Introduction
The genus Arthothelium A. Massal. encompasses a group of chiefly tropical or subtropical corticolous lichens that is distinguished from the large genus Arthonia Ach. by the muriform (rather than transversely septate) ascospores. Other characters that unite these species (and some taxa of Arthonia s. lat.) are the ecorticate crustose thallus with a trentepohlioid photobiont, the peculiar apothecioid ascomata with a highly reduced or excluded exciple, frequently yielding a ‘rubbed down’ appearance, the branched and anastomosed paraphyses, and the broadly clavate to subglobose, hemiamyloid, Arthonia-type asci (Coppins Reference Coppins, Smith, Aptroot, Coppins, Fletcher, Gilbert, James and Wolseley2009).
The status of the genus remains unclear. The cladistic study of Tehler (Reference Tehler1990) suggested that at least the type species, A. spectabile Flot. ex A. Massal., is distinct from Arthonia in the broad sense. The relationship between the two genera was also discussed briefly by Sundin & Tehler (Reference Sundin and Tehler1998) and Grube (Reference Grube1998), with the former demonstrating that Arthonia and Arthothelium are both paraphyletic. Subsequently, using molecular data, Ertz & Tehler (Reference Ertz and Tehler2010) also concluded that Arthothelium is paraphyletic, with at least some species belonging in Arthonia; the same conclusion was also reached by Frisch et al. (Reference Frisch, Thor, Ertz and Grube2014). Certainly ascospore septation alone has been found increasingly to be of limited taxonomic value, as seen, for example, in the Graphidaceae where the traditional spore-based genera have been almost entirely dismantled (Staiger Reference Staiger2002). Thus the continued application of the name Arthothelium as it is currently understood is probably limited in time, and some recent ‘flora’ accounts now treat Arthothelium species under Arthonia (e.g. Grube Reference Grube, Nash TH, Gries and Bungartz2007; McCune Reference McCune2017). Nevertheless, Arthothelium remains in use with other authors (e.g. Coppins Reference Coppins, Smith, Aptroot, Coppins, Fletcher, Gilbert, James and Wolseley2009; Gupta & Sinha Reference Gupta and Sinha2018; Cannon et al. Reference Cannon, Ertz, Frisch, Aptroot, Chambers, Coppins, Sanderson, Simkin and Wolseley2020) and until its broader phylogenetic position is resolved, the genus will continue to serve as a practical placeholder for a group of unequivocally related species. This approach is retained here.
In the Australasian region, the genus remains largely unstudied. New species have been described by Knight & Mitten (Reference Knight and Mitten1860), Nylander (Reference Nylander1867, Reference Nylander1888), Krempelhuber (Reference Krempelhuber1876), Knight (Reference Knight1883), Shirley (Reference Shirley1889), Müller (Reference Müller1893, Reference Müller1895a, Reference Müllerb), Kantvilas & Vĕzda (Reference Kantvilas and Vězda1992), Makhija & Patwardhan (Reference Makhija and Patwardhan1995b), Ertz & Fryday (Reference Ertz and Fryday2017) and others; many of these species were described initially in Arthonia and only later combined into Arthothelium (Müller Reference Müller1894; Zahlbruckner Reference Zahlbruckner1924). Chiefly on the basis of these sources, McCarthy (Reference McCarthy2020) records 12 species for Australia (five in Tasmania) whereas Galloway (Reference Galloway2007) treats nine species for New Zealand. Whilst no systematic revision has been undertaken, the accounts by Galloway (Reference Galloway1985, Reference Galloway2007) remain extremely useful entry points to the genus in the region. However, because of the widespread nature of many of the species, regional treatments, especially from other temperate areas, can also be highly relevant and useful. These include the Mediterranean (Grube & Giralt Reference Grube and Giralt1996), the British Isles (Coppins Reference Coppins, Smith, Aptroot, Coppins, Fletcher, Gilbert, James and Wolseley2009; Cannon et al. Reference Cannon, Ertz, Frisch, Aptroot, Chambers, Coppins, Sanderson, Simkin and Wolseley2020), North America (Fink Reference Fink1935) and western Europe (Clauzade & Roux Reference Clauzade and Roux1985). Much relevant information is also provided by Willey (Reference Willey1890) although, due to the nature of this pivotal text with its lack of keys, it is not particularly user-friendly. The comprehensive revision of the genus in India by Makhija & Patwardhan (Reference Makhija and Patwardhan1995a) also includes many general observations of the genus as a whole.
The present account deals with 10 species of the genus in Tasmania, derived from nearly four decades of fieldwork throughout the island. Five of these are new to science, with the remainder having broader, chiefly temperate Australasian distributions.
Materials and Methods
The study is based chiefly on the author's collections, housed in the Tasmanian Herbarium (HO), and other Australian material held in the Australian National Herbarium (CANB) and the National Herbarium of Victoria (MEL). Type and other historical collections in London's Natural History Museum (BM), the Finnish Museum of Natural History (H) and the Museum of New Zealand Te Papa Tongarewa (WELT) were also studied. Investigations were undertaken on hand-cut sections of the thallus and apothecia, using standard methods, reagents and stains: water, 10% KOH (K), 50% HNO3 (N), commercial bleach solution (C), lactophenol cotton blue (LCB), Lugol's iodine (I) and concentrated HCl (H). Calcium oxalate was detected by eluting hand-cut sections in 20% H2SO4.
Measurements of ascospores are based on at least 50 observations of each taxon and are presented in the format 5th percentile–average–95th percentile, with outlying values in brackets and n signifying the number of observations. Routine thin-layer chromatographic analysis (TLC) was performed using standard methods, with solvent A as the preferred medium (Orange et al. Reference Orange, James and White2010).
Taxonomic Characters in Tasmanian Arthothelium
The major characters underpinning species-level classification in Arthothelium are the morphology of the apothecia, apothecial pigments, and ascospore morphology and dimensions. A summary of these features in the species studied is presented in Table 1. The thallus of Arthothelium is invariably crustose and ecorticate, and, in the case of the saxicolous species in particular, it may be extremely reduced to the area immediately adjacent to or beneath the apothecia. Whereas Makhija & Patwardhan (Reference Makhija and Patwardhan1995a) considered thallus colour and texture useful taxonomic characters, these features appear to be of limited application in the Tasmanian species. Variation in colour from whitish to some shade of dingy grey or brown, and whether the thallus is smooth, wrinkled or rimose, are considered to be mainly a function of exposure or substratum texture. The photobiont in all species studied is trentepohlioid, although there are taxa elsewhere where the photobiont is chlorococcoid (e.g. Grube & Giralt Reference Grube and Giralt1996) or absent (e.g. Coppins Reference Coppins, Smith, Aptroot, Coppins, Fletcher, Gilbert, James and Wolseley2009). No Tasmanian species has been found to contain lichen substances, although a wide range of compounds, including xanthones, fatty acids, depsides and depsidones, has been detected in taxa from other areas (Makhija & Patwardhan Reference Makhija and Patwardhan1995a; Aptroot & Wirth Reference Aptroot and Wirth2006). Most species are corticolous or lignicolous but saxicolous species are not uncommon in the genus. One of the Tasmanian taxa described below is lichenicolous.
Salient features of Tasmanian species of Arthothelium.
The ‘typical’ Arthothelium apothecium is immarginate with a rather thin, ‘rubbed down’ appearance, and ranges from applanate to adnate and slightly convex. Such apothecia are seen in the Tasmanian species, although in a small number of taxa, notably A. insolitum, A. bacidinum and A. suffusum, the apothecia are markedly convex to subglobose, and more typical of a species of Micarea. Although there are members of the genus with variously coloured apothecia, including whitish, yellowish or reddish, the apothecia of all Tasmanian species are brown-black to black, with the exception of A. macounioides where they are dark reddish brown.
The Arthothelium-type ascus has been referred to in the literature by numerous authors (e.g. Grube & Giralt Reference Grube and Giralt1996; Grube Reference Grube1998; Sparrius Reference Sparrius2009). It is subglobose to broadly obovoid, hemiamyloid, with a thick, non-amyloid wall and a well-developed KI+ blue tholus with a small, darker-staining ring and beak-like ocular chamber (Fig. 1). These features are best observed in younger asci and, as the ascospores mature, the tholus and ring structure become compressed between the ascus wall and the ascoplasm. Eventually they become imperceptible.
The Arthothelium-type ascus as exemplified by A. magenteum: young ascus (A), mature ascus (B), with amyloid parts stippled. Scale = 20 μm.
The ascospores of Tasmanian species fall into one of two main types (Fig. 2). In the first, the spores are broadly ellipsoid and muriform throughout. The longitudinal and transverse septa may occur at right angles to each other, yielding a neat arrangement of relatively regular, rectangular locules like bricks in a wall. Alternatively, the septation may be relatively haphazard. The second type of ascospore is macrocephalic, with a large, undivided proximal cell and a septate ‘tail’; both transverse and longitudinal septa may be frequent, or the latter may be few, with some transverse segments undivided. The ascospores of A. interveniens are unusual in that they have a large, undivided central cell.
Ascospores of Tasmanian species of Arthothelium. A, A. ampliatum. B, A. bacidinum. C, A. endoaurantiacum. D, A. insolitum. E, A. interveniens. F, A. macounioides. G, A. magenteum. H, A. subtectum. I, A. suffusum. J, A. velatium. Scale = 10 μm.
