A NEW SPECIES OF TRICHIA (MYXOMYCETES) FROM SINGAPORE

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1 A NEW SPECIES OF TRICHIA (MYXOMYCETES) FROM SINGAPORE 1 G. MORENO, 2 D.W. MITCHELL, 3 W.C. ROSING & 4 S.L. STEPHENSON 1 Departamento Biología Vegetal (Botánica), Universidad de Alcalá, Alcalá de Henares, Madrid. España 2 Walton Cottage, Upper Hartfield, East Sussex, England, TN7 4AN 3 Department of Biology, Middle Tennessee State University Murfreesboro, Tennessee 37132, USA 4 Department of Biological Sciences, University of Arkansas, Fayetteville, Arkansas 72701, USA. Resumen. MORENO, G., D.W. MITCHELL, W.C. ROSING & S.L. STEPHENSON (2013). Una nueva especie de Trichia (Myxomycetes) de Singapur. Bol. Soc. Micol. Madrid 37: Se describe una nueva especie de Trichia (T. nodosa) de material recogido en Singapur, Los esporocarpos se han desarrollado en trozos de corteza en cámara húmeda. Se incorporan fotografias al MO y MEB para ilustrar las características distintivas de esta nueva especie. Palabras clave: Amebozoa, hongos mucosos, MEB, myxobiota, taxonomía, Trichiales. Summary. MORENO, G., D.W. MITCHELL, W.C. ROSING & S.L. STEPHENSON (2013). A new species of Trichia (Myxomycetes) from Singapore. Bol. Soc. Micol. Madrid 37: A new species of Trichia (T. nodosa) is described from material collected in Singapore, where it formed well developed sporocarps on pieces of dead bark in moist chamber culture. Both LM and SEM photomicrographs are provided to illustrate the distinguishing features of this new species. Key words: Amebozoa, myxobiota, slime moulds, Trichiales, SEM, taxonomy. INTRODUCTION Trichia Haller is the type genus of the order Trichiales T. Macbr. in the family Trichiaceae Chevall. Members of the genus can be distinguished by their stalked or sessile (or more rarely subplasmodiocarpous) sporocarps; elastic capillitium consisting of free, simple or sparsely branched elaters with acuminate (sometimes elongated) tips and marked with 2 to 5 (rarely more) spiral bands; and spores that are yellow, yellow-brown or reddish in mass but hyaline, tinted by transmitted light (MARTIN & ALEXOPOULOS, 1969). The current concept Bol. Soc. Micol. Madrid

