Brahmaculus moonlighticus P.R.Johnst.

From Hindu mythology, named after Brahma, the four-headed creator god, reflecting the multiple heads of the apothecia, and the masculine diminutive -culus.

Phylogenetically Chlorociboriaceae, distinguished from Chlorociboria by its terrestrial habitat, and apothecium with stipe branched near apex, each branch with an apothecial cup.

Apothecia stipitate, yellow rhizomorphs at base of stipe, the stipe branched apically several times, each branch holding an apothecial cup. Receptacle and stipe densely covered with short hairs. Hairs more or less straight, cylindric, thin walled, with a few septa, pale brown intracellular pigment, externally densely encrusted with yellowish material, encrusting material dissolving in KOH + Melzer’s reagent. The hymenium within each apothecial cup is typically divided into smaller segments, with areas comprising asci and paraphyses separated by clumps of hair-like elements. Excipulum comprises cylindric cells arranged more or less parallel to the surface, cells mostly long-cylindric, but sometimes with outermost 1–2 layers of cells short and broad-cylindric, cell walls slightly thickened, hyaline, cells near base of hairs with pale brown vacuolar pigment. Asci with wall thickened at apex, amyloid pore extending through the wall, flaring near the inside and especially toward outside of the wall, 8-spored, with croziers. Paraphyses simple or tapering to apex, of similar length as asci. Ascospores oblong-elliptic, 0-septate, hyaline.

The four species described below are phylogenetically distinct but remarkably similar morphologically. There appear to be small differences in size and colour of the apothecia and shape of the paraphyses and hairs but having only a single specimen available for each species makes the significance of these differences difficult to assess. The rhizomorphs at the base of the stipe appear to be associated with tree roots. Based on the collecting sites, in South America and New Zealand the roots are likely to be Nothofagaceae, in Australia they may also be Nothofagaceae but Eucalyptus species were also growing in the vicinity. Observations from the South American specimens showed a loose weft of mycelium around the Nothofagaceae roots but there was no clear evidence of a mantle or ectomycorrhizal association. It is possible that these fungi are root endophytes, or perhaps parasites of Nothofagaceae-associated ectomycorrhizal fungi.

Chile – Magallanes • Puente San Pedro, south of Punta Arenas, stream near the end of the road, Nothofagus betuloides forest; -53.6993, 70.9695; Alija Mujic (MES2454) leg.; 5 Apr 2017; SGO – Holotype; FLAS-F-65086 – isotype; PDD 116650 – isotype.

Refers to the Magellanic forests of the type locality.

Phylogenetically distinct from other known Brahmaculus spp., apothecia 3–8 × 1–2.5 mm, paraphyses undifferentiated to rounded apex, ascospores 5.5–9 × 1.5–2 µm (average 7.3 × 1.7 µm).

Apothecia 3–8 mm high, stipe 0.4–0.6 mm wide, cap 1–2.5 mm wide, the more or less globose cap comprising several closely packed apothecial cups, these arising from short, branches at the top of the stipe, hymenium pale yellow, hymenial areas broken into smaller segments by groups of bright yellow, hair-like elements amongst the fertile parts of the hymenium. Receptacle densely covered with stiff, bright yellow hairs, stipe with shorter hairs. Hairs 45–70 × 2.5–4.5 µm, straight, cylindric, tapering gradually in apical half toward small, rounded apex, thin-walled, sparsely septate, pale brown vacuolar pigment, densely encrusted with coarse, bright yellow crystals that dissolve in KOH + Melzers. Ectal excipulum comprising narrow-cylindric cells 8–20 × 2.5–3 µm oriented at low angle to receptacle surface, wall slightly thickened, mostly hyaline except cells at the base of hairs have pale brown vacuolar pigment. Medullary excipulum similar in structure but cells wider, 4.5–8 µm diam. Paraphyses 1.5–2.5 µm diam., undifferentiated at rounded apex, about same length as asci. Asci 40–55 × 4–5 µm, cylindric, apex rounded, wall thickened, amyloid pore extends through the wall, diffuse and flaring slightly towards the outside of the wall, crozier at base, 8–spored. Ascospores 5.5–9 × 1.5–2 µm (average 7.3 × 1.7 µm, n = 12), oblong elliptic, ends rounded, flattened on one side, straight to slightly curved, 0-septate, hyaline.

