Lichenologist 32(2): 105–128 (2000) doi:10.1006/lich.1999.0251 Available online at http://www.idealibrary.com on NOTES ON SOME CORTICOLOUS AND LIGNICOLOUS BUELLIA SPECIES FROM THE IBERIAN PENINSULA Mireia GIRALT*, Mercedes BARBERO* and John A. ELIX‡ Abstract: The morphology, anatomy, chemistry and ecology of some corticolous and lignicolous species of Buellia from the lberian Peninsula that contain secondary lichen compounds are discussed, including B. chloroleuca, B. disciformis, B. erubescens, B. hyperbolica and B. leptoclinoides. These species are compared with other corticolous Buellia species that occur outside the study area, including B. arnoldii, B. insignis, B. regularis and B. sanguinolenta. The misunderstood B. zahlbruckneri J. Steiner and B. jorgei, are synonyms of B. erubescens, and the invalid taxon B. zahlbruckneri sensu Schauer non Steiner and B. poeltii, are synonyms of B. chloroleuca. New chemical data on several taxa are reported. A key to the species and illustrations of their ascospores are provided.  2000 The British Lichen Society Introduction The present study is a further contribution towards extending the knowledge of the Physciaceae in the lberian Peninsula. The aim of this investigation was to reappraise all those corticolous and lignicolous Buellia species that contain secondary lichen substances. Although the emphasis here is on the lberian species, many of these taxa are also found elsewhere in Europe. Furthermore, several species that do not occur in the study area have also been investigated for comparative purposes. The corticolous and lignicolous species that have vegetative propagules, polyspored asci or triseptate ascospores, as well as secondary metabolites, are not included in this paper but will be treated in future studies. Furthermore, several of the specimens examined that have 8-spored asci and secondary chemistry like the species discussed in this paper will be described as new in a forthcoming paper. It should be pointed out that this work should be regarded as a first step towards a better understanding of these corticolous/lignicolous Buellia species. It has been undertaken for a small part of the world and it is based on a relatively small number of specimens. For this reason, we have not attempted to examine all type specimens and until a world-wide taxonomic revision of the genus Buellia is carried out many problems will remain unresolved. *Department de Biologia Vegetal (Botànica), Facultat de Biologia, Universitat de Barcelona, Diagonal 645, 08028 Barcelona, Spain. ‡Department of Chemistry, Faculty of Sciences, The Australian National University, Canberra ACT 0200, Australia. 0024–2829/00/020105+24 r35.00/0  2000 The British Lichen Society 106 THE LICHENOLOGIST Vol. 32 In the present treatment all taxa are retained in the genus Buellia pending further data to confirm whether the type species of the genus Buellia is B. parasema de Not., as proposed by Kalb & Elix (1998), or B. aethalea (Ach.) Th. Fr., as more recently proposed by Moberg et al. (1998). Characters of the species A comprehensive account of the comparative morphology and anatomy of the Buellia species studied is not included. However, the taxonomic value of some characters requires discussion. According to our observations, the characters of greatest taxonomic value in the genus Buellia are the development and structure of the ascospores, the presence of hymenial oil droplets, the degree of development of the excipulum and its reaction with K, and the presence of particular lichen compounds. Other characters such as thalline and apothecial morphology and the degree of swelling of the apical cells of the paraphyses are important features but of little diagnostic significance. The ascospores of the species treated in this paper are of the following types: the Callispora-type or similar (compare Kalb 1986; Mayrhofer 1984; Scheidegger 1993; Pusswald et al. 1994), with the inner ascospore wall thickened subapically and at the septum (B. arnoldii, B. disciformis, B. leptoclinoides, B. regularis and B. sanguinolenta). In addition, two species have ascospores that are intermediate between the Callispora- and the Buellia-types (B. chloroleuca and B. insignis). Finally, B. erubescens and B. hyperbolica possess ascospores that are intermediate between the Physconia- and the Buellia-type (see Scheidegger 1993), with the inner ascospore wall thickened at the septum in the early stages of their ontogeny but lacking any wall thickening when mature. All the present species have ascospores that originate via a type A ontogeny: in young and hyaline ascospores the septum is inserted before apical internal wall-thickenings become distinct (cf. Giralt 1994; Giralt & Mayrhofer 1994, 1995). Two types of ascospore ornamentation can easily be recognized with light microscopy, at least after addition of 10% KOH: the weakly and the roughly microrugulate ornamentation [type 1 and 2 of Nordin (1996), respectively]. The spore ornamentation is weak in B. leptoclinoides, rough in B. chloroleuca, B. erubescens, B. hyperbolica and B. insignis and smooth in B. disciformis and in the B. arnoldii-group. The thallus morphology of all the species treated is rather variable and may range from thin and continuous to thick and rimose. Well-developed thalli and that of lignicolous specimens may often develop a granulose to verrucose surface. The prothallus is especially visible when adjoining thalli of other species. The apothecia are lecideine in all the species treated except in B. hyperbolica in which they very soon become biatorine. The excipulum is more or less well developed in all the species except in B. hyperbolica where it is very thin. The brownish pigmentation gives a yellow to yellow-brownish solution with K in 2000 Iberian Buellia—Giralt et al. 107 B. chloroleuca, B. erubescens, B. hyperbolica and B. insignis (compare Scheidegger 1993 and Nordin 1996). The hymenium is interspersed with numerous oil droplets (opaque even in thin sections) in B. arnoldii, B. disciformis, B. leptoclinoides, B. regularis and B. sanguinolenta, but oil droplets are absent in the other species treated. The asci are 8-spored, clavate and of the typical Bacidia-type (Rambold et al. 1994) in all the present species. Spermogonia are rare and have been found only in B. chloroleuca, B. disciformis and B. erubescens. The spermatia are always hyaline, bacilliform, straight, single-celled and thin walled, of 4·5–9
m long and 0·8–1
