Taxonomic Investigations of Lecanora Strobilina and L. Symmicta (Lecanoraceae, Lecanorales) in Northeastern North America

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Taxonomic investigations of Lecanora strobilina and L. symmicta (Lecanoraceae, Lecanorales) in northeastern North America

Article in The Bryologist · August 2013 DOI: 10.1639/0007-2745-116.3.287

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The user has requested enhancement of the downloaded file. Taxonomic investigations of Lecanora strobilina and L. symmicta (Lecanoraceae, Lecanorales) in northeastern North America Author(s): Scott LaGreca and H. Thorsten Lumbsch Source: The Bryologist, 116(3):287-295. 2013. Published By: The American Bryological and Lichenological Society, Inc. DOI: http://dx.doi.org/10.1639/0007-2745-116.3.287 URL: http://www.bioone.org/doi/full/10.1639/0007-2745-116.3.287

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BioOne sees sustainable scholarly publishing as an inherently collaborative enterprise connecting authors, nonprofit publishers, academic institutions, research libraries, and research funders in the common goal of maximizing access to critical research. Taxonomic investigations of Lecanora strobilina and L. symmicta (Lecanoraceae, Lecanorales) in northeastern North America

Scott LaGreca1,2 and H. Thorsten Lumbsch3

2 Plant Pathology Herbarium, 334 Plant Science Building, Cornell University, Ithaca, NY 14853-5904, U.S.A.; 3 Department of Botany, The Field Museum, 1400 South Lake Shore Drive, Chicago, IL 60605-2496, U.S.A.

ABSTRACT. Two members of the Lecanora varia group (sensu Eigler), L. strobilina and L. symmicta, can be difficult to distinguish in northeastern North America. Morphological and chemical investigation of 277 specimens recognizes two species in northeastern North America, both fitting European concepts of L. strobilina and L. symmicta. The only reliable character for separating them is the size of the exciple (consistently larger in L. symmicta). Within each of these two species, previously unknown chemical variation is revealed—L. strobilina comprises three distinct chemotypes, while L. symmicta includes two. An isotype of L. symmictera is shown to contain the thiophanic acid chemosyndrome by HPLC, and hence the name L. symmictera cannot be applied to the xanthone-deficient chemotype of L. symmicta. Chemistry is not correlated with morphology or geographic distribution in either L. strobilina or L. symmicta,sowe regard the chemotypes of these two species as intraspecific variation.

KEYWORDS. Chemotaxonomy, Lecanora varia group, New England.

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The genus Lecanora is a large group of crustose lichens defined by Eigler (1969) as species containing usnic traditionally characterized by lecanorine apothecia, green- acid as a major secondary product, completely lacking algal photobionts and colorless, non-septate spores in atranorin, and having a corticolous or lignicolous eight-spored, Lecanora-type asci. Lecanora has been the habit. The North American species were treated by subject of numerous investigative studies over the past S´liwa & Wetmore (2000) and Printzen (2001), with a century (for an excellent synopsis, see Pe´rez-Ortega number of species new to science proposed in the latter et al. 2010), but for the most part, the genus is still publication. As part of ongoing, collaborative efforts to regarded as an artificial assemblage of species by many further document the lichens of northeastern North modern workers. Currently within the genus, infrageneric America, we have been investigating the L. varia group ‘‘species groups’’ (e.g., the ‘‘L. subfusca group’’; the in this region (LaGreca & Lumbsch 2001; LaGreca & ‘‘L. subcarnea group’’) based primarily on chemistry Stutzman 2005). Based on Lumbsch & Brodo (2000), and apothecial anatomy are recognized. Some keys to S´liwa & Wetmore (2000), herbarium records and our Lecanora are subdivided into sections that treat particular own observations, six species of the L. varia group occur species groups (e.g., Ryan 1997). Molecular phylogenetic here—Lecanora conizaeoides Nyl. ex Crombie, L. cupressi studies over the past 15 years (Arup & Grube 1998, 2000; Tuck., L. expallens Ach., L. orae-frigidae R. Sant., L. Papong et al. 2013; Pe´rez-Ortega et al. 2010) have strobilina (Spreng.) Kieffer and L. symmicta (Ach.) Ach. demonstrated that some of these species groups, while Most are easily diagnosed on the basis of morphology, convenient for organizing identification keys, are