Figure 3. 

Brahmaculus magellanicus (PDD 116650) A, B fresh apothecia C dried apothecia D ascoma in vertical section, showing multiple apothecial cups on short branches E ascoma in vertical section showing excipular tissue and hairs, and a clump of hairs within the hymenium F squash mount showing hairs in KOH G asci H paraphyses I ascospores. Scale bars: 1 mm (A–C); 100 µm (D); 10 µm (E–I).

The two Chilean species differ macroscopically, B. magellanicus having noticeably thinner stipes than B. osornoensis. The only known collection of B. magellanicus is from Magellanic subpolar forest in Patagonia that is dominated by Nothofagus betuloides. It is possible that this Brahmaculus species is restricted to these sub Antarctic cold southern forests but more specimens are needed to determine the range of the species.

New Zealand – Buller • Moonlight Creek; -42.2713, 171.4587; on soil under Nothofagaceae; A. Chinn leg.; 10 May 2018; PDD 112225 – holotype.

From the type locality. Historically important as a gold mining area (where T.H. Chinn, the great-great grandfather of the collector of the type specimen, prospected for gold in the 1880’s), the name also reflects the deep golden colour of the apothecia of this fungus.

Phylogenetically distinct from other known Brahmaculus spp., apothecia 1.5–3 × 1–1.8 mm, paraphyses taper slightly to rounded apex, ascospores 6.5–8.5 × 1.5–2(–3) µm (average 7.7 × 1.9 µm).

Apothecia 1.5–3 mm high, stipe 0.25–0.5 mm wide, cap 1–1.8 mm wide, bright golden-yellow when fresh, consistently with four short branches arising from top of stipe, each branch with its own apothecial cup, hymenium pale yellow, divided into a complex pattern with hymenial areas separated by narrow groups of golden yellow hair-like elements. Receptacle densely covered with stiff, bright yellow hairs. Hairs 40–60 × 3–4 µm, straight, cylindric, tapering slightly towards rounded apex, pale brown vacuolar pigment, wall smooth, encrusted with coarse yellow-brown crystals that dissolve in KOH + Melzer’s, few-septate. Ectal excipulum comprising long-cylindric cells 15–25 × 3–5 µm, but with the outermost 1–2 layers of cells short and broad-cylindric, 6–8 µm diam., cell walls slightly thickened, hyaline, cells near base of hairs with pale brown vacuolar pigment. Medullary excipulum comprising partly tangled hyphae 3–4 µm diam. with walls thin, hyaline. Paraphyses 2–3 µm diam., tapering slightly towards rounded apex, about same length as asci. Asci 45–55 × 5.5–6 µm, cylindric, tapering slightly to broad subtruncate apex, wall uniformly thickened across apex, amyloid pore extending through wall, flaring slightly towards both inside and outside of wall. Ascospores 6.5–8.5 × 1.5–2 (–3) µm (average 7.7 × 1.9 µm, n = 50), oblong-elliptic to subfusoid, sometimes tapering suddenly to a narrower lower half, ends rounded, flat one side in side view, sometimes slightly curved or sigmoid, 0-septate, hyaline.

Figure 4. 

Brahmaculus moonlighticus (PDD 112225) A fresh apothecia (dried apothecium inset) B detail, fresh apothecia C ascoma in vertical section, showing multiple apothecial cups D ascoma in vertical section showing excipular tissue and hairs E hairs in squash mount in KOH F asci and ascospores G asci and paraphyses H, I ascospores. Scale bars: 1 mm (A, B); 100 µm (C); 10 µm (D–I).

Brahmaculus moonlighticus has a stipe that consistently has 4 distinct branches near the apex. The other species have several separate hymenial cups, but these are held on very short branches arising from across the apex of the stipe, the margins of these cups superficially forming a more or less continuous layer.

Chile • Parque Nacional Vicente Perez Rosales, Volcan Osorno, on the road to the ski area just above Mirador el Bosque, Nothofagus dombeyi forest; -41.1382, 72.5370; Matthew Smith and Alija Mujic (MES2942) leg.; 17 April 2017; SGO – holotype; FLAS-F-65492 – isotype; PDD 116649 – isotype.