Apothecial pigments are very useful in lichen taxonomy (Meyer & Printzen Reference Meyer and Printzen2000) and of critical importance in many crustose groups, such as Megalaria (Kantvilas Reference Kantvilas2016), Micarea (Kantvilas & Coppins Reference Kantvilas and Coppins2019) and Mycoblastus (Kantvilas Reference Kantvilas2009). Arthothelium is no exception. Three main pigments were observed in the species studied and are named and characterized here:
Endoaurantiacum-gold: golden yellow in water, or orange-brown to reddish brown when very highly concentrated; brownish orange in polarized light; K+ crimson; C+ pink (fading); unchanged in H and N (Fig. 3A). This pigment is present in high concentrations in four Tasmanian species (A. bacidinum, A. endoaurantiacum, A. macounioides and A. subtectum). It also appears to be present in the New Zealand species, A. spadiceum, although in lower concentrations. Likewise in the specimen of A. macounii seen from the Northern Hemisphere, the pigment is rather dilute, brownish and limited to the upper parts of the apothecia, but its reactions with all the reagents tested is the same.
Apothecial pigments in Tasmanian Arthothelium species, in water (left) and 10% KOH (right). A, Endoaurantiacum-gold, illustrated by A. endoaurantiacum and also seen in A. bacidinum, A. macounioides and A. subtectum. B, Magenteum-red, illustrated by A. magenteum. C, Interveniens-brown, illustrated by A. interveniens and also seen in A. ampliatum, A. insolitum, A. suffusum and A. velatium.
Magenteum-red: dark maroon-red in water, fluorescing red-pink in polarized light; K+ vivid magenta-pink; C+ orange-brown; N+ orange-brown; H− (Fig. 3B). Found in A. magenteum.
Interveniens-brown: brown to red-brown in water, not reacting in polarized light; K+ olive green; C+ olive green; ±intensifying orange-brown in N and H (Fig. 3C). In Tasmania, it is found in A. ampliatum, A. insolitum, A. interveniens and A. suffusum. The same pigment also occurs in species from other regions, for example the tropical A. macrothecum. It may well be the same as Atra-brown of Meyer & Printzen (Reference Meyer and Printzen2000), albeit in consistently low concentrations and therefore never appearing opaque blackish as in many taxa of Arthoniales, such as species of Opegrapha.
Key to the Arthothelium species
1 Ascospores longer than 50 μm, entirely muriform; apothecia strongly convex to subglobose………A. insolitum
Ascospores mostly shorter than 40 μm, entirely muriform or macrocephalic, with an undivided terminal cell; apothecia usually applanate or flattened, and only rarely strongly convex………2
2(1) Ascospores macrocephalic, with a conspicuously enlarged, undivided proximal cell and a muriform ‘tail’………3
Ascospores not macrocephalic, muriform throughout .………5
3(2) Apothecia internally with a brownish, K+ olive pigment .………A. ampliatum
Apothecia internally golden yellow, K+ crimson………4
4(3) Ascospores with 4−5 transverse and 0−3 longitudinal septa; on sheltered rocks in inland areas of moderate elevation……… ………A. subtectum
Ascospores with 6−8 transverse and 2−4 longitudinal septa; corticolous in wet forest………A. macounioides
5(2) Apothecia internally with brownish, K+ olive pigment only………. 6
Apothecia with golden yellow or reddish pigments yielding crimson or vivid pink reactions in K………8
6(5) Apothecia markedly convex and basally constricted, coarsely white pruinose at least when young; on the dry, rough bark of mature rainforest trees………A. suffusum
Apothecia applanate or plane, adnate, always epruinose; on smooth bark………7
7(6) Apothecia to 2 mm wide; ascospores sparingly muriform, with 4−8 transverse and 0–1(–2) longitudinal septa, the central, widest cells typically undivided .………A. interveniens
Apothecia rarely > 0.6 mm wide; ascospores densely muriform throughout, with 6−10 transverse and 2−3 longitudinal septa ………A. velatium
8(5) Apothecial pigment dark maroon-red, K+ vivid magenta-pink………A. magenteum
Apothecial pigment golden yellow, K+ crimson .………9
9(8) Apothecia emergent, plane and adnate; corticolous in coastal scrub and woodland………A. endoaurantiacum
Apothecia superficial, subglobose and basally constricted; on littoral rocks………A. bacidinum
Arthothelium ampliatum (C. Knight & Mitten) Müll. Arg.
Bull. Herb. Boissier 2, App.1, 85 (1894).—Arthonia ampliata C. Knight & Mitten, Trans. Linn. Soc. Lond. 23, 106 (1860); type: New Zealand, ?Auckland, 1858, C. Knight 276 (lectotype, fide Galloway (Reference Galloway1985)—BM!; isolectotype—WELT!).
Arthothelium obtusulum (Nyl.) Müll. Arg., Bull. Herb. Boissier 2, App.1, 84 (1894).—Arthonia obtusula Nyl., Lich. Nov. Zel., 123 (1888); type: New Zealand, sine loco [prob. Wellington], 1882, Charles Knight s. n. (lectotype, fide Makhija & Patwardan (Reference Makhija and Patwardhan1995b)—H-NYL 5728!).
Arthothelium pellucidum (C. Knight) Müll. Arg., Bull. Herb. Boissier 2, App.1, 85 (1894).—Arthonia pellucida C. Knight, Trans. N. Z. Inst. 15, 352 (1883); type: New Zealand, sine loco [prob. Wellington], 22.xi.[18]77, Charles Knight s. n. (holotype—WELT 006972!).
Arthothelium polycarpum Müll. Arg., Bull. Herb. Boissier 3, 323 (1895); type: Australia, Brisbane, F. M. Bailey 682 (lectotype, fide Makhija & Patwardan (Reference Makhija and Patwardhan1995b)—G).
Morphology of Tasmanian species of Arthothelium I. A, A. ampliatum, showing the ‘rubbed down’ apothecial morphology typical of the genus. B, A. bacidinum. C, A. endoaurantiacum. D, A. insolitum, lichenicolous on the grey, squamulose thallus of an unknown lichen. E, A. interveniens. F, A. macounioides. Scales = 1 mm. In colour online.
Thallus very thin and patchy, mottled dingy grey to beige-grey to whitish, in section I+ reddish brown, KI+ blue, usually forming irregular, undelimited patches to c. 25 mm wide amongst other lichens; photobiont trentepohlioid, with cells globose to ellipsoid, 10−20 μm wide, in short chains or scattered in clusters.
Apothecia abundant, scattered, 0.3−1(−1.4) mm wide, 100−140 μm thick, black, epruinose, applanate to slightly convex, roundish, ellipsoid or irregularly elongate. Proper exciple excluded. Hypothecium hyaline to pale brown, K+ olive, barely differentiated from the hymenium, mostly to 20−50 μm thick. Hymenium 50−90 μm thick, hyaline, I+ orange-red, KI+ blue, overlain by a brown, K+ olive epithecium 10−20 μm thick; asci of the Arthothelium-type, 50−65 × 32−46 μm, 8-spored; paraphyses branched and anastomosing, 1.5−3 μm wide, brown in the uppermost part, with the apices not expanded. Ascospores broadly ellipsoid to oblong, straight or occasionally slightly curved, hyaline at first, becoming brownish grey and rugulose when old, (24−)25−29.8−37(−39) × (9−)10−11.8−14(−15) μm (n = 85), macrocephalic with an enlarged, undivided upper cell and muriform ‘tail’ with 5−6 transverse and 0−3 longitudinal septa.
Conidiomata not found.
Chemistry
No substances detected by TLC.
Remarks
This species is readily recognized by the combination of macrocephalic ascospores with an enlarged, undivided, proximal cell and sparsely septate ‘tail’, and the brown, K+ olive apothecial pigment (Interveniens-brown). In the Tasmanian flora, similar macrocephalic ascospores are found in A. macounioides and A. subtectum, both of which contain golden yellow, K+ crimson, Endoaurantiacum-gold apothecial pigment. Interveniens-brown is widely distributed in the genus worldwide and occurs in several Tasmanian species, but none of these has macrocephalic spores. An exception is Arthonia ilicina Taylor, which is ±identical macroscopically, occurs sympatrically, and essentially differs only by having macrocephalic ascospores that lack any longitudinal septa. In that regard, distinguishing these two species should be undertaken with care and be based on multiple observations, since young spores of A. ampliatum may lack longitudinal septa and hence be mistaken for Arthonia ilicina.
Arthothelium pellucidum, based on a New Zealand type that is in very poor condition, is placed here into synonymy with A. ampliatum. Galloway (Reference Galloway2007) distinguished these species based on A. pellucidum having effigurate apothecia and somewhat smaller ascospores ((20−)22−25 × 8−10 μm). An examination of the type specimen of this taxon indicated that the ascospores can be as long as 30 μm, and fall comfortably within the range of those of A. ampliatum. The effigurate apothecia are considered to be a function of age and substratum texture.
A further new synonym is the New Zealand species A. obtusulum (Nyl.) Müll. Arg. According to Makhija & Patwardhan (Reference Makhija and Patwardhan1995b) and Galloway (Reference Galloway2007), this differs from A. ampliatum only by having smaller apothecia, 0.3−0.5 μm wide, but these fall within the size range of A. ampliatum as understood in the present work. The type specimen of A. polycarpum (from Queensland, Australia) was not studied, and this synonymy is derived from Makhija & Patwardhan (Reference Makhija and Patwardhan1995b).