m wide. The spermatophores are branched, resembling the Type VI of Vobis & Hawksworth (1981). The species treated in this paper are characterized by the presence of various secondary lichen compounds, which can be detected after the application of reagents (spot tests) and include: atranorin, fumarprotocetraric acid, protocetraric acid, 6-O-methylarthothelin (=granulosin) and norstictic acid (with connorstictic acid). Atranorin reacts K+yellow and it is present (at least in traces) in all the species treated; fumarprotocetraric and protocetraric acids react PD+orange-red and are present in B. hyperbolica; 6-O-methylarthothelin reacts K+and C+yellowish and KC+orange and is present in B. chloroleuca and B. insignis, whereas norstictic acid reacts K+red (with the formation of red crystals under the microscope) and is present in B. erubescens and B. sanguinolenta. All these colour reactions are usually strong and are visible under the stereomicroscope. Other secondary lichen compounds present in the species treated in this paper (e.g. zeorin) that do not give spot test reactions or are present only in traces (e.g. gyrophoric acid) must be detected by thin layer chromatography (TLC) or high performance liquid chromatography (HPLC). The following lichen compounds have been detected for the first time in the genus Buellia: protocetraric, confumarprotocetraric, convirensic and hyposalazinic acids, and fulgoicin. Further, an unknown present in B. disciformis (tentatively identified as norfulgoicin) and an unknown secalonic acid derivative present in B. erubescens [also detected in B. multispora and Gassicurtia coccinea (Kalb & Elix 1998)] have yet to be fully characterized chemically. For the most part, notes on the ecology and distribution of the species refer only to the study area. Materials and Methods All the morphological descriptions are based on our personal examinations. Current mycological terminology has been employed and generally follows that of Hawksworth et al. (1995). The terminology used for the asci follows that of Rambold et al. (1994), while for the ascospore-types the terminology of Kalb (1986), Mayrhofer (1982, 1984), Scheidegger (1993), Giralt (1994) and Giralt & Mayrhofer (1994, 1995) is followed. The standardized methods of TLC (e.g. Culberson & Ammann 1979; Culberson et al. 1981; Culberson & Johnson 1982) and HPLC (Feige et al. 1993) for the identification of lichen substances were employed. As most of the species considered have previously been described in detail in other revisions, including those of Sheard (1964), Schauer (1965) and Orange et al. (1992), the descriptions given here emphasize characters of value for species identification but do not repeat features common to all of the species. 108 THE LICHENOLOGIST Vol. 32 Key to the species 1 Thallus K+yellow-orange then red; containing norstictic acid . . 2 Thallus K+yellow, yellowish or yellowish brown, never K+yelloworange then red; norstictic acid absent . . . . . . . . . . . . . 3 2(1) Hymenium with numerous oil droplets. Excipulum K
. Ascospores (21–)26–32(–35)(11–)12–13(–16)
m, of the Callispora-type . . . . . . . . . . . . . . . . . . . . . . Buellia sanguinolenta Hymenium without numerous oil droplets. Excipulum K+yellow (microscope slide). Ascospores (11–)13–16(–18)(6–)7–8·5 (–9·5)
m, Physconia-type when young, then Buellia-type when mature . . . . . . . . . . . Buellia erubescens (Chemotype I) 3(1) Thallus PD+orange-red; fumarprotocetraric and protocetraric acids present. Apothecia rapidly becoming biatorine, 0·2–0·3(–0·4) mm diam. . . . . . . . . . . . . . . . . . . . . Buellia hyperbolica Thallus PD
; fumarprotocetraric and protocetraric acids absent. Apothecia rarely becoming biatorine, significantly larger . . . . 4 4(3) Hymenium with numerous oil droplets . . . . . . . . . . . . . . 5 Hymenium without numerous oil droplets . . . . . . . . . . . . . 7 5(4) Ascospores 15–20(–23)(6·5–)8–9(–9·5)
m, placodiolic acid present . . . . . . . . . . . . . . . . . . Buellia leptoclinoides Ascospores significantly larger, placodiolic acid absent . . . . . . 6 6(5) Ascospores (23–)26–29(–31)(10–)12–14(–16·5)
m, thallus containing only atranorin . . . . Buellia arnoldii (incl. B. regularis) Ascospores (16–)19–25(–28)7·5–9(–11)
m, thallus containing atranorin, fulgidin and fulgoicin . . . . . . Buellia disciformis 7(4) Thallus KC+and UV+orange, section C+orange (microscope slide), containing 6-O-methylarthothelin. Ascospores intermediate between the Callispora and the Buellia-type . . . . . . . . . . . 8 Thallus KC
and UV
, section C
, 6-O-methylarthothelin absent. Thallus K+yellow, containing atranorin and traces (not always) of several homologues of norstictic acid. Ascospores Physconia-type when young, then Buellia-type when mature, (11–) 13–16(–18)(6–)7–8·5(–9·5)
m . . . . . . . . . . . . . . . Buellia erubescens (Chemotype II) 8(7) Ascospores (19–)21–25(–29)(8–)9–11(–12)
m . . . . . . . . . . . . . . . . . . . . . . . . . . Buellia insignis Ascospores smaller, (13–)16–19(–22)(6–)7–9
m . . . . . . . . . . . . . . . . . . . . . . . . Buellia chloroleuca 2000 Iberian Buellia—Giralt et al. 109 F. 1. Ascospore ontogeny. A, Buellia choroleuca (Lichenes Alpium 192, W); B, B. insignis (G—lectotype). Buellia-type ascospores showing tendencies to the Callispora-type (arrows). Scale=10
m. The Species Buellia chloroleuca Körb. Parerg. Lich.: 191 (1860); type: Polen: Schlesien, Sudeten, an Fichten, Körber (L!, M!—isotypes?). New synonyms: Buellia poeltii T. Schauer, Mitt. Bot. München 5: 616 (1965); type: Germany: Oberbayern, Lahnenwies-Graben prope Garmisch, in cortice Aceris pseudoplatani, 1470 m, 4 September 1963, Th. Schauer (M!—holotype). (Fig. 1A) Thallus whitish, whitish grey or cream, rarely thin and smooth to rimose, usually moderately thick, warted, continuous and well developed, covering large areas, not delimited by a blackish prothallus. Apothecia (0·3–)0·5–0·8 (–1) mm diam., abundant and confluent, adnate, rarely becoming sessile. Disc initially plane, rapidly subconvex to strongly convex, epruinose. Proper margin initially thick, becoming thinner and finally excluded. Excipulum 60–80
m wide, K+deep yellow-brownish (microscope slide). Hymenium without oil droplets, 80–100
m high. Hypothecium interspersed with oil droplets. Apical cells of the paraphyses strongly swollen, 4–6(–7)
m wide, brown pigmented. Ascospores (13–)16–19(–22)(6–)7–9
m, when young showing tendencies to the Callispora-type, when mature of the Buellia-type, 110 THE LICHENOLOGIST Vol. 32 apices pointed, straight to slightly curved, roughly ornamented. Spermatia bacilliform, straight, 4·5–5·5(–6)1
m. Chemistry. Thallus UV+orange, K+and C+yellowish, KC+orange; thallus section C+orange. Containing 6-O-methylarthothelin (=granulosin) (major), arthothelin (minor) and atranorin (traces) by TLC and HPLC. Traces of several additional unidentified substances have been detected in some of the specimens analysed by TLC. Depending on the concentration of 6-Omethylarthothelin (with arthothelin), the spot tests mentioned above are more or less clearly visible. According to Wirth (1988), Buellia poeltii (=B. chloroleuca) contains placodiolic acid. This report is not in agreement with our TLC and HPLC analyses. The xanthone 6-O-methylarthothelin has previously been reported for this species (as B. zahlbruckneri) by Huneck et al. (1992) and by Kalb & Elix (1998). Within the genus Buellia, it is known also from B. granulosa (Leuckert & Mathey 1975), B. insignis, B. melanochlora, B. mediospora and B. parasema (Kalb & Elix 1998). Taxonomic remarks. Schauer (1965) incorrectly neotypified Buellia zahlbruckneri J. Steiner and he chose as neotype the specimen Arnold: Lich. Exs. 1589 from M. The specimens of Arnold: Lich. Exs. 1589 (DUKE, FH, UPS) investigated (invalid ‘isoneotypes’) belong to B. chloroleuca. Thus, the invalid taxon B. zahlbruckneri in the sense of Schauer (1965) is a ‘synonym’ of B. chloroleuca. See also the taxonomic remarks under B. erubescens. Discussion. Buellia chloroleuca is characterized by the presence of 6-Omethylarthothelin. Other distinguishing characters of this species include the typically thick and warted thallus, which usually covers wide areas, the abundant, confluent and adnate apothecia, the K+deep yellow-brownish reaction of the rather well developed