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Soil ascomycetes from Spain. XIII. Two new species of Apiosordaria

     Instituto de Investigaciones Fundamentales en Agricultura Tropical "Alejandro de Humbolt", calle 1 esq.2, Santiago de las Vegas, Boyeros, C. de La Habana, Cuba

A. M. Stchigel
J. Guarro ¹

     Facultat de Medicina i Ciéncies de la Salut, Universitat Rovira i Virgili, C/ Sant Llorenç 21, 43201, Reus, Tarragona, Spain

	ABSTRACT

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Apiosordaria hispanica sp. nov. and Apiosordaria globosa sp. nov. isolated from soil samples collected in Tarragona, Catalonia, Spain, are described and illustrated. Both species are morphologically close to A. otanii. The ascospores of A. hispanica have tuberculate walls, while those of A. otanii have small warts. Apiosordaria globosa differs from those species by the globose upper cell of the ascospores, which has a small apical protrusion with sub-apical germ pore when young. In A. hispanica and A. globosa the lower cells of the ascospores are slightly warted, while in A. otanii the lower cell of the ascospores is smooth-walled.

Key words: Ascomycota, Lasiosphaeriaceae, Mediterranean region, soil borne fungi

	INTRODUCTION

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The genus Apiosordaria Arx & Gams (Lasiosphaeriaceae, Ascomycota) is characterized by dark ascomata, non-amyloid asci and two-celled ascospores (Arx and Gams 1967). These are composed of a dark and ornamented upper cell and a hyaline lower cell (Krug et al 1983). The presence of spines, warts or pits in the upper cell of the ascospores is the principal character that differentiates this genus from other related genera of the Lasiosphaeriaceae. Triangularia Boedijn and Zopfiella Winter both have a smooth upper cell, although they also have unequal two-celled ascospores (Guarro and Cano 1988). The genus Apiosordaria comprises 21 species, including mainly soil-borne and coprophilous species (Krug et al 1983, Stchigel et al 2000).

During a continuous survey of soil ascomycetes from Spain, two interesting species of Apiosordaria were isolated. Their morphological characteristics differentiate them from all previously described species of the genus (Krug et al 1983, Guarro and Cano 1988, Stchigel et al 2000). In the present report these species are fully described, illustrated, and proposed as new.

	MATERIALS AND METHODS

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The soil samples were collected from several places in Els Gorgs de la Febró, Tarragona, Spain. The vegetation is mostly composed of Quercus ilex L. and Pinus halepensis Mill. The area has a Mediterranean climate with an annual average temperature of 15–36 C and an annual precipitation up to 600 mm. Soil was mainly collected from the A₀ horizon with sterile polyethylene bags. These were sealed with a rubber band, and labeled. On returning to the laboratory samples were stored at 4–7 C until used. Approx 1 g of the sample was treated with 2% (p/v) phenol (Phenol crystal, Panreac, Barcelona, Spain) for 10 min. The supernatant was discarded and the solid phase resuspended in 10 mL of sterilized water. The suspensions were cultured on potato carrot agar (PCA, potato 20 g, carrot 20 g, agar 20 g) at 15 C in the dark. Strains were grown on oatmeal agar (OA; Difco, Detroit, Michigan, USA), PCA and malt extract agar (MEA, Difco, Detroit, Michigan, USA). Cultures were made and structures were measured as described previously by Stchigel et al (1998). Color notations in parentheses are from Kornerup and Wansher (1984). Photomicrographs were obtained with a Leitz Dialux 20 EB microscope. Scanning electron microscopy techniques were described previously by Figueras and Guarro (1988).

	TAXONOMY

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Apiosordaria hispanica D. García, Stchigel et Guarro, sp. nov. Figs. 1–9

View larger version (46K): [in this window] [in a new window]    FIGS. 1–2. Apiosordaria hispanica FMR 7398. 1. Ascus with ascospores. 2. Ascospores. Scale bars: 1= 20 µm, 2= 10. µm.

