (Luer 1997). The same year Ames described Pleurothallis navarrensis Ames from plants collected by P.C. Standley at Navarro de El Muñeco in Cartago, now deposited at AMES and US (Ames 1925; Fig. 17). Ames compared this species with Pleurothallis abjecta Ames [today considered a synonym of Lankesteriana barbulata (Lindl.) Karremans, Karremans (2014)], from which it differs mainly by the abbreviate peduncle and the fimbriate, lanceolate petals. Schweinfurth (1937) described Pleurothallis rotundata C.Schweinf. on the basis of a plant found by Hunter and Allen in the mountains of La Pintada, Coclé, Panama (Fig. 18). The description was based on a flower in advanced state of anthesis. In 1981, Luer described Trichosalpinx operculata Luer, also from Panama, distinguishing it from T. rotundata (C.Schweinf.) Luer by the lip that is not fimbriate in the last third, as opposed to T. rotundata. Nevertheless, after rehydrating the flowers of the type of T. rotundata, Luer reduced his T. operculata to the synonymy of the former species (Luer 1997). In one of his many collecting expeditions around San Ramón de Alajuela, Brenes found a small plant with inflorescences of few pale flowers, which he sent to Harvard University. There, Schweinfurth described it as Pleurothallis membraniflora C.Schweinf. (Schweinfurth 1938; Fig. 19). In 1940 Schweinfurth described P. broadwayi var. tricarinata C.Schweinf. (Fig. 20) from a plant collected in Palmira, Alajuela by A. Smith. More recently, Luer (1977) recognized this variety at the species level but the epithet tricarinata had been previously used for a species in Pleurothallis, and it was necessary to propose a new name. Luer named it Pleurothallis carinilabia (Luer) [now Trichosalpinx carinilabia (Luer) Luer].   15   In 1996, Luer described Trichosalpinx fruticosa Luer, T. lankesteriana Luer, T. ringens Luer and T. todziae Luer (Figs. 21–24). The first, apparently rare, was found at Cerro de la Muerte (type locality), on the highlands of the Cordillera de Talamanca in Costa Rica (Luer 1996), as well as in Guatemala and El Salvador. Trichosalpinx lankesteriana was described from a plant in cultivation at Lankester Botanical Garden with no collecting data. In the description, Luer indicated that T. lankesteriana is barely something more than a geographical variation of T. blaisdellii (Luer 1996), which presents ample distribution from Mexico to Panama. Trichosalpinx blaisdellii has been considered a morphologically very variable species, thus it is not surprising that the few morphological differences of T. lankesteriana had made the author reconsider the species and add it to the long list of synonyms of T. blaisdellii just a year later (Luer 1997). In 1997 Trichosalpinx parsonsii Luer & Dod was described by Luer and Dod based upon a plant collected in 1990 by Grantham and Parsons at Santa María de Dota, on the Pacific side of the Cordillera de Talamanca (Luer 1997). The type was the only specimen known, until a plant from San Ramón de Alajuela was recently found and preserved at JBL.   16   IV. CURRENT PHYLOGENETIC CIRCUMSCRIPTION In their phylogenetic study of Pleurothallidinae based on molecular evidence, Pridgeon and colleagues (2001) carried out separate and combine analyses of nuclear and plastid DNA sequences. Four species of Trichosalpinx, T. arbuscula (subgenus Tubella), T. berlineri, T. blaisdellii and T. orbicularis (Lindl.) Luer (subgenus Trichosalpinx) were incorporated in the survey, although only the last three were included in the combined tree. According to the results, Trichosalpinx belongs to the Lepanthes clade, which includes Frondaria, Lepanthes, Lepanthopsis, Pleurothallis subgenus Acuminatia Luer and Specklinia section Muscosae Lindl., and Zootrophion. This clade had 100% bootstrap support in the combined tree. According to the authors, a unique feature identifies the clade: the existence of imbricating or sclerotic sheaths covering the stem (Pridgeon et al. 2001). In the particular case, the results show that Trichosalpinx is a polyphyletic group. The tree from the ITS matrix shows T. arbuscula isolated from the other four Trichosalpinx species included in the analysis, and placed next to members of Pleurothallis subgenus Acuminatia [=Pleurothallis section Acuminatae Lindl. (Lindley 1859)], presently classified in the genus Anathallis (Pridgeon & Chase 2001). Lindley had already recognized the affinities of T. arbuscula with members of Anathallis, having placed both groups in Pleurothallis section Acuminatae. According to Lindley, these affinities are based upon the presence of racemes longer than the leaf and acuminated sepals (Lindley 1859, Pridgeon et al. 2001). Furthermore, the tree from the ITS matrix shows T. orbicularis and T. blaisdelli together in a subclade of their own, with complete bootstrap support in the combined tree. For a   17   trained eye, the morphological affinities of T. orbicularis, T. blaisdellii and T. berlineri are evident, with all three species having caespitose habits with no prolific ramicauls, racemes shorter than the leaf, relatively fleshy, ciliate sepals, and an oblong lip with a central callus. However, in the study of Pridgeon and colleagues (2001), T. berlineri forms a subclade (although weakly supported in the combined tree) along with members of Zootrophion (a genus with which Trichosalpinx shares