Ramalina asahinae W. Culberson & C. Culberson

Marcano, Vicente, Méndez, Antonio Morales & Prü, Ernesto Palacios, 2021, The genus Ramalina Acharius (Ascomycota, Lecanoromycetes, Ramalinaceae) in northern South America, Phytotaxa 504 (1), pp. 1-77 : 30-35

5. Ramalina asahinae W. Culberson & C. Culberson

J. Jap. Bot. 51: 374 (1976). Type:― MEXICO. Chiapas: 11 km west of San Cristobal Las Casas , 2154 m (holotype DUKE).

Ramalina grumosa Kashiwadani , in H. Inoue (ed.), Stud. Crypt. South. Perú, Tokai Univ. Pres., 134 (1987). Type:― PERÚ. Pasco: Oxapampa, around Laguna El Oconal, on tree bark, 1500 m, H . Kashiwadani 22541 (holotype TNS) .

Thallus corticolous, fruticose, whitish green in field, pale yellow in herbaria, growing from a narrow holdfast, up to 2.0 mm in thick, branching dichotomously anisotomic. Branches solid, irregular flat, 5–10 cm long, 0.25–1.25 mm wide, surface smooth, slender or broad, soft or stiff. Pseudocyphellae ellipsoid to short linear, 0.5–4 mm wide, lateral. Isidial structures granular or cylindrical, sometimes absent. Soralia granular, ellipsoid to labriform. Cortical tissue distinct. Chondroid tissue weakly, or not, cracked. Medulla white, compact. Pycnidia and apothecia not seen.

Chemistry (TLC, HPTLC): This species presents 8 strains which are summarized in the Table 4. Strain 1. Galbinic (tr.), consalazinic, salazinic and protocetraric acids (Vareschi 6208). Strain 2. Salazinic and protocetraric acids (Vareschi 6212). Strain 3. Homosekikaic, sekikaic, boninic, succinprotocetraric and protocetraric acids (Vareschi 3495). Strain 4. Protocetraric acid (Marcano 171–92; Morales 138, Marcano 150–92, 151–92, 152–92, 154–92; Morales 88). Strain 5. Sekikaic and protocetraric acids (Marcano 161–92). Strain 6. Sekikaic and boninic acids (Aguirre & Sipman 5727–C). Strain 7. Sekikaic, ramalinolic and boninic acids (Valencia & Boekhout 164–A; Sipman et al. 23613–A, Florschütz 3618–A). Strain 8. Salazinic, consalazinic acids and 1 unknown compound (Vareschi 6174).

Ecology and distribution: This species grows on trees dead trunks tree in pastures at 950–3500 m. It is known from México and South America ( Brazil, Ecuador, Perú, Colombia, Venezuela, Paraguay and Uruguay).

Remarks: Ramalina grumosa is considered to be a synonym of R. asahinae as both species have branchlets which often change into granular or cylindrical isidial structures and/or show granular soralia in its ending. Field observations usually revealed the presence of individuals (Morales 88, 138; Marcano 154–89) from a single population having isidiform-granular structures, others without isidial structures, and an irregularly continuous chondroid tissue (Fig. 25). Likewise, these individuals showed several chemosyndromes. Ramalina asahinae is a chemically variable species with sekikaic, boninic and ramalinolic acids (strains 3, 6, 7) and/or salazinic, protocetraric acids and aggregates (strains 1, 2, 4, 5, 8) as major metabolites ( Table 4). Kashiwadani & Kalb (1993) showed that R. asahinae from Brazil contained boninic acid as a major chemical substance, accompanied by 2– O –methylsekikaic and 4– O –methylpaludosic acids as minor compounds. The same substances were reported by these authors from specimens of R. grumosa . Chester & Elix (1978) identified these substances for first time from the type of R. asahinae . However, 2– O –methylsekikaic and 4– O –methylpaludosic acids were not found in the specimens of this species from Colombia and Venezuela. More recently Gumboski (2016) reported only boninic acid in R. asahinae from Brazil.