Given the widespread nature of A. ampliatum and its broad ecological amplitude, it is likely to have further, hitherto overlooked synonyms. For example, published descriptions of the European species A. dictysporum (Coppins & P. James) Coppins (Coppins & James Reference Coppins and James1979; Coppins Reference Coppins, Smith, Aptroot, Coppins, Fletcher, Gilbert, James and Wolseley2009; Cannon et al. Reference Cannon, Ertz, Frisch, Aptroot, Chambers, Coppins, Sanderson, Simkin and Wolseley2020) suggest that this species is very similar to A. ampliatum. Closer investigation of the relationships between these taxa is warranted, although A. ampliatum is the oldest name.
Distribution and ecology
Arthothelium ampliatum is frequently collected in Tasmania although it is rarely common and is usually seen as small thalli scattered within diverse assemblages of epiphytic lichens. It has a wide ecological amplitude. Most Tasmanian collections are from coastal woodland, where it grows on shrubs such as Acacia sophorae and Leucopogon parviflorus. It has also been found in coastal Melaleuca ericifolia swamps, either on the papery bark of Melaleuca itself or on subdominant shrubs, and on subdominant trees and shrubs in open eucalypt forests. Less commonly it occurs in wet eucalypt forests where it colonizes smooth-barked hosts such as Bursaria spinosa, Pittosporum bicolor or Pomaderris apetala. It is also known from mainland Australia and New Zealand.
Selected specimens examined
Australia: Tasmania: Strickland Ave, Hobart, 42°55′S, 147°16′E, 180 m, 1906, W. A. Weymouth (HO); Deal Island, 39°28′S, 147°19′E, 5 m, 1971, J. S. Whinray s. n. (HO, MEL); Maria Island, 1.5 km NW of Mt Maria, 350 m, 1981, G. Kantvilas 167/81 (HO); Denium Hill, 40°45′S, 144°53′E, 5 m, 1993, G. Kantvilas 130/93 & J. Elix (HO); Stumpys Bay, 40°52′S, 148°13′E, 5 m, 1995, G. Kantvilas 57/95 (HO); Wielangta Road, 42°43′S, 147°51′E, 260 m, 1996, G. Kantvilas s. n. (HO); Flinders Island, Memana Road along Patriarch River, 39°57′S, 148°10′E, 10 m, 2006, G. Kantvilas 67/06 (HO); South Sister, 41°32′S, 148°10′E, 750 m, 2006, G. Kantvilas 312/06 (HO); W of Tahune Bridge in the Warra SST, 43°06′S, 146°41′E, 100 m, 2008, G. Kantvilas 364/08 (HO); Stony Point, 40°45′S, 144°59′E, 2 m, 2016, G. Kantvilas 280/16 (HO); Cape Portland, Musselroe Wind Farm, Tregaron Lagoons, 40°46′55″S, 147°58′09″E, 2 m, 2018, G. Kantvilas 339/18 (HO); Lichen Hill, 43°04′S, 147°56′E, 560 m, 2020, G. Kantvilas 336/20 (HO); Stony Head MTA, Ryans Hill, SE of summit, 41°01′05″S, 147°01′43″E, 210 m, 2020, G. Kantvilas 215/20 (HO); Pine Creek, N of Marion Bay, 42°45′S, 147°53′E, 30 m, 2021, G. Kantvilas 2/21 (HO); Stony Head MTA, Quarry Rd, 41°02′26″S, 146°59′40″E, 50 m, 2021, G. Kantvilas 84/21 (HO).
Also examined
Australia: Victoria: Lake Tyres, 1888, F. R. M. Wilson (MEL 11320). New South Wales: Gloucester Tops, 32°04′S, 151°34′E, 1150 m, 1988, G. Kantvilas 418/88 (HO, NSW).
Arthothelium bacidinum Kantvilas sp. nov.
MycoBank No.: MB 841467
Arthothelio endoaurantiaco simile et item apotheciis pigmentum aurantiacum, in kalio vivide carmesinum continentibus et ascosporis ubique muriformibus, 25−40 μm longis, 11−19 μm latis, sed habitu saxicola et apotheciis convexis vel subglobosis, basi constrictis differt.
Typus: Australia, Tasmania, Lime Bay Nature Reserve, c. 1 km N of Plunket Point, 42°59′S, 147°43′E, 2 m, on littoral sandstone boulders, 15 October 2016, G. Kantvilas 254/16 (holotypus—HO; isotypi—E, H).
Thallus inapparent, endolithic, at most visible as a dingy grey discoloration of the substratum, or as minute, scurfy, greyish areoles at the base of the apothecia, forming extensive undelimited patches to c. 150 mm wide; subapothecial medulla orange-red; photobiont trentepohlioid, with cells globose to ellipsoid, 10−19 μm wide, in short chains.
Apothecia very abundant, 0.3−1(−1.7) mm wide, 200−400(−500) μm thick, single and scattered or confluent in clusters, black, epruinose, smooth or minutely verruculose, strongly convex to subglobose, adnate or basally constricted, in section infused entirely with a golden yellow pigment, K+ crimson, N−, C+ transient pink; pigment most heavily concentrated in the outer parts of the apothecia and then deep orange-brown to red-brown, in the innermost parts bright golden yellow. Proper exciple excluded. Hypothecium poorly differentiated from the hymenium, 100−280 μm thick, intensely orange, or becoming massive and orange-red. Hymenium 75−120 μm thick, intensely golden yellow, K+ crimson, I+ orange-red, KI+ blue, overlain by a thick, lumpy, red-brown epithecium, 25−50 μm thick; asci of the Arthothelium-type, 60−80 × 40−60 μm, 8-spored but often with as many as five spores aborted at maturity; paraphyses branched and anastomosing, 1−1.5 μm wide, with the apices greyish and expanded to 2−4 μm wide. Ascospores broadly ellipsoid to oblong, sometimes a little sole-shaped, mostly straight, hyaline at first, very soon becoming brown but remaining smooth-walled, 25−32.3−39(−40) × 11−14.8−18(−19) μm (n = 60), muriform throughout with 6−9 transverse and 2−4 longitudinal septa; apices frequently with oblique septa; brown, overmature spores generally somewhat larger, to 45 × 20 μm.
Conidiomata not found.
Chemistry
No substances detected by TLC.
Etymology
The specific epithet alludes to the subglobose or markedly convex apothecia which are more reminiscent of a species of Bacidia (especially B. littoralis Kantvilas, which occurs in similar habitats) or other member of the Lecanoromycetes than of a member of the Arthoniaceae.
Remarks
This species is well characterized by the combination of the golden yellow, K+ crimson pigmented apothecia (Endoaurantiacum-gold) and the entirely muriform ascospores. The latter readily distinguishes it from the saxicolous A. subtectum and the corticolous A. macounioides, both of which contain the same pigment but have macrocephalic spores with an undivided proximal cell. Anatomically, the most similar species to A. bacidinum is the corticolous A. endoaurantiacum, which has entirely muriform ascospores and an identical pigment. Yet the two species look nothing like each other (emergent, generally plane apothecia in the latter, superficial, subglobose apothecia in the former) and although these differences may be a function of habitat, regarding the two taxa as distinct seemed, after careful consideration, justified. In addition to the morphological differences discussed, the two species also have subtly different ascospores, with those of A. bacidinum being slightly larger and slightly more muriform, an impression formed when comparing the two in section. The strongly basally constricted apothecia of the new species are suggestive of any number of lichens (such as Bacidia s. lat.) rather than an Arthothelium. When first collected and sectioned, to discover it was a species of this genus rather than a member of the Lecanoromycetes was a genuine surprise, especially given its habitat! The subglobose, basally constricted apothecia contrast starkly with the ‘typical’ plane, ‘rubbed down’ appearance of most corticolous species of the genus, although there are some corticolous species, for example A. insolitum and A. suffusum, that have such apothecia.
Littoral, saxicolous species in the genus are not unusual. For example, there is the subantarctic species, A. diffluens (Nyl.) Imshaug & Fryday (Fryday Reference Fryday2002), and Follman (Reference Follman1968) records three further taxa from Chile, but none share any salient characters with A. bacidinum.
Distribution and ecology
Arthothelium bacidinum is known from a single locality, where it was extremely abundant on the sheltered faces of huge sandstone boulders along a sheltered seashore. It formed extensive thalli, and the habitat yielded several other unusual (for Tasmania) lichens, including Bacidia curvispora Coppins & Fryday and Catillaria glaucogrisea Fryday. Triassic sandstone is a common rock type in south-eastern Tasmania but is infrequent at the coast. Several areas with this geology have been searched for the new species without success.
Additional specimens examined
Australia: Tasmania: Lime Bay Nature Reserve, c. 1 km N of Plunket Point, 42°59′S, 147°43′E (type locality), 2 m, 2016, G. Kantvilas 240/16, 244/16 (CANB, HO).
Arthothelium endoaurantiacum Makhija & Patw.
Trop. Bryol. 10, 208 (1995); type: New Zealand, sine loco [prob. Wellington], 25.xi.[18]77, Charles Knight s. n. (holotype—WELT 006970!).
Thallus pale grey, rimose, 50−100(−180) μm thick, in section I+ red, KI+ pale blue, or inapparent and perceived as a pale dingy grey to whitish grey discoloration of the substratum, forming irregular patches to 20−40 mm wide, mostly delimited by a thin, black prothallus; photobiont trentepohlioid, with cells globose to ellipsoid, 7−18 μm wide, in short chains or scattered in clusters.