few morphological traits), and isolated from the group formed by T. blaisdellii and T. orbicularis (Pridgeon & Chase 2001). A recent analysis carried out by Karremans (2014) including five species of Trichosalpinx confirms Pridgeon et al.’s (2001) results and shows that Trichosalpinx is, at least, biphyletic, and probably triphyletic. A first clade formed by T. blaisdellii and T. orbicularis is found (with low bootstrap support) to be sister to Anathallis, in turn sister to the clade formed by Lepanthes, Frondaria and Lepanthopsis. A second clade formed by T. berlineri and T. dependens (Luer) Luer is sister to the recently proposed genus Lankesteriana Karremans, and in turn sister to Zootrophion. An accession of T. arbuscula is sister to all clades except for the outgroup species. The need of a better representation of Trichosalpinx taxa in phylogenetic analysis to correctly assess their evolutionary position is evident, but it is almost certain that Trichosalpinx, as currently circumscribed, is polyphyletic.   18   V. MORPHOLOGY Roots Roots are cylindrical, measuring 1–3 mm in diameter, developing from the nodes of the rhizome, with greenish or yellowish tips (Fig. 26). The formation of aerial roots was frequently observed in species of subgenus Tubella (Fig. 26-A). They anchor the plant to the substrate and entangle with tufts of lichens and mosses, serving an important structural function. In species of subgenus Trichosalpinx, only a few plants of T. memor and T. blaisdellii produced aerial roots. Pridgeon (1982) made several observations on the root, stem and leaf anatomy of Trichosalpinx. In the transverse section of T. carinilabia he found that the velamen is uniseriate with elliptical or rectangular cells; the exodermis is uniseriate with long, rectangular, polygonal or elliptical cells with U-shaped wall thickenings (no details to the identity of the wall are mentioned), while the passage cells have thin walls. The cortical parenchyma cells are isodiametric to elliptical in cross section, and the endodermis is uniseriate with O- or U-shaped periclinal wall thickenings. The pericycle is also uniseriate; the stele is tetrarch and the pith sclerotic (Pridgeon 1982, Pridgeon et al. 2005). Root morphology showed no special value in the taxonomy of the group at the specific level. Characters evaluated (color, length and diameter) varied regardless of the species. However, production of aerial roots was representative of subgenus Tubella. Stems   19   Although there is some agreement in the application of the term rhizome to designate the horizontal portion of the stem, the application of a name to dessignate the vertical portion that bears leaves and inflorescences is more controversial. It has been called a “secondary stem” (), but the term condradicts the concept that both the horizontal and vertical portions are formed by a succession of nodes and internodes, and thus, refers to a single organ, a unique stem (Dressler 1981). Stern and Pridgeon (1984) proposed the term “ramicaul” to distinguish it from the rhizome; nevertheless, the word itself has been considered inappropriate because the first part refers to a (not necessarily present) branching property of the term (Rasmussen 1985). Soto Arenas (1987) used “stem” to name the vertical portion after considering that it was impossible for it to be confused with any other organ of the plant. Here, I use the terms rhizome and ramicaul, the latter defined as the part of the stem above of the last reiteration bud in Pleurothallids. This term has been used extensively in the most thorough taxonomic treatments of Trichosalpinx (Luer 1983, 1986, 1994), which are baselines for this systematic study. The first internodes of each sympodial unit are somewhat thickened and grow horizontally, and make up the segment of stem called the rhizome. In this portion of the stem, meristems produce new shoots and roots. The ramicauls produced are terete, from 7 mm (as in T. parsonsii) to 20 cm long (as in T. minutipetala), mostly slendmnmmer and elongated (Fig. 27). In subgenus Tubella prolific stems are present in all the species except for T. pergrata. The ramicauls are frequently longer than the leaves, and have three or more internodes. The presence of a conspicuous annulus is consistent in all species. Although production of new shoots from the apical meristems is characteristic in subgenus Tubella (Fig. 28A), the same phenomenon was observed in some plants of subgen.   20   Trichosalpinx in the wild and in cultivation. Producing proliferations may enhance the plant photosinthetic capability. The ramicaul cortex is composed of sclerified subepidermal layers and parenchymatous inner cells. The vascular bundles are colateral and arranged in one or two concentric rings, with sclerenchyma bundle caps mostly developed at the outer (phloem) flank. The outermost ring of vascular bundles is continuous with the sclerenchyma sheath, and there can be up to eight cell layers between bundles (Pridgeon 1982). Ramicaul size and shape are diagnostic for some Costa Rican taxa. The fractiflex ramicaul of Trichosalpinx ciliaris is unique to that species. The ramicauls of the prolific T. arbuscula, which forms long chains of superposed ramicauls, reach as long as 23 cm altogether, and easily exceed the length of all other Costa Rican species. In T. pergrata, the abscence