non- substrate and/or chemistry (Brodo & Va¨nska¨ 1984; monophyletic. Lumbsch & Brodo 2000; S´liwa & Wetmore 2000). Two One such species group is the well-known Lecanora of them, however, L. strobilina and L. symmicta, can be varia group, recently shown to be non-monophyletic difficult to distinguish from each other in northeastern by Pe´rez-Ortega et al. (2010). The group was broadly North America. The similarity of these two species has been mentioned by several authors, including Brodo 1 Corresponding author’s e-mail: [email protected] (1967), Brodo et al. (2001) and S´liwa & Wetmore (2000). DOI: 10.1639/0007-2745-116.3.287 The latter authors even placed them in their own group

The Bryologist 116(3), pp. 287–295 Published online: September 19, 2013 0007-2745/13/$1.05/0 Copyright E2013 by The American Bryological and Lichenological Society, Inc. 288 The Bryologist 116(3): 2013 within the L. varia group, the ‘‘L. strobilina group.’’ More Sonoran Desert material. However, S´liwa & Wetmore recently, Pe´rez-Ortega et al. (2010) recognized a ‘‘L. (2000) reported a variable chemistry for L. strobilina, symmicta group,’’ containing L. symmicta and seven with only some specimens reported as containing other species, in their phylogenetic tree. They did not decarboxysquamatic acid (which they called ‘‘LSTR’’, include L. strobilina in their analysis, however, so its as this depside was, at that time, unnamed), sometimes relationship to L. symmicta and other members of the L. with traces of squamatic acid. Our initial chemical symmicta group (sensu Pe´rez-Ortega et al.) is unknown. analyses of L. strobilina indicate that specimens from In northeastern North America, specimens of our region are chemically variable in a fashion similar Lecanora strobilina and L. symmicta can usually be to the report of S´liwa & Wetmore (2000). separated by the anatomy of their young apothecia—L. The objective of the present study is to clarify the strobilina has a leprose, ecorticate, thalline amphithe- species within the Lecanora strobilina group sensu S´liwa cium, while L. symmicta possesses a biatorine apothe- & Wetmore (2000) in northeastern North America; it cial margin (Brodo 1967; Laundon 1976; Printzen represents the first intensive taxonomic study of this 2001; S´liwa & Wetmore 2000). However, in both group in this region. The questions we were trying to species, the apothecial margins become excluded with answer are: (i) how many species occur in this group in age, so specimens without young apothecia are difficult New York, New England and adjacent temperate and to separate on this basis. According to the literature, L. boreal Canada (ii) do they match the species concepts strobilina might also be separated from L. symmicta by of L. strobilina and L. symmicta, which were described virtue of a granular, often ecorticate thallus in the from Europe—or are additional concepts applicable, former species vs. a smooth thallus in the latter. This e.g., the new species described in Printzen (2001) from character appears to be sufficient to distinguish many southwestern North America? specimens from our region, but we have noted some variation within both species—some specimens are MATERIALS AND METHODS granular, while others are completely smooth. In A total of 277 specimens of Lecanora strobilina, addition, L. symmicta also shows variation with regards L. symmicta and L. symmictera from the following to apothecial color—specimens in our region vary herbaria were utilized in this study: CANL, FH, MAINE, from yellow-green to bluish, orange or yellowish. They MSC, NY, and the personal collections of D. Greene, also vary with regards to fusion of apothecia—some S. LaGreca, E. Lay, and P. May. The study area includ- specimens possess individual apothecia, while others ed the following states and provinces: Connecticut, have apothecia which are fused together. Maine, Massachusetts, New Brunswick, New Hamp- Chemically, the Lecanora strobilina group (sensu shire, New York, Newfoundland, Nova Scotia, Ontario, S´liwa & Wetmore) consistently produces usnic acid Quebec, Rhode Island and Vermont. In order to limit and usually zeorin. When present in abundance, zeorin the study area to temperate and boreal phytogeo- often crystallizes on the surfaces of old specimens of graphic zones, an arbitrary line of 50u N latitude was both L. strobilina and L. symmicta, giving them a used; specimens collected north of this latitude were ‘‘fuzzy’’ appearance. Within L. symmicta, some authors not included in this study. For comparative purposes, have differentiated