Apothecia scattered, 0.3−1 mm wide, 100−220 μm thick, black, emergent and sometimes retaining an incomplete ‘veil’ of vestigial thalline tissue that becomes abraded and may appear pruina-like, adnate, plane to convex, sometimes markedly so, roundish to irregularly elongate-ellipsoid, in section entirely infused with a golden yellow, K+ crimson, C+ pink (±fleeting), N− pigment, most heavily concentrated in the outer parts of the apothecia and then deep orange-brown to red-brown, in the innermost parts bright golden yellow. Proper exciple reduced or excluded, to 10−15 μm thick, deep orange-brown, K+ crimson, sometimes with additional brownish, K+ olive-grey pigment, comprised of anastomosed, entangled hyphae 2−3 μm wide. Hypothecium 20−90 μm thick, intensely golden yellow, sometimes also with additional brownish pigment. Hymenium 70−100 μm thick, intensely golden yellow, I+ orange-red, KI+ blue, K+ crimson, overlain by a more intensely pigmented epithecial layer to 20 μm thick; asci of the Arthothelium-type, 58−80 × 35−52 μm, 8-spored; paraphyses branched and anastomosing, 1−1.5 μm wide in the lower part, to 2−3 μm wide and brownish in the uppermost part, with the apices sometimes slightly expanded. Ascospores broadly ellipsoid, mostly straight, hyaline at first, becoming brown but remaining smooth-walled when old, (21−)25−30.1−36(−37) × 11−13.8−16 μm (n = 65), densely muriform throughout, with 5−10 transverse and 1−4 longitudinal septa.
Conidiomata not found.
Chemistry
No substances detected by TLC.
Remarks
This species is very well characterized by the combination of golden yellow, K+ crimson pigment, here termed Endoaurantiacum-gold, and the ascospores, which are muriform throughout and do not have an undivided terminal cell. The type specimen is annotated by the collector, Charles Knight, as ‘A. ampliatum’, a completely different and very distinctive taxon (see above). Three other Tasmanian species have this pigment. Arthothelium macounioides and A. subtectum are easily distinguished by their macrocephalic spores. Separation from A. bacidinum is more subtle and discussed under that species, which is best recognized by its saxicolous habit and strongly convex to subglobose apothecia. Also containing this pigment is the New Zealand species A. spadiceum (C. Knight) Müll. Arg., which has ascospores 33−46 × 13−25 μm, with 4−10 transverse and 1−5 longitudinal septa, and so larger and more septate than those of A. endoaurantiacum. In addition, the spores of A. spadiceum are rather oblong and the septa and locules are regularly arranged like bricks in a wall. The type specimens of this species (in BM and WELT, both examined) also have pale red-brown to yellow-brown apothecia and the golden yellow pigment is less concentrated when viewed in microscope sections; this is most likely due to environmental factors.
Distribution and ecology
This species appears to be uncommon (or overlooked) in Tasmania. It has been collected in the drier northern and eastern parts of the island in coastal scrub, open Eucalyptus-dominated woodland and Melaleuca-dominated swamp, growing on the twigs and branches of Acacia verticillata, Bursaria spinosa, Leucopogon parviflorus, Melaleuca ericifolia and Pomaderris apetala. It is also known from New Zealand.
Additional specimens examined
Australia: Tasmania: Cape Portland, Musselroe Bay Conservation Area, Abalone Rocks, 40°47′26″S, 148°06′08″E, 2018, G. Kantvilas 387/18 (HO); Cape Portland, northern end of Musselroe Bay, 40°49′36″S, 148°06′41″E, sea level, 2019, G. Kantvilas 236/19 (HO); Stony Head MTA, 250 m W of Black Rock Point, 40°59′18″S, 147°03′19″E, 20 m, 2020, G. Kantvilas 363/20 (HO); Canoe Bay, 43°08′S, 147°58′E, 5 m, 2020, G. Kantvilas 347/20 (HO); Tasman Track, between Canoe Bay and Bivouac Bay, 43°07′S, 147°58′E, 20 m, 2020, G. Kantvilas 350/20 (HO).
Arthothelium insolitum Kantvilas sp. nov.
MycoBank No.: MB 841468
Thallus mox lichenicola, lichenum ignotum corticolum squamulosum incolens, apotheciis 0.2−0.7 mm latis, valde convexis vel subglobosis, pigmentum brunneum, in kalio olivaceum infusis, ascosporibus late ellipsoideis vel oblongis, ubique muriformibus, 50−80 μm longis, 22−40 μm longis.
Typus: Australia, Tasmania, Rebecca Creek, 41°13′S, 144°47′E, 120 m, on Nematolepis squamea in mature mixed forest, 9 December 1993, G. Kantvilas 323/93 (holotypus—HO; isotypus—E).
Thallus initially autonomous, very thin, patchy, whitish, to c. 50 μm thick, I+ reddish, KI+ patchily pale bluish, with short chains of subglobose to ellipsoid, trentepohlioid cells 10−19 × 8−13 μm wide clustered beneath the apothecia, soon disappearing and the lichen evident only as apothecia overgrowing an unidentified squamulose lichen with a chlorococcoid photobiont with subglobose cells 8−12 × 6−9 μm.
Apothecia scattered, abundant, 0.2−0.7 mm wide, 250–420 µm thick, brown-black to black, epruinose, markedly convex to subglobose, adnate or basally constricted. Proper exciple ±excluded, at most seen as a layer of brown, K+ olive pigmented tissue to 30 μm thick at the sides of the apothecia. Hypothecium poorly differentiated from adjacent tissues, hyaline to pale yellowish, K−, mostly c. 80−150(−250) μm thick. Hymenium 90−180 μm thick, hyaline, I+ red, KI+ blue, inspersed with oil droplets when well developed, overlain by a brown, K+ olive, C+ olive epithecium 5−10 μm thick; asci relatively few and widely dispersed within a network of paraphyses, of the Arthothelium-type, 90−150 × 60−110 μm, mostly 8-spored at least when young but with as many as six spores aborted at maturity; paraphyses 1−1.5 μm wide, sometimes with the apices expanded to 3 μm and pigmented when extending into the epithecium. Ascospores broadly ellipsoid to oblong, straight or rarely a little curved, hyaline at first, soon becoming yellow-brown and then brown-grey, minutely densely papillate when old, (50−)53−68.6−80 × (22−)23.5−30.6−37(−40) μm (n = 45), not macrocephalic, densely muriform throughout, with 9−16 transverse and 4−8 longitudinal septa across the central cells.
Conidiomata not found.
Chemistry
No substances detected by TLC in the lichen; the host lichen reacts K−, KC+ red, C−, P−, UV+ white and contains 2′-O-methylperlatolic acid.
Etymology
The specific epithet alludes to the perplexing and unusual characters of this remarkable lichen.
Remarks
This new species is known from a single, rich specimen. It displays very perplexing characters and its biology has been difficult to interpret. The best developed apothecia occur on a relatively common (in rainforest), as yet unnamed, sterile squamulose lichen of unknown taxonomic affinities that has a chlorococcoid photobiont and contains 2′-O-methylperlatolic acid. In such cases, no or very few strands or clusters of trentepohlioid algae, the characteristic photobiont of Arthothelium, can be observed and then only at the very base of the apothecia. In other parts of the large collection, apothecia occur on an effuse whitish thallus which is interpreted as belonging to the Arthothelium because it contains a trentepohlioid photobiont and displays the anticipated reactions in iodine. Although seemingly well-developed apothecia are very abundant, most consist only of a dense network of paraphyses and contain no asci. Asci are rarely seen and are found seemingly by chance, scattered within the network of paraphyses and usually as few as 2−4 per section. Well-formed ascospores are likewise rare; overmature, distorted, brown ascospores a little less so.
The anatomy of this lichen offers no reason to believe it is misplaced in Arthothelium, even if the convex apothecia and strange disposition of the asci are unusual. At first sight, it would most likely be considered a species of Micarea or Bryobilimbia, both of which have representative taxa in such habitats. The lichenicolous biology of the species is unusual for the genus, although Ertz & Fryday (Reference Ertz and Fryday2017) described the remarkable A. hymeniicola from the apothecia of a species of Bacidia. It is unfortunate that at present the identity of the host of the new lichen cannot be established.
The very large ascospores (the largest of any species in Tasmania), convex apothecia and unique biology mean that it is unlikely to be confused with any other Tasmanian member of the genus. Similarly large, highly septate, non-macrocephalic spores occur in two species that occur on the mainland of Australia, A. huegelii (Nyl.) Zahlbr. and A. macrothecum (Fée) A. Massal., but both have applanate, adnate apothecia more typical of the genus as a whole.
Distribution and ecology
This new species is known from a single collection from the bark of Nematolepis squamea (formerly Phebalium squameum) in mature mixed forest dominated by Eucalyptus obliqua. Nematolepis has a smooth aromatic bark favoured by a wide range of macrolichens as well as crustose species. At this site, the new species was associated with Arthonia ilicina Taylor, Pseudocyphellaria bratti D. J. Galloway & Kantvilas and Sarrameana albidoplumbea (Hook. f. & Taylor) Farkas. Since the collection was made, the site has been logged and burned. The area is remote and searching for additional material in the same general vicinity has not been possible.
Arthothelium interveniens (Nyl.) Zahlbr.
Cat. Lich. Univers. 2, 127 (1924).—Arthonia interveniens Nyl., Acta Soc. Sci. Fenn. 7, 482 (1863); type: America septentrionalis ad cortices laeves, Carolina, ex hb. E. Tuckerman (lectotype, fide Patwardan & Makhija ined.—H-NYL 5470!).