excipulum, the strongly swollen apical cells of the paraphyses and the relatively small Buellia-type ascospores, which show Callispora-type tendencies. Among the species treated, B. chloroleuca is closely related to the corticolous/muscicolous B. insignis, which also contains the xanthone 6-O-methylarthothelin and has similar morphological characters (a rather thick and warted thallus, adnate and abundant apothecia, lack of hymenial oil droplets, large apical cells of the paraphyses, etc). However, the latter species is well separated by the larger ascospores (see comments under B. insignis). As indicated above (see Chemistry), the intensity of the spot test with UV, K, C and KC may vary depending on the concentration of 6-Omethylarthothelin in the thallus. When these test reactions are doubful, B. chloroleuca could be confused with the chemotype of B. erubescens, which lacks or has low concentrations of norstictic acid. Apart from TLC analyses (which give a rapid solution), the following morphological characters should be carefully studied: the degree of development of thallus, the size of apothecia, the thickness of the excipulum and its reaction with K, the size of the apical 2000 Iberian Buellia—Giralt et al. 111 cells of the paraphyses, the size of the ascospores and the ascospore-ontogeny (see comments under this taxon). Buellia poeltii was first described by Schauer (1965). The holotype and other specimens examined give a K+pale yellow thalline reaction, which was considered by Schauer (op. cit.) to be negative. For this reason, he compared this species with other Buellia taxa that lack secondary lichen compounds, such as Amandinea punctata (as B. punctata) and B. schaereri, rather than with species with K+yellow thalli, such as B. chloroleuca or B. disciformis. Consequently B. poeltii was described as new and distinguished from A. punctata and B. schaereri on the basis of its larger ascospores. Our morphological and chemical studies on several isotypes and one additional specimen of B. chloroleuca established that they differ from specimens of B. poeltii (holotype included) only by their more granulose thalli, a character that we consider of little taxonomic value. For this reason we synonymize B. poeltii with B. chloroleuca. The same thalline character separates the ‘isoneotypes’ and other specimens of the invalid taxon B. zahlbruckneri sensu Schauer examined from B. chloroleuca. But as Schauer’s neotypification of B. zahlbruckneri is totally invalid no synonymy between B. chloroleuca and B. zahlbruckneri sensu Schauer must be considered (more details are given under the taxonomic remarks on B. erubescens). Ecology and distribution. In the study area B. chloroleuca occurs at high altitudes in the Pyrenees. Overall, the European distribution of B. chloroleuca is poorly known due to confusion of this species with B. zahlbruckneri sensu Schauer. Nevertheless, it seems to be a montane to alpine species, rather common in the Alps where it occurs mainly on lignum. Many records of B. zahlbruckneri growing on lignum may refer to this species. In the area studied, only the record in Etayo (1989b) corresponds to B. chloroleuca. The specimen reporded as B. zahlbruckneri in Boqueras (1993) could also belong to B. chloroleuca but it is so severely parasitized by the lichenicolous fungus Lichenodiplis lecanorae that its identity could not be confirmed. Other Iberian records of B. zahlbruckneri checked refer to other Buellia taxa but not to B. chloroleuca [e.g. to B. erubescens. Etayo (1989a) or to Buellia sp., Marcos (1985)]. Exsiccatae examined: Anzi, Lich. exs. minus rari Ital. sup. 296, 297, 299 (M, as Buellia parasema v. saprophila). Hepp, Fl. Eur. 150 (M, as Lecidea punctata v. saprophila). Rabenhorst, Lich. Eur. 729 (M, as Buellia parasema v. saprophila). Roumeguère, Lich. Gall. exs. 191 (M, as Lecidea disciformis f. ecrustacea). Schaerer, Lich. Helv. exs. ed. I. 198 (M, as Lecidea punctata v. saprophila). Vězda, Lich. sel. exs. 897 (W, as B. zahlbruckneri) and 1793 (as Buellia disciformis, BCC). Lichenes Alpinum 192 (W, M, as B. zahlbruckneri). Selected additional specimens examined: Spain:. Navarra: Larra, 1800 m, on dry twigs of Pinus uncinata, J. Etayo (hb. Etayo).—Austria: Carinthia: Turracher Höhe ’supra lacum Scwarzsee, 1840–1900 m, ad lignum et corticem Pini cembrae’, 1985, A. Vězda (MUB-6999); Nockgebiet, Huberalpe, östl. St. Lorenzen bei Ebene Reichenau, on lignum, 1984, S. Wagner (KL); Gailtaler Alpen, Kerschdorfer Alm (östl. der Windischen Höhe), on lignum, 19 vi 1984, S. Wagner (KL).—France: Alpes Maritimes: oberh. Casterine, westl. St. Dalmas de Tende, 1600 m, an Larix, 1963, Th. Schauer (M).—Germany: Oberbayern: Wildsee am Krottenkopf bei Garmish, 1400 m, an Sorbus aucuparia, 1962, Th. Schauer (M); Sägertalbach bei Linderhof, 1400 m, 112 THE LICHENOLOGIST Vol. 32 an Fichtenstumpf, 1962, Th. Schauer (M).—Italy: Südtirol, Belluno, Pinus cembra-Äste mi Valgrande nahe Schluderbach in den Ampezzaner Alpen, 1874, F. Arnold (M); Bozen, P. cembra-Strunk mi Damersthale auf dem Shlem, 1867, F. Arnold (M); Trento, Dolomiten, Dürre Fichten-Zweige bei Paneveggio, 1878, F. Arnold (M).—In Herbarium F. Arnold: locality not indicated (M). Buellia disciformis (Fr.) Mudd Manual Brit. Lich.: 216 (1861) Lecidea parasema de Not. var. disciformis Fr., Nov. Sched. Critic.: 9 (1826); type: Fries, Lich. Suec. Exs. 215A (UPS!—lectotype). (Fig. 2A) Thallus whitish to whitish grey, varying from thin and smooth to rather thick and cracked-areolate, continuous, often delimited by a blackish prothallus. Apothecia (0·5–)0·7–1·1(1·3) mm diam., usually scattered, adnate to sessile. Disc plane to subconvex, rarely strongly convex, epruinose. Proper margin initially thick, entire to flexuose, prominent, becoming thinner, usually persistent. Excipulum (50–)60–80(–100)
m wide, K
(microscope slide). Hymenium with numerous oil droplets, 100–120
m high. Apical cells of the paraphyses slightly swollen, 2·5–3·5(–4·5)
m wide, brown pigmented. Ascospores (16–)19–25(–28)7·5–9(–11)
m, similar to the Callispora-type, with inner wall clearly thickened subapically, but not or only slightly thickened at septum, often curved and becoming triseptate, smooth. Spermatia elongate bacilliform, straight, 5·5–7·5(–9)0·8–1
m. Chemistry. Thallus K+yellow. Atranorin (major) and usually traces of fulgidin (minor), fulgoicin (minor or trace), and of an additional unknown lichen substance () (tentatively identified as norfulgoicin), by TLC and HPLC. Fulgoicin is reported for the first time from the genus Buellia. Fulgidin is also known from B. parasema de Not., the type species of the genus proposed by Kalb & Elix (1998). Discussion. The presence of atranorin, fulgidin and fulgoicin together with the large apothecia, the hymenial oil droplets and the near Callispora-type ascospores are diagnostic characters of B. disciformis. Morphologically B. disciformis comes close to B. leptoclinoides but is clearly distinguished chemically. Further distinguishing features of B. leptoclinoides (absent in B. disciformis) include the thicker, often yellowish thallus due to the presence of placodiolic acid, the more crowded and convex apothecia possessing large crystals in the outer part of the excipulum, and the significantly smaller ascospores. These taxa also exhibit ecological differences. Buellia arnoldii is also similar morphologically but it contains only atranorin and has larger ascospores (see also the discussion under that species). Buellia disciformis is a relatively uniform species despite the fact that the size and septation of the ascospores may vary. Whereas the average spore length in the type material is 21