Refers to the type locality, Volcan Osorno.

Phylogenetically distinct from other known Brahmaculus spp., apothecia 3–6 × 1–2.5 mm, paraphyses taper slightly to rounded apex, ascospores 6.5–10(–11) × 1.5–2 µm (average 8.3 × 2 µm).

Apothecia 3–6 mm high, stipe 0.5–1 mm wide, cap 1–2.5 mm wide, the more or less globose cap comprising several closely packed apothecial cups, these arising from short, branches at the top of the stipe, hymenium pale yellow, hymenial areas broken into smaller segments by groups of bright yellow, hair-like elements amongst the fertile parts of the hymenium. Receptacle densely covered with stiff, bright yellow hairs, stipe with shorter hairs. Hairs 50–85 × 2.5–4 µm, straight, with a broad basal cell then cylindric, apically tapering suddenly to the narrow-rounded apex, thin-walled, sparsely septate, densely encrusted with coarse, bright yellow crystals that dissolve in KOH + Melzers. In squash mount, excipular cells broad-cylindric, about 15–30 × 8–12 µm, wall slightly thickened, hyaline. Paraphyses 2–2.5 µm, tapering slightly to rounded apex, about same length as asci. Asci 40–50 × 4–4.5 µm, cylindric, apex rounded, wall thickened, amyloid pore extends through the wall, flaring slightly towards the outside. Ascospores 6.5–10(–11) × 1.5–2.5 µm (average 8.3 × 2.0 µm, n = 50), oblong elliptic, ends rounded, one side flat in side view, sometimes slightly curved, 0-septate, hyaline.

Figure 5. 

Brahmaculus osornoensis (PDD 116649) A fresh apothecia (dried apothecia inset) B squash mount, excipulum and hairs in 3% KOH + Melzer’s reagent C squash mount, hairs in water showing encrusting crystals D paraphyses and asci E ascospores. Scale bars: 1 mm (A); 10 µm (B–E).

The two Chilean species differ macroscopically, Brahmaculus osornoensis having noticeably broader stipes than B. magellanicus and slightly longer ascospores. B. osornoensis is known only from Nothofagus dombeyi forest in northern Patagonia on Volcan Osorno in the Vicente Perez Rosales National Park. It is possible that this Brahmaculus species is restricted to the wetter and warmer forests in northern Patagonia, but more specimens are needed to determine the range of the species.

Australia – Tasmania • Geeveston District, Hermons Rd; -43.2652, 146.8613; J.M. Packham (6/R6/26) leg.; 5 June 1995; MEL 2363173 – holotype; PDD 117311 – isotype.

Named after the late Jillian (“Jill”) Mary Packham whose assiduous collecting activities detected the type collection.

Phylogenetically distinct from other known Brahmaculus spp., apothecia up to 11 × 2.5 mm, paraphyses undifferentiated to rounded apex, ascospores 5.5–8.5 × 1.5–2.5 µm (average 7.2 × 1.8 µm).

Apothecia up to 11 mm high, stipe up to 0.8 mm wide, cap up to 2.5 mm wide, the cap comprising several closely packed apothecial cups, these arising from short branches at the top of the stipe, hymenium white when fresh. Receptacle densely covered with stiff, bright yellow hairs, stipe with shorter hairs, yellow rhizomorphs at base. Hairs 40–60 × 2.5–3.5 µm, straight, narrow flask-shaped, broad near base then tapering suddenly to narrow-cylindric apical part, apex rounded, thin-walled, 1–2 septate near the base, densely encrusted with coarse, bright yellow crystals, that dissolve in KOH + Melzers. Ectal excipulum comprising cylindric cells 8–15 × 3–5 µm, oriented at a low angle to the receptacle surface, walls slightly thickened, hyaline. Medullary excipulum comprises partly tangled hyphae 3–5 µm diam. with walls thin, hyaline. Paraphyses 2–2.5 µm diam., undifferentiated at the rounded apex, about the same length as the asci. Asci 35–45 × 4.5–5.5 µm, cylindric, tapering slightly to broad, subtruncate apex, wall thickened across apex, amyloid pore extending through wall, flaring toward outside of wall. Ascospores 5.5–8.5 × 1.5–2.5 µm (average 7.2 × 1.8 µm, n = 20), oblong-elliptic, tapering slightly to rounded ends, one side flat in side view, sometimes slightly curved, 0-septate, hyaline.