Arthothelium ferax Müll. Arg., J. Linn. Soc., Bot. 30, 462 (1895); type: Tasmania, Cheshunt, W. Archer (holotype—G!; isotype—BM!).
Thallus very thin and patchy, pale beige to brownish grey, to 40−100 μm thick, in section I+ reddish brown, KI+ blue, forming irregular patches to c. 40 mm wide, usually delimited by a thin, black prothallus, especially when contiguous with other lichens; photobiont trentepohlioid, with cells globose to ellipsoid, 6−15 μm wide, in short chains or scattered in clusters.
Apothecia abundant, scattered, 0.3−2 mm wide, 60−120 μm thick, black, epruinose, applanate, roundish, or irregularly effigurate. Proper exciple excluded. Hypothecium dark red-brown, K+ olive, 10−50 μm thick. Hymenium 50−80 μm thick, hyaline to dilutely yellow-brown or red-brown, K+ olive, I+ orange-red, KI+ blue, overlain by a deep red-brown, K+ olive epithecium 12−20 μm thick; asci of the Arthothelium-type, 40−60 × 28−45 μm, 8-spored; paraphyses indistinct. Ascospores broadly ellipsoid, straight or slightly curved, hyaline at first, at most pale greyish and minutely rugulose when old, (23−)24−30.8−35(−36) × (8−)8.5−11.1−12 μm (n = 60), not macrocephalic, with 4−8 transverse and 0−1(−2) longitudinal septa; central cells mostly undivided; apices rounded.
Conidiomata not found.
Chemistry
No substances detected by TLC.
Remarks
Macroscopically this species is recognized by its relatively large, effigurate, very thin, applanate apothecia, which contrast with the usually slightly convex, roundish and generally smaller fruiting bodies of most other Tasmanian species of the genus. However, examination of its ascospores is essential for unequivocal identification. These are very distinctive, especially in the combination of being non-macrocephalic, with rounded apices, typically one (or at most two) longitudinal septa and, critically, that the central, largest spore locules are usually undivided. This feature is unusual in that in most species of the genus as a whole, it is normal to have the greatest number of longitudinal septa across the widest (and therefore central) cells. Occasional spores will have no longitudinal divisions at all. Similar ascospores are illustrated by Lee & Hur (Reference Lee and Hur2019) for the Korean species A. ulleungdoensis Lee & Hur, which may well be a later synonym.
The most similar species in the Tasmanian flora is A. velatium, which contains the same apothecial pigment but has smaller, more convex apothecia, and which differs most clearly by its ascospores that have more longitudinal septa, including across all the larger, central segments, and have the cells arranged ±regularly like bricks in a wall. There is another similar but as yet unidentified species in the rainforests of New South Wales which differs chiefly by having ascospores with rather more pointed apices (Kantvilas Reference Kantvilas1990). The brown, K+ olive apothecial pigment, here termed Interveniens-brown, occurs commonly in the genera Arthothelium and Arthonia, and is seen in A. ampliatum (which differs by having macrocephalic ascospores) and other species from Tasmania and elsewhere.
Distribution and ecology
This species is known from the south-eastern Australian mainland (New South Wales and Victoria), New Zealand and, remarkably, from the eastern United States, the provenance of the type collection. In Tasmania, it is a frequent component of the rich, crustose lichen flora that occurs in the understorey of callidendrous rainforests (nomenclature after Jarman et al. (Reference Jarman, Kantvilas and Brown1994)), especially on the smooth bark of Atherosperma moschatum, and on the understorey twigs of various other trees and shrubs (Eucryphia lucida, Nothofagus cunninghamii, Phyllocladus aspleniifolius, Tasmannia lanceolata). It can also be found in wet scrub and wet sclerophyll forest communities which are interpreted as seral stages in the development of callidendrous rainforest. This species was particularly common in the extensive callidendrous rainforests of north-west Tasmania, which have since been cleared and the land converted to eucalypt plantations largely devoid of epiphytic lichens.
Selected specimens examined
Australia: Tasmania: Sumac Road, Spur 2, S of Arthur River, 41°08′S, 145°02′E, 170 m, 1981, G. Kantvilas 342/81 (BM, HO); Simons Road, near Ben Nevis, 830 m, 1981, G. Kantvilas 1080/81 (BM, HO); Flannel Road, Arthur River, 430 m, 1982, G. Kantvilas 16/82 (BM, G, HO); Pieman Road near Huskisson River, 41°44′S, 145°29′E, 180 m, 1989, G. Kantvilas 18/89 (HO); Bun Hill, Forestier Peninsula, 42°58′S, 147°56′E, 320 m, 1989, G. Kantvilas 372/89 (HO); Anthony Road, 41°47′S, 145°37′E, 390 m, 1990, G. Kantvilas 192/90 (HO); Saxons Road, c. 1.5 km NW of Parrawe, 41°17′S, 145°34′E, 560 m, 1992, G. Kantvilas 52/92, B. Fuhrer & J. Jarman (HO); Buckland Military Training Area, S of Bluestone Tier, 42°30′S, 147°48′E, 300 m, 2003, G. Kantvilas 331/03 (HO); Savage River Pipeline Road near Rapid River, 41°16′S, 145°20′E, 440 m, 2015, G. Kantvilas 223/15 (HO); Rabalga Track, 41°05′S, 145°22′E, 280 m, 2019, G. Kantvilas 96/19 (HO); MacGregor Peak, 42°59′S, 147°57′E, 590 m, 2020, G. Kantvilas 149/20 (HO); Stony Head MTA, Ryans Hill, SE of summit, 41°01′05″S, 147°01′43″E, 210 m, 2020, G. Kantvilas 186/20 (HO).
Selected comparative material also examined
Australia: Victoria: Errinundra Plateau, Hensleigh Creek Road, 37°16′50″S, 148°58′00″E, 1020 m, 1999, S. Ford (MEL); Yarra Ranges NP, The Beeches Rainforest Walk, 37°29′25″S, 145°50′24″E, 800 m, 2008, V. Stajsic 4715 (HO, MEL). New South Wales: Mt William, Barrington Tops NP, 32°04′30″S, 151°28′00″E, 1400 m, 1988, G. Kantvilas 284/88 (HO, NSW); Gloucester Tops, 32°04′S, 151°34′E, 1150 m, 1988, G. Kantvilas 417/88 (HO, NSW).—USA: Louisiana: East Feliciana, Felixville, on parish road 432, c. 8 miles NE of Clinton, 1968, S. Tucker 7715 (HO). South Carolina: Berkeley Co., Francis Marion National Forest, 33°11′43″N, 79°31′22″W, 21 ft, 2013, J. C. Lendemer 40862 (HO, NY).
Arthothelium macounioides Kantvilas sp. nov.
MycoBank No.: MB 841469
Arthothelio macounii similis et item apotheciis pigmentum in kalio vivide carmesinum infusis et ascosporis macrocephalicis sed ascosporis maioribus, 29−42 μm longis, 11−18 μm latis, et cauda ascosporum septatior, 6−8-plo transverse et 2−4-plo longitudinaliter, differt.
Typus: Australia, Tasmania, Wielangta Road, 42°43′S, 147°51′E, 260 m, on Acacia riceana in Eucalyptus globulus-dominated wet forest, 20 March 1996, G. Kantvilas s. n. (holotypus—HO).
Thallus very thin, ±inapparent or dull pale greyish, scurfy, discontinuous, forming undelimited patches to c. 30 mm wide, I−, KI+ very patchily bluish; photobiont trentepohlioid, with cells subglobose to oblong-ellipsoid, 10−23 × 6−18 μm wide, forming chains.
Apothecia abundant, scattered, roundish, 0.4−0.8 mm wide, 220−400 μm thick, dark reddish brown, epruinose, adnate or basally constricted, markedly convex, in section entirely infused with a golden yellow pigment, K+ crimson, N−, C+ transient pink; pigment most heavily concentrated in the outer parts of the apothecium and then deep orange-brown to red-brown, in the innermost parts bright golden yellow. Proper exciple reduced, mostly excluded. Hypothecium poorly differentiated from the hymenium, c. 60−120 μm thick. Hymenium 90−150 μm thick, golden yellow, K+ crimson, I+ red, KI+ blue; asci of the Arthothelium-type, 60−90 × 45−60 μm, 8-spored; paraphyses branched and anastomosed, relatively stout, 1.5−3 μm wide, often greyish in the upper part but with the apices not expanded. Ascospores broadly ellipsoid to oblong, straight or slightly curved, hyaline, becoming brown and rugulose when old, (29−)30−34.4−40(−42) × 11−14.0−17(−18) μm (n = 55), macrocephalic, with an enlarged, undivided upper cell and muriform ‘tail’ with 6−8 transverse and 2−4 longitudinal septa.
Conidiomata not found.
Chemistry
No substances detected by TLC.
Etymology
The species epithet refers to the similarity of the new species to A. macounii.
Remarks
The golden yellow, K+ crimson apothecial pigment (Endoaurantiacum-gold) of this species is shared by three other Tasmanian species (A. bacidinum, A. endoaurantiacum and A. subtectum), but only the saxicolous A. subtectum has macrocephalic ascospores similar to A. macounioides, although these are shorter and have fewer septa. The new species is superficially very similar to the Northern Hemisphere's A. macounii (G. Merr.) W. J. Noble, which also has macrocephalic spores and contains a K+ red apothecial pigment. However, the ascospores of that species are somewhat smaller (20−38 × 10−14 μm; Coppins Reference Coppins, Smith, Aptroot, Coppins, Fletcher, Gilbert, James and Wolseley2009), with 4−6 transverse and 1−2 longitudinal septa. Furthermore, in A. macounii it is not uncommon to have some transverse spore sections with no longitudinal divisions, whereas in A. macounioides each transverse segment is always divided, as much as four times. At first glance the pigments of the two species appear different but their profiles in a range of reagents are identical and the perceived differences are possibly due to concentration levels.