Populations of R. asahinae from Venezuela and Colombia, and Brazil could represent different ecological and geographical varieties such as occur in R. reducta and R. canaguensis . Therefore, molecular studies including specimens from those three countries are necessary in order to determine the significance of its morphological variation and distribution pattern.

Specimens examined: COLOMBIA: Cundinamarca: Between Cogua and San Cayetano, near Laguna Seca , 3450 m, 13 September 1972, P. A . Florschütz 3618– A ( B); Municipio San Francisco, vereda Sabaneta, near Cueva Grande, 2500 m, 17 July 1986, H . Sipman , H . Cardozo & M . Ballestreros 23613– A ( B, COL) . Tolima: Municipio Santa Isabel, El Ochoral, near quebrada Las Damas , 3130 m, 25 march 1988, H . Valencia & T . Boekhout 166– A ( B, COL) . Huila: Municipio La Plata, E-side of Cordillera Central, vereda La Candelaria , Finca Merenberg , 2400 m, 30 September 1984, J . Aguirre & H . Sipman 5727– C ( B, COL). Valle: Municipio Zarzal, Hacienda El Medio, between la Paila and Zarzal , 950 m, 17 February 1988, P. A . Silverstone-Sopkin 3577 ( B) . VENEZUELA: Miranda: Near Los Guayabitos, 1 August 1954, Vareschi 3495 ( VEN); Los Guayabitos, 8 January 1956, Vareschi 6208, 6212 ( VEN); Los Guayabitos, 8 January 1957, Vareschi 6174 ( VEN) . Mérida: La Culata massif, Paramo de los Conejos road, 2600 m, 6 april 1989, V . Marcano 154–89 ( MER), Santo Domingo, Paramo de Mucubají , 3500 M , March 1992, V . Marcano 150–92, 151–92, 152–92, 161–92, 171–92, 174– B – 92 ( MER); Hacienda Los Topes, San Juanito, Chiguará, 1200 m, 9 April 1983, A . Morales 88 ( B, MERF); near Chiguará, Los Topes, A . Morales 138 ( MER) .

. Ramalina aspera Räsänen

Ann. Bot. Soc. Zool. Bot. Fenn. Vanamo 20: 5 (1944). Type :― PARAGUAY. Chaco : Rio Verde, Villa Hayes, W . G . Herter 3 (holotype H!) .

Thallus corticolous, rugose, 1–2 cm high, pale yellow, sparingly branched. Branches solid, flat, dorsiventral Pseudocyphellae tuberculated, laminal or marginal, 0.1–0.3 mm wide. Cortex yellowish, chondroid tissue not cracked, discontinuous, medulla white, algal layer continuous. Soredia absent. Pycnidia not seen. Apothecia rounded, subapical to apical, laminal or marginal, concave to flat, weakly pruinous. Ascospores 1–septate, ellipsoid, 10–12 x 3.5–4 µm.

Chemistry (TLC,HPTLC):Strain1.Protocetraric acid(Vareschi3323).Strain2.Divaricatic and cryptochlorophaeic acids (Marcano 1–92; Vareschi 6156).

Ecology and distribution: This species was found growing over Citrus sp. and Cactaceae in dry forests at 1200– 1900 m. It is known from East Africa ( Kenya) and South America (the Galápagos Islands, Brazil, Paraguay and Venezuela).

Remarks: This species may be confused with R. caracasana , but the latter species is distinguished by possessing fusiform spores (17–20 µm long) ( Krog & Swinscow 1975). Kashiwadani & Kalb (1993) reported the spores to be 12–14 µm long in R. aspera from Brazil. In East Africa, R. aspera exhibits two chemical strains, one with boninic acid, the other with cryptochlorophaeic acid ( Krog & Swinscow 1975, Swinscow & Krog 1988), whereas the Brazilian specimens contain divaricatic acid (strain 1) and cryptochlorophaeic acid (strain 2) ( Kashiwadani & Kalb 1993, Gumboski 2016). Kashiwadani & Kalb (1993) and Aptroot & Bungartz (2007) claimed that R. aspera might be confused with R. complanata , which differs in having more flattened lobes containing salazinic acid, protocetraric acid and its aggregates and lacking cryptochlorophaeic acid.

