The lichen family Graphidaceae is widely distributed in the tropical and subtropical regions of the world. Prior to 2002, the schematic generic concepts based on apothecial and ascospore types first established by Müller Argoviensis (1880 Reference Müller Argoviensisa , Reference Müller Argoviensisb ) were followed to describe and classify species of the family. Staiger (Reference Staiger2002) provided a new systematic revision based on structure of the thallus and exciple, type of hymenium and ascospore, and secondary chemistry. Later, Frisch et al. (Reference Frisch, Kalb and Grube2006), Mangold et al. (Reference Mangold, Elix and Lumbsch2009), Nelsen et al. (Reference Nelsen, Lücking, Rivas Plata and Mbatchou2010), Lücking et al. (Reference Lücking, Rivas Plata, Kalb, Common, Barcenas-Peña and Duya2011, Reference Lücking, Parnmen and Lumbsch2012, Reference Lücking, Tehler, Bungartz, Rivas Plata and Lumbsch2013), Rivas Plata et al. (Reference Rivas Plata, Kalb and Frisch2010, 2012 Reference Rivas Plata, Lücking and Lumbscha , Reference Rivas Plata, Lücking and Lumbschb , Reference Rivas Plata, Parnmen, Staiger, Mangold, Frisch, Weerakoon, Hernández, J., Cáceres, Kalb and Sipman2013), Cáceres et al. (Reference Cáceres, Rivas Plata and Lücking2012, Reference Cáceres, Aptroot, Parnmen and Lücking2014), Parnmen et al. (Reference Parnmen, Lücking and Lumbsch2012, Reference Parnmen, Cáceres, Lücking and Lumbsch2013), Sipman et al. (Reference Sipman, Lücking, Aptroot, Chaves, Kalb and Umaña Tenorio2012), Kraichak et al. (Reference Kraichak, Parnmen, Lücking and Lumbsch2014) and Mercado-Diaz et al. (Reference Mercado-Diaz, Lücking and Parnmen2014) established many new genera in the family based on molecular data. These changes have resulted in the transfer of many species from earlier established genera, such as Graphis, Phaeographis, Graphina and Phaeographina, to other genera. As such, Graphina is now a synonym of Graphis (Staiger Reference Staiger2002) and Phaeographina an illegitimate name (Lücking et al. Reference Lücking, Kalb, Staiger and McNeill2007). These changes in the generic concept of the family have necessitated the rearrangement of a number of Indian taxa.
In an earlier publication, Singh & Swarnlatha (Reference Singh and Swarnlatha2009) made eight new combinations. As a result of our continuing study on Indian Graphidaceae and examination of more type specimens preserved at Agarkar Research Institute, Pune, India (AMH), Botanical Survey of India, Allahabad (BSA), Botanische Staatssammlung München (M), British Museum of Natural History, London (BM), and the National Botanical Research Institute, Lucknow, India (LWG), we further propose six new combinations, namely Diorygma aeolum, D. spilotum, Kalbographa hypoglaucoides, Pallidogramme awasthii, P. divaricoides and Phaeographis firmula. In addition, Diorygma megasporum, Phaeographis albolabiata and Acanthothecis subconsocians are synonymized with Diorygma aeolum, Nitidochapsa leprieurii and Pallidogramme divaricoides, respectively.
Type specimens deposited in AMH, BM and LWG-LWU herbaria were examined morphologically, anatomically and chemically. Morphological characters of thalli (reproductive structures, colour, size and shape) were examined using a Nikon SMZ 1500 stereomicroscope. Thin hand-cut sections of thalli and ascomata, mounted in an aqueous solution of KOH, were examined. All anatomical measurements were made in water mounts and examined using a Nikon Eclipse 50i compound microscope. Ascospores were stained with Lugol’s solution to check the amyloid reaction and measured in water. Secondary metabolites were identified by thin-layer chromatography (TLC) using solvent A (180 toluene: 45 dioxane: 5 acetic acid), following Orange et al. (Reference Orange, James and White2001).
New Combinations
Diorygma aeolum (Stirt.) Pushpi Singh & Kr. P. Singh comb. nov.
MycoBank No.: MB 819997
Graphis aeola Stirt., Proc. Roy. Soc. Glasgow 11: 316 (1879).—Graphina aeola (Stirt.) Zahlbr., Cat. Lich. Univ. 2: 394 (1923); type: India, Tamil Nadu, ‘Nelghiri’ (Nilgiri hills), G. Watt (BM 001097383!—lectotype).
Diorygma megasporum Kalb, Staiger & Elix, Symb. Bot. Upsal. 34(1): 160 (2004); type: Myanmar, Yomah, Khaboung Pass, 2000 ft, 21.01.1868, S. Kurz, s. n (M 0102172!—holotype).
(Fig. 1A)
Fig. 1 Habits of species studied. A, Diorygma aeolum (lectotype); B, Diorygma spilotum (lectotype); C, Pallidogramme awasthii (holotype); D, Pallidogramme divaricoides (holotype); E, Phaeographis firmula (lectotype); F, Nitidochapsa leprieurii (holotype of Phaeographis albolabiata). Scales=1 mm. In colour online.
Thallus corticolous, whitish to pale grey, ecorticate, basal layer not carbonaceous.
Apothecia lirelliform, immersed, irregularly branched; disc closed to slit-like, ±pruinose; exciple uncarbonized, convergent to slightly divergent; hymenium not inspersed, weakly to distinctly I+ blue in lateral parts; asci 4–6-spored; ascospores colourless, ellipsoidal, muriform with all locules of equal size, (76–)90–120×26–40 µm, I+ violet; stictic acid (major) and constictic acid (minor) present.