Figure 6. 

Brahmaculus packhamiae (PDD 117311) A, B dried apothecia C detail of head of dried apothecium D hymenial surface of rehydrated apothecium, showing multiple separate apothecial cups E squash mount showing excipular cells and hairs in KOH + Melzer’s reagent F ascoma in vertical section G paraphyses, asci, and ascospores H ascospores. Scale bars: 1 mm (A–C); 0.1 mm (D); 10 µm (E–H).

Brahmaculus packhamiae is macroscopically and microscopically similar to the Chilean B. magellanicus, both species having relatively long and narrow stipes. Notes with the specimen, indicate that when fresh the ascomata “seem to be attached to roots”.

New Zealand – Bay of Plenty • vic. Rotorua, Tarawera Falls (-38.1573, 176.5193); on fallen leaves Metrosideros excelsa; P.R. Johnston (D2565) leg.; 16 May 2019; PDD 116740 – holotype; ICMP 23410 – ex type culture.

Refers to the host substrate of the known specimens.

Phylogenetically a Chlorociboria, differs in developing on dead leaves rather than wood and in the asci being 4-spored when mature.

Apothecia developing on partly decomposed fallen leaves, not associated with pigmentation of substrate. Apothecia less than 1 mm diam., sessile, with short, matted hairs around the margin, hymenium yellow. Hairs 20–45 × 4 µm, cylindric, walls thin, roughened. Apothecium in vertical section with ectal excipulum 30–40 µm wide, comprising short, broad-cylindric cells 5–7.5 µm diam., with walls hyaline, slightly thickened, rows of cells arranged at a high angle to the receptacle surface near the base of the cup, more parallel to the surface near edge of cup; medullary excipulum of narrow-cylindric cells with thin walls. Paraphyses 2–3 µm diam., taper slightly and gradually to rounded apex, extending 5–10 µm beyond asci. Asci 40–55 × 5.5–7 µm, cylindric, tapering slightly to broad, subtruncate apex, wall thickened at apex with amyloid pore extending as two narrow, parallel bands extending through the wall, initially with 8 spores, 4 spores aborting and 4–spored at maturity, crozier present. Ascospores 7.5–9.5 × 2.5–3.5 µm (average 8.3 × 3.1 µm, n = 20), oblong-elliptic, tapering to rounded ends, one side flat in side view, widest point towards one end, 0–septate, hyaline.

Figure 7. 

Chlorociboria metrosideri (PDD 116740) A fresh apothecia B margin of receptacle in vertical section C surface of receptacle in squash mount showing rough-walled hairs D apex of asci and paraphyses E immature ascus with 8 spores, and mature asci with 4 spores. Scale bars: 0.1 mm (A); 20 µm (B, C); 10 µm (D, E).

New Zealand – Auckland • Rangitoto Island, Kidney Fern Glen; -36.805544, 174.860064; on fallen, partly rotten Metrosideros excelsa leaves; P.R. Johnston (D2329) leg.; 23 Apr 2012; PDD 102723.

The substrate in both specimens was partly rotted leaves. It is possible that this fungus has a broader host range as most host-specialised, leaf-inhabiting Leotiomycetes are found on recently fallen leaves of their preferred host. Cultures are slow growing (on PDA, 9 mm after 8 weeks) with sparse mycelium and pale brownish pigmentation, remaining sterile.

New Zealand – Fiordland • Kepler Track, control gates; -45.4396, 167.6822; on Nothofagaceae sp. dead wood; P.R. Johnston (D1484), R.E. Beever, S.R. Pennycook, R. Leschen, T. Lebel leg.; 10 May 2000; PDD 77447 – holotype; ICMP 18766 – ex type culture.

Refers to the country of origin, in contrast to Argentina and the morphologically similar C. argentinensis, with which C. novae-zelandiae was previously confused.

Similar to Chlorociboria argentinensis in having small, allantoid ascospores and lacking tomentum hyphae, but phylogenetically distinct and with smaller ascospores and narrower asci.

