New data on Sculptolumina japonica (Physciaceae) MIREIA GIRALT Departament de Bioquı́mica i Biotecnologia (Àrea de Botànica), Facultat d’Enologia de Tarragona, Universitat Rovira i Virgili, Marcel?lı́ Domingo s/n, 43007, Tarragona, Spain e-mail: mireia.giralt@urv.cat GRACIELA PAZ-BERMÚDEZ E.U.E.T. Forestal, Campus A Xunqueira, Universidade de Vigo, 36005 Pontevedra, Spain e-mail: graciela@uvigo.es JOHN A. ELIX Department of Chemistry, Building 33, Australian National University, Canberra, ACT 0200, Australia e-mail: John.Elix@anu.edu.au ABSTRACT. Sculptolumina japonica (Tuck.) Marbach, a species growing on lignum, bark or decaying plants and hitherto known from subtropical and tropical areas and one locality in North America, is now reported from Europe (northern Portugal) and the Canary Islands (La Gomera). This species has been found to possess filiform conidia. As a consequence we propose to amend the genus Sculptolumina to accommodate species that have long filiform conidia in addition to hymenial oil droplets and ascospores with internal wall thickenings. New chemical data are reported for S. japonica, the world distribution of this species is mapped and illustrations of the ascospores and conidia are provided. KEYWORDS. Sculptolumina, S. japonica, morphology, distribution, taxonomy, Physciaceae, Portugal, Canary Islands. ¤ A peculiar and very distinctive species of Buellia s. lat. was discovered among Sampaio’s collections in PO and among J. Etayo’s collections from the Canary Islands. The specimens were labelled as Buellia disciformis and Rinodina sp., respectively. A detailed morphological and chemical study of the specimens as well as additional material (including the type) for comparison purposes, has confirmed that this represents Sculptolumina japonica (Tuck.) Marbach, a species found on lignum, bark or decaying plants, ¤ ¤ which is known to be widely distributed in subtropical and tropical areas of Africa, America, Asia and Australia (Marbach 2000) and a single locality in North America (Sheard et al. 2008). MATERIALS AND METHODS This study was based on herbarium material from FH, PO and from the private herbaria of J. Etayo (Navarra, Spain) and K. Kalb (Neumarkt, Germany). Lichen morphology was examined by standard The Bryologist 112(2), pp. 397–403 Copyright E2009 by The American Bryological and Lichenological Society, Inc. 0007-2745/09/$0.85/0 398 THE BRYOLOGIST 112(2): 2009 Figure 1. Ascospore ontogeny of type B of Sculptolumina japonica (holotype, FH). Scale 5 10 mm. techniques using stereo and compound microscopes. Current mycological terminology generally follows Kirk et al. (2001). Only free ascospores and conidia lying outside the asci and pycnidia have been measured. Measurements were made in water at 10003 magnification. The terminology used for the asci follows Rambold et al. (1994), while that of ascospore-type and ontogeny follows Giralt (2001) and Foucard et al. (2002). Chemical constituents were identified by high performance liquid chromatography (HPLC) (Elix et al. 2003; Feige et al. 1993). THE SPECIES Sculptolumina japonica (Tuck.) Marbach, Bibliotheca Lichenologica 74: 297. 2000; Lecidea japonica Tuck., Proc. Amer. Acad. Arts. 5: 421. 1862; Buellia japonica (Tuck.) Tuck., Lichens of California, Oregon and the Rocky Mountains 25. 