of prolific ramicauls separates this species from other Tubellas. Ramicaul sheaths Ramicauls are covered with lepanthiform sheaths, a term used to describe the sheaths of several taxa including Draconanthes, Lepanthes, Lepanthopsis and Trichosalpinx. Lepanthiform sheaths are tubular, more or less imbricate, longitudinally ribbed with oblique ostia, frequently dilated and thickened at the margins; the ribs and margins can be ciliate, scabrous or rarely glabrous. When young, sheaths are glumaceous and frequently densely covered with purplish cilia. When old, sheaths turn dry and fibrous with less numerous, harder cilia. The apical sheath of the ramicaul is longer and wider than the rest (although this is less evident in Tubella),   21   and the apex is often elongated to cover both the petiole and the base of the leaf blade. The distalmost sheath tissue can be broken by the emergence of new organs, like leaves, inflorescences and roots. Taxonomically, sheath morphology is only significant to distinguish the two subgenera of Trichosalpinx present in Costa Rica. In subgenus Trichosalpinx, the sheaths are mostly funnel-shaped, and the ostia has particularly thickened margins; plants of T. minutipetala bear amply dilated ostia. In species like T. blaisdellii, T. memor and T. minutipetala the distalmost sheath of a ramicaul is reticulated (Fig. 28C). In subgenus Tubella the sheaths are more tubular and adpressed. The longitudinal ribs are more evident when the sheaths have recently developed (Fig. 28B). Leaves Most Trichosalpinx species show well developed, conduplicate, erect to suberect, thin to fleshy or coriaceous, glabrous, ovate to elliptic, acute to obtuse leaves. Members of subgenus Trichosalpinx bear mainly coriaceous, fleshy, ovate to elliptic, acute leaves with short (5–15 mm) petioles (Fig. 27). In this subgenus, overall leaf size is greater than in members of Tubella. The leaves of some species (as T. blaisdellii, T. memor and T. rotundata) are suffused with purple beneath, more intensively along the veins. Some leaf characters are of diagnostic use. Only plants of T. ciliaris, T. minutipetala and T. reflexa develop narrowly elliptical, acute leaves. Along with distribution data and the shape of ramicaul sheaths, the coriaceous, convex, orbicular leaves of T. caudata, T. orbicularis and T. rotundata are helpful when determining specimens.   22   On the other hand, species of Tubella bear ascending to erect, coriaceous, glabrous, orbicular to ovate, acute to subacute leaves, with short petioles 0.3–5.5 cm long. The leaves of this group tend to be smaller than those of subgenus Trichosalpinx (1.2–8.1 cm). Young leaves frequently show purple-stained veins, more evident abaxially, as in T. cedralensis, T. dura and T. pusilla (Kunth) Luer. This peculiarity is also observed in some species of Lepanthes and Zootrophion. Iridescence was observed on the adaxial leaf surface of several individuals of T. blaisdellii and T. memor, frequently found in the main trunk of trees and bushes at understory level. This phenomenon may be an adaptation for the capture of photosynthetically active light in leaves that have grown in low light conditions (Glover & Whitney 2010). As summarized in Pridgeon et al. (2005), there are hairs present on both surfaces (glandular, sunken, consisting of two or three cells), while the stomata are restricted to the abaxial surface. The epidermal cells are polygonal, rectangular or elliptical, usually with conspicuous primary pit-fields. In the transverse sections the cuticle is of variable thickness, often minutely papillose; the epidermal cells are elliptical, rectangular, or peg- shaped, and adaxial cells are larger than abaxial cells. The stomata are flushed with the abaxial surface. The hypodermis presents spiral thickenings; the chlorenchyma is heterogeneous, the pallisade mesophyl 1-3 layers, spongy 4-10 layers; spirally thickened idioblasts are present. Vascular bundles are collateral and arranged in a single row in the centre of the mesophyll; the largest bundles with sclerenchyma caps of 1–2 files of fibers adaxially and up to four files of fibers abaxially. Fiber bundles are absent. For more details on the anatomical structure of Trichosalpinx, see Pridgeon (1982) and Pridgeon et al. (2005).   23   Inflorescences Inflorescences of Trichosalpinx are alternate, distichous, mostly secund, erect, descending or pendulous, terminal racemes (except for the fascicles produced by T. ringens and T. sanctuarii). The structure is remarkably similar throughout the examined species. Peduncles are produced at the apex of the ramicaul near the base of the adaxial face of the leaf from a conduplicate, papyraceous, ovate to triangular, acute spathe, 1–2 mm long. Although most examined inflorescences grow adpressed against the abaxial leaf surface, the peduncles occasionally turned around the petiole and grow facing the adaxial leaf surface. The rachis is terete, glabrous and persistent, and is covered with small, papyraceous floral bracts. Ovaries and pedicels are terete, glabrous, from 1 to 3.5 mm long and exhibit no ornamentation. Ovaries are shallowly, longitudinally channeled. In most species, peduncles shift direction, positioning flowers in a secund or subsecund acommodation. After flowering, the pedicels remain photosynthetic and are persistent until they become dry. With the exceptions of the few-flowered Trichosalpinx pergrata, T. ringens and T. sanctuarii, inflorescences of Costa Rican Trichosalpinx species produce three or more flowers per inflorescence (Fig. 29). Furthermore, one to three racemes can develop simultaneously from the same ramicaul. As noted by Luer (1997) inflorescence/leaf length ratio is diagnostic at the subgeneric level. Inflorescences of the nominal subgenus are mostly shorter than the leaf, while in species of Tubella the inflorescence is commonly longer than the leaf, except in T. pergrata.   