Specimens examined: VENEZUELA: Lara: Near to Barquisimeto, on xerophytic vegetation, 1200 m, Vareschi 3323 ( VEN) . Miranda: Los Guayabitos, on lemon tree, 1 January 1957, Vareschi 6156 ( VEN) . Mérida: Near Bailadores, Las Playitas , 1900 m, 13 April 1992, V . Marcano 1–92 ( MER) .

. Ramalina asperula Krempelhuber

Verhandl. Zool. Bot. Gesellsch Wien. 26: 441 (1876) . Type:― PERÚ. Lima: leg. Dr Barranca, comm. Dr Wawra (lectotype in M, fide Krog & Swinscow 1975).

Thallus solid, 1–3 cm long, moderately branched. Branches pale yellow or greenish, dorsiventral, irregular flat, rugose or reticulately wrinkled. Pseudocyphellae punctiform, plane or depressed, laminal, on both sides of the branch. Soredia not seen. Pycnidia not seen. Apothecia lateral, subapical, concave; thalline exciple pseudocyphellate, reticulately wrinkled. Ascospores 1–septate, ellipsoid, 9–12 x 3–3.5 µm.

Chemistry (TLC, HPTLC): Strain 1. Divaricatic and protocetraric acids (Vareschi 2089–A, 2291). Strain 2. Divaricatic, salazinic and protocetraric acids (Morales 155).

Ecology and distribution: This species is corticolous, growing over Cordia sp. in submontane and montane forests at 200–1200 m. It is known from East Africa ( Kenya, Tanzania) and South America ( Perú, Venezuela).

Remarks: Ramalina asperula can be confused with R. africana , but the latter species has tuberculate pseudocyphellae and larger spores (11–14 µm long). In Colombia and Venezuela R. asperula can be distinguished from R. aspera chemically by the occurrence of divaricatic acid without accompanying cryptochlorophaeic acid. Salazinic and protocetraric acids, which are present in chemical strain 2 from Venezuela, have not been reported in specimens from other regions e.g. Perú, East Africa ( Krog & Swinscow 1975, Kashiwadani 1987, Swinscow & Krog 1988).

Specimens examined: VENEZUELA: Bolívar: Caroni Falls , 1 November 1952, Vareschi 2089a ( VEN) . Dto. Federal: Cerro Casquillo, Caracas-La Guaira road, km 27, 20 January 1947, T . Lasser 2291 ( VEN) . Mérida: Near Chiguará, Los Topes , 1200 m, A . Morales 155 ( MER) .

. Ramalina bogotensis Nylander

Prodrom. Florae Nov. Granat., Lichen.: 16 (1864). Type:― COLOMBIA. Bogotá: 2700 m, In sylvis et ramis arborum longe pendula, A. Lindig 2752 (holotype H –NYL! 37512) .

Thallus corticolous, pendulous, contorted, yellowish to whitish, densely branched, up to 40 cm long. Branches flattened. Soralia variable, apical, having abundant granular soredia. Cortex distinct, yellowish, 5–10 μm thick, chondroid tissue distinctly cracked, continuous. Pycnidia not seen. Apothecia marginal or laminal, up to 0.8 mm diameter. Ascospores long-fusiform, with pseudosepta, 18–20 x 2.5–3 µm.

Chemistry (TLC, HPTLC): Strain 1. Divaricatic, 4ˊ– O –demethylsekikaic, boninic, 2ˊ– O –methylsekikaic, sekikaic, fumarprotocetraric and protocetraric acids (Mägdefrau 674). Strain 2. Sekikaic and 4ˊ– O –demethylsekikaic acids (Aguirre & Sipman 5811).

Ecology and distribution: This species is corticolous on shrubs in low secondary and montane forests at 10–2300 m. It is known only from northern South America ( Colombia and Venezuela).