1866. TYPE: JAPAN: U.S. Navy Pacif. Explor. Exped., C. Wright (holotype: FH!). Figs. 1–3 Further synonyms are given in Marbach (2000) and Sheard et al. (2008) Figure 2. Filiform conidia of Sculptolumina japonica (holotype, FH). Scale 5 10 mm. Figure 3. Ascospore ontogeny of type B and ascospore variability of Sculptolumina japonica (PO). Scale 5 10 mm. Description. Thallus crustose, continuous, indeterminate, inconspicuous to thin and smooth or thicker and leprose-granulose, gray to olive brown, minutely orange-spotted in places, under the microscope the cortical layer of these orange spots with an amorphous yellow-orange K+ purple pigment; medulla I-, totally interspersed with small yellowish to orange crystals (polarized light), dissolving in K and C, giving an intense yellow and red solution, respectively (microscope slide). Apothecia lecideine, black; excipulum poorly developed, 15–35 mm wide; hymenium to 140 mm high, inspersed with abundant oil droplets to 7 mm diam; hypothecium dark brown, to 120 mm deep; asci Bacidia-type, often containing less than 8 spores (see Marbach 2000: p. 28, Abb. 10C). Ascospores Mischoblastia-type, (18–)19–25(–31) 3 9–13 mm; torus absent or poorly developed; spore-wall clearly ornamented; ontogeny of type B (Figs. 1, 3). Conidia filiform, straight, slightly curved or irregularly curved, (7–)11–21 3 1 mm (type material) (Fig. 2). Chemistry. Thallus K+ brownish or K+ purple, C+ pink to orange, P-, UV+ dark orange in part, Giralt et al.: Sculptolumina japonica Figure 4. Distribution of Sculptolumina japonica. n New records. medulla I-. Flavo-obscurin B1 [major], flavoobscurin B2 [major], flavo-obscurin A [minor or trace], 7-chloroemodin [minor or trace], skyrin [trace or absent], two unknown anthraquinones [minor], one or two unknown flavo-obscurin derivatives [minor] by HPLC. The analyses were carried out on three specimens (FH-holotype, Kalb 28556 and PO 1472) and supplements the chemical constituents identified by Kalb and Elix (1998) and Marbach (2000). We wish to emphasize that the secondary chemistry of this species is unique and that, within the Physciaceae, the anthraquinones flavo-obscurin A, B1, B2 and 7-chloroemodin have only been detected in Heterodermia obscurata and several other species of Heterodermia (Cohen & Towers 1995; Yosioka et al. 1968a–c). Distribution. Sculptolumina japonica is currently known from a single locality in Europe (northern Portugal) and one in the Canary Islands (La Gomera). These new records extend the distribution of this species from the tropics and subtropics north to the Macaronesian region and to the Eurosiberian region under oceanic influence (Fig. 4). Whereas this 399 N Literature records. species was collected on the bark of an old oak tree at the European locality, in La Gomera it was found growing on decaying plants. Characterization. This species is characterized by the crustose thallus, the minutely orange-spotted upper surface (anthraquinones), the lecideine apothecia with a poorly developed proper exciple, inspersed hymenium and brown hypothecium, the Bacidia-type asci, the large Mischoblastia-type ascospores with type-B ontogeny and the filiform conidia. Observations. A detailed study of the holotype material led to the discovery of conidia in this specimen. The long, filiform conidia were straight or slightly curved, and (7–)11–21 3 1 mm. Thus additional distinctive features of Sculptolumina japonica reported here for the first time include the long, filiform conidia, the discontinuous, amorphous yellow-orange, K+ purple pigment located in some parts of the thalline cortex, the medullary crystals which give K+ intense yellow, and C+ red solutions and the type-B ascospore-ontogeny. Presently the genus Sculptolumina includes only two species, the type species, S. japonica, and S. 400 THE BRYOLOGIST 112(2): 2009 serotina (Malme) Marbach. Both grow on bark, lignum or decaying plants in subtropical to tropical areas. Sculptolumina serotina differs from S. japonica in having Pachysporaria-type rather than