24   Flowers Dorsal sepals are mostly broadly ovate to narrowly triangular with acuminate to attenuate apices. Lateral sepals frequently form a mentum at the base to accommodate the column- foot. The sepals of subgenus Trichosalpinx are fleshy, ciliate and colorful (dark vinaceous, red, purple), with a white base and discolored, thickened apices. The lateral sepals in this subgenus are fully or partially connate, and sometimes form a concave synsepal or bifid laminae, as in T. ciliaris and T. blaisdellii, respectively. In subgenus Tubella, the sepals are membranous, free to partially free, carinate, glabrous and mostly unicolor, white to yellowish or greenish, except for the fuchsia to pink coloration of T. pergrata. (Fig. 30). Petals are very similar among species; they are consistently straight, flat, membranous (except for the fleshy petals of T. pergrata, T. ringens and T. sanctuarii), 1-veined, mostly ovate to elliptical, oblique in shape, and acute to acuminate or obtuse, often with fimbriate or denticulate apical margins. Lips are ovate to oblong with the disc broadened in, at least, half of the species, but simpler or more complex lips also occur. Lips in subgenus Trichosalpinx are mostly brown- purple to dark purple, marginally ciliate, oblong, orbicular or triangular, from 2 to 4 mm long. The disc (in this treatment, the central area along the basal half of the lip) usually bears a keel or callus from the base to the middle where it disappears or forks in two. The base presents two retrorse auricles; the apex is usually not thickened, softly descendent. The lip is attached to the column-foot by a flexible, membranous hinge (except in T. ringens and T. sanctuarii, where the lip is rigidly attached to the column foot) that enables it to move up and down with the help of wind or pressure, as in some species of Bulbophyllum (Teixeira et al. 2004) and Lankesteriana (Karremans 2014). In subgenus   25   Tubella, lips are yellow or white, glabrous, elliptical-oblong, flat to slightly arcuate; the base is frequently broadened, with erect, lateral lobes; the apex is thickened in most species. The presence and shape of a longitudinal callus in the disc area, as well as the thickening of the midvein and apex are of diagnostic value in species of both subgenera. Although they belong to subgenus Trichosalpinx, T. ringens and T. sanctuarii have a general flower architecture that differ significantly from the rest of the group. The dorsal sepals are narrowly ovate, thick and erect, and the veins are profusely stained. Petals are ciliate, thick and 3-veined. The lips are fleshy, arcuate and the adaxial surface is partially pillose; they lack the basal, retrorse lobules and the flexible hinge present in other species, which suggests a different pollination syndrome (Fernández & Bogarín 2013). In general terms, columns are white, straight, terete, with a variously irregular apex, and with a projected basal portion forming a conspicuous column foot, to which the lip attaches by a membranous articulation. The apical portion of the column can be provided with lateral appendages called wings, more frequently in subgenus Trichosalpinx. The anther is apical and the stigma ventral in subgenus Trichosalpinx, while both are mostly ventral in subgenus Tubella (Fig. 31). The anther cap is incumbent, ovate and emarginate. Pollinaria Pollinaria are formed by two ovoid to pyriform pollinia. Their structure can be of three basic kinds: a) pollinia with a flat pair of suborbicular to ovate, basally adjoined caudicles (“whale-tail” pollinaria sensu Karremans et al. 2013), as in subgenus Trichosalpinx (Fig. 32A); b) pollinia free or adjoined by the celullar, oval caudicles, as in subgenus Tubella (Fig. 32B); c) pollinia free with no evident joining structure upon removal from the anther,   26   as in T. sanctuarii. As in other orchids lacking a viscidium, the viscous, hyaline substance present in the rostellum presumably allows the attachment of the pollinia to the back of the pollinator, enabling their removal when the pollinator recedes to leave the flower (Fernández & Bogarín 2013). Fruits Fruits are fleshy, ovoid, 3-valved, dehiscent capsules with parietal placentation. The external surface is smooth, shiny and not ornamented; in subgenus Trichosalpinx it is usually pigmented with pink or fuchsia, while in species of Tubella it remains green. Dehiscence is loculicidal, but the capsules split along only two functional longitudinal slits, as observed in most pleurothallids (Fernández et al., in prep.). The dehiscence lines are incomplete, so that the two valves of the capsule remain united at both ends. Perianth remnants are persistent at the apex of the fruit (Fig. 33). Hygroscopic endocarpic trichomes occur in the fruits of Trichosalpinx. The trichomes are unicelular, borne in two rows on either side of each carpel midrib. The capillitium (the aggregation of the trichomes) is responsible for dispersing seeds when they twist and recoil with changes of humidity as soon as the fruit opens (Dunsterville 1969, Blanco et al. 2006, Rasmussen & Johansen 2006). The seeds form masses that are densely attached around the prolongations of the placentae. Each mass is interspersed by several trichomes, but not all trichomes intersperse masses of seeds. The seeds of Trichosalpinx subgenus consist of an oval embryo that occupies nearly a fourth of the total length of the fusiform testa. The testa bears subquadrate to linear periclinal walls. The seeds of species in subgenus Tubella are unknown.   