Remarks: This rare species is usually confused in several herbaria with R. usnea . Both species have a contorted thallus, fusiform spores and are very similar chemically. However, R. bogotensis is distinguished by the presence of spores with pseudosepta and apical soralia producing abundant coarse granules which are absent in R. usnea . In R. bogotensis the major substances present are sekikaic acid and its aggregates, whereas in R. usnea the major substances are divaricatic acid and substances of the protocetraric acid complex. Ramalina bogotensis could be confused with R. tenaensis but the latter can be distinguished by the branches with verruculose surface, and lacks of soralia, pseudocyphellae and protocetraric acids and its aggregates.

Specimens examined: COLOMBIA: Huila: Municipio La Plata, vereda La Candelaria, E-side of Cordillera Central, headwaters of Rio La Candelaria , 2300 m, 1 October 1984, J . Aguirre & H. J. Sipman 5811 (B, COL). VENEZUELA: Carabobo: Near to Puerto Cabello, 6 April 1958, K . Mägdefrau 674 (VEN).

. Ramalina calcarata Krog & Swinscow

Norw. J. Bot. 21: 115 (1974). Type:― KENYA. Rift Valley Prov. : Uasin Gishu District, 5 km NW of Timboroa Summit, 2650 m , H. Krog & T. D. V. Swinscow no. 2k19/12y (holotype O, isotype BM).

Thallus corticolous, fistulose, subdichotomously or irregularly branched, up to 4 cm long. Branches partly compressed, dorsiventral, 0.6–2.2 mm thick, surface smooth, having coalescing perforations, orbicular or irregular, separated by strands of chondroid tissue, fibrose-reticulate. Chondroid tissue not cracked, continuous. Medulla white, continuous. Pseudocyphellae and soredia not seen. Pycnidia not seen. Apothecia numerous, subapical or apical, spurred, plane or convex, up to 1 cm diameter, yellowish brown. Ascospores 1–septate, long ellipsoid, straight or curved, 12–15 x 5–7 µm.

Chemistry (TLC, HPTLC): Strain 1. Triterpenes, divaricatic, 4ˊ– O –demethylsekikaic, salazinic and protocetraric acids (Vareschi 6161–A). Strain 2. Divaricatic and salazinic (tr.) acids (Morales 141–89). Strain 3. Salazinic acid (Morales 92–84). Strain 4. Divaricatic acid (Marcano 158–92). Strain 5. Divaricatic and protocetraric acids (Marcano 6108), the latter sometimes only in traces (Marcano 6116–B; Sipman et al. 34116–A).

Ecology and distribution: This species is corticolous in rainforests and exposed subparamo as well as in Colombia on Fraxinus in pastures at 1200–3200 m. It is known from East Africa ( Ethiopia, Kenya, Tanzania, Uganda) and South America ( Brazil, Colombia and Venezuela).

Remarks: This species closely resembles R. subcalcarata and R. pusiola . All three species are fistulose, but differ in several important characters: R. subcalcarata has short ellipsoid ascospores (8–12 x 3–4 µm) whereas R. pusiola has ellipsoid-fusiform ascospores (15–17 x 5–7 µm) and R. calcarata long ellipsoid ascospores (12–15 x 5–7 µm). Ramalina pusiola contains substances in the sekikaic acid and protocetraric acid complexes. In Brazil and East Africa Ramalina calcarata produces only divaricatic and salazinic acids ( Krog & Swinscow 1974, Kashiwadani & Kalb 1993, Gumboski 2016).

Specimens examined: COLOMBIA: Antioquia: Municipio El Retiro, along road Medellin-La Ceja , between Las Palmas and El Retiro, 2050 m, 6 July 1986, H . Sipman, M . Escobar & J . Rubiano 34116 ( B) . VENEZUELA: Mérida: Las Playitas, near Bailadores , 1950 m, 14 April 1992, V . Marcano 6100, 6103 ( MER); Paramo de La Culata , 2800– 3200 m, January 1992, V . Marcano 158–92 ( MER); near Chiguará, Los Topes , 1200 m, A . Morales 92, 141 ( MER) . Miranda: Los Guayabitos, 1 January 1957, Vareschi 6161 ( VEN) . Táchira: Garcia lake, near Pregonero , 1900 m, 14 April 1992, V . Marcano 6108 ( MER); Near Villa Paez, Delicias , 1900–2100 m, 13 April 1992, V . Marcano 6116– B ( MER) .