Notes. Examination of the type material of Graphis aeola, borrowed from BM, revealed that it belongs to the genus Diorygma. The type of Diorygma megasporum Kalb et al., borrowed from M, originates from Myanmar (Burma) and not from India as reported earlier (Kalb et al. Reference Kalb, Staiger and Elix2004). It agrees with that of D. aeolum in morphology, anatomy, the 4–6-spored asci, ascospore size and chemistry. Therefore, D. megasporum is synonymized with D. aeolum. Morphologically, Diorygma aeolum resembles D. junghuhnii (Mont. & Bosch) Zahlbr. which has 1-spored asci and norstictic acid. In chemistry, it also closely resembles D. albovirescens Makhija et al., which has smaller, 66–99×12–36 µm ascospores (Makhija et al. Reference Makhija, Chitale and Sharma2009). So far, D. aeolum is known from India and Myanmar and widely distributed in hilly regions.
Diorygma spilotum (Stirt.) Pushpi Singh & Kr. P. Singh comb. nov.
MycoBank No.: MB 819998
Graphis spilota Stirt., Proc. Roy. Soc. Glasgow 13: 187 (1881).—Graphina spilota (Stirt.) Zahlbr., Cat. Lich. Univ. 2: 425 (1923); type: India, Assam, 1879, A. Watt, s. n. (BM 001097478! —lectotype).
(Fig. 1B)
Thallus corticolous, pale or creamish to whitish-greyish, ecorticate, basal layer not carbonaceous.
Apothecia lirelliform, simple, immersed, brownish black; disc exposed, brownish, epruinose; exciple uncarbonized, divergent; hymenium not inspersed, laterally weak I+ blue; asci 1-spored; ascospores colourless, muriform with all locules ± of equal size, 90–110(–140)×30–45 µm, I− or I+ faintly violet; wall thickening I−; norstictic acid present.
Notes. Examination of the type material of Graphis spilota, borrowed from BM, revealed that it belongs to the genus Diorygma. In its chemistry and 1-spored asci, D. spilotum resembles Diorygma junghuhnii (Mont. & Bosch) Kalb et al., D. tuberculosum (Stirt.) Kalb et al. and D. soozanum (Zahlbr.) M. Nakan. & Kashiw. Diorygma junghuhnii differs by its erumpent to prominent lirellae and completely I+ blue hymenium, while D. tuberculosum differs by its lirellae being distinctly raised from the thallus surface and its peripheral ascospore locules being distinctly smaller than central ones. Diorygma soozanum is distinguished by its erumpent lirellae with a thick whitish pruinose disc (Kalb et al. Reference Kalb, Staiger and Elix2004). The species is endemic to India and so far known only from the type locality.
Kalbographa hypoglaucoides (Kr. P. Singh & D. D. Awasthi) Kr. P. Singh & Pushpi Singh comb. nov.
MycoBank No.: MB 819999
Phaeographis hypoglaucoides Kr. P. Singh & D. D. Awasthi, Bull. Bot. Surv. India 21(1–4): 109 (1979); type: India, Tamil Nadu, Palni Hills, on way to Thandikuddi, alt. 1350 m, D. D. Awasthi & K. P. Singh 70.459 (LWG-LWU!—holotype).
Fig. 2 Kalbographa hypoglaucoides (holotype). A, habit; B, cross-section of apothecia; C, mature ascospores. Scales: A=1 mm; B=100 μm; C=20 μm. In colour online.
Thallus saxicolous, grey to brown, smooth to cracked; cortex indistinct.
Apothecia lirelliform, immersed, simple to bifurcate; disc exposed, plane to slightly concave, black, covered by faintly whitish pruina; exciple uncarbonized; hymenium not inspersed; asci 8-spored; ascospores brown, ovoid-ellipsoid, thin-walled, 3-septate, 10–13×5·0–7·5 µm, I−; no lichen substances present.
Notes. Examination of the type material of Phaeographis hypoglaucoides, from LWG-LWU, revealed that it belongs to the genus Kalbographa. So far, four species of Kalbographa are known, viz. K. caracasana (Müll. Arg.) Lücking, K. lobata Lücking, K. lueckingii Kalb and K. miniata Lücking. All are characterized by their corticolous habit, in addition to submuriform ascospores in K. caracasana, K. lobata and K. miniata (Lücking Reference Lücking2007). The remaining species, K. lueckingii, produces 5(–6)-septate, 18–23 µm long ascospores, and norstictic and connorstictic acids as secondary substances (Kalb et al. Reference Kalb, Buaruang, Papong and Boonpragob2009). Kalbographa should not be confused with Phaeographopsis, which also has the same type of ascospores but differs in having an ecorticate thallus and a pseudomazaedium.
Pallidogramme awasthii (Patw. & C. R. Kulk.) Kr. P. Singh & Pushpi Singh comb. nov.
MycoBank No.: MB 820010
Phaeographina awasthii Patw. & C. R. Kulk., Norw. J. Bot. 26: 49 (1979); type: India, Kerala, Idukki District, Anamalai Hills, near Munnar, Chinnar, C. R. Kulkarni & A. V. Prabhu 76.462 (AMH!—holotype).
(Fig. 1C)
Thallus corticolous, brown, glossy, smooth, corticate.
Apothecia lirelliform, emergent to prominent, ashy white to dull brownish, simple to flexuous; disc narrow slit-like; exciple uncarbonized, brownish, striate; hymenium inspersed; asci 8-spored; ascospores brown, muriform, 80–105×20–30 µm, I+ reddish brown; lichen substances absent.
Notes. Examination of the type material of Phaeographina awasthii, from LWG-LWU, revealed that it belongs to the genus Pallidogramme. In morphology, it closely resembles Pallidogramme bengalense B