Mycelium composed of pale brown to pale olive-brown, septate, branched, smooth-walled hyphae, 1–6 µm diam. Colonies on PCA growing rapidly, attaining 84–86 mm diam in 14 d at room temperature (22–25 C), flat, thin, consisting of submerged mycelium and sparse aerial hyphae, zonate and slightly radiate, brown (M. 5E6); ascomata produced after one mo; reverse brown (M. 5F5). Ascomata immersed to superficial, ostiolate, subglobose to pyriform, dark brown to black, 350–420 x 265–330 µm, tomentose, covered with long, slender, pale yellowish-brown, smooth, septate, unbranched or inconspicuously branched hypha-like hairs, 2–3 µm diam, with a conspicuous neck; neck 78–88 (–113) µm long, 127–137 µm wide at the base, with brown, papillate cells disposed around the ostiole. Peridium membranaceous, pilose, 6–9-layered, 17–20 µm thick, brownish-orange to brown; outer layer with textura intricata, composed by brown cells, 2–5 µm diam; inner layer composed by thin-walled polygonal cells, 9–15 µm diam. Asci 8-spored, fasciculate, cylindrical, thick-walled when young, becoming thin-walled at maturity, 146–232 x 15–22 µm, rounded at the apex, with a thin apical ring, stipitate, evanescent. Paraphyses hyaline, filiform, septate, 0.5–2 µm wide. Periphyses hyaline, filiform, septate, unbranched, 0.5–1 µm wide. Ascospores uniseriate, clavate, hyaline, and 1-celled when young, becoming two-celled by the formation of a transverse septum; upper cell obovoid, truncate at the base and with a slightly acuminate apex, brown, thick-walled, 23–28 (–30) x 18–22 (–23) µm, uniformly ornamented with numerous rounded warts of 0.5–2 µm diam, with an apical to sub-apical germ pore, 0.5–2 µm diam; lower cell sub-hyaline, conical and slightly warted, 1–5 (–6) µm long. Anamorph unknown.

Colonies on OA and MEA growing rapidly, attaining 55–70 mm diam in 14 d at room temperature (22–25 C), zonate and slightly radiate, cottony, brown to olive-brown (M. 5E6 to M. 4E8); reverse dark olive brown. Ascomata produced after one mo on OA; on MEA, ascomata not produced. At 15 and 35 C growing rapidly on PCA and OA; on MEA colonies restricted, ascomata not produced.

Specimen examined SPAIN, TARRAGONA: Vilaplana, Els Gorgs de la Febró, soil, 25-IX-1996, col. AM Stchigel and M Calduch (HOLOTYPE: FMR 7398).

Apiosordaria hispanica is morphologically similar to Apiosordaria vermicularis (Morinaga et al) Krug, Udagawa & Jeng (Morinaga et al 1978), Apiosordaria yaeyamensis Morinaga, Utatsu & Minoura (Morinaga et al 1979), Apiosordaria otanii Udagawa (Udagawa 1990) and Apiosordaria microcarpa Udagawa & Muroi (Udagawa and Muroi 1981). All have 8-spored asci and the upper cell of the ascospores more or less spinulose. Apiosordaria vermicularis can be easily differentiated from A. hispanica by its non-ostiolate ascomata and longer lower cell of the ascospores (12–20 µm), which is frequently 1-septate (Morinaga et al 1978). Apiosordaria microcarpa and A. yaeyamensis have ostiolate ascomata, but the upper cell of the ascospores is smaller (15–19 x 10–12.5 µm and 10–12.5 x 6–8 µm, respectively) (Udagawa and Muroi 1981, Morinaga et al 1979). In Apiosordaria otanii, the upper cell of the ascospores has very small warts and the lower cell is smooth-walled (Udagawa 1990), while A. hispanica has ascospores with the upper cell ornamented with large tubercles and the lower cell slightly warted.

Apiosordaria globosa D. García, Stchigel et Guarro, sp. nov. Figs. 10–20

View larger version (45K): [in this window] [in a new window]    FIGS. 10–11. Apiosordaria globosa FMR 7400. 10. Ascus with ascospores. 11. Ascospores. Scale bars: 10= 20 µm, 11 = 10 µm.

Mycelium composed of pale brown to pale olive-brown, septate, branched, smooth-walled hyphae, 1–6 µm diam. Colonies on PCA growing rapidly, attaining 80–86 mm diam in 14 d at room temperature, flat, thin, consisting of submerged mycelium and sparse aerial hyphae, zonate and slightly radiate, brown (M. 5E6); ascomata produced after one mo; reverse brown (M. 5F5). Ascomata immersed to superficial, ostiolate, subglobose to pyriform, dark brown to black, 600–770 x 320–540 µm, tomentose, covered with long, slender, pale brownish-yellow, smooth, septate, hypha-like hairs, 1–3 µm diam, unbranched or inconspicuously branched, with a conspicuous neck; neck 220–280 µm long, 210–250 µm wide at the base, with brown, papillate cells around the ostiole. Peridium membranaceous, pilose, 6–9-layered, 15–19 µm thick, brownish orange to brown; outer layer with textura intricata, composed of brown irregular cells, 2–5 µm diam; inner layer composed of thin-walled polygonal cells, 9–17 µm diam. Asci 8-spored, fasciculate, cylindrical, thick-walled when young becoming thin-walled when mature, 195–250 x 24–28 µm, rounded at the apex, with a thin apical ring, stipitate, evanescent. Paraphyses hyaline, filiform, septate, unbranched, 0.5–2 µm wide. Periphyses hyaline, filiform, septate, 0.5–1 µm wide. Ascospores uniseriate, broadly fusiform, hyaline and 1-celled when young, becoming two-celled by the formation of a transverse septum; upper cell globose, dark brown to black and opaque, with a small subapical protuberance, which disappears when ascospores mature, thick-walled, 25–27 (–29) x (22–) 23–27 µm, uniformly ornamented with small warts; lower cell sub-hyaline, conical, (1–) 4–6 µm long, slightly warted; germ pore sub-apical, 1–2 µm diam. Anamorph unknown.