27   Fruits have been uncommonly found in the wild. However, plants of T. blaisdellii, T. memor and T. reflexa cultivated in the greenhouses at Lankester Botanical Garden produce fruits abundantly. Diagnostic fruit characters at the specific level were not found during the course of this study, especially because of the lack of fruit production in most species. However, some tendencies can be inferred at the subgeneric level. Fruits of subgenus Trichosalpinx tend to be bigger, ellipsoid and suffused with red or pink externally. In subgenus Tubella fruits are smaller, spheroid and green with no red stains. Further studies are needed to evaluate their diagnostic use in taxonomy.   28   VI. SYSTEMATIC TREATMENT In 1983, Luer created the genus Trichosalpinx to allocate species with lepanthiform sheaths, which are also present in Draconanthes, Lepanthes and Lepanthopsis. Nevertheless, Trichosalpinx is differentiated by the longer rather than broader petals (transversely bilobed in Lepanthes), the lip with an elongate mid lobe (absent or turned into a small appendix in Lepanthes), trilobed, usually membranous petals (simple in Lepanthopsis, fleshy in Draconanthes), and an elongate column with an elliptical stigma (short, with a transversally bilobed stigma in Lepanthopsis) usually provided with a foot (without a foot in Lepanthes, Lepanthopsis and Draconanthes; Luer 1997, Luer & Thoerle 2010). According to the classification of Luer (1986, 1997), the genus is subdivided in four subgenera: Trichosalpinx, Tubella Luer, Pseudolepanthes Luer and Xenia Luer. Subgenus Trichosalpinx is composed of species of caespitose habit with non-prolific ramicauls and usually coarse lepanthiform sheaths, racemes shorter than the leaf (except for T. pringlei), petals that are frequently ciliate, denticulate, erose, fimbriate or serrate, a lip with two basal, retrorse auricles (except for T. pringlei), and a straight column with a foot (Luer 1997). Species belonging to subgenera Tubella are mostly provided with prolific stems, racemes commonly longer than the leaf, an oblong, ovate, entire or trilobed lip with 2 or 3 longitudinal calli and no basal auricles, and a column foot, to which the lip is attached. Subgenus Pseudolepanthes is characterized by the non-prolific stems, racemes longer than the leaf, sepals more or less spiculate, a thick, wide, basally unguiculate lip with a conspicuous, verrucose callus, and a footless column (Luer 1997). Finally, subgenus Xenia clusters species with no resemblance to the other three subgenera. Few are the   29   characteristics shared among the species of subgenus Xenia, including non-prolific ramicauls, racemes longer than the leaf, glabrous sepals, a variously lobed lip with a callus, and a footless column (Luer 1997). All known species belonging to this two subgenera are found in South American countries, with emphasis in Colombia and Ecuador. While some Trichosalpinx species present low morphological variability, others (generally those with ample distribution) appear to form complexes characterized by numerous morphological variations. In Costa Rica, may be observed among populations of T. blaisdellii, T. dura, T. memor and T. orbicularis. Luer emphasized his opinion that the recognition of these variations as different taxa, far from clarifying relationships, can lead to a major confusion in the taxonomy of the group (Luer 1997). Nonetheless, the systematic study of morphological, anatomical biogeographical and molecular data sets, supported by an ample sampling of these species complexes, can offer relevant information about the variation allowing, in some cases, the recognition of well-defined entities. Archila (2000) elevated subgenera Tubella (to which he also added species from subgenus Xenia) and Pseudolepanthes to the generic level. These transfers were based on morphological differences –some of which previously recognized by Luer (1997)– including the inflorescence/leaf length ratio in relation to that of the leaf, ramicaul branching degree, presence of roots growing from the base of proliferating ramicauls, anther position, and shape and size of the rostellum and caudicle. To further support his proposal, Archila cited differences in the type of leaf venation and the composition of flavonoids, but he did not indicate which species and how many samples per species were   30   analyzed. In the present treatment, the traditional circumscription of Trichosalpinx is provisionally accepted (i.e., including subgenus Tubella and Pseudolepanthes). Trichosalpinx Luer, Phytologia 54(5): 393–394. 1983. TYPE SPECIES: Trichosalpinx ciliaris (Lindl.) Luer, Phytologia 54(5): 395. 