between L. symmicta and L. some material from western North America, the Great symmictera Nyl. on the basis of xanthone presence Lakes states and Europe were examined. A complete and absence, respectively (e.g., Harris 1977), which list of specimens examined for this study can be found corresponds to the original, European concepts for in Supplementary online Table S1. these taxa by Acharius (1810) and by Nylander (1872), Types were not included in our examinations, respectively. Still other authors have applied the names except for a type of Lecanora symmictera, which L. symmicta and L. symmictera interchangeably to happened to be deposited in the herbarium where we North American specimens (e.g., Brodo 1988; Ryan initiated this study (FH). Instead of types, species 1997). Most modern authors simply use the older delimitations were based on papers by Lumbsch et al. name (L. symmicta) to include all members of the L. (1995), Printzen (2001) and S´liwa & Wetmore (2000), strobilina group which do not correspond to L. as well as authoritatively identified collections and strobilina (e.g., Gowan & Brodo 1988; Lumbsch & exsiccati. Authorities for names follow Esslinger (2013) Brodo 2000), effectively synonymizing L. symmictera and Kirk (2013). with L. symmicta (Esslinger 2011; Nimis & Martello Both dissecting and compound microscopes were 2008). Lecanora strobilina was differentiated chemically used in morphological studies. Two gross morpholog- from L. symmicta by the invariable presence of ical characters were recorded for all specimens using a decarboxysquamatic acid by Printzen (2001) in dissecting scope: thallus structure (smooth vs. granular) LaGreca & Lumbsch: Lecanora in northeastern North America 289 and apothecial color. Excipular tissues of representative specimens were hand-sectioned, then examined in water with an Olympus BH-2 compound microscope. Representative sections were stained with Congo Red (Lillie 1977) and/or 10% KOH solution. When possible, both young and mature apothecia were examined. Epihymenium granule solubility in 10% KOH, 50% KOH and concentrated HNO3 was also checked. Spore measurements are given as ‘‘(smallest single measurement–) smallest mean–largest mean (–largest single measurement).’’ All specimens were analyzed with thin-layer chromatography (TLC) using pre-gelled silica plates and the following solvents: A, B9, and C (Culberson & Ammann 1979; Culberson & Johnson 1982). Com- pounds were identified by running with known standards on the same plate. HPLC confirmation of suspected xanthones was performed for 15 specimens [including three from NY], including an isotype of L. symmictera (Hepp Flecht. Eur. Exs. 68), as described in Figure 1. Morphology of apothecia in Lecanora strobilina examined in the Lumbsch (2002). HPLC was also used to confirm the present study. Scale bar 5 0.50 mm. Apothecia of various ages (May s.n., chemistry of seven additional specimens of L. strobilina Katama site 3, FH). and L. symmicta. All specimens which were analyzed S´liwa & Wetmore (2000) maintain that L. symmicta has with HPLC are noted in the Discussion. shorter spores; while this is suggested by the mean measurements reported here, spore sizes are not RESULTS AND DISCUSSION entirely reliable diagnostic characters because there is Morphological investigations. Among the 277 so much overlap between the two species, at least in specimens examined for this study, thallus structure northeastern North American material. (granular vs. smooth) was variable in Lecanora As noted by Printzen (2001), the structure of the strobilina, but always smooth in L. symmicta. Apothe- apothecium provides the most useful characters for cial color was variable in L. symmicta, but was invariably beige in L. strobilina. Apothecia of some specimens of L. symmicta were fused together, but those of L. strobilina were always separate. As noted above, the apothecial margins of young apothecia of L. strobilina are leprose, ecorticate and thalline (top of Fig. 1; a modification of the ‘‘varia’’-type sensu Eigler 1969), while those of L. symmicta are biatorine (Fig. 2; ‘‘symmicta’’-type sensu Eigler 1969). This distinction, however, disappears in older apothecia of both species, as the margins are often excluded with age (middle of Fig. 1). In the herbarium material we examined, crystals were absent in the apothecial margins of both species. The epihymenium of both species was covered, and inspersed with, brown granules, which did not dissolve in either concentrated HNO3 or 10% KOH, but did dissolve in 50% KOH. Paraphyses of both species were indistinguishable as well, being 1–2 mm thick, non-pigmented, non-capped, and sometimes anastomosing. Spore measurements for Lecanora strobilina were Figure 2. Morphology of apothecia in Lecanora symmicta examined in the (7–)11–13(–15) 3 (3–)3.5–4.0(–5) mm, and for L. present study. Scale bar 5 0.50 mm. Young apothecium (Greene s.n., symmicta they were (9–)10–11(–15) 3 (3–)4(–6) mm. Nottingham, Rt. 152, FH). 