2 G. MORENO, D.W. MITCHELL, W.C. ROSING & S.L. STEPHENSON used for the genus encompasses a relatively large number of species, thus making it one of the more important genera within the myxomycetes. KIRK & al. (2008) recognized 35 species of Trichia, LADO ( ) recognized 34 species, and MITCHELL (2013) listed 33 taxa and 8 infraspecific taxa. The purpose of this paper is to describe a new species of Trichia, based on material appearing in moist chamber culture on pieces of the dead bark of Fagraea fragrans Roxb. collected from Labrador Park in Singapore. The species can be distinguished from other similar members of the genus by a combination of macro- and microscopic characters, which are described herein. This work completes the previous contribution of ROSING & al. (2011). MATERIALS AND METHODS The material upon which the description of this new species is based was studied in the Department of Plant Biology at the University of Alcalá de Madrid (AH), where small portions of the type material (2 slides) are preserved as isotypes. Sporocarps for detailed study were mounted in Hoyer s medium following the method described by KOSKE & TESSIER (1983) and SCHNITTLER & NOVOZHILOV (1996). Spores were measured, including surface structures such as spines or warts, with the use of an oil immersion lens. Light microscopy (LM) was carried out with a Nikon microscope equipped with an automatic photographic system. The SEM micrographs were produced with a Zeiss DSM- 950 microscope. For ultramicroscopic studies, the material to be examined was rehydrated in concentrated ammonium hydroxide (28 30%) for 30 minutes, dehydrated in aqueous ethanol (70%) for 30 minutes, fixed for 2 hours in pure ethylene glycol dimethyl ether (= 1,2 dimethoxymethane) and finally immersed in pure acetone for at least 2 hours. This was followed by critical point drying and sputtering with gold-palladium. This technique allows the use of very little material (i.e., part of a single sporocarp or sometimes no more than a few spores). Terminology used to describe spore ornamentation follows that of RAMMELOO (1974, 1975). DESCRIPTION Trichia nodosa G. Moreno, D.W. Mitch., W.C. Rosing & S.L. Stephenson, sp. nov. Figs MycoBank MB Etymology: nodosa derived from latin nodosus with nodes in the capillitium. Specimen studied: Republic of Singapore, Singapore, Labrador Park (1º 16 0 N, 103º E), appearing on dead bark of Fagraea fragrans Roxb. in moist chamber culture, leg. W. C. Rosing, 10-III-2009, WCR in AH Holotype. Sporocarps gregarious, stipate. Sporotheca mm, globose to subglobose, sometime fused in pairs, dark purplish-brown to dark brown, with yellow-brown markings that indicate where dehiscence will occur (Fig. 1). Stipe mm long, blackish, rough longitudinally, longer than the sporotheca (Fig. 1). Peridium single, membranous, thickened with dark brown areolae of refuse material, pale yellow-brown and somewhat glossy between the areolae. Dehiscence areolate, the areolae appearing as dark purplish brown patches by LM (Fig. 2). Hypothallus membranous and poorly developed, dark brown. Plasmodium unknown. Spores 7-8 µm in diam., globose to subglobose, yellowish, verrucose (Fig. 9). Capillitium abundant, dense, consisting of yellowish elaters 5-6 µm wide and with abundant free ends and nodes (Figs. 3-8). The ends are short, acutely pointed and µm in length (Figs. 3-4). The nodules are globose to ellipsoidal and covered with spines 5-12 µm in length (Figs. 6-8). Sometimes, the nodules are terminal and replace the acute tips. By SEM the filaments of the capillitium are seen to be covered by sinuous spirals, which cause the wall to be decorated with a reticulum (Figs ) which is not visible by LM. The nodules have spines of variable dimensions (Figs ). 2 Bol. Soc. Micol. Madrid

3 A NEW SPECIES OF TRICHIA (MYXOMYCETES) FROM SINGAPORE Figs Trichia nodosa holotype. 1. Scattered sporocarps. 2. Detail of peridium tessellation Free ends of elaters. 5. Capillitium Detail showing spinose nodules of capillitium by LM. 9. Spores. 10. Detail of peridium and insertion of capillitium by SEM Detail of spinose nodules of capillitium by SEM Free ends of elaters. 18. Sinuose ornamentation of elater spiral. 19. Detail of the partial retículum on the elater wall Spores by SEM. Bars: 1 = 0.5 mm, 2 = 100 μm, 3-4 = 10 μm, 5 = 25 µm, 6-9 = 10 µm, 10 = 2 µm, = 5 μm, = 2 μm. Bol. Soc. Micol. Madrid