m, it can reach 25
m in other specimens examined. The specimens that possess larger ascospores are also those that contain a 2000 Iberian Buellia—Giralt et al. 113 F. 2. Ascospore ontogeny. A, Buellia disciformis (Spain, Las Batuecas, 1989, Marcos, SALAF1577); B, B. leptoclinoides (Portugal, 14 km of Lagos, 1993, P. v. d. Boom, Boom 14670). Typical Callispora-type ascospores. Scale =10
m. higher number of triseptate ascospores. Several records of B. lauricassiae from the study area refer to this morphotype of B. disciformis. Buellia parasema de Not. resembles B. disciformis in having a hymenium inspersed with oil drops and callisporoid ascospores with tendencies to 114 THE LICHENOLOGIST Vol. 32 become four-celled. This species is mainly distinguished from B. disciformis by its alternative chemistry since it contains atranorin, as the major substance, and norstictic acid, dissectic acid, methyl -orsellinate, 6-Omethylarthothelin, brialmontin 1, brialmontin 2 and fulgidin, as minor substances (cf. Kalb & Elix 1998). Ecology and distribution: Buellia disciformis seems to be a rather common species, widely distributed in Portugal and in northern Spain except for coastal localities. It grows from low to middle altitudes, mainly on the smooth bark of deciduous trees. In the Mediterranean area of the Iberian Peninsula, B. disciformis is restricted to the montane forests of Fagus and has been reported many times from the study area. Additional specimens examined. Portugal: Algarve: Serra de Monchique, road Casais to Marmelete near Rua Nova, 400 m, on Quercus suber, 1993, P.v.d. Boom, (hb. Boom 14621). Alentejo: SSW of Évora, SE of Alvito, S side of Barragem Odivelas, 100 m, on Q. ilex, 1995, P.v.d. Boom (hb. Boom 17185). Beira Litoral: 8 km SW of Oliveira do Hospital, Venda da Galizes to Ponte das Tres Entradas, 350 m, on Pinus, 1993, P.v.d. Boom (hb. Boom 14505). Minho: Serra do Gerez-Pedra Bela, on Betula, 1947, C. Tavares 2014, 2023 (LISU, UPS); Serra do Gerez-Caldas, junto a Vacaria, on Betula, 1944, C. Tavares 82 (LISU); Serra do Gerez-Vacaria, no viveiro, 450 m, on Fagus, 1948, C. Tavares 2541 (LISU). Tras os Montes: EN of Villa Real, N of road to Murça, S of Escarao, 700 m, on Castanea, 1995, P.v.d. Boom (hb. Boom 16977); Villa Real, EN side of Village, Parque forestal, 700 m, on Q. rubra, 1995, P.v.d. Boom (hb. Boom 16939).—Spain: Andalucía: Huelva, Galaroza, Sierra de Navahermosa, La Suerte, 680 m, on Q. suber, 1992, S. Fos (VAB-8445). Aragón: 14 km WNW of Bielsa, Ordesa, NW of Parador, 1350 m, on Betula, 1992, P.v.d. Boom (hb. Boom 13599). Castilla-León: León, Puerto del Pando, 1300 m, on Q. robur and Q. petraea, J. Etayo (hb. Etayo 5676). Salamanca, Montemayor, on Castanea, 1981, B. Marcos (SALAF-1360); Salamanca, Casillas de Flores, on Q. pyrenaica, 1982, B. Marcos (SALAF-1368); Salamanca, Monsagro, on Castanea, 1984, B. Marcos (SALAF-1404); Salamanca, Herguijuela de la Sierra, on Fagus sylvatica, 1982, B. Marcos (SALAF-1381); Salamanca, Las Batuecas, La Alberca, on Ailanthus altissima, 1989, B. Marcos (SALAF-1577); Palencia, Espigëte, Cardaño de Abajo, 1400 m, on Q. pyrenaica, 1990, A. Terrón & A. Ubeda (LEB-205). Catalunya: Vallès Oriental, Montseny, Turó de l’Areny, 900 m, on F. sylvatica, 1978, A. Gómez-Bolea (BCC-3340); Font de Passaret, 1200 m, on F. sylvatica, 1979, A. Gómez-Bolea (BCC-3338); Turó de I’Home, on F. sylvatica, 1978, A. Gómez-Bolea (BCC-3337). Vall d’Aran von Kursiva, Viella, Bosc de Conangles, on cortex, 1984, A. Gómez-Bolea (BCC-3075, 3322); Artiga de Lin, 1500 m, on F. sylvatica, 1979, A. Gómez-Bolea (BCC-3339); Montsianès non Kursiva, Serra dels Ports, La Sènia, Barranc del Retaule, 1000–1100 m, on F. sylvatica, 1987, M. Boqueras (BCC-5866); ibid, 1986, V. Atienza (VAB-945). Galicia: La Coruña, Grañas, Mañón, on Q. robur, 1988, A. Garcia (SANT-6388); Rebolás, Ortigueria, on Q. robur, 1988, A. Garcia (SANT-6385); Ancares, on Betula, 1992, G. Paz Bermúdez et al. (SANT-8707). Navarra: Zugarramundi, 209 m, on Q. robur, 1988, J. Etayo (hb. Etayo 4423); Goizueta, 200 m, Q. robur, 1986, J. Etayo (hb. Etayo 4125). Buellia erubescens Arnold Verhandl. zool.-bot. Gesellsch. 25: 493 (1875); type: Austria, Tirol, im Walde zwischen Passthurm and Kirchamwald, c. 1070–1274 m, on Alnus incana, viii 1873, F. Arnold (M!—holotype). New synonyms: Buellia zahlbruckneri J. Steiner, Annal. Naturhist. Hofmuseums Wien 23: 122 (1909); type: Asia Minor, (Sandschack) districtus Trapenzunti: ‘in catena montium ad septentrionem fluminis Charshut inter vicos Fol et Eseli prope alpem Kisyl Ali-Jaila, 1850 m, cortice Fagi et Aceris’, 16 vii 1907, Handel-Mazzetti 1040 (W!—lectotype, selected here, WU!isolectotype). Buellia jorgei Samp., Bolet. Soc. Broterian. 2(2): 177 (1923); type: Portugal, Minho, Ponte do Lima: Sá (na veiga), ix 1922, G. Sampaio (LISU!—syntype). 2000 Iberian Buellia—Giralt et al. 115 F. 3. Ascospore ontogeny. A, Buellia erubescens (M—holotype); B, B. hyperbolica (Fl—holotype). Buellia-type ascospores showing tendencies to the Physconia-type (arrows). Scale =10
m. (Fig. 3A) Thallus whitish to whitish grey, thin, smooth to rimose, continuous, usually delimited by a black prothallus. Apothecia (0·3–)0·8·1(–1·5) mm diam., sessile, scattered. Disc usually plane, more rarely subconvex to convex, often pruinose, pruina when present usually K+red crystals (microscope slide). Proper margin thick, entire, prominent, becoming thinner with age, usually persistent. Excipulum very well developed, expanding to 80–125
m below, K+yellow (microscope slide). Hymenium without oil droplets, (80–)100– 110
m high. Apical cells of the paraphyses slightly swollen, 2–3·5(–4)
m wide, brown pigmented. Ascospores (11–)13–16(–18)(6–)7–8·5(–9·5)
m, some when yoúng of the Physconia-type, with inner wall slightly thickened at septum, soon becoming of the Buellia-type, with walls uniformly thin, apices rounded, straight, roughly ornamented. Spermatia bacilliform, 6–7·50·8– 1
m. Chemistry. Chemotype I: Thallus K+yellow-orange then red, PDyelloworange; thallus section and often also epihymenium crystals K+deep yellow followed by the formation of red crystals (microscope slide). Atranorin (major), norstictic acid (major), connorstictic acid (trace), hyposalazinic acid (trace),zeorin and unknown secalonic acid derivative (minor), by TLC and HPLC. 116 THE LICHENOLOGIST Vol. 32 Chemotype II: Thallus K+yellow and PD+yellowish. Atranorin, traces of norstictic and connorstictic acids () and traces of secalonic acid derivative () by TLC. Hyposalazinic acid is recorded here for the first time from the genus Buellia. This substance was also detected, by TLC and HPLC, in a chemotype of the saxicolous B. sequax, which lacks norstictic acid. In this chemotype, hyposalazinic acid co-occurs with hypostictic acid. Hypostictic acid has been recorded previously from ‘Buellia’ lindingeri (Kalb & Hafellner 1992). Inoue (1993) mentions an unidentified minor constituent in B. subfrigida and B. frigida. According to the chromatograms published by Inoue (1993: 21) this unidentified chemical could well be hypostictic acid. Taxonomic remarks. As a consequence of Schauer’s