Distribution and ecology
This distinctive species is known only from the type collection. It grew in deep shade on the bark of the small understorey tree Acacia riceana, in wet sclerophyll forest that was logged and regenerated approximately a century ago. This forest type is not known for being a hot spot of lichen diversity or noteworthiness, and hence has been little studied; thus the new species may well be more widespread.
Comparative material of A. macounii examined
Great Britain: Scotland: V.C. 97, West Inverness-shire, Ardnamurchan, hazelwood near Sonachan Hotel, NM 451664, 52 m, J. R. Douglass (HO).
Arthothelium magenteum Kantvilas sp. nov.
MycoBank No.: MB 841470
Species corticola, ascosporis ubique muriformibus, 25−55 μm longis, 11−22 μm latis, pigmento singulari, marronino-rubro, in kalio vivide magento recognita.
Typus: Australia, Tasmania, W of Tahune Bridge in the Warra SST, Coupe 1E, 43°06′S, 146°41′E, 100 m elevation, on Nematolepis squamea in Eucalyptus obliqua wet forest, 24 February 2004, G. Kantvilas 94/04 (holotypus—HO; isotypus—E).
Morphology of Tasmanian species of Arthothelium II. A, A. magenteum. B, A. subtectum. C, A. suffusum, with basally constricted, subglobose, pruinose apothecia. D, A. velatium. Scales = 1 mm. In colour online.
Thallus effuse, continuous or patchy, 20−60(−120) μm thick, dull grey, pale whitish, brownish grey or olive-brown, forming usually undelimited, irregular patches to c. 60 mm wide, in section ecorticate, I+ brownish red, KI+ pale blue, sometimes overlain with a patchy, dark maroon-red, K+ magenta-pink pigment as in the apothecia; photobiont trentepohlioid, with cells globose to broadly ellipsoid, 10−16 × 9−13 μm, in short chains or clusters.
Apothecia abundant, scattered, 0.2−1(−1.5) mm wide, 100−200 μm thick, black to brown-black, epruinose, applanate or slightly to markedly convex, adnate, roundish to ellipsoid or fleck-like when young, when old becoming more irregular, lobate and sometimes confluent. Proper exciple lacking or extremely reduced, at most comprised of a hyaline to pale brown layer to c. 20 μm thick of entangled, anastomosing hyphae 1−1.5 μm wide. Hypothecium barely differentiated from the hymenium, 10−50 μm thick, hyaline or, more typically, infused with or subtended by dark maroon-red pigment. Hymenium 110−180 μm thick, hyaline to pale yellowish brown, I+ orange-red, KI+ mottled orange-red and blue, with the latter reaction confined mainly to the asci, overlain by a thick, lumpy layer of dark maroon-red pigment which suffuses downwards, sometimes through the entire hymenium; pigment in irregular lumps that congeal the hymenium, K+ vivid magenta-pink, C+ orange-brown, N+ orange-brown, H−, fluorescing red-pink in polarized light; asci of the Arthothelium-type, 50−90 × 36−60 μm, 8-spored; paraphyses branched and anastomosing, embedded in a gel matrix, 1−1.5(−2) μm wide, with the apices not expanded. Ascospores ellipsoid to ovoid, sometimes a little curved, hyaline at first, becoming brown with age, (25−)33.5−41.4−53.5(−55) × (11−)12.5−17.7−22 μm (n = 55), not macrocephalic, densely muriform throughout, with 7−15 transverse and 1−5 longitudinal septa.
Conidiomata not found; conidia rarely seen within the apothecia, arising from the degeneration of overmature spores, bacilliform, 4.5−6 × 0.8 μm.
Chemistry
No substances detected by TLC.
Etymology
The specific epithet refers to the colour reaction in K of the apothecial pigment that characterizes this species.
Remarks
The unique apothecial pigment, termed here Magenteum-red, characterizes this species. In some specimens, especially those from coastal areas, longer elution in K also produces a diffuse blue coloration. It remains unclear whether an additional pigment is present or whether this effect represents a breakdown of the one pigment.
There are several different pigments in Arthothelium that produce a shade of pink or red with K, but none has the same characteristic profile as Magenteum-red in the various standard reagents or in visible or polarized light. In the Tasmanian flora, A. bacidinum, A. endoaurantiacum, A. macounioides and A. subtectum all contain Endoaurantiacum-gold, which reacts K+ crimson. Brownish, K+ crimson pigments are found in the Northern Hemisphere's A. macounii and in the New Zealand species, A. spadiceum; these are likely to be Endoaurantiacum-gold as well, albeit in lower concentrations than those seen in the Tasmanian taxa. There is also a brownish, K+ reddish pigment in the Northern Hemisphere's A. spectabile which is different again. In addition to apothecial pigments, the ascospores of the species mentioned also differ from those of A. magenteum, being either macrocephalic, or muriform throughout but with the septa parallel and the locules ±even-sized. In contrast, the ascospores of A. magenteum are frequently a little attenuated at one end, sometimes curved, and the septa and locule size are irregular. A further species studied in this context is A. homoeophanum (Nyl.) Zahlbr. (type specimen in H examined), described from New Caledonia (Nylander Reference Nylander1863). Here the greyish green pigment is Sedifolia-grey (Meyer & Printzen Reference Meyer and Printzen2000), reacting K+ violet, C+ violet. Although not known from other species of Arthothelium, it is not uncommon in the Lecanoromycetes, being found, for example, in species of Micarea, Rimularia and Thalloidima.
The record of A. macrothecum (Fée) A. Massal. from Tasmania (Jatta Reference Jatta1911) is based solely on a specimen of A. magenteum (held in NAP). This American, essentially tropical species does not occur in Tasmania, and differs starkly from A. magenteum in many characters, including its black, applanate apothecia, Interveniens-brown pigmentation, and large ascospores (46−70 × 17−28 μm) with as many as 17 transverse and 7 longitudinal septa (Fink Reference Fink1935; this study). Specimens from tropical Queensland, held in CANB, confirm the occurrence of this species in Australia.
Distribution and ecology
Arthothelium magenteum is known only from Tasmania where it is a common epiphyte on smooth bark, mostly in the wetter, western parts of the island but also in locally wetter, coastal parts of the south-east. It is found mainly in rainforest, especially in communities of the thamnic type (nomenclature after Jarman et al. (Reference Jarman, Kantvilas and Brown1994)), colonizing the bark of hosts such as Anodopetalum biglandulosum, Eucryphia lucida, Lagarostrobos franklinii, young Nothofagus cunninghamii, N. gunnii and Pittosporum bicolor, either in moderately shaded gaps in the understorey, or on young branches within the forest canopy. It is also found in old-growth wet eucalypt forests which represent successional stages towards thamnic (as distinct from callidendrous) rainforest. In such communities, the bark of mature understorey trees, notably Nematolepis squamea, is a preferred habitat. It is found at subalpine to lowland elevations and has also been collected in wet, coastal scrub.
Selected specimens examined
Australia: Tasmania: near Geeveston, 1100 ft [330 m], W. A. Weymouth s. n. (NAP); Mueller Road, 500 m, 1981, G. Kantvilas 1036/81 (BM, HO); Weindorfers Forest, 41°38′S, 145°56′E, 1000 m, 1988, G. Kantvilas 10/88 (BM, H, HO); Anthony Road, 41°50′S, 145°38′E, 560 m, 1989, G. Kantvilas 11/89 (HO); Badger Creek, c. 2.5 km S of Greystone Bluff, 43°06′S, 146°02′E, 280 m, 1989, G. Kantvilas 72/89 (HO); 1 km W of Mt Charter, 41°37′S, 145°40′E, 780 m, 1989, G. Kantvilas 160/89 (HO); 4 km N of Precipitous Bluff, 43°25′30″S, 146°36′30″E, 730 m, 1990, G. Kantvilas 111/90 (HO); Olga River at Line 7, 42°51′S, 145°50′E, 70 m, 1990, G. Kantvilas 174/90 (HO); c. 3 km S of Teepookana, 42°13′S, 145°26′E, 220 m, 1990, G. Kantvilas 649/90 (HO); Green Head, 43°06′S, 146°04′E, 750 m, 1991, G. Kantvilas 79/91 (HO); Pelion Plains, 41°50′S, 146°02′E, 900 m, 1992, G. Kantvilas 220/92 (HO); W of Tahune Bridge in the Warra SST, 43°06′S, 146°41′E, 90 m, 1997, G. Kantvilas 273/97 (CANB, HO); ibid., 200 m, 1998, G. Kantvilas s. n. (HO, UPS); Windy Moor, 42°40′S, 146°38′E, 1170 m, 2013, G. Kantvilas 8/13 (HO); Crest Range, 43°17′28″S, 146°30′26″E, 980 m, 2016, G. Kantvilas 229/16 (HO); Tasman Track, W of O'Hara Bluff, 43°05′S, 147°57′E, 380 m, 2020, G. Kantvilas 273/20 (HO); Bivouac Bay, 43°07′S, 147°58′E, 5 m, 2020, G. Kantvilas 345/20 (HO).