4 G. MORENO, D.W. MITCHELL, W.C. ROSING & S.L. STEPHENSON The free ends have spirals reaching to the apex (Figs ). By SEM the sporal ornamentation consists of densely spaced bacula with an irregular distribution (Figs ). Other material examined: Hemitrichia spinifera isotype, Colombia, 104 km from Bucaramanga, on the Bucaramanga-Pamplona Rd. Dpto., north of Santander, on wood, leg. K.F. Dumont, M.A. Sherwood & L.F. Velásquez, 21-VIII-1976, The New York Botanical Garden, Dumont-Co. 7006, BPI Hemitrichia spinosa holotype: Venezuela, Anden, Estado de Merida, Páramo de Mucubaji, Umgebung der Laguna Negra, 3500 m, 16-III-1969, leg. B. & F. Oberwinkler, Herbarium F. Oberwinkler nº b, det. M.L. Farr as Hemitrichia intorta (Lister) Lister, National Fungus Collections 1040, BPI DISCUSSION Sporocarps of Trichia nodosa are characterized by their stipitate habit and globose to subglobose areolate sporothecae. The base of the stalk is blackish, and is rougher than the sporotheca, whereas the spores are 7-8 µm in diameter, globose, yellowish and verrucose. The capillitium consists of individual elaters; these are yellowish, 5-6 µm wide, have short free ends and globose to ellipsoidal spinose nodules. The capillitial spirals are covered by spines, which gives the overall ornamentation a denticulate appearance. This species is known only from the type. Two species described originally from the Americas have a conspicuously spiny capillitium and have been assigned to the genus Hemitrichia Rostaf. These are H. spinifera M.L. Farr and H. spinosa Neubert & Nann.-Bremek. Both these species possess a branched capillitium without free elaters (Table 1). Hemitrichia spinifera (Figs ), was described by FARR (1979) from a single specimen collected in Colombia. This species is characterized by its stipitate sporocarps, Trichia nodosa Hemitrichia spinifera Hemitrichia spinosa Sporothecae Peridium Stalk Capillitium Spores globose to subglose, dark purplish-brown with light areolae well developed turbinate to subglobose, dark purplishbrown with light yellow-brown reticulations that form areolae pyriform, brown, ± opaque dark brown areolae readily apparent areolae readily apparent well developed, 50(-65%) of the overall height, reddish-brown to dark purplishbrown nonareolate well developed, blackish, longer than the sporotheca poorly developed, about 0.6 mm high, 0.4 mm wide 5-6 µm diam., with spiral bands, and abundant free ends, spiny nodules 7-8 µm in diam. (5-)6-8 µm diam., densely spiny spirals without free ends, reticulate 4-7 µm diam., densely spiny spirals, with few free ends, rounded, without striae 7-8 µm 8-10 µm µm Table 1. Differences that exist between Trichia nodosa, Hemitrichia spinifera and H. spinosa. 4 Bol. Soc. Micol. Madrid

5 A NEW SPECIES OF TRICHIA (MYXOMYCETES) FROM SINGAPORE Figs Hemitrichia spinifera isotype. 24. Label on the box lid in which the collection was conserved. 25. Mature scattered and united sporocarps after conservation showing tessellated peridium. 26. Sclerotium. 27. Detail of hemitrichioid capillitial tessellation by LM Detail of capillitial spines by LM Capillitial spines by SEM Detail of spirals with abundant spines by SEM Spores by SEM. 38. Detail of sporal ornamentation by SEM. Scale bars: = 1 mm, 27 = 100 µm, = 10 µm, = 5 µm, = 2 µm, 38 = 1 µm. Bol. Soc. Micol. Madrid

6 G. MORENO, D.W. MITCHELL, W.C. ROSING & S.L. STEPHENSON Figs Hemitrichia spinosa holotype. 39. Label on the box in which the collection was conserved. 40. Sporocarps. 41. Detail of peridium Detail of spines of capillitium by LM. 45. Spores by LM. 46. Detail of internal surface of peridium by SEM. 47. Capillitial spines by SEM. 48. Detail of the spirals with abundant spines and the cross-linked raised bands that form a reticulation on the surface of the elater wall by SEM Spores by SEM. 52. Detail of the spore ornamentation by SEM. Scale bars: = 1 mm, = 10 µm, 46 = 1 µm, 47 = 5 µm, 48 = 2 µm, = 2 µm, 52 = 1 µm. 6 Bol. Soc. Micol. Madrid