0. Ramalina camptospora Nylander

Bull. Soc. Linn. Normandie, sér. 2, 4: 120 (1870) . Type :― CUBA, C. Wright (holotype H–Nyl. 37248).

Thallus caespitose, corticolous, branching dichotomous or irregular, up to 7 cm long. Branches flat or subterete, irregular canaliculate, surface smooth to rarely rugose, having globuliform protuberances (which are ecorticate), 0.8– 0.1 mm thick. Pseudocyphellae continuous, concave, marginal. Isidial structures globuliform, marginal. Soralia ellipsoid or linear, scattered, concave. Pycnidia not seen. Apothecia apical or subapical, margin entire, up to 5.5 mm diameter. Ascospores 1–septate, curved or sigmoid, 12–14 x 4–4.5 µm.

Chemistry (TLC, HPTLC): Nil (Vareschi 5930, 7604–A–1) or usnic acid only (Sipman & Valencia 10300).

Ecology and distribution: This species is found growing on branches of shrubs in rain forests and paramo at 1200–3400 m. It is known from the Caribbean ( Cuba, Jamaica and Haiti), Central America ( Guatemala, Nicaragua, Costa Rica and Panama) and South America (the Galápagos Islands, Colombia, Brazil and Venezuela).

Remarks: This species could be confused with R. canaguensis var. canaguensis , but the latter can be distinguished by the dense coralliform isidial structures on branchlets, as well as the multiseptate, shorter ascospores (10–11 µm). Ramalina rectangularis is also similar, but it has punctiform pseudocyphellae and contains salazinic acid. The collections of R. camptospora from Venezuela and Colombia lacked medullary compounds. Kashiwadani & Kalb (1993) and Gumboski (2016) reported that in Brazil this species also produces usnic acid.

Specimens examined: COLOMBIA: Cundinamarca: Between Bogotá and Fusagasuga , c 5 km SW of roadtoll, 2700 m, 31 January 1979, H . Sipman & H . Valencia 10200 ( B, COL, U) . VENEZUELA: Mérida: Laguna Coromoto , 3400 m, 13 January 1957, V . Vareschi 5930 ( VEN) . Miranda: Los Guayabitos , 1200–1400 m, 1 July 1962, Vareschi 7604a–1 ( VEN)

. Ramalina canaguensis var. canaguensis V. Marcano & A. Morales

Ernstia 3: 101 (1993) . Type:― VENEZUELA. Mérida: El Molino, between Estanquez and Canaguá, 1500–2000 m, 5 November 1991, A. Morales 400 (holotype MERF!) .

Thallus corticolous, pendulous, dichotomous, up to 6 cm long. Branches solid, complanate or broadly canaliculate, pale yellow, 0.3–0.4 mm wide, with a smooth surface. Pseudocyphellae marginal, shortly linear to ellipsoid. Soralia rare, marginal, also laminal, producing coarse granules. Isidial structures numerous, forming a coralliform cylinder, marginal, originating from pseudocyphellae and soralia, up to 1 mm high. Pycnidia not seen. Apothecia rare, marginal, subterminal, disc concave or convex, 1.4–1.6 um diameter. Ascospores 1–3 septate, sigmoid to ellipsoid-curved, 10– 10.5 x 5–5.5 μm.

Chemistry (TLC, HPTLC): Nil (Vareschi 535–D; 3102–I; Wolf 793, 953; Cleef 6051–B) or traces of usnic acid only (Marcano 164–92; Vareschi 5356–A, 5767–A).