Colonies on OA growing rapidly, attaining 80–86 mm diam in 14 d at room temperature, zonate and slightly radiate, cottony, with immersed and sparse aerial mycelium, olive-brown (M. 4E5); reverse dark brown; ascomata produced after one mo. Colonies on MEA restricted, elevated, cottony and radiately wrinkled, olive-brown (M. 4E5); reverse olive-brown (M. 4E8), ascomata not produced. At 15 and 35 C, the growth rate is similar to that at room temperature on all media used; ascomata not produced.

Specimen examined. SPAIN, TARRAGONA: Vilaplana, Els Gorgs de la Febró, from soil, 25-IX-1996, col. AM Stchigel, M Calduch (HOLOTYPE: FMR 7400).

Apiosordaria globosa is very similar to A. otanii (Udagawa 1990). Both have cylindrical, 8-spored asci and small warts on the upper cell of the ascospores. However, they can be differentiated by the shape and size of the upper cell of the ascospores, which are broadly ellipsoidal with a truncate base, 20–24 x 15–18 µm in A. otanii, and globose, measuring 25–27 (–29) x 23–27 µm in the new taxon. In addition, A. otanii produces a Chrysosporium-like anamorph, which is lacking in the new species, and the lower cell of the ascospores is smooth, while in A. globosa it is slightly warted.

View larger version (135K): [in this window] [in a new window]    FIGS. 3–9. Apiosordaria hispanica FMR 7398. 3. Ascoma. 4–6. Asci. 7. Detail of ascospore wall. 8, 9. Ascospores. Scale bars. 3 = 200 µm, 4 = 40 µm, 5–7= 20 µm, 8, 9 = 10 µm

View larger version (148K): [in this window] [in a new window]    FIGS. 12–20. Apiosordaria globosa FMR 7400 12. Ascoma. 13. Asci. 14, 15. Detail of ascus. 16. Detail and optical section of ascospored wall. 17, 18. Ascospores. 19, 20. Ascospores (SEM). Scale bars: 12 = 200 µm, 13 and 15= 20 µm, 14 and 16–20 = 10 µm

	ACKNOWLEDGMENTS

	FOOTNOTES

 
¹ Corresponding author, umb{at}fmcs.urv.es

Accepted for publication June 17, 2002.

	LITERATURE CITED

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS TAXONOMY LITERATURE CITED

 
Arx JA von, Gams W., 1967 Über Pleurage verruculosa und die zugehörige Cladorrhinum-Konidienform. Nova Hedwigia 13:199-208

Figueras MJ, Guarro J., 1988 A scanning electron microscopic study of ascoma development in Chaetomium malaysiense. Mycologia 80:298-306

Guarro J, Cano J., 1988 The genus Triangularia. Trans Br Mycol Soc 91:587-591

Kornerup A, Wanscher JH., 1984 Methuen handbook of colour. 3rd ed. London: Eyre Methuen. 252 p

Krug JC, Udagawa S-I, Jeng RS., 1983 The genus Apiosordaria. Mycotaxon 17:533-549

Morinaga T, Minoura K, Udagawa S-I., 1978 New species of microfungi from Southeast Asian soil. Trans Mycol Soc Japan 19:135-148

———, Utatsu I, Minoura K., 1979 Notes on filamentous fungi found in Japan. II. Trans Mycol Soc Japan 20:493-498

Stchigel AM, Cano J, Guarro J., 1998 A new species of Gelasinospora from Argentinian soil. Mycol Res 102:1405-1408

———, ———, ———, Gugnani HC., 2000 A new Apiosordaria from Nigeria, with a key to the soil-borne species. Mycologia 92:1206-1209

Udagawa S-I., 1990 A new species of Apiosordaria and some interesting ascomycetes from Nepal. In: Cryptogams of the Himalayas. Vol. 2. Tsukuba, Japan: National Science Museum. p 73–84

———, Muroi T., 1981 Notes on some Japanese Ascomycetes. XVI. Trans Mycol Soc Japan 22:11-26

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