Mischoblastia-type ascospores and in containing lobaric acid rather than anthraquinones (Kalb & Elix 1998; Marbach 2000). In Portugal and the Canary Islands, S. japonica could only be mistaken for species of Buellia with oil droplets in the hymenium (i.e., Hafellia species, see Gams 2004) including, Buellia disciformis (Fr.) Mudd, B. arnoldii Servı́t, the corticolous form of B. leptoclinoides (Nyl.) Steiner, B. sanguinolenta Schauer, Hafellia alisioae Etayo & Marbach and H. gomerana Etayo & Marbach (recently described by Etayo & Marbach 2003). However, the Callisporatype ascospores clearly distinguish these species (see Etayo & Marbach 2003; Giralt et al. 2000). European specimen examined. PORTUGAL. MINHO: Ponte de Lima, Santa Comba, carvalhos vellos, 11 Oct 1917, Sampaio 1472L (PO). Macaronesian specimen examined. CANARY ISLANDS. LA GOMERA: Vallehermosos, crtra. Vallehermoso-Valle Gran Rey, afloramientos, 665 m, 5 Aug 1994, Etayo (HB. ETAYO) Additional specimens examined for comparison (HB. KALB). AUSTRALIA. QUEENSLAND: Oberhalb von Lake Placid, wenige km südlich von Kuranda (etwa 20 km NW von Cairns), in einem tropischen Regenwald, 450 m, 16u509S, 145u389E, 27 Aug 1988, Kalb 21321; Mount Nebo Road, ca. 40 km W von Brisbane, Mt. Glorious in einem subtropischem Regenwald, 550 m, 27u239S, 152u479E, 29 Aug 1995, Kalb 29640. BRAZIL. BAHIA: Chapada Diamantina, Serra do Tombador; zwischen Mundo Novo und Morro do Chapeú, 1000 m, 18–20 Jul 1980, an Vellozia, Kalb 13664, an Holzzaun, Kalb 13658. MINAS GERAIS: Serra do Espinhaço, Serra do Caraça, Ungebung des Klosters Caraça (Hauptsammelgebiet von Vainio), an vertrockneter Vellozia, 1250 m, 8 Jul 1978, Kalb 13665. SÃO PAULO: Bezirk Rio Claro, etwa 2 km westlich von Rio Claro, 31 May 1980, Kalb 28566; etwa 25 km ostnordöstlich von São José do Rio Preto, an einem ungestürzten Laubbaum in einem Cerrado, 500 m, 14 Oct 1979, Kalb 28571; Serra de Mantiqueira, Campos do Jordão, 150 km nordöstlich von São Paulo in einem hellen, feuchten Urwald, 1700 m, 25 May 1978, Kalb 28971; Serra de Mantiqueira, Westanstieg zur Pedra do Bau, oberhalb von São Bento do Sapicaı́, an einem Holzstumpf, 1400 m, 24 Feb 1980, Kalb 10539; Horto Forestal von São Paulo, 800 m, 16 Aug 1980, Kalb 28570. MASCARENE ISLANDS. REUNIÓN: Zwischen le Brûlé (S von St-Denis) und Plaine des Cnicots, Tropisher Regenwald mit Nastus borbonicus, Acacia heterophylla, Cyathea borbonica, Philippia montana etc., 1400–1600 m, 20u579S, 55u279E, 15 Aug 1991, Kalb 26138; Fahrstrasse zwischen le Vingt-Septième (Bourg Mourat) und dem Piton de la Furnaise, Kleiner Bestand aus Sophora denudata und Philippia montana, 2250 m, 21u139S, 55u399E, 29 Aug 1991, Kalb 26310, 26317. MEXICO. CHIAPAS: bei Jitotol, an der Nationalstrasse 195, in einem lichten, trockenen Kiefernwald, 1600 m, 21 Jan 1979, Kalb 28567. PHILIPPINES. NORD LUZÓN: Prov. Baguio, in einem lichten Wald aus Pinus sp., 1400 m, 10 Aug 1983, Kalb & Schrögl. VENEZUELA. MÉRIDA: Distr. Sucre, oberhalb der Hacienda ‘‘Los Topes,’’ San Juanito, wenige km ENE von Chiguara, 8u309N, 71u309W, 1500 m, 3 Aug 1989, Kalb 24016. DISCUSSION The genus Sculptolumina was segregated from Buellia by Marbach (2000). According to him, the genus was characterized by the following combination of characters: crustose thallus, lecideine apothecia with epruinose discs and an excipulum lacking secondary lichen substances, hymenium inspersed with oil droplets, paraphyses with long and weakly expanded apical cells and ascospores with small, funnel-shaped or rounded lumina (with thickened inner walls, of Mischoblastia- or Pachysporaria-types). Within the family Physciaceae the form and length of the conidia is of great taxonomic