Comparative material of A. macrothecum examined
Australia: Queensland: Mossman, Mt Molloy Rd, 16°32′05″S, 145°22′59″E, 390 m, 2006, J. A. Elix 36903 (CANB).—Brazil: Sao Paolo: Ilha de Santo Amaro, 1978, K. Kalb & G. Plöbst (K. Kalb: Lichenes Neotropici 353) (CANB).
Arthothelium subtectum Kantvilas sp. nov.
MycoBank No.: MB 841471
Apotheciis pigmento aurantiaco, in kalio vivide carmesino infusis et ascosporis macrocephalicis, 22−36 μm longis, 9−14 μm latis, Arthothelio macounii simile sed habitu saxicola et apotheciis parvioribus, plerumque ad 0.5 mm latis, convexis, sessilibus vel leviter basi constrictis differt.
Typus: Australia, Tasmania, Three Thumbs summit area, 42°36′S, 147°52′E, 540 m elevation, on the undersides of large, inclined dolerite boulders in a damp, sheltered aspect in dry sclerophyll forest, 15 August 2019, G. Kantvilas 186/19 (holotypus—HO; isotypus—E).
Thallus inapparent, perceived mainly as a discoloration of the substratum, occasionally unevenly scurfy-verruculose and dull grey or brown, forming extensive undelimited patches to c. 50 mm wide; photobiont trentepohlioid, with cells globose to ellipsoid, 8−22 μm wide, in short chains or scattered in clusters.
Apothecia very abundant, scattered, 0.2−0.5(−0.8) mm wide, 160−320 μm thick, black, epruinose, adnate or slightly basally constricted, markedly convex, irregularly roundish or ellipsoid, in section entirely infused with a golden yellow pigment, K+ crimson, N−, C+ transient pink; pigment most heavily concentrated in the outer parts of the apothecia and then deep orange-brown to red-brown, in the innermost parts bright golden yellow. Proper exciple generally reduced and reflexed, in younger apothecia 10−25 μm thick, deep orange-brown, comprised of rather amorphous, anastomosed hyphae c. 1 μm wide inspersed with minute granules. Hypothecium barely differentiated from the hymenium, mostly to 80−200 μm thick, intense golden yellow. Hymenium 70−120 μm thick, intensely golden yellow, K+ crimson, I+ orange-red, KI+ blue, overlain by a thick, lumpy layer of pigment 10−20 μm thick; asci of the Arthothelium-type, 55−75 × 20−45 μm, 8-spored; paraphyses branched and anastomosed, 1 μm wide, usually brownish in the uppermost part, with the apices expanded to 4 μm. Ascospores broadly ellipsoid to oblong, straight, hyaline at first, soon becoming brown and, when old, minutely rugulose, 22−27.1−33(−36) × 9−11.6−14 μm (n = 55), macrocephalic with an enlarged, undivided upper cell and muriform ‘tail’ with 4−5 transverse and 0−3 longitudinal septa.
Conidiomata not found.
Chemistry
No substances detected by TLC.
Etymology
The specific epithet is derived from the Latin sub, meaning ‘under’ and tectum, meaning ‘roof’, and alludes to the habitat of the new species under rocky overhangs.
Remarks
Arthothelium subtectum is characterized by the saxicolous habit, the convex, sessile or slightly basally constricted apothecia, heavily infused with Endoaurantiacum-gold pigment, and the macrocephalic ascospores, 22−36 × 9−14 μm. Only one other saxicolous species of Arthothelium is known in Tasmania, and whereas both contain the same apothecial pigment, A. bacidinum is readily distinguished by its larger, non-macrocephalic ascospores. These two species also have a different habitat ecology, with A. subtectum being restricted to upland elevations whereas A. bacidinum is coastal. Macrocephalic ascospores, in combination with Endoaurantiacum-gold, are also seen in the corticolous species A. macounioides, which differs in having the muriform tail of its ascospores more divided, with up to four longitudinal septa across the widest part (compare Fig. 2F and 2H). Also with macrocephalic ascospores is A. ampliatum, but that species is corticolous, has a brownish, K+ olive green apothecial pigment, and the spores are somewhat larger. The most similar taxon to A. subtectum is the Northern Hemisphere species A. macounii (G. Merr.) W. J. Noble. This lichen has ascospores of very similar dimensions (Coppins & James Reference Coppins and James1979, as A. ilicinum var. reagens; Coppins Reference Coppins, Smith, Aptroot, Coppins, Fletcher, Gilbert, James and Wolseley2009) but is exclusively corticolous on smooth bark and has adnate, applanate apothecia. The record of A. macounii from Tasmania by Kantvilas et al. (Reference Kantvilas, Elix and Jarman2008) refers to A. subtectum. In the field, the strongly convex, black apothecia and the saxicolous habitat of A. subtectum are initially more suggestive of a species of Brianaria or Micarea, all of which have an entirely different apothecial anatomy and ascospore morphology.
Distribution and ecology
Arthothelium subtectum has a very distinctive and narrow habitat ecology, being restricted to relatively dry, sheltered, deeply shaded underhangs of large dolerite outcrops in wet eucalypt forest and scrub. It is also noteworthy for apparently occurring mainly in the mid-elevation peaks of the eastern half of Tasmania and has not been found on western and central Tasmanian peaks, despite extensive lichen collecting having been undertaken there. This distribution pattern is not unique in Tasmania, and a small number of lichens display a similar, restricted pattern, viz. Chiodecton montanum Thor, Cresponea graemeannae Kantvilas, Lecanactis scopulicola Kantvilas, Lobaria scrobiculata (Scop.) DC. and Nephroma rufum (C. Bab.) P. James; all, like Arthothelium subtectum, are representatives of lichen genera usually associated with old-growth forest vegetation and stable, continuous ecological conditions, and their occurrence on rocks in more dynamic and disturbed vegetation types can be considered unusual.
Specimens examined
Australia: Tasmania: Hartz Mtns, 43°13′S, 146°46′E, 1963, P. W. James s. n. (BM, HO); South Sister, 41°32′S, 148°10′E, 750 m, 2004, G. Kantvilas 431/04 (HO); Mt Hobbs summit, 42°30′S, 147°35′E, 820 m, 2018, G. Kantvilas 59/18 (HO); Mt Forestier, 42°55′S, 147°51′E, 315 m, 2020, G. Kantvilas 86/20 (HO); Quoin Mtn, 42°33′S, 147°17′E, 870 m, 2020, G. Kantvilas 130/20 (HO).
Arthothelium suffusum (C. Knight) Müll. Arg.
Bull. Herb. Boissier 2, App.1, 84 (1894).—Arthonia suffusa C. Knight, Trans. N. Z. Inst. 15, 353 (1883); type: New Zealand, sine loco [prob. Wellington], 23.xi.[18]77, Charles Knight s. n. (holotype—WELT 006974!; isotype—BM!).
Arthothelium subspectabile Vĕzda & Kantvilas, Telopea 4, 665 (1992); type: Australia, Tasmania, 3 km SW of Mt Agnew, 41°55′S, 145°11.5′E, on dry trunk of mature Nothofagus cunninghamii in rainforest, 190 m, 6 April 1989, G. Kantvilas 135/89 (holotype—HO!; isotype—PRA-V!).
Thallus very thin, grey-white to dingy olive-grey, continuous, smooth, faintly rimose, I−, KI+ patchily pale bluish, forming irregular, undelimited patches to 100 mm across; photobiont trentepohlioid, with cells subglobose to ellipsoid-oblong, 10−15 μm wide, in short chains or clusters.
Apothecia scattered, abundant, roundish, 0.25−0.4(−0.6) mm wide, 180–250 µm thick, black, coarsely granular whitish pruinose with calcium oxalate crystals, epruinose when overmature, initially semi-immersed, soon becoming basally constricted and markedly convex. Proper exciple reduced to a layer of red-brown, K+ olive pigmented tissue 10−15 μm thick at the sides of the apothecia and extending continuously in a band beneath the hypothecium. Hypothecium poorly differentiated, hyaline to pale yellow-brown to brown, K+ olive, mostly c. 30−50 μm thick. Hymenium 110−200 μm thick, hyaline in the lower part, red-brown, K+ olive in the upper part, I+ red, KI+ blue, densely inspersed oil droplets and with rhomboidal, hyaline granules to 5−8 μm wide, soluble in K; asci of the Arthothelium-type, 60−85 × 40−55 μm, mostly 8-spored; paraphyses 1−1.2 μm wide, rather indistinct, with the apices not expanded. Ascospores broadly ellipsoid to ovoid, hyaline, becoming brown and rough-walled when old, (27−)28−33.9−42(−43) × (13.5−)14−17.1−22(−24) μm (n = 55), not macrocephalic, with 7−9(−11) transverse and 2−5(−7) longitudinal septa.
Conidiomata not found.
Chemistry
No substances detected by TLC.
Remarks
This species is very well characterized by its coarsely white-pruinose, convex, basally constricted apothecia, with a hymenium heavily inspersed with crystals and oil droplets. The pruina is composed of calcium oxalate, an unusual occurrence in Arthothelium, whereas the hymenial crystals are of an unknown composition. It has no confusing species in the Tasmanian flora, and indeed no other members of the genus with similar characteristics are known. The brown, K+ olive pigment is Interveniens-brown, which is found in many other species, both in Tasmania and elsewhere. Illustrations and further data are provided by Kantvilas & Vĕzda (Reference Kantvilas and Vězda1992).