Ecology and distribution: Ramalina canaguensis var. canaguensis occurs in very moist, rainforests, growing on trees and shrubs. It seems to prefer exposed parts of the trunk. It is also found in paramo. In Colombia, R. canaguensis var. canaguensis grows on Weinmannia tolimensis Cuatrecasas and Neurolepis at 1410–3370 m. It is known only from northern South America ( Colombia and Venezuela).

Remarks: R. canaguensis is somewhat similar to R. usnea , but it differs in having disrupted pseudocyphellae, coralliform isidial structures (Fig. 26) and in lacking lichen substances. In R. usnea the thallus is contorted, has complanate laciniae and in areas where the two species overlap, R. usnea usually contains sekikaic and ramalinolic acids. Furthermore, R. usnea has long fusiform spores and lacks soralia whereas R. canaguensis has sigmoid or curvedellipsoid spores and soralia.

Ramalina canaguensis shows four chemical races each with several chemical strains. The populations of this species are morphologically indistinguishable and but they occupy distinct ecological niches according to the acids present. In this case the chemical races are regarded as taxonomically significant ( Elix 1982) and thus are here recognized at the varietal level.

Additional specimens examined: Colombia: Cundinamarca: San Cayetano, Hacienda Portugal, 2750 m, 8 November 1972, A . Cleef 6051– B ( B). Riseralda: Municipio Santa Rosa, camino real between Termales of Santa Rosa and hacienda La Sierra , 3370 m, 25 February 1986, J . Wolf 793 ( B, COL, U); municipio Santa Rosa, camino real between Termales of Santa Rosa and hacienda La Sierra, 3370 m, 24 April 1986, J . Wolf 953 ( B, COL, U) . Venezuela: Miranda: Los Guayabitos, 1410 m, 9 December 1956, V . Vareschi 535b– A , 5767– A ( VEN); Los Guayabitos, El Volcán , 1420 m, 28 December 1951, V . Vareschi 535 ( VEN) . Trujillo: Paramo del Jabón , near to Carache, 1981, M . Lopez- Figueiras et H . Rodriguez 26404 ( MERF) ; Táchira: El Portachuelo, near to Paramo El Zumbador, 21 April 1992, V . Marcano 164–92, 4285, 4286 ( MERF) .

. Ramalina canaguensis var. colombiana V. Marcano & A. Morales var. nov.

Thallus ut in Ramalina canaguensis var. canaguensis sed acidum divaricaticum et acidum protocetraricum continente differt. Type:― COLOMBIA. Cundinamarca: Municipio San Francisco, vereda Sabaneta, near Quebrada Cueva Grande , 2500 m, 17 July 1986 ,

H. Sipman 23625 (holotype B).

Chemistry (TLC, HPTLC): Divaricatic and protocetraric acids.

Ecology and distribution: Ramalina canaguensis var. colombiana occurs on trees along; track through pastures in strongly modified Quercus forest remnants at 2500 m. It is known only from Colombia.

Remarks: Ramalina canaguensis var. colombiana is known only from the type locality. It is mainly characterized by the presence of one orcinol para -depside (divaricatic acid). The other varieties show β-orcinol depsidones or usnic acid only.

The name of this variety refers to the type locality.

. Ramalina canaguensis var. guascasensis V. Marcano & A. Morales var. nov.

Thallus ut in Ramalina canaguensis var. canaguensis sed acidum salazinicum et acidum sticticum continente differt. Type:― COLOMBIA. Cundinamarca: Municipio Guasca, Paramo de Guasca, Cordillera Pena Negra , along road Guasca-Guacheta , W-

slope, valley of El Chusqual, 3200 m, 6 September 1984, J . Aguirre & H. J. M . Sipman 5147 (holotype B) .

Chemistry (TLC, HPTLC): Salazinic and stictic acids.

Ecology and distribution: Ramalina canaguensis var. guascasensis occurs as an epiphyte in mixed subparamo woodland containing Brunellia , Drimys , Miconia , Oreopanax and Clusia at 3200 m. It is known only from Colombia.

Remarks: Ramalina canaguensis var. guascasensis is known only from the type locality and is characterized by the presence of salazinic and stictic acids.

The name of this variety refers to the type locality.