1983. BASIONYM: Specklinia ciliaris Lindl. Edwards's Bot. Reg. 24: Misc. 31. 1838. Trichosalpinx subgenus Trichosalpinx Luer, Monogr. Syst. Bot. Missouri Bot. Gard. 15: 1–81. 1986. = Pleurothallis section Lepanthiformes (Lindl.) Cogn., Flora Bras. 3(4): 579. 1896, in part. BASIONYM: Pleurothallis section Brachystachyae subsection Lepanthiformes Lindl., Fol. Orch., Pleurothallis 25. 1859. LECTOTYPE SPECIES, designated by Luer, Mongr. Syst. Bot. Missouri Bot. Gard. 15: 65. 1986: Specklinia ciliaris Lindl., Edwards's Bot. Reg. 24: Misc. 31. 1838. = Pleurothallis section Bipaleolatae Pabst, Orchid. Bras. 1: 162. 1975. TYPE SPECIES: Specklinia orbicularis Lindl., Edwards's Bot. Reg. 24: 31. 1838. Trichosalpinx subgenus Tubella Luer, Monogr. Syst. Bot. 15: 66. 1986. TYPE SPECIES: Pleurothallis acremona Luer, Selbyana 5(2): 157. 1979. = Pleurothallis sect. Elongatae subsect. Lepanthiformes Lindl., Fol. Orch., Pleurothallis 26. 1859. LECTOTYPE SPECIES, designated by Luer, Monogr. Syst. Bot. Missouri Bot. Gard. 15: 65. 1986: Pleurothallis chamaelepanthes Rchb.f., Bonplandia 3: 240. 1855.   31   = Pleurothallis sect. Acuminatae subsect. Lepanthiformes Lindl., Fol. Orch. Pleuroth. 32. 1859. LECTOTYPE SPECIES, designated by Luer, Monogr. Syst. Bot. Missouri Bot. Gard. 15: 65. 1986: Pleurothallis arbuscula Lindl. Edwards's Bot. Reg. 28: Misc. 72-73. 1842. = Trichosalpinx subgen. Tubella sect. Tubellae Luer, Monogr. Syst. Bot. Missouri Bot. Gard. 15: 68. 1986. = Tubella Archila, Revista Guatemalensis 3(1): 46. 2000. TYPE SPECIES: Pleurothallis acremona Luer, Selbyana 5: 157. 1979. Trichosalpinx subgen. Pseudolepanthes Luer, Monogr. Syst. Bot. Missouri Bot. Gard. 64: 5. 1997. TYPE SPECIES: Trichosalpinx pseudolepanthes Luer & Escobar, Orquideología 16(2): 183. 1984. = Trichosalpinx subgen. Tubella sect. Pseudolepanthes Luer, Monogr. Syst. Bot. Missouri Bot. Gard. 15: 68. 1986. TYPE SPECIES: Trichosalpinx pseudolepanthes Luer & R.Escobar, Orquideología 16(2): 183. 1984. = Psedulepanthes Archila, Revista Guatemalensis 3(1): 76. 2000. TYPE SPECIES: Trichosalpinx pseudolepanthes Luer & R.Escobar, Orquideología 16(2): 183. 1984. Trichosalpinx subgen. Xenia Luer, Monogr. Syst. Bot. Missouri Bot. Gard. 64: 6. 1997. TYPE SPECIES: Trichosalpinx escobarii Luer, Monogr. Syst. Bot. Missouri Bot. Gard. 64: 114. 1997. = Tubella Archila, Revista Guatemalensis 3(1): 46. 2000. TYPE SPECIES: Trichosalpinx escobarii Luer, Monogr. Syst. Bot. Missouri Bot. Gard. 64: 114. 1997.   32   1. Key to the Costa Rican subgenera and species of Trichosalpinx Out of the four subgenera, only subgen. Trichosalpinx and Tubella have representatives in Mesoamerica. Based on the morphological differences given in this treatment, three keys to the Costa Rican Trichosalpinx are presented. Species not found during the course of this study were inluded in the keys based on the protologue and exsiccata material, and marked with an asterisk (*). Key to the subgenera of Costa Rican Trichosalpinx 1 Ramicauls not prolific, with coarse lepanthiform sheaths; racemes shorter than the leaf; petals frequently ciliate, denticulate, or fimbriate; lip entire, the base with two auricles, the disc with one central callus ................................................. subgenus Trichosalpinx 1' Ramicauls prolific (except in T. pergrata), with papyraceous to coarse lepanthiform sheaths; racemes longer than the leaf (except in T. pergrata); petals entire; lip entire or trilobed, the base without auricles, the disc with or without a prominent midvein, and frequently with two lateral, longitudinal calli .............................. subgenus Tubella   33   Key to the Costa Rican species of subgenus Trichosalpinx 1 Leaves ovate, orbicular or elliptic ................................................................................. 2 2 Inflorescence a succesive 1- or 2- simultaneously flowered fascicle, present on the adaxial leaf surface ........................................................................................................ 3 3 Sepals longitudinally veined, yellow with brown-red along the veins; petals translucent with red stripes; lip trilobed, with a central callus that forks in two below the middle, gently arcuate, with no plates ........................................... T. sanctuarii 3’ Sepals purple-brown and pale yellow toward the apices; petals entirely brown-red; lip entire with no callus, deeply arcuate, with a pair densely ciliate lamellate plates ................................................................................................................. T. ringens 2’ Inflorescence a several-flowered raceme, presented on the abaxial leaf surface ..... 4 4 Sepals connate to near their apex, forming a concave synsepal .......................... 5 5 Lip apex broadened, without a longitudinal keel .......................... T. trachystoma 5’ Lip apex acute to obtuse, with a longitudinal, apically bifid keel from the base to near the middle .................................................................................... T. memor 4’ Sepals connate to the middle of their length or less, forming a bifid blade ........... 6 6 Sepals glabrous ................................................................................................ 