C. novae-zelandiae : New Zealand – Fiordland. • Borland Lodge nature trail; on Nothofagaceae sp. dead wood; P.R. Johnston (D1471.2) leg.; 9 May 2000; PDD 77446 – North Canterbury. • Mt Thomas Forest, Richardson Track; on Nothofagaceae sp. dead wood; P.R. Johnston (D679) leg.; 15 Mar 1991; PDD 58574, ICMP 15616 – Nelson. • Arthur Range, Graham Valley Rd, track from Flora car park to Mt Arthur Hut; P.R. Johnston (D993) leg.; 6 May 1994; PDD 77444.

C. argentinensis : Argentina – Tierra del Fuego • Lago Fagnano, vic. Kosobo, road to hot springs; on Nothofagus pumilio fallen wood; P.R. Johnston (SA86), L. Lorenzo leg.; 22 Mar 1996; PDD 92026; ICMP 16994 – Patagonia. • Rio Negro, Nahuel Huapai National Park, path from Puerto Blest to Los Cantaros; on Nothofagus dombeyi fallen wood; P.R. Johnston (SA 188), I. Gamundí, C. Brion leg.; 2 Apr 1996; PDD 92027; ICMP 16995.

Johnston and Park (2005: 690–693) provided a description of C. novae-zelandiae, from New Zealand specimens reported under the name C. argentinensis. Subsequent DNA sequencing of specimens from Argentina identified as C. argentinensis, showed that the New Zealand species is phylogenetically distinct. Morphologically, the two species are similar, both with an excipulum comprising highly gelatinous textura intricata, the apothecia lacking hair-like tomentum hyphae, and with small, allantoid ascospores. The New Zealand species has somewhat smaller ascospores (average 7.0 × 1.5 µm versus 9.9 × 1.9 µm) and narrower asci (3.5–4.5 µm versus 4–5.5 µm) compared with the Argentinian specimens recognised here as C. argentinensis. The Argentinian specimens match closely the description of Dixon (1975).

New Zealand – Fiordland • Fiordland National park, Kepler Track, Rainbow Reach; -45.4429, 167.6802; on Fuscopora solandri fallen leaves; P.R. Johnston (D686) leg.; 17 Mar 1991; PDD 58580 – holotype; ICMP 23686 – ex-type culture.

Refers to the host substrate of the holotype.

Phylogenetically a Chlorociboria, developing on fallen leaves rather than wood, differs from Chlorociboria metrosideri in having flexuous, coiled hairs and lanceolate paraphyses.

Apothecia developing on fallen leaves, not associated with any pigmentation of substrate. Apothecia less than 1 mm diam., short-stipitate, receptacle densely covered with short, white hairs, hymenium pale yellow. Hairs 30–40 × 3–4 µm, short-cylindric, undifferentiated to apex, septate, thin-walled, roughened all over, flexuous, coiled and tangled. Apothecia in vertical section with ectal excipulum 45 µm wide, comprising short-cylindric to subglobose cells 5–8 µm diam. oriented at high angle to receptacle surface, with walls hyaline, thickened, agglutinated, amyloid in some specimens. Medullary excipulum non-gelatinous, comprising narrow-cylindric hyphae with thin walls. Tissue at base of stipe of gelatinous textura intricata. Paraphyses up to 5 µm diam., lanceolate, tapering to narrow rounded apex, extending 20–30 µm beyond asci, wall distinctively thickened at base, amyloid in some specimens. Asci 40–55 × 4.5–5.5 µm, cylindric, tapering gradually to small, subtruncate apex, wall thickened at apex, amyloid pore extending through wall, flaring towards outside of wall, crozier present, 8–spored. Ascospores 8–11.5 × 1.5–2 µm (average 10.0 × 1.7 µm, n = 20), oblong-elliptic to subfusoid, widest point slightly towards the upper end, taper to narrow-rounded ends, 0–septate, hyaline.

Figure 8. 

Chlorociboria solandri A dried apothecium B fresh apothecium C margin of receptacle in vertical section D ascospores E paraphyses F amyloid paraphysis G base of paraphysis with thick wall H details of coiling, rough-walled excipular hairs. Images: PDD 58580 (A, C, D, F–H); PDD 61833 (B, E). Scale bars: 0.1 mm (A, B); 20 µm (C); 10 µm (D–H).