290 The Bryologist 116(3): 2013 diagnosing species in the Lecanora varia group. In the material we examined, while the sizes of many of the apothecial tissues overlap considerably, Lecanora strobilina consistently has a much smaller, less developed parathecium (exciple) than L. symmicta when apothecia of roughly the same age are compared. This is true even in older apothecia in which the margins become Figure 3. Proposed structures for (a) decarboxysquamatic acid and (b) excluded. Printzen’s measurements for this tissue show barbatic acid, two closely related depsides produced by the acetyl- great overlap for L. strobilina and L. symmicta (15–45 mm polymalonyl pathway in Lecanora strobilina. The arrow in (a) indicates and 20–70 mm, respectively); this may be the result of where a carboxyl group would be in squamatic acid. including immature apothecia in his study. This overlap ´ is not true for northeastern North American material, sequence. Chemotype 3 was reported by Sliwa & when comparing apothecia of approximately the same Wetmore (2000), and is also known from Colorado age. It is remarkable that while the gross morphology of (C. Printzen, pers. comm.). It is the second-most the thallus is variable in both species, the apothecial common chemotype in northeastern North America, anatomy is consistent; indeed, the size of the para- and is probably produced by a loss mutation that thecium is the only reliable character for telling these completely turns off depsidone production via the two species apart in northeastern North America. acetyl-polymalonyl pathway. A molecular phylogenetic Chemical variation. TLC analysis revealed previ- analysis, including many samples of each chemotype, ously unknown chemical variation in both Lecanora could explore whether chemotypes 2 and 3 are strobilina and L. symmicta. Lecanora strobilina includes polyphyletic—a likely scenario, since these two chemo- three distinct chemotypes: (1) usnic acid, zeorin and types would, in theory, be easy to achieve via loss decarboxysquamatic acid as the major secondary mutations in any number of different genes controlling products (45 specimens); (2) usnic acid, zeorin and the acetyl-polymalonyl pathway. barbatic acid as the major secondary products (one Lecanora symmicta comprises two chemotypes: (1) specimen); and (3) usnic acid and zeorin only (16 usnic acid, zeorin and chlorinated xanthones of the specimens). Chemotype 1, the most common chemo- thiophanic acid chemosyndrome (ca. 44 specimens) type in the northeast, was first reported by LaGreca et al. and (2) usnic acid and zeorin only (ca. 171 specimens). (2000) after Jack Elix (Canberra, Australia) identified As noted by Printzen (2001), it is not always possible to decarboxysquamatic acid for us. It was also reported by detect xanthones with spot tests or TLC; HPLC is both S´liwa & Wetmore (2000) and Printzen (2001). required. That is why we have qualified the number of Lecanora strobilina is the only lichen species known to specimens per chemotype (with ‘‘ca.,’’ circa): it was possess decarboxysquamatic acid as its major product. impossible to confirm the presence (or absence) of This depside is otherwise only found as a minor/trace xanthones with HPLC for each and every specimen. compound in some specimens of the squamatic acid Lecanora symmicta chemotype 2 corresponds to L. chemotpe of Cladonia capitellata (Hook.f. & Taylor) symmictera s. auct., a species described by Nylander C. Bab., where squamatic acid is the major secondary (1872) as differing from L. symmicta only in its C- product (J. Elix, pers. comm.). The presence of reaction (which we interpret as a lack of xanthones). squamatic, baeomycesic and barbatic acids in small However, an isotype of L. symmictera analyzed as part amounts in some specimens of chemotype 1 supports of the present study was found to contain the the postulation by Culberson et al. (1979) that these thiophanic acid chemosyndrome by HPLC, even depsidones are produced by simple stepwise oxidation though these xanthones were undetectable by TLC, at the 3-a position in the sequence: barbatic R 3-a- and the thallus and apothecia of this