7 7 Dorsal sepal narrowly triangular; lateral sepals fleshy, thickened at the apex ....................................................................................................... T. caudata 7’ Dorsal sepal ovate; lateral sepals membranous, not thickened at the apex ................................................................................................... T. orbicularis 6’ Sepals pubescent ............................................................................................. 8 8 Petals almost as long as sepals; lip with the margin ciliate or fimbriate, the adaxial surface glandular-pubescent .................................. T. navarrensis*   34   8’ Petals half or less as long as sepals; lip with the margin entire, the surface smooth ........................................................................................................ 9 9 Lip yellow, narrowly triangular ................................................. T. rotundata 9’ Lip purple, oblong ................................................................... T. blaisdellii 1’ Leaves narrowly elliptic to Ianceolate ........................................................................ 10 10 Synsepal reflexed ..................................................................... T. reflexa 10’ Synsepal concave ............................................................................... 11 11 Petals quadrate; lip sharply reflexed ......................... T. minutipetala 11’ Petals oblong; lip straight ................................................... T. ciliaris   35   Key to the Costa Rican species of subgenus Tubella. 1 Ramicauls not prolific, with the sheaths densely hispid along ribs and margins; sepals fuchsia-rose to dark-rose (exceptionally yellow) ............................................. T. pergrata 1’ Ramicauls prolific, with the sheaths minutely ciliate along ribs and margins; sepals yellow or white ................................................................................................................. 2 2 Leaves fleshy, obovate to obelliptic, surface convex, dark green to purple .. T. pusilla 2’ Leaves coriaceous, elliptic, flat, green or stained with purple only along the veins ... 3 3 Lateral sepals reflexed ca. 90 degrees at the base; petals crenate .. T. carinilabia 3’ Lateral sepals straight or slightly reflexed in the apical half; petals entire or occasionally serrate ............................................................................................. 4 4 Petals ovate ….................................................................................... T. fruticosa 4’ Petals obovate or elliptical .................................................................................. 5 5 Lip apex not thickened ...................................................................................... 6 6 Lip without lateral lobes ............................................................... T. arbuscula 6 Lip with lateral lobes ............................................................ T. membraniflora 5’ Lip apex thickened ........................................................................................... 7 7 Lip with lateral, erect lobes, the disc without calli or thickened midvein .................................................................................................... T. cedralensis 7’ Lip with no lobes or subtle lateral lobes, these not erect ............................... 8 8 Lip disc flat ................................................................................... T. parsonsii 8 Lip disc with a central calli or thickened midvein ............................................ 9 9 Lip with two submarginal, longitudinal calli and a thickened midvein; column broadly winged apically .............................................................. T. dura   36   9’ Lip with two shallow thickenings at the forward angle between the lateral lobes and the papillous inferior lobe; column obtusely winged apically .............................................................................................................. 10 10 Plants erect; pedicels 1.5-4.0 mm; sepals long-attenuate; lip flat or inconspicuously thickened apically ........................................ T. todziae 10’ Plants descendent or pendulous; pedicels 2.5-6.0 mm long; sepals acute; lip conspicuously thickened apically ............................... T. nana   37   2. Species description and discussion The most recent published catalogue of Costa Rican orchid species (Ossenbach et al. 2007) recorded twenty species of Trichosalpinx for the country. Since then, two new species have been added (Fernández & Bogarín 2011, 2013) and another one, previously considered a synonym, has been shown to be a good species (Fernández 2011), all as part of the present study. In the following account, species are presented alphabetically by their currently accepted name. Square brackets [ ] indicate entries made by the author that do not necessarily reflect data present on the specimen labels or the protologue. 1. Trichosalpinx arbuscula (Lindl.) Luer, Phytologia 54(5): 394. 1983. BASIONYM: Pleurothallis arbuscula Lindl., Edwards's Bot. Reg. 28: Misc. 72−73. 1842. Humboltia arbuscula (Lindl.) Kuntze, Revis. Gen. Pl. 2: 667. 1891, nom. illeg. TYPE: Perú [Ecuador] found near Loja, Hartweg s.n. (holotype, K-79691; isotype, AMES-118556; photo of type, AMES, LD, W). Fig. 34. Trichosalpinx dinotherii (Rchb.f.) Luer, Phytologia 54: 395. 