7 A NEW SPECIES OF TRICHIA (MYXOMYCETES) FROM SINGAPORE turbinate to subglobose sporothecae, with dark purplish-brown areolae with light yellow-brown reticulations. Stalk stout, sulcate, reddishbrown to purplish-brown, often darker above. Peridium areolate, single and membranous (Fig. 25). Dehiscence areolate (Fig. 27). Capillitium abundant, dense, without free ends, reticulate, the tubules yellowish to reddish brown, (5-)6-8 µm diam., not including the ornamentation of three loosely-wound spirals and the dense covering of (4-)6-8 µm long spines (Figs ). By SEM, spines are arranged along the elater spirals and ornamentation is not observed between the spirals (Figs ). Spores light yellow-brown, 8-10 µm diam., densely verruculose (Fig. 30). By SEM spore ornamentation consisting of bacula densely and irregularly distributed. (Figs ). Hemitrichia spinosa Neubert & Nann.- Bremek. (Figs ), was described originally by NEUBERT & NANNENGA-BREMEKAMP (1977) from material collected in Venezuela and is characterized by its stipitate sporocarps and pyriform sporothecae (Fig. 40). Peridium opening by irregular dehiscence of the upper part, the lower half remaining as a deep cup (Fig. 41). Stalk very under-developed, about 0.6 mm high and 0.4 mm wide. Peridium brown, not areolate, with the inner surface presenting SEM crests which merge into a more-or-less reticulate structure (Fig. 46). Capillitium elastic, 4-7 µm thick with 4-7 spiral bands, pale yellow-brown to dark brown, densely covered with spines, these up to 7 µm long (Figs ). By SEM the spines are arranged along the spirals of the elaters (Fig. 47) and there is a small ornamentation crosslinked between the spirals (Fig. 48). Spores pale yellow-brown, finely warted, µm diam. (Fig. 45). By SEM spore ornamentation consists of irregularly distributed and densely spaced bacula (Figs ). ACKNOWLEDGEMENTS We wish to express our gratitude to Mr. A. Priego and Mr. J.A. Pérez of the Electron Microscopy Service of the University of Alcalá de Henares for their invaluable help with the SEM. We also thank L. Monje and A. Pueblas of the Department of Drawing and Scientific Photography at the Alcalá University for his help in the digital preparation of the photographs, to Dr. J. Rejos, curator of the AH herbarium for his assistance with the specimens examined in the present study. W.C.R. extends his thanks the Singapore Botanic Gardens for the award of a Research Fellowship in BIBLIOGRAPHY FARR M.L. (1979). Notes on Myxomycetes II. New taxa and records. Nova Hedwigia 31: KIRK, P.M., P.F. CANNON, D.W. MINTER & J.A. STALPERS (2008). Dictionary of the Fungi Ainsworth & Bisby s Tenth edition. CABI Europe-UK. 771 pp. KOSKE, R.E. & B. TESSIER (1983). A convenient permanent slide mounting medium. News Lett. Mycol. Soc. Amer. 34: 59. LADO, C. ( ). An on line nomenclatural information system of Eumycetozoa. (5-I-2013). MARTIN, G.W. & C.J. ALEXOPOULOS (1969). The Myxomycetes. Iowa City. University of Iowa Press. MITCHELL, D.W. (2013). Myxomycetes DVD of Keys and Inventory of all world species. Private publication by the author, continuously updated. NEUBERT H. & N.E. NANNENGA- BREMEKAMP (1977). Drei neue Arten aus der Klasse Myxomycetes. Zeitsch. f. Pilzkunde 43: RAMMELOO, J. (1974). Structure of the epispore in the Trichiaceae (Trichiales, Myxomycetes) as seen with the scanning electron microscope. Bull. Soc. Roy. Bot. Belgique 107: RAMMELOO, J. (1975). Structure of the epispore in the Stemonitales (Myxomycetes) as seen with the scanning electron microscope. Bull. Jard. Bot. Belgique 45: Bol. Soc. Micol. Madrid

8 G. MORENO, D.W. MITCHELL, W.C. ROSING & S.L. STEPHENSON ROSING, W.C., D. W. MITCHELL, G. MORENO & S. L. STEPHENSON (2011). Additions to the myxomycetes of Singapore. Pacific Science 65: SCHNITTLER, M. & Y. NOVOZHILOV (1996). The Myxomycetes of boreal woodlands in Russian northern Karelia: a preliminary report. Karstenia 36: Bol. Soc. Micol. Madrid

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