specimen were C-. hydroxybarbatic R baeomycesic R squamatic (co- Therefore, the name L. symmictera cannot actually be occurrence of these same compounds also occurs in applied to chemotype 2, even if we recognized this some other lichens, e.g., Cladonia strepsilis and Xantho- chemotype at the species-level; the name L. symmictera parmelia moctezumensis). Subsequently, decarboxysqua- should be synonymized with L. symmicta. Incidentally, matic acid is probably derived from squamatic acid by xanthones are apparently absent in L. strobilina;we decarboxylation at the 1-b position (Fig. 3). Chemotype extrapolate this from HPLC analysis of five specimens 2—represented by only one specimen, collected from (see ‘‘Selected Specimens Examined’’, below). inland Maine—is reported here for the first time. This Contrary to the suggestion of S´liwa & Wetmore chemotype is probably the result of a loss mutation (2000), chemistry is not correlated with the granular that eliminates the oxidase catalyzing the oxidation quality of the thallus in Lecanora strobilina. In fact, we LaGreca & Lumbsch: Lecanora in northeastern North America 291

Figure 4. Distribution of chemotypes of Lecanora strobilina in northeastern North America. Black dots 5 chemotype 1; white dot 5 chemotype 2; gray dots 5 chemotype 3. could not find any morphological features that examined; in northeastern North America, L. strobilina correlated with chemistry in either L. strobilina or L. and L. symmicta correspond to European concepts of symmicta. The assertion of Harris (1977) that L. these species. symmicta chemotype 1 only occurs along the coast is Lecanora strobilina (Sprengel) Kieffer, Bull. Soc. Hist. not supported by our observations. Indeed, Figures 4 Nat. Metz 19: p. 74. 1895. Fig. 1. &5show no pattern in the geographic distribution of MYCOBANK MB 389285 any of the chemotypes of either L. strobilina or L. Lecanora strobilina Ach., Synops. Lich. p. 171. 1814. symmicta. [It should be noted that, with regards to the nom. illeg. (see Laundon 1976). map for L. symmicta (Fig. 5, only specimens whose chemistry was verified by HPLC are plotted.] This lack of Parmelia strobilina Sprengel, Syst. Veg. 4: p. 300. 1827. correlation of chemistry with morphology and geogra- TYPE:FRANCE. Gallia merid[ionalis], Pin[us] marit[ima], phy—as well as the fact that the secondary products of [Dufour?] 240 (H-ACH 1141a, holotype, not seen). chemotypes1and2ofL. strobilina are produced from Discussion. Lecanora strobilina—often erroneously the same pathway (Fig. 3)—leads us to conclude that the called L. conizaea (Ach.) Nyl. in the older North chemotypes of these two species are intraspecific American literature (e.g., Brodo 1967, 1968), as pointed variants, unworthy of recognition at species-level. out by Laundon (1976)—is widespread in temperate North America, including southern Canada. It is found TAXONOMY from the lowland coasts of California (Printzen 2001) The species. In recent decades, a number of species to New England, and many states in between (S´liwa & have been segregated from both Lecanora strobilina and Wetmore 2000). This species grows on both hardwood L. symmicta based on minor morphological and and softwood trees, most often on twigs; in our region, chemical differences (Brodo 1981; Giralt & Go´mez- it is also commonly found on old, worked wood and Bolea 1991; Kantvilas 2012; Kantvilas & LaGreca 2008; pinecone scales. Interestingly, L. strobilina is now rare in Padro´n et al. 1991; Printzen 2001; Prinzten & May Europe, although herbarium specimens indicate it was 2002; Santesson 1984). For example, Lecanora sub- once more widespread there (Laundon 1976; H.T. strobilina Printzen differs from L. strobilina in having a Lumbsch pers. obs.). In Europe, this species seems to different exciple, broader spores and xanthones; L. lack depsides. We did not, however, chemically analyze subtecta (Stirton) Kantvilas & LaGreca differs from L. many European specimens; a thorough chemical study symmicta in its heavily yellow-pruinose apothecia. We of European populations is necessary to assess the full found no such character differences in the material we extent of chemical variation there. 292 The Bryologist 116(3): 2013

Figure 5. Distribution of chemotypes of Lecanora symmicta in northeastern North America. Black dots 5 chemotype 1; gray dots 5 chemotype 2. Note: only specimens for which chemistry has been verified by HPLC are shown.