treatment (1965: 623), the synonymy of B. erubescens Arnold and B. zahlbruckneri J. Steiner has remained in doubt. Some authors consider that these taxa are conspecific (Orange et al. 1992) while others remained unconvinced (Santesson 1993). Buellia zahlbruckneri was described by Steiner (1909) and after the Latin diagnosis he included the following information: ‘An Buchen- und Ahornrinde bei der Kisil Ali-Jaila, 1850 m, Handel-Mazzetti nr 1040’. Thus, the specimens so labelled should be considered the type specimens of B. zahlbruckneri. We have checked two specimens labelled as such by Steiner (1909), one stored in W and the other one in WU. The former (W) corresponds to the chemotype of B. erubescens that contains norstictic acid (thallus K+red) and the second (WU) to the chemotype that possesses only traces (detected by TLC) of this substance (thallus K+yellow). Given their analogous morphology, we believe that B. zahlbruckneri J. Steiner is a synonym of B. erubescens Arnold. In the original description of B. zahlbruckneri, Steiner (1909) discussed the differences between the new species and B. disciformis but he did not compare it with B. erubescens. Several specimens of B. erubescens in WU were identified by Steiner as B. zahlbruckneri J. Steiner var. erubescens Arnold. This shows that Steiner knew of B. erubescens Arnold but that he considered it to be a variety (characterized by the K+red thallus reaction) of his new taxon (see Zahlbruckner 1931: 431). When Schauer (1965) observed that the type material of B. zahlbruckneri (probably the specimen from W) gave a K+red thallus reaction, he thought that this specimen was a false type and concluded that there was not type material of B. zahlbruckneri (‘Das erwähnte Exemplar nr. 1040 muß falsch bestimmt sein, so daß für B. zahlbruckneri J. Steiner, kein Typus existiert’). Consequently, Schauer (1965) incorrectly neotypified B. zahlbruckneri and, on the basis of the discussion of Malme (1927: 258), he chose as neotype the specimen Arnold: Lich. Exs 1589 from M. As indicated before (see taxonomic remarks under B. chloroleuca), the specimens (invalid ‘isoneotypes’) of Arnold: Lich. Exs. 1589 investigated (DUKE, FH, UPS) belong to B. chloroleuca. Thus, the invalid taxon B. zahlbruckneri in the sense of Schauer (1965) is a ‘synonym’ of B. chloroleuca. Discussion. In the study area, B. erubescens is a rather uniform species, normally well characterized by the K+red thallus, the large and often pruinose apothecia, which always possess a well developed K+yellow excipulum, the 2000 Iberian Buellia—Giralt et al. 117 lack of hymenial oil droplets and the small Buellia-type ascospores. Although several of the specimens investigated lacked the K+red thallus reaction (norstictic acid), traces of this substance could nearly always be detected by TLC. Besides the diagnostic characters cited above, a further feature that can assist in identifying specimens of B. erubescens that lack the K+red thallus reaction is the K+strong yellow reaction of the excipulum (microscope slide). A similar reaction can be observed in the excipulum of B. chloroleuca, B. hyperbolica and B. insignis. However, these species are well separated from B. erubescens by their alternative chemistry, larger ascospores and significantly different ecology (see comments under those taxa). In the study area, B. erubescens could be confused with an undescribed Buellia species, a species that also contains norstictic acid, lacks hymenial oil droplets and has small ascospores. However, the new species is well separated by smaller apothecia with a poorly developed excipulum, larger ascospores that show clear Callispora-type tendencies and alternative ecology. Buellia sanguinolenta, a further species that contains norstictic acid, is readily distinguished from B. erubescens by the presence of numerous oil droplets in the hymenium and much larger Callispora-type ascospores (see also comments under B. sanguinolenta). No significant differences have been observed between the type of B. jorgei, a taxon originally described by Sampaio (1923) from Portugal, and the specimens of B. erubescens examined (including the holotype). For this reason we synonymyze B. jorgei with B. erubescens. Ecology and distribution. In the Atlantic regions of the Iberian Peninsula, B. erubescens seems to be a common species that grows from low to middle altitudes, usually on the smooth bark of deciduous trees. From these areas it extends to the Pyrenees but does not reach the Mediterranean mountains. Buellia erubescens has been reported many times from the study area. Indeed, most records of B. jorgei also refer to this species. The Mediterranean records of B. erubescens refer to the undescribed Buellia species mentioned above. Exsiccata examined. Merrill, Lich. exs. 165 (as B. parasema, W). Additional specimens examined. Portugal: Beira Litoral: 12 km S of Oliveira do Hospital, Ponte das Tres Entradas, 350 m, on Alnus, 1993, P. v.d. Boom (hb. Boom 14288); Serra da Lousa, Quinta de Alfocheira, na Mata, on Tilia, 1943, C. Tavares (LISU); Serra do Caramulo, Paredes do Guardao, próximo da Ponte dos Amores, on Prunus, 1942, C. Tavares (LISU). Minho: Serra do Gerez-Pedra Bela, 750 m, on Betula, 1947, C. Tavares (LISU-2013, 2040, UPS); Serra do Gerez, próximo da Bouça da Mó, on llex, 1948, C. Tavares (LISU-2688). Tras os Montes: Villa Real, EN side of Village, Parque Forestal, 700 m, on Quercus rubra, 1938, P. v.d. Boom (hb. Boom 16938).—Spain: Castilla-León: Salamanca, Las Batuecas, on Q. rotundifolia, 1981, B. Marcos (SALAF-1356); on Alnus, 1983, B. Marcos (SALAF-1355). Galicia: La Coruña, Taboada, Monfero, on Q. robur, 1988, A. Garcia (SANT-6387); Sarandons, Abegondo, on Q. robur, 1989, A. Garcia (SANT-4824); Trazo, on Q. robur, 1989, A. Garcia (SANT-4823); Mairame, Cercede, on Q. robur, 1989, A. García (SANT-4822); Fene, on Robinia pseudoacacia, 1987, A. García (SANT-5966); Igrexafreda, S. Sadurniño, on Q. robur, 1988, A. Garcia (SANT-4823); Presa de Vilagudún, Ordes, 1989, A. Garcia (SALAF-6308); Caaveiro, Pontedeume, on Q. robur, 1986, J. Alvarez & M.E. López de Silanes (SANT-2423, 2421); ibid. on Castanea and Alnus, 1986, J. Alvarez & M.E. López de Silanes (SANT-2422); Santiago de Compostela, on Q. robur, 1982 118 THE LICHENOLOGIST Vol. 32 (SANT-18); Lugo, Busir, Guitriz, on Q. robur, 1988, A. Garcia (SANT-4823); Carretera de Pambre, on Betula celtiberica, J. Etayo (hb. Etayo-5637); Pontevedra, Capital, on Q. robur 1987, A. Garcia (SANT-7037); Lourizán, on C. sativa, Q. robur, 1982 (SANT-19, 20). Navarra: Refugio de Belagua, 900–1000 m, on Fagus, 1985, J. Etayo (hb. Etayo-3708); Guerendiain, Viscarret, 500 m, 1986, J. Etayo (hb. Etayo); Alcoz, 560 m, on Q. robur, 1986, J. Etayo (hb. Etayo); Lizardoya, 800–100 m, 1985, J. Etayo (hb. Etayo 4313); Bertiz, 200 m, on Fagus, 1985, J. Etayo (hb. Etayo 4341); Irati, 800–100 m, on Fagus, J. Etayo (hb. Etayo 3202); Oroquieta, 580 m, on Fagus, 1986 J. Etayo (hb. Etayo). Buellia hyperbolica Bagl Nuov. Giorn. Botan. Ital. 3: 266 (1871); type: Italy, Toscana: ‘Sulle vecchie ceppaie di castagno al Mt. Pisano’, Beccari (FI!—holotype). (Fig. 3B) Thallus of whitish, whitish grey to yellowish green granules, continuous or not, usually covering a wide area. Apothecia lecideine, rapidly becoming biatorine, 0·2–0·3(–0·4) mm diam., adnate, abundant and confluent. Disc plane at first, soon hemispherical. Proper margin very thin, and even when juvenile almost totally excluded. Excipulum poorly developed, up to 20