value and has been used for delimiting genera for some time [e.g., Physcia and Physciopsis (5 Hyperphyscia) (Choisy 1950; Hafellner et al. 1979); Mobergia (Mayrhofer et al. 1992); Amandinea and Buellia (Scheidegger 1993; Australiaena (Matzer et al. 1997)]. Thus, the long, filiform conidia found in the type of Sculptolumina represent an important diagnostic character for the genus. Thus Sculptolumina can be recircumscribed to include species that have long Giralt et al.: Sculptolumina japonica filiform conidia in addition to lecideine apothecia, brown hypothecia, inspersed hymenia and ascospores with thickened inner walls. In our opinion, this combination of characters better define the genus Sculptolumina and more readily distinguish it from other members of the Physciaceae, especially from those belonging to the very large crustose genera Rinodina and Buellia s. lat., including Buellia s. str. (5 Hafellia, see Gams 2004), Amandinea or Tetramelas. Unfortunately, the conidia of the only other species assigned to Sculptolumina (S. serotina) are unknown at present. According to Aptroot (2002) the genus Sculptolumina is indistinguishable from Rinodina in the current sense, and he regards them as being synonymous. Although the lecideine apothecia, brown hypothecia and Bacidia-type asci present in Sculptolumina distinguish it from most Rinodina species, the long filiform conidia totally exclude this synonymy. At present, the only species provisionally retained in Rinodina (Giralt 2000, 2001; Giralt & Matzer 1994) with lecideine apothecia, brown hypothecia and Bacidia-type asci are R. ericina (Nyl.) Giralt, R. insularis (Arnold) Hafellner and R. kalbii Giralt & Matzer (5 R. ericina-group). Like all other Rinodina species, two of these species have short bacilliform conidia (unknown in R. insularis), to 8 mm long. If the presence of lecideine apothecia, brown hypothecia and Bacidia-type asci makes the generic position of the species of R. insularis group questionable (Giralt & Matzer 1994; Helms et al. 2003; Rambold et al. 1994), the inclusion of Sculptolumina within Rinodina is even more unlikely given that in addition to these features it has long filiform conidia, an important taxonomic character in the Physciaceae. Although molecular studies have yet to include either the R. ericina-group or Sculptolumina, these taxa clearly exhibit characters which distinguish them from Buellia and Rinodina (Helms et al. 2003). Atproot (2002) further suggested that the genus Sculptolumina is closely related to the Rinodina oxydata-group which has identical Mischoblastia-type ascospores. Aspects of this view are defended by Sheard et al. (2008). Although this recent study emphasizes the significant interest of Buellia japonica because it shares typical characters of Rinodina 401 (Mischoblastia ascospores) and Buellia (lecideine apothecia and brown hypothecia), it gives four reasons for not recognizing Sculptolumina and for maintaining S. japonica within Buellia. Like Aptroot (2002), Sheard et al. (2008) referred to the possibility that given its Mischoblastia-type ascospores, B. japonica could be closely related to the species of the R. oxydata-group, and advocated the possible inclusion of B. japonica and R. oxydata group within Mischoblastia Massal. (Massalongo 1852; Rambold et al. 1994), dependent upon further studies. Rambold et al. (1994) were tempted to reëstablish the genus Mischoblastia to include the species of the R. oxydatagroup based mainly on the fact that the ‘‘asci were intermediate between the Lecanora and the Bacidiatypes.’’ Further correlating characters mentioned by these authors were the presence of Bagliettoana-green pigment and the tendency of the lecanorine apothecia to become pseudolecanorine (see also Matzer & Mayrhofer 1996: 22). These three characters, together with the colorless hypothecia, the hymenia lacking oil droplets and the presence of atranorin and bacilliform conidia (present in the species of R. oxydata-group or Mischoblastia, if accepted) are all absent in Sculptolumina. Thus we are confident that S. japonica is unrelated to the R. oxydata-group because they only share one key character, the presence of Mischoblastia-type ascospores. Moreover, in contrast to R. oxydatagroup, where the Mischoblastia-type ascospores develop with type-A ontogeny, the ascospores of S. japonica develop with type-B ontogeny. Although Aptroot (2002) did not consider the second species of Sculptolumina (S. serotina), with Pachysporaria type ascospores (Marbach 2000), Sheard et al. (2008) used the fact that the two species assigned to the genus have alternative types of spores as a further argument for rejecting the genus Sculptolumina. However, within the Physciaceae, genera often include species which exhibit different ascospore-types [e.g., Amandinea and Buellia (Buellia- and Physconia-types); Hyperphyscia, Phaeophyscia and Physcia (Physcia- and Pachysporaria-types); Rinodina (many different ascospore-types)] whereas different genera often exhibit the same ascospore-types [e.g., Buellia-type (Amandinea, Buellia, Phaeorryza); Dirinaria-type 402 THE BRYOLOGIST 112(2): 2009 (Australiaena, Diploicia, Pyxine, Rinodina, etc.); Physcia-type (Mobergia, Rinodina, etc.); Physconiatype (Anaptychia, Tornabea, etc.)]. According to Helms et al. (2003), most species of Rinodina, including those of the R. oxydata-group, would be included in the family Physciaceae, whereas the species of Sculptolumina and those of the R. ericina-group, would belong to the Caliciaceae or Buelliaceae. Indeed, ascospores with inner wall thickenings are not exclusive to the Physciaceae since all genera in the Buelliaceae with Callispora, Dirinaria and Physconia-type ascospores possess the same feature (e.g., Hafellia, Pyxine, Buellia). Not unexpectedly, the Physciaceae include at least one species (‘‘Buellia’’ parvula) and one genus (Phaeorriza) with Buellia-type ascopores which lack internal wall thickenings. Sheard et al. (2008) maintained S. japonica within Buellia s.l. However, when it is well known that Buellia is polyphylethic and when genera previously segregated or excluded from it are widely accepted [e.g., the genera Hafellia (5 Buellia s. str., see Gams 2004), Tetramelas, Australiaena], it seems inconsistent not to accommodate B. japonica within the well-circumscribed genus Sculptolumina. In our opinion, the joint occurrence of filiform conidia, inspersed hymenium and ascospores with inner wall thickenings clearly delimit the genus Sculptolumina and distinguishes it, not only from Rinodina (including the R. oxydata- and R. ericinagroups), Amandinea, Buellia, Hafellia and Tetramelas but also from all genera hitherto described in the Physciaceae (including the Buelliaceae). ACKNOWLEDGMENTS The authors are indebted to the keepers of FH and PO and to Dr. Klaus Kalb (Neumarkt, Germany) and J. Etayo (Pamplona, Spain) for the loan of material used in this study. The first author thanks the Comissionat per a la Recerca (Catalan Government) and the second the University of Vigo for financial support. LITERATURE CITED Aptroot, A. 2002. Note 3551. Sculptolumina Marbach. In O. E. Eriksson, H. O. Baral, R. S. Currah, K. Hansen, C. P. Kurtzman, T. Laessøe & G. Rambold (eds.), Notes on ascomycete systematics. Nos. 3304–3579. Myconet 8: 1–54. 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