In northeastern North America, there are no other (350 nm) UV light, we tested whether the related lichens that can be readily confused with Lecanora compound decarboxsquamatic acid shares its fluo- strobilina, aside from Lecanora symmicta. Sometimes, rescent properties. No specimens of chemotype 1 however, the granular surface of L. strobilina is so fluoresced, however—in other words, decarboxysqua- well-developed that it may appear sorediate. When this matic acid does not fluoresce. Chemotype 1 is happens, Lecanora strobilina mightbeconfusedatfirst indistinguishable in this regard from specimens of with truly sorediate Lecanora species, such as L. chemotypes 2 and 3. conizaeoides Nyl. ex Crombie. The latter species, however, Selected specimens examined. [chemotype 1:] has corticate, non-sorediate margins (at least, in North CANADA. NOVA SCOTIA. Shelburne Co. Tobeatic Wilder- America; see LaGreca & Stutzman 2005—contrary to ness Area, Lay 99-0143 (hb. Lay; HPLC’d). ONTARIO.St. Brodo et al. 2001, p. 388). Furthermore, the thallus of L. Lawrence Islands National Park, Gorden Island, Wong conizaeoides always contains fumarprotocetraric acid 3462 (CANL 86168). Hastings Co., Bellville, Macoun 1993 (which is never found in L. strobilina) in the medulla, (CANL). U.S.A. MAINE. Oxford Co., Buckfield, Parlin 7831 which reacts P+ red. Another sorediate Lecanora species (FH; HPLC’d). MASSACHUSETTS. Berkshire Co., Windsor, that may be superficially confused with L. strobilina in Windsor State Forest, LaGreca 2065 (CUP). Dukes Co., northeastern North America is L. expallens Ach.; it differs Martha’s Vineyard, Katama, site 3, May s.n. (FH; from L. strobilina in possessing true soredia, pruinose HPLC’d). NEW HAMPSHIRE. Rockingham Co., Notting- apothecia, xanthones and sometimes also a bluish-black ham, Rt. 152, Greene s.n. (FH; HPLC’d). [chemotype 2:] prothallus. U.S.A. MAINE. Oxford Co., West Paris, Lay 00-0024 (hb. Contrary to S´liwa & Wetmore (2000), Lecanora Lay). [chemotype 3:] CANADA. NOVA SCOTIA. Kejimkujik strobilina and L. symmicta cannot always be separated National Park, Brodo 18929 (CANL). QUEBEC. Mun. Re´g. by chemistry, as chemotype 3 of L. strobilina is the de Cte´ de Papineau, Lac Clair, Brodo 23581 (CANL 93688; same as chemotype 2 of L. symmicta. As mentioned HPLC’d). U.S.A. MAINE. Knox Co., Rockland, ex hb. previously, however, one can always check the exciple L.W. Riddle, Merrill s.n. (FH). MASSACHUSETTS. Bristol Co., to tell these two species apart. Within L. strobilina,we New Bedford, no collector,exhb. H. Willey (FH). tested whether the three chemotypes were distinguish- Nantucket Co., Nantucket Island, between Nantucket able by spot tests; however, all tests (K, C and P) & Siasconset on Siasconset Road, Brodo 4067 (CANL). performed on a few specimens of each chemotype NEW HAMPSHIRE. Carroll Co., Chatham, no collector, July produced negative results. In addition, since squamatic 1905,exhb. L.W. Riddle (FH). NEW YORK. Hamilton Co.: acid fluoresces bright blue-white under long wave Forked Lake, Brodo 25922 (CANL). LaGreca & Lumbsch: Lecanora in northeastern North America 293

LECANORA SYMMICTA (Ach.) Ach., Syn. Lich.: p. 340. Another species that could be confused with L. 1814. Fig. 2. symmicta is L. confusa Almborn. The two species share MYCOBANK MB 121491 the same general morphology, and L. confusa also has Lecanora varia q symmicta Ach., Lich. Univ. p. 379. the same chemistry as chemotype 2 of L. symmicta 1810. (S´liwa & Wetmore 2000). Our investigations of the TYPE: Suecia [SWEDEN], in sepimentis ligneis Sueciae (H- specimens cited here did not reveal L. confusa in ACH 1134A, lectotype, selected by Brodo & Va¨nska¨, northeastern North America. This mostly agrees with Lichenologist 16: p. 46. 1984). the view of S´liwa & Wetmore (2000) who, aside from Lecidea symmicta (Ach.) Ach., Syn. Lich.: p. 36. 1814. a casual mention of specimens of L. confusa from Biatora symmicta (Ach.) A. Massal., Mem. Lich.: p. 128. Ontario (no actual specimens cited from there), treat it 1853. as a species of western North America and Europe. Lecanora symmictera Nyl., Flora 55: p. 249. 