m wide, K+yellowish (microscope slide). Hymenium without oil drops, 70–90 (–100)
m high. Hypothecium interspersed with abundant oil droplets, 100–120
m deep, K+yellow (microscope slide). Apical cells of the paraphyses strongly swollen (3·5–)4–5
m wide, pigmented dark brown. Ascospores (19–)21–24(–29)(7·5)8–9·5(–11)
m, of the Physconia-type when young, with inner wall thickened at septum, Buellia-type when mature, with walls uniformly thin; apices pointed, often slightly curved, roughly ornamented. Spermatia not seen. Chemistry. Thallus K+yellowish brown, PD+orange-red. Atranorin (traces), fumarprotocetraric acid (major), protocetraric acid (minor), confumarprotocetraric acid (minor) and convirensic acid (traces), by TLC and HPLC. Hitherto, no chemical data were available for this species. Furthermore, this is the first record of protocetraric, convirensic and confumarprotocetraric acids from the genus Buellia. Discussion. The chemistry readily distinguishes B. hyperbolica from the other species treated in the present paper. Other diagnostic characters of this species are the granulose thallus, the small, biatorine apothecia showing a poorly developed excipulum, the lack of hymenial oil droplets and the large ascospores. According to the various revisions of this genus (e.g. Imshaug 1951; Sheard 1964; Harris 1988; Scheidegger 1993), B. dialyta (Nyl.) Tuck. and B. rubifaciens R.C. Harris, are the only species of the genus reported to contain fumarprotocetraric acid. Buellia rubifaciens, a corticolous species from Florida, differs from B. hyperbolica in having smaller ascospores, 13–16·56·5–7·5
m (Harris 1988). Buellia dialyta, a corticolous species originally described from California, also reported from several localities in the northeastern United 2000 Iberian Buellia—Giralt et al. 119 States (Imshaug 1951), would appear to be closely related if not conspecific with B. hyperbolica, as both taxa have the same diagnostic characters. Nevertheless we prefer to maintain the name B. hyperbolica until the type specimen of B. dialyta is available for study. According to Marbach (1999), B. dialyta and B. rubifaciens belong to the new genus Chrismofulvea Marbach, mainly characterized by the presence of small apothecia, fumarprotocetraric acid and Physconia-type ascospores. He also includes in Chrismofulvea two additional species, C. omalia Marbach and C. pinastri (Erichsen) Marbach. The former species is separated from Buellia hyperbolica by its sorediate thallus and the latter by its hymenium inspersed with oil droplets. Ecology and distribution. Buellia hyperbolica is a corticolous/lignicolous species that grows mainly on mature trunks and wood of Castanea and Quercus. It seems to be locally abundant in relatively undisturbed Castanea and Quercus forests of the Mediterranean and southern Atlantic regions of the Iberian peninsula. Hitherto the European distribution of this species was poorly known as it has been reported only from the type locality in Italy, from Portugal, (Boom et al. 1990), and from Spain (Boom & Gómez-Bolea 1991). An additional record in Alvarez & Carballal (1992) from Galicia (Sierra de Caurel, Lugo) has not been verified. The species has also been collected in Windsor Great Park (London), on an ancient Quercus, and in Tuscany (Italy), on Castanea (Coppins pers. com.). Additional specimens examined. Portugal: Minho: 32·5 km of Braga, Parque Natural de Gerês, N of Gerês, on wood of Quercus, P. v.d. Boom (hb. Boom 6334, 6339).—Spain: Asturias: 35 km E of Cangas de Onis, 10 km of Panes, 0·7 km S of Trescares, on Castanea, 250 m, 1989, P. v.d. Boom (hb. Boom 8947); 28 km E of Cangas de Onis, 2 km E of Arenas de Cabrales, S of road to Panes, 200 m, on wood of Castanea, 1989, P. v.d. Boom (hb. Boom 8918, BCC-5116). Cantabria: Santander, Picos de Europa, 4 km W of Potes, Argüebanes, 500 m, on wood of Castanea, 1987, P. v.d. Boom (hb. Boom 6066). Navarra: Roncesvalles, 1000 m, on Castanea wood, 1986, J. Etayo (hb. Etayo 3631). Buellia leptoclinoides (Nyl.) J. Steiner Verhandl. zool.-bot. Gesellsch. Wien 57: 357 (1907); type: France, Pyrénées-orientales, Port-Vendres, 6 vii 1887, Nylander (H-Nyl 10541—lectotype, not seen). (Fig. 2B) Thallus whitish, pale yellowish grey to ochraceous, thick, rimose to crackedareolate, continuous, sometimes delimited by a blackish prothallus. Apothecia (0·3–)0·5–0·8(–1) mm diam., scattered to confluent, adnate to sessile. Disc plane to convex, epruinose. Proper margin initially moderately thick, becoming thinner, finally excluded. Excipulum (30–)40–60
m wide, K
(microscope slide). Hymenium with numerous oil droplets, 80–120
m high. Apical cells of the paraphyses slightly swollen, 2·5–3·5
m wide, dark brown pigmented. Ascospores 15–20(–23)(6·5–)8–9(–9·5)
m, of typical Callispora-type, with inner wall clearly thickened subapically and at 120 THE LICHENOLOGIST Vol. 32 the septum, sometimes slightly curved and becoming triseptate, weakly ornamented. Spermatia 4·5–61
m, bacilliform, straight. Chemistry. Thallus K+yellow. Atranorin (minor) with chloratranorin (traces) and placodiolic acid (major), by TLC and HPLC. Discussion. Buellia leptoclinoides is characterized by the thick, pale yellowishgrey to ochraceous, cracked-areolate thallus, which contains atranorin and placodiolic acid, the abundant and often crowded apothecia with convex discs, the hymenium with numerous oil droplets, and the typical Callispora-type ascospores. Among the species treated, B. leptoclinoides is unique in containing placodiolic acid. In the study area only B. arnoldii and B. disciformis, which also possess hymenial oil droplets and Callispora- or near Callispora-type ascospores, could be mistaken for B. leptoclinoides. However, both taxa are well separated by the significantly larger ascospores, the alternative chemistry and ecology. A further distinguishing feature of B. leptoclinoides, which is absent in B. arnoldii and B. disciformis, is the presence of large crystal complexes in the thallus and outer part of the excipulum (placodiolic acid?). The size of the spermatia also distinguishes B. disciformis from B. leptoclinoides. This is the first time that B. leptoclinoides has been reported as growing on bark. For the most part the different substratum separates these collections from the more typical, saxicolous specimens of B. leptoclinoides. An additional feature that seems to distinguish both ecotypes has been observed in the spore ornamentation, which is smooth in the saxicolous specimens (Scheidegger 1993; Nordin 1998) but is weak in those specimens growing on bark. However, in our opinion, this character is of little taxonomic significance to separate a new taxon. According to the literature (e.g. Kalb 1986; Scheidegger 1993; Sheard 1992; Pusswald et al. 1994), B. leptoclinoides is clearly distinguished from all other corticolous and lignicolous Buellia species previously included in the genus Hafellia (see Kalb & Elix 1998; Marbach 1999) by its smaller ascospores. Only B. bahiana (=Hafellia bahiana), has ascospores of similar size (cf. Sheard 1992). However, the presence of norstictic acid makes this North American species clearly distinct from B. leptoclinoides. Buellia leptoclinoides differs from all other Buellia species previously included in the genus Hafellia, by the presence of placodiolic acid. Ecology and distribution. In the study area the corticolous specimens of B. leptoclinoides have been collected only in coastal localities of central and southern Portugal. It seems to be confined to the coast, occurring in warm places subject to humid coastal winds as well as in the salt-spray belt. It grows mainly on the bark of maritime trees such as Ceratonia and Ficus. Specimens examined. Portugal: Algarve: N of Albufeira, 1 km N of Alte, 400 m, on Ceratonia, 1993, P.v.d. Boom (hb. Boom 14925); W of Lagos, road Vale de Boi to Barao de San Miquel, 25 m, on Prunus dulcis, 1993, P. v.d. Boom (hb. Boom 14571); 14 km W of Lagos, along road to Salema, 50 m, on Ceratonia, 1993, P.v.d. Boom (hb. Boom 14670); Alvor, zwischen Lagos und Portimao, on Ficus, 1988, O. Breuss (hb. Breuss 5288). Estremadura: 25 km W of Setubal, Aldiea do Meco, 40 m, on Ficus, 1987, P.v.d. Boom (hb. Boom 6600). 