New Zealand – Taupo • Kaimanawa Forest Park, Tree Trunk Gorge; on Fuscopora solandri fallen leaves; P.R. Johnston (D877), I. Gamundí leg.; 1 Feb 1993; PDD 61833 – Mid Canterbury • Craigieburn, Cave Stream; on Fuscopora solandi fallen leaves; E. Horak leg.; 31 Mar 1983; PDD 92925.

Chlorociboria solandri is micromorphologically distinctive in having scattered, large, lanceolate paraphyses, short-cylindric to more or less globose, thick-walled excipular cells, excipular tissue reacting either blue or red to Melzer’s reagent, and coiling, rough-walled hairs. Known from two specimens from Fuscopora solandri leaves. A third specimen in poor condition, PDD 92925, could be the same species; it is morphologically similar but has longer hairs than the other two specimens. Cultures on agar are very slow growing (10 cm after 4 weeks), have little aerial mycelium and pale yellow brown pigments, remaining sterile.

New Zealand – Westland • Haast Pass Summit, Lookout Track; -44.1063, 169.3519; on fallen leaves Dracophyllum sp.; P.R. Johnston (D2515), M. Padamsee leg.; 16 May 2018; PDD 112247 – holotype.

From subtilis (delicate) referring to the stature of the apothecia.

Blue-green apothecia on blue-green stained fallen, partly decomposed leaves, hairs on receptacle rough-walled, somewhat flexuous, ascospores filiform, 45–55 × 1 µm.

Apothecia erumpent from blue-green stained leaf tissue. Apothecia less than 1 mm diam., cupulate with short, broad stipe, receptacle pale blue-green with tangled, white hairs, especially near the edge of the cup. Hairs 55–75 × 3–4 µm, somewhat flexuous, wall roughened. Apothecia in vertical section with ectal excipulum up to 30 µm wide, cells 6–10 µm diam., short-cylindric to square, walls thick, cells arranged in rows with a high angle to the receptacle surface; medullary excipulum poorly developed, two or three rows of narrow-cylindric cells, walls encrusted with blue-green material; stipe with thick-walled textura intricata. Paraphyses 1.5–2 µm diam., slightly wider towards the apex, often branched in the upper 20–30 µm, extending 15 µm beyond asci. Asci 85–105 µm × 5.5–6.5 µm cylindric, tapering gradually to small, truncate apex, wall thickened at apex, amyloid pore in inner half of wall, reaction most intense on inner edge of wall, pore appears more or less U-shaped, sloping outwards slightly through the wall, 8-spored, spores confined to the upper 60–100 µm of ascus, crozier present. Ascospores 45–55 × 1 µm, filiform, straight, 0–septate, hyaline.

Figure 9. 

Chlorociboria subtilus (PDD 112247) A dried apothecium B apothecium in vertical section C detail of margin of receptacle in vertical section D asci, ascospores and paraphyses E detail of apex of paraphyses and asci F excipular hairs (squash mount). Scale bars: 0.1 mm (A); 100 µm (B); 10 µm (C–F).

New Zealand – Nelson • Arthur Range, Graham River Valley Rd, track from Flora car park to Mt Arthur Hut; on Dracophyllum pyrimidale fallen leaves; P.R. Johnston (D990) leg.; 6 May 1994; PDD 105292 – Central Otago •vic. Dunedin, Great Moss Swamp; on Dracophyllum uniflorum fallen leaves; P.R. Johnston (D82) leg.; 12 May 1984; PDD 105293 – Mid Canterbury • Craigieburn, Cave Stream; on Dracophyllum uniflorum fallen leaves; P.R. Johnston (D248) leg.; 23 Feb 1988; PDD 105294 – Taupo • Tongariro National Park, Ohakune Mountain Road, Blyth Track; on Fuscopora cliffortioides fallen leaves; P.R. Johnston (D353) leg.; 20 May 1989; PDD 55523 • Rangitoto Station, Ranginui Summit; on Dracophyllum pyrimidale fallen leaves; P.R. Johnston (D1622), S.R. Whitton leg.; 6 May 2001; PDD 117584.

Most specimens are on fallen leaves of Dracophyllum spp., but the host range may be more extensive. A specimen on Fuscopora cliffortioides (PDD 55523) is morphologically similar, but perhaps with longer ascospores.