1872. Members of the Lecanora saligna group sensu S´liwa TYPE: [Switzerland?], no locality information, no date, & Wetmore (2000) can also sometimes be confused for Hepp Flecht. Eur. Exs. 68 (isotype: FH!). L. symmicta (and to a lesser degree, L. strobilina). In northeastern North America, there are three such Discussion. Lecanora symmicta ranges throughout species: L. albellula Nyl., L. saligna (Schrad.) Zahlbr. the northern United States, and most of temperate and and L. subintricata (Nyl.) Th. Fr. The first two of these boreal Canada (Brodo et al. 2001). In North America, usually have smaller, darker brown apothecia than L. this lichen grows on both hardwoods and conifers symmicta. They can also be consistently excluded by in a variety of light regimes (shaded to exposed) and the presence of isousnic acid in addition to usnic acid elevations. Because of its worldwide distribution and (a chemistry that defines the L. saligna group); they extensive morphological variation, it is perhaps best will not be considered further here except to say that L. to regard Lecanora symmicta as a ‘‘species complex’’ saligna can be locally common in certain parts of New comprising many species worldwide (see Kantvilas & England where it is often sterile but bears abundant LaGreca 2008 for a discussion). pycnidia which produce characteristic, crescent-moon- In northeastern North America there are a number shaped macroconidia (e.g., the Boston metropolitan of lichens, besides Lecanora strobilina, that can be area; S. LaGreca pers. obs.). confused with L. symmicta. Lecanora filamentosa Lecanora subintricata, however, does not produce (Stirt.) Elix & Palice is nearly identical to it, but differs isousnic acid, although it sometimes produces, in in having shorter conidia and more frequently addition to usnic acid, a compound called brialmon- branched and anastamosing paraphyses; it also some- tin-1, which has been mistakenly reported as isousnic times possesses a thalline margin, which is visible in acid in the past (van den Boom & Brand 2008). younger apothecia, before the margins are excluded Therefore, TLC cannot be used reliably to separate this (Printzen & May 2002). In addition, L. filamentosa species from L. symmicta. Specimens of L. subintricata occurs almost exclusively on conifer trees (usually that only produce usnic acid can be separated by the twigs) in boreal habitats. The most reliable way to following suite of characters: (i) smaller, often dark diagnose L. symmicta from L. filamentosa is with brown apothecia, always with thalline margins (also, TLC—the latter species always contains atranorin as older apothecia with excluded margins remain dis- the major secondary product, with usnic acid produced tinctly marginate, unlike in L. symmicta; see Printzen in lesser amounts. The newly-described, nearly iden- 2001); (ii) narrower ascospores; and (iii) a completely tical L. schizochromatica Pe´rez-Ortega, T. Sprib. & endophloeodal (immersed) thallus. Based on our Printzen has not yet been found in northeastern North observations of the material cited here, we consider America; its separation from L. filamentosa is based L. subintricata to be rare in northeastern North mainly on the results of a molecular phylogenetic America. This species was recently placed in the L. analysis (the minor spore size and apothecial pigment polytropa group sensu Pe´rez-Ortega et al. (2010); differences cited by them are not diagnostic; see Pe´rez- clearly, the taxonomy of the L. saligna group as a Ortega et al. 2010). Neither of these two species was whole requires attention (see van den Boom & Brand considered by Eigler (1969), but their production of 2008 for a recent revision of the European species). atranorin excludes them from his L. varia group. This As in Lecanora strobilina, spot tests cannot be used is in fact supported by the molecular phylogenetic to separate chemotypes within L. symmicta. We tested analysis of Pe´rez-Ortega et al. (2010), who place them multiple specimens of each chemotype with K, C and in their own group within Lecanora, the L. filamentosa P. All tests were negative except for a few specimens of group (their Clade 1). chemotype 1 of L. symmicta, the apothecia of which 294 The Bryologist 116(3): 2013 yielded a C+ orange color due to the presence of Acharius, E. 1814. Synopsis Methodica Lichenum. Lund. thiophanic acid. This compound was verified by HPLC Arup, U. & M. Grube. 1998. Molecular systematics of Lecanora in some specimens—contrary to S´liwa and Wetmore subgenus Placodium. Lichenologist 30: 415–425. Brodo, I. M. 1967. Lichens collected in Wisconsin on the 1965 foray of (2000), who stated that this compound is not pro- the American Bryological Society. The Bryologist 70: 208–227. duced by L. symmicta. As mentioned above, however, Brodo, I. M. 1968. The Lichens of Long Island, New York: A the C+ orange reaction is unreliable within this che- Vegetational and Floristic Analysis. Bulletin 410, New York State motype, however. Some apothecia of specimens which Museum & Science Service. The University of the State of New definitely contained thiophanic acid (verifed by HPLC) York, Albany. reacted C-, probably because thiophanic acid is pro- Brodo, I. M. 1981. Lecanora luteovernalis, a new species of the L. symmicta complex from the Canadian Arctic. The Bryologist 84: duced in low amounts in these specimens. In other 521–526. words, as stated by Printzen (2001), HPLC is the only Brodo, I. M. 1988. Lichens of the Ottawa Region (2nd edition). Special reliable way to tell chemotypes apart within L. symmicta; Publication No. 3. Ottawa Field-Naturalists’ Club and the National spot tests (and TLC) are unreliable. Museum of Natural Sciences, Ottawa. Selected specimens examined. [chemotype 1:] Brodo, I. M., S. D. Sharnoff & S. Sharnoff. 