2000 Iberian Buellia—Giralt et al. 121 F. 4. Ascospore ontogeny. A, Buellia arnoldii (Italy, Südtitol, Arnold, PRM—syntype); B, B. regularis (Kalb 23182—isotype); C, B. sanguinolenta (M—holotype). Typical Callispora-type ascospores. Scale =10
m. Other corticolous Buellia species examined that do not occur in the study area Buellia arnoldii Servít Vest. Král. Spol. Nauk. Tr. 2: 39 (1931); types: Austria: Tirol, auf dem dickerer Äste von Taxus im Walde östl. von Plansee, Arnold (M!, PRM!—syntypes).—Germany: Oberbayern, Wetterstein, dürre Zirbenäste auf dem Schachen bei Partenkirchen, Arnold (M!, PRM!—syntypes).—Italy: Südtirol, auf Rinde abgestorbener Fichtenzweige im Latemarwald-Karerpass, Arnold (M!, PRM!—syntypes). (Fig. 4A) Thallus whitish to whitish grey, thin, smooth to rimose, continuous. Apothecia 0·4–0·8 mm diam, scattered, sessile. Disc flat to subconvex. Proper margin thin, persistent. Excipulum 60–90
m wide, K
(microscope slide). Hymenium 100–150
m high, inspersed with numerous oil droplets. Apical cells of the paraphyses slightly swollen, (2·5–)3–4
m wide, brown pigmented. 122 THE LICHENOLOGIST Vol. 32 Ascospores (23–)26–29(–31)(10–)12–14(–16·5), Callispora-type, with inner ascospore wall thickened subapically and at the septum, apices pointed, often curved, smooth. Spermatia not seen. Chemistry. Thallus K+yellow. Atranorin, by TLC, according to our own analyses and Orange et al. (1992). Due to the scanty material of B. arnoldii, no type specimen has been analysed by TLC and no lectotype selected. Discussion. Buellia arnoldii is characterized by the interspersed hymenium, the large Callispora-type ascospores and the presenće of atranorin. Among the other species studied, B. arnoldii could be mistaken only for B. sanguinolenta or B. regularis, both of which have inspersed hymenia and Callispora-type ascospores. Buellia sanguinolenta is distinguished from B. arnoldii by its alternative chemistry as it contains norstictic and traces of connorstictic acids in addition to atranorin (). Our investigations have indicated that in the genus Buellia, specimens that differ only by the presence/absence of norstictic acid should not be considered different taxa, but only different chemotypes. Scheidegger (1993) and Nordin (1996) came to a similar conclusion where, for example, norstictic acid was found in only one of the two chemotypes of B. sequax and of B. alboatra and B. venusta, respectively. In the present paper, no taxonomic significance has been given to the presence or absence of norstictic acid in B. erubescens. According to Schauer (1965) and Orange et al. (1992), an additional distinguishing feature between these two taxa can be found in the size of the apical cells of the paraphyses, being 4–4·5
m wide in B. arnoldii and only 2–3
m wide in B. sanguinolenta. However, according to our observations, these cells measure (2·5–)3–4
m in the types of B. arnoldii and 2·5–3·5(–4)
m in that of B. sanguinolenta. The recently described B. regularis (Kalb & Hafellner 1992) is closely related if not conspecific with B. arnoldii. It is said to be distinguished by (significantly?) shorter and wider ascospores (25–2913–16·5
m), that furthermore, are generally strongly curved [viz. (23–)26–29(–31)(10–) 12–14(–16·5)
m, often curved ascospores, in B. arnoldii]. In our opinion, the characters used for the delimitation of B. sanguinolenta and B. regularis fall within the variability of B. arnoldii. Furthermore, according to the literature, the three taxa show very similar ecological preferences (see Ecology and distribution). Nevertheless, until a thorough revision of the B. arnoldii-group is carried out, we refrain from placing B. sanguinolenta and B. regularis into synonymy with B. arnoldii. Additional studies on the secondary lichen substances and the spermatia would probably help in the delimitation of the species of B. arnoldii-group. Unfortunately, spermatia were not found in any of the specimens examined (types included). Further, TLC analyses could not be carried out on the type specimens of B. arnoldii or B. regularis because of the scanty type material. Literature reports on the spermatia and the chemistry of B. arnoldii-group are inconsistent. According to Harris (1977) and Sheard (1992), the spermatia of B. arnoldii are elongate-ellipsoid (6·5–7·51
m), whereas Orange et al. (1992) reported them to be narrowly ellipsoid to oblong (3–3·51
m). Only Orange et al. (1992) have described the spermatia of B. sanguinolenta, these 2000 Iberian Buellia—Giralt et al. 123 being oblong-ellipsoid (3–41–1·5
m). In the original description of B. regularis (Kalb & Hafellner 1992) there is no mention of its spermatia. As far as the secondary lichen substances are concerned, B. arnoldii and B. regularis have been reported to contain atranorin (Orange et al. 1992; Kalb & Hafellner 1993) and B. sanguinolenta to contain norstictic acid (Orange et al. 1992). The latter result is not in complete agreement with our TLC analyses, since we have shown that the holotype of B. sanguinolenta possesses traces of atranorin and connorstictic acid in addition to norstictic acid. Another species present in the study area that could be mistaken for B. arnoldii is B. disciformis. However, it is well separated by the better developed thallus, larger apothecia, smaller ascospores and significantly different chemistry and ecology (see also the discussion under B. disciformis). Ecology and distribution. Buellia arnoldii seems to be an oceanic species that is widespread in western Europe. It has also been reported from a few localities scattered throughout the Alps (Servı́t & Nádvornik 1931; Schauer 1965), from Sweden (Fritz 1998) and from Macaronesia (Breuss 1988). The species also occurs in western Scotland (Coppins pers. comm.). It usually grows on conifers, in the montane belt. The single record of B. arnoldii from the Iberian Peninsula in Paz Bermudez (1993) refers to B. disciformis. Additional specimens examined. Austria: Tirol: Salzburg, Baumleichen des Hochmoorgebiets der Gerlosplatte, an Fichte, 1640 m, 1963, Th. Schauer (M).—France: Mittelfranken, Dürre Fichtenäste im Walde des Affentals bei Eichstätt, 1873, F. Arnold (M). Buellia insignis (Nägeli ex Hepp.) Th. Fr. Nova Acta Reg. Soc. Scient. Upsal. 3(3): 327 (1861); type: Hepp, Fl. Eur. 39a (G!—lectotype, selected here; G!, BM!, PRM!—isolectotypes). (Fig. 1B) Thallus whitish, grey, cream to ochraceous, usually thick and warted, rarely smooth, continuous and well developed, not delimited by a blackish prothallus. Apothecia (0·5–)0·7–1(–1·3) mm diam., abundant and confluent, subimmersed to adnate, rarely becoming sessile. Disc initially plane to convex, epruinose. Proper margin initially thick, rapidly becoming thinner and finally excluded. Excipulum well developed, 70–100