1983. BASIONYM: Pleurothallis dinotherii Rchb.f & Warsc., Bonplandia 2: 114. 1854. Humboldtia dinotherii (Rchb.f.) Kuntze, Rev. Gen. Pl. 2: 667. 1891, nom. illeg. TYPE: Perú, Warscewicz s.n. (holotype, K; isotype, AMES; drawing of type, AMES-38011). Pleurothallis diptera Lindl., Fol. Orchid. Pleurothallis 44. 1859. Humboldtia diptera (Lindl.) Kuntze, Rev. Gen. Pl. 2: 667. 1891, nom. illeg. TYPE: Ecuador: Pichincha, forest on the road to Nanegal, Jameson s.n. (holotype, K; drawing of type, AMES).   38   Trichosalpinx moschata (Rchb.f.) Luer, Phytologia 54: 396. 1983. BASIONYM: Pleurothallis moschata Rchb.f., Xenia Orchid. 3(2): 42–43, t. 217. 1881. TYPE: Costa Rica: Cultivated at Hamburger Botanischen Garten, collected by Endres in Costa Rica, Endres s.n. (holotype, W). Pleurothallis lepanthoides Schltr., Repert. Spec. Nov. Regni Veg. Beih. 7: 106. 1920. TYPE: Colombia: Cauca, M. Madero s.n. (holotype, B, destroyed; neotype designated by Luer (1997, as “lectotype”), AMES-28789, illustration of type). Pleurothallis tricaudata Schltr., Repert. Spec. Nov. Regni Veg. Beih. 9: 77. 1921. TYPE: Peru: Huánuco, Huánuco, mountains between Chinchao and Acomayo, Sept 1913, 3100-3200 m, Weberbauer 6827 [holotype, B, destroyed; lectotype, selected by Christenson (1996), MOL; isolectotypes, AMES, MOL, US]. Trichosalpinx arbusculoides (Hashimoto) Luer, Phytologia 54: 394. 1983. BASIONYM: Pleurothallis arbusculoides Hashimoto, Bull. Natl. Sci. Mus. 4: 9. 1978. TYPE: Bolivia: Pampatambo, Chapare, division to Ende Planta Corani, 68 km northeast of Cochabamba, 7 Dec 1974, Nishida, Ono, Hashimoto & Ohga 01043 (holotype, TNS). Herb epiphytic, caespitose, erect, up to 33 cm tall including the inflorescence. Roots slender, flexuous, to 1 mm in diameter. Ramicauls erect to ascendent, slender to stout, prolific, 2–7 cm long, enclosed by 3–9 tubular, papyraceous, ribbed, microscopically ciliate, lepanthiform sheaths, 4–15 mm long, pale to dark brown, basally adpressed, obtuse and dilated apically. Leaves elliptical to obovate, apiculate, erect, coriaceous, 0.7–3.5 × 0.4– 1.5 cm, basally cuneate, narrowing into a petiole up to 1 cm long. Inflorescence a 9–13   39   successively flowered, distichous raceme, 6–10 cm long including the peduncle 2–4 cm long, produced at the apex of the ramicauls from a conduplicate, ovate, acute spathe. Floral bracts cuneiform, conduplicate, papyraceous, 1–2 × 2 mm. Pedicels 2–6 mm long, persistent. Ovary cylindric, 1 mm long. Flowers with the sepals basally white, green towards the apex, occasionally suffused with pink-purple along the veins; petals translucent white; the lip brown at the base and greenish-yellow towards the apex, the column green, the column-foot brown, the anther yellow. Dorsal sepal narrowly ovate, basally concave, acute, attenuate, glabrous, carinate, 7–10 × 2–3 mm, 3-veined. Lateral sepals narrowly ovate to triangular, basally concave, oblique, glabrous, carinate, 7.0–9.6 × 1.3–2.0 mm, 3-veined. Petals elliptic to obovate, attenuate, obtuse, entire, repand towards the apex, 1.8–2.3 × 0.6–1.0 mm. Lip oblong, obtuse, arcuate, glabrous, 2.0–2.5 × 1 mm; disc with two longitudinal, lateral calli; midvein thickened; submarginal surface papillose. Column semiterete, broadly and deeply winged above the middle, 1.2–1.8 mm long; clinandrium apical; stigma ventral. Anther cap incumbent, broadly ovate, emarginate. Pollinia 2, pyriform, on a pair of cellular, ovate caudicles. Fruit not seen. Distribution. Costa Rica, Panama, Colombia, Venezuela, Ecuador, Peru and Bolivia (Luer, 1997). Additional material examined. Costa Rica. – Alajuela: San Ramón, Piedades Sur, San Miguel (La Palma), en El Alto despúes de San Bosco, 10º07’43.7”N 84º33’13.5”W, 1289 m, en árboles solitarios en potrero, 21 Dic 2010, A. Karremans 3303, J.A.J. Karremans & M. Contreras (JBL-D4775); A. Karremans 3304, J.A.J. Karremans & M. Contreras (JBL- D4774); – San Ramón, Reserva Biológica A. M. Brenes, 900 m, 23 May 1998, flowered in cultivation at Jardin Botánico Lankester, Oct 2001, M.A. Blanco 872 (USJ-80003); – San   40   Ramón, Angeles, Reserva Biológica A.M. Brenes, alrededores de la Estación Biológica del Río San Lorencito, 900-1100 m, 27-28 agosto 2011, M. Blanco 4047 (JBL-D6219); M. Blanco 4048, J. Abarca, M. Díaz, M. Chavarría & J. Gómez-Laurito (JBL-D5757); – M. Blanco 4049, J. Abarca, M. Díaz, M. Chavarría & J. Gómez-Laurito (JBL-D5758); – M. Blanco 4050, J. Abarca, M. Díaz, M. Chavarría & J. Gómez-Laurito (JBL-D5759); – M. Blanco 4052, J. Abarca, M. Díaz, M. Chavarría & J. Gómez-Laurito (JBL-D5763). – Cartago: Cartago, San Francisco, Muñeco, Finca Loma Verde y Jilguero, camino a Alto Belén, entre Río Sombrero y Quebrada Patarrá, 9°46’50.3”N 83°54’21.1”W, 1542 m, bosque pluvial premontano, epífitas en bosque secundario y árboles en zonas abiertas, 22 abr 2008, D. Bogarín 4500, A. Gaillard, R. Gómez, Y. Kisel, R. Phillips & R. Trejos (JBL- D3831); – Paraíso, Orosi, P.N. Tapantí, estribaciones Sur, Alto Peralta, 9.75 -83.783, 1400 m, 21 Dic 1992, G. Herrera 5816 (CR-169702, USJ-050992); – Turrialba, Tayutic, Moravia de Chirripó, unos 3 Km después de un albergue en Hacienda Moravia, siguiendo la calle hacia platanillo, 500 m más arriba de la única pulpería que hay en el lugar, sobre unos trechos al lado de la calle, bosque perturbado, 9°50’1