2001. Lichens of North America. Yale University Press, New Haven. CANADA. NEW BRUNSWICK. Albert Co., Fundy National Brodo, I. M. & H. Va¨nska¨. 1984. Notes on the maritime, lignicolous Park, Gowan 2064 (CANL; not TLC’d, but C+ orange lichen Lecanora orae-frigidae. Lichenologist 16: 45–51. apothecia). NOVA SCOTIA. Shelburne Co., Tobeatic Wil- Culberson, C. F. & K. Ammann. 1979. Standardmethode zur derness Area, Buck 35737 (NY; HPLC’d). QUEBEC.Quebec Du¨nnschichtchromatographie von Flechtensubstanzen. Herzogia Co., Stoneham, Lay 97-0436 (hb. Lay; HPLC’d). U.S.A. 5: 1–24. CONNECTICUT. Litchfield Co., Canaan, Lay 91-0324 (hb. Culberson, C. F. & A. Johnson. 1982. Substitution of methyl tert-butyl ether for diethyl ether in the standardized thin-layer chromatographic Lay; HPLC’d). MAINE. Knox Co., Rockland, Merrill s.n., method for lichen products. Journal of Chromatography 128: 10 October 1909 (FH; HPLC’d). Sagadahoc Co., 6 mi. N. 253–259. or Woolwich, Harris 20879 (NY; HPLC’d). Washington Culberson, C. F., T. H. Nash III & A. Johnson. 1979. 3-a- Co., Great Wass Island Preserve, Lay 93-0479 (hb. Lay; hydroxybarbatic acid, a new depside in chemosyndromes of some HPLC’d). MASSACHUSETTS. Hampshire Co., Granby, Lay Xanthoparmeliae with b-orcinol depsides. The Bryologist 82: 94-0839 (hb. Lay; HPLC’d). NEW HAMPSHIRE.CarrollCo., 154–161. Dobson, F. S. 2011. Lichens: An Illustrated Guide to the British and Irish Center Sandwich, Lay 89-0047 (hb. Lay; HPLC’d). NEW Species (6th edition). Richmond Publishing Co., Slough, England. YORK. Ulster Co., Olive, Lay 94-0411 (hb. Lay; HPLC’d). Eigler, G. 1969. Studien zur Gliederung der Flechtengattung Lecanora. Ulster Co., Slide Mountain, near Winnisook, Harris Dissertationes Botanicae 4: 1–195. 13494 (NY, HPLC’d). [chemotype 2:] U.S.A. MAINE. Esslinger, T. L. 2013. A cumulative checklist for the lichen-forming, Washington Co., Milbridge, May 565A (FH; HPLC’d). lichenicolous and allied fungi of the continental United States and Penobscot Co., Orono, no collector, July 1907,exhb. L.W. Canada. North Dakota State University: http://www.ndsu.edu/ pubweb/,esslinge/chcklst/chcklst7.htm (first posted 1 December Riddle (FH; HPLC’d). MASSACHUSETTS. Middlesex Co., 1997, most recent version (#18) posted 13 December 2012), Fargo, FH Reading, Town Forest, Greene s.n. ( ; HPLC’d). North Dakota [accessed March 2013]. Giralt, M. & A. Go´mez-Bolea. 1991. Lecanora strobilinoides, a new ACKNOWLEDGMENTS lichen species from north-eastern Spain. Lichenologist 23: 107–112. We dedicate this paper to the memory of Doug Greene, who performed Gowan, S. P. & I. M. Brodo. 1988. The lichens of Fundy National Park, many of the morphological and chemical analyses. Elisabeth Lay New Brunswick, Canada. The Bryologist 91: 255–325. (Boston) is thanked for originally suggesting this research project. Jack Harris, R. C. 1977. Lichens of the Straits Counties, Michigan. Published Elix (CANB) and Christian Printzen (FR) are thanked for helpful by the author, Ann Arbor. discussions of the chemistry and morphology of Lecanora strobilina. Kantvilas, G. 2012. Lecanora coppinsiarum, a new Tasmanian lichen We also thank Keith Babuszczak (Ithaca), Bob Dirig (CUP), and Kathie related to Lecanora symmicta. Lichenologist 44: 247–251. Hodge (CUP) for logistical support. Accommodations for SLG during Kantvilas, G. & S. LaGreca. 2008. Lecanora subtecta, an Australian his many visits to FH were generously provided by Tony Mercadante. species in the Lecanora symmicta group (Lecanorales). Muelleria 26: Genevieve Lewis-Gentry and Michaela Schmull kindly provided 72–76. assistance while working at FH. We are especially grateful to Donald Kirk, P. M. 2013. Index Fungorum Search Page. CABI: http://www. Pfister (FH) for a Geneva Sayre award to HTL, which made the indexfungorum.org/Names/Names.asp (accessed March 2013), inception of this project at FH possible. 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