m wide, K+yellow (microscope slide). Hymenium without oil droplets, 100–130
m high. Hypothecium with oil droplets. Apical cells of the paraphyses strongly swollen, 4–6(–7)
m wide, brown pigmented. Ascospores (19–)21–25(–29)(8–)9–11(–12)
m, resembling the Callispora-type when young, Buellia-type when mature, apices pointed, often curved, roughly ornamented. Spermatia not seen. Chemistry. Thallus UV+orange, K+yellowish, KC+orange; thallus section C+orange. Atranorin (traces), 6-O-methylarthothelin (major),arthothelin (traces) and gyrophoric acid (traces), by TLC and HPLC. The clarity of the spot tests are dependent upon the concentration of 6-O-methylarthothelin (similar variation was also observed with B. chloroleuca). This is the first report of 6-O-methylarthothelin, arthothelin and gyrophoric acid in B. insignis. 124 THE LICHENOLOGIST Vol. 32 The chemistry of B. insignis is close to that of several species of Pertusaria, subgenus Pionospora, for example, P. bryontha and P. dispar (Archer 1993), which also contain a combination of a chlorinated xanthone and an orcinol tridepside (6-O-methylarthothelin and gyrophoric acid, respectively) as major compounds. Discussion. Buellia insignis is well characterized by its chemistry, the lack of hymenial oil droplets and the large Buellia-type ascospores, which show Callispora-type tendencies when young. The description given in Sheard (1964) and Orange et al. (1992) is, at least in part, based on one specimen that, in our opinion, does not correspond to B. insignis. This specimen (Vězda: Lich. sel. exs. 621, BM) is readily distinguished from B. insignis by possessing a thallus composed of subhemisphaerical areolae, smaller apothecia with pruinose discs and hymenial oil droplets. Imshaug (1951) considered that B. papillata (Sommerf.) Tuck. was the correct name for B. insignis but according to Sheard (1964), Schauer (1965), Wirth (1988) and Galloway et. al. (1998), these taxa are not conspecific. This problem can be solved only by studying the type specimen of B. papillata. Ecology and distribution. Buellia insignis is an arctic-alpine species that occurs on mosses, plant debris and more rarely on coniferous bark. It has not been reported from the Iberian Peninsula. For its European distribution see Orange et al. (1992) and Nimis (1993). Exsiccatae examined. Fellman, Lichenes arctici colleti aestate 1863 201 (BM). Selected specimens examined. Austria: ‘Ad terram et muscos in montibus altioribus et alpibus prov. Sondriensis’ (BM).—Norway: Berleraag, 1664, Th. Fries (BM).—Sweden: Lule Lappmark, Kvikkjokk par., 3 km SSW of Kvikkjokk, SE slope of Nammatj, 500–525 m, Picea forest, 1977, B. J. Coppins & L. Tibell (BM).—Switzerland: Berner Oberland, Iffigenfall mi Simmebtal bei Lenk, 1450 m. on Picea, 1964 Frey & Schauer (M).—‘In terra muscosa alpis Sujarrak Lapponiae Lulensis’, 1871, J. Hellbom (BM). Buellia regularis Kalb Herzogia 9: 56 (1992); type: Portugal (Macaronesia), Madeira, an der Levada da Serra do Faial entlang, etwas SW von Santo da Serra in Richtung Camacha. In letzten Resten eines alten, jedoch stark gestörten, NW-exponierten Eucalyptus-Waldes. An Erica arborea, 750 m, 13 viii 1990, K. & A. Kalb (hb. Kalb 23183—holotype, 23182!—isotype). (Fig. 4B) Thallus whitish grey, thin, smooth, continuous, often delimited by a black prothallus. Apothecia 0·3–0·7 mm diam. Disc flat to subconvex. Proper margin thin, persistent. Excipulum 60–80
m wide, K
(microscope slide). Hymenium 100–150
m high, interspersed with numerous oil droplets. Apical cells of the paraphyses 2·5–3·5
m wide, brown pigmented. Ascospores 25– 2913–16·5
m, Callispora-type, with inner ascospore wall thickened subapically and at the septum; apices pointed, usually strongly curved, smooth. Spermatia not seen. 2000 Iberian Buellia—Giralt et al. 125 Chemistry. Thallus K+yellow. Atranorin by TLC (Kalb & Hafellner 1992). Discussion. In our opinion, B. regularis is simply a form of B. arnoldii with shorter, thicker and more strongly curved ascospores and which grows on Erica arborea bark instead of on coniferous bark (including Juniperus). See also comments under B. arnoldii. Ecology and distribution. Buellia regularis is a submontane species originally reported from Madeira where it grows on Erica arborea (Kalb & Hafellner 1992). It is also widespread in the Canary Islands, where it occurs mostly on conifers, Erica and the twigs of Laurus (Hafellner 1995). It has not been reported from the Iberian Peninsula. See also comments on the ecology and distribution of B. arnoldii. Buellia sanguinolenta T. Schauer Mitt. Bot. München 5: 620 (1965); type: Austria, Niederösterreich, Gr. Urwald bei Lunz, in cortice Abietis albae, 1100 m, 221962, Th. Schauer (M!—holotype). (Fig. 4C) Thallus thin, whitish, smooth, continuous or discontinuous and then composed of granules. Apothecia 0·3–0·6 mm diam., scattered, sessile. Disc plane to convex epruinose. Proper margin thin, becoming excluded. Excipulum 60–70
m wide, K
(microscope slide). Hymenium up to 140
m high, interspersed with numerous oil droplets. Apical cells of the paraphyses slightly swollen, 2·5–3·5(–4)
m wide, brownish pigmented. Ascospores (21–)26–32 (–35)(11–)12–13(–16)
m, of the Callispora-type, with inner ascospore wall thickened subapically and at the septum at all ontogenetic stages; apices pointed, often curved, some spores becoming triseptate, smooth. Spermatia not seen. Chemistry. Thallus K+yellow-orange, at first, to finally red, PDyelloworange; thallus section K+deep yellow followed by the formation of red crystals (microscope slide). Atranorin (traces), norstictic and connorstictic acids by TLC. Discussion. Buellia sanguinolenta is well characterized by the presence of atranorin () and norstictic (with connorstictic) acid in the thallus, the hymenium inspersed with numerous oil droplets and the large Callispora-type ascospores. The presence of norstictic acid separates B. sanguinolenta from B. arnoldii. In our opinion, B. sanguinolenta may be just one of the two chemotypes of B. arnoldii, characterized by the occurrence of this lichen substance (see also comments under B. arnoldii and B. regularis). Buellia erubescens shows analogous chemical races. Specimens of B. erubescens that contain norstictic acid are mainly distinguished from B. sanguinolenta by the lack of oil droplets in the hymenium and smaller ascospores (see also comments under B. sanguinolenta). 126 THE LICHENOLOGIST Vol. 32 Ecology and distribution. According to Schauer (1965) and Orange et al. (1992), B. sanguinolenta is an oceanic species, known from Scotland and from three localities in the western Alps situated between 1100 and 1350 m altitude. In Scotland it grows on smooth bark of Sorbus and in Austria on Abies. The record of B. sanguinolenta from the Iberian Peninsula in Atienza (1991) corresponds to the same undescribed Buellia species mentioned in the discussion of B. erubescens (see above). Additional specimen examined: Germany: Oberbayern, Garmisch, Lahnenwies-Graven, 1350 m, an Tanne, 1963, Schauer (GZU). 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