Distribution and ecology
In Tasmania, Arthothelium suffusum has a very distinctive habitat, being confined to the dry, sheltered, deeply shaded faces of the oldest, largest trees in cool temperate rainforest. Lagarostrobos franklinii and Nothofagus cunninghamii appear to be the preferred hosts, both trees being capable of massive stature and producing flaking, coarse bark when old. This lichen is part of the Lecanactis abietina-L. mollis association (Kantvilas Reference Kantvilas1988) and occurs together with calicioid lichens, Arthonia spp., Bactrospora arthonioides Egea & Torrente, Chrysothrix candelaris (L.) J. R. Laundon and other small crustose lichens. It was first described from New Zealand where it appears to have a similar habitat ecology.
Specimens examined
Australia: Tasmania: Weldborough, 41°12′S, 147°54′E, 640 m, 1981, G. Kantvilas 1127/81 (BM, HO); Badger Creek, c. 2.5 km S of Greystone Bluff, 43°06′S, 146°02′E, 280 m, 1989, G. Kantvilas 78/89 (HO); c. 3 km S of Teepookana, 42°13′S, 145°26′E, 220 m, 1990, G. Kantvilas 663/90 (HO); Sumac Road, Spur 2, S of Arthur River, 41°08′S, 145°02′E, 170 m, 1993, G. Kantvilas 306/93 (HO); Weldborough Pass, 41°13′S, 147°57′E, 1995, G. Kantvilas 58/95 (HO); Rabalga Track, 41°05′S, 145°23′E, 280 m, 2019, G. Kantvilas 88/19 (HO).
Also examined
New Zealand: North Island: Gisborne, Urewera NP, 1.5 km S of Lake Waikereiti, 38°44′S, 177°09′E, 1981, L. Tibell 12993 (MEL, UPS).
Arthothelium velatium Müll. Arg.
Bull. Herb. Boissier 1, 60 (1893); type: Victoria, Oakleigh, on Casuarina, 8 August 1892, F. R. M. Wilson 1586 (lectotype, fide Kantvilas (Reference Kantvilas1990)—G!; isolectotypes—MEL!, NSW!).
Thallus very thin and patchy, pale cream to whitish grey, in section I+ red, KI+ patchily bluish, forming irregular, usually undelimited patches to 10−50 mm wide; photobiont trentepohlioid, with cells globose to ellipsoid, 8−16 μm wide, in short chains or scattered in clusters.
Apothecia scattered, 0.3−0.6(−1) mm wide, 60−180(−250) μm thick, black to brown-black, epruinose, adnate or sometimes a little immersed and covered by a thin veil of thalline tissue, plane to a little convex, roundish, fleck-like or irregularly effigurate. Proper exciple excluded or reduced to a layer of brown-pigmented tissue c. 10−15 μm thick. Hypothecium pale brown to brown, K+ olive or olive-yellow, mostly c. 20−40 μm thick. Hymenium 70−80 μm thick, hyaline to dilutely brownish, frequently inspersed with occasional oil droplets, I+ orange-red, KI+ blue, overlain by a deeper brown to red-brown, rather granular, K+ olive-grey to olive-yellow, rarely rather strongly although transiently K+ yellow, N− or intensifying brown, epithecium to 30 μm thick; asci of the Arthothelium-type, 40−75 × 30−50 μm, 8-spored; paraphyses 1–1.5 μm wide, sometimes greyish-pigmented and expanded to 2.5 μm in the uppermost part. Ascospores broadly ellipsoid, broader at one end, straight or slightly curved, hyaline at first, becoming brown when old, (26−)30−35.5−41 × (11−)12−14.8−18(−20) μm (n = 80), not macrocephalic, with 6−10 transverse and 2−3 longitudinal septa.
Conidiomata not found.
Chemistry
No substances detected by TLC.
Nomenclatural note
Müller (Reference Müller1893) coined the epithet ‘velatius’ and made reference to the affinities of his new species to ‘A. velato Müll. Arg.’ which appears to be a nomen nudum. The correct inflexion of ‘velatius’ (indicating affinity to ‘velato’) should be the neuter ‘velatium’.
Remarks
This species is distinguished by the combination of its particular ascospore type and the presence of brownish, K+ olive apothecial pigment (Interveniens-brown). The same pigment occurs in several other species, notably A. interveniens and A. ampliatum, although in A. velatium it is comparatively dilute and the reactions can be rather weak. In the two coastal specimens seen, the hymenium and hypothecium yield a more yellowish reaction in K which gradually fades to olive; this is presumed to be a function of concentration rather than the result of some additional substance being present. In older specimens, the reaction tends towards a brownish olive colour. Further collections will be required to determine whether these varying colour reactions are correlated with any other taxonomic characters.
The ascospores of A. velatium are distinctive in that they tend to be wider at the proximal end, are not macrocephalic, and the cells are ±square or rectangular and arranged neatly like bricks in a wall, at least in the central parts of the spore. In an earlier study, Kantvilas (Reference Kantvilas1990) cited the ascospores as being (16.5−)26−37 × 8−13 μm, with 8−11 transverse and 2−3(−4) longitudinal septa. Examination of further type material suggested that these dimensions were an underestimate, probably based on immature spores; the isolectotype in MEL has ascospores of 32−36 × 10−14 μm, with (7−)9−10 transverse and 3 longitudinal septa. These ascospores are rather more septate and narrower in comparison to the Tasmanian specimens described above, but in general this name offers a ‘good fit’.
Also of relevance in this investigation is the Northern Hemisphere taxon A. norvegicum Coppins & T. Tønsb. The description of this species by Coppins & Tønsberg (Reference Coppins and Tønsberg1984) also matches the Tasmanian collections very well, and it is possible that it is closely related if not synonymous with A. velatium. Coppins & Tønsberg (Reference Coppins and Tønsberg1984) also discuss the distinction between A. norvegicum and A. spectabile Flot. ex A. Massal., noting that the latter has similar but slightly shorter ascospores (26−36 × 12−15 μm) with fewer (5−7) transverse septa, a brown, K+ reddish brown pigment in the hymenium and epithecium, and a thallus that reacts I+ blue in section. An examination of herbarium specimens of A. spectabile with an I+ blue thallus, chiefly from North America, revealed ascospores 30−45(−50) × 14−19(−20) μm (n = 35) with (5−)6−8(−9) transverse septa and 1−3 longitudinal septa, and a red-brown hymenium that is ±unchanged or reddish in K. A single Tasmanian specimen has these apothecial characters but its thallus is I− and therefore it remains unplaced.
Arthothelium velatium is also somewhat similar to the Australian taxon A. huegelii (Nyl.) Zahlbr., which has larger and more divided ascospores, 41−60 × 20−27 μm, with 9−14 transverse and 4−7 longitudinal septa (type material in H examined).
Distribution and ecology
Arthothelium velatium is known from Tasmania, Victoria and New South Wales. In Tasmania, it is never common, the few herbarium collections are fortuitous, and consequently it is difficult to form a picture of its ecological requirements. It has been recorded from the smooth-barked foliage twigs of Atherosperma moschatum in the understorey of cool temperate rainforest, on Beyeria viscosa in wet eucalypt forest, and on shrubs in wet coastal scrub.
Specimens examined
Australia: Tasmania: Anthony Road, 41°50′S, 145°38′E, 560 m, 1989, G. Kantvilas 12/89 (HO); ibid., 41°49′S, 145°38′E, 480 m, 1992, G. Kantvilas 516/92 (HO); Pelion Plains, c. 1 km SW of Pelion Hut, 41°50′S, 146°02′E, 900 m, 1992, G. Kantvilas 209/92 (HO); Wielangta Road, 42°43′S, 147°51′E, 260 m, 1996, G. Kantvilas s. n. (HO); South Cape Bay, 43°37′S, 146°50′E, 50 m, 2006, G. Kantvilas 354/06 (HO); Bivouac Bay, 43°07′S, 147°58′E, 5 m, 2020, G. Kantvilas 343/20 (HO).
Comparative material also examined
Arthothelium norvegicum. Norway: Nord-Trøndelag, Namdalseid, south of Lake Altvatn, 80−120 m, 1983, T. Tønsberg 8254a (BG, HO).
Arthothelium spectabile. Georgia: Distr. Soči, 1982, A. Vĕzda (A. Vĕzda: Lich. Sel. Exsicc. 1851 (CANB).—USA: North Carolina: Haywood County, Great Smoky Mtns NP, Cataloochie Divide Trail, 35°35′02″N, 83°04′39″W, 2011, E. Tripp et al. 2175 (Lichens of Eastern North America Exsiccati 464) (CANB, HO). Massachusetts: Essex, 1849 (Reliquiae Tuckermanniae 7) (CANB). Iowa: White Pine Hollow State Forest Preserve, 1965, C. M. Wetmore 13727 (CANB, MEL).
Acknowledgements
I thank Jean Jarman for the habit photographs that illustrate this paper, and for preparing the other illustrations for publication. Matthew Baker assisted with photomicroscopy. John Douglass and Brian Coppins kindly provided a reference specimen of A. macounii, and I am much indebted to Brian for his ‘helpful hints’ during the course of the work. The hospitality and assistance of various herbaria, either via specimen loans or hosting my visits, is much appreciated; I particularly thank the curators of BM, CANB, H, MEL and WELT. Special thanks go to Leena Myllys and Saara Velmala (H), and Kim Hill (HO) for hastily arranging a last-minute loan amidst Covid restrictions. For their patience and companionship in the field in quest of Arthothelium specimens, I thank Brigitte de Villiers and Jean Jarman. This study was in part financially supported by the Australian Biological Resources Study through its Bush Blitz Programme.
Author ORCID
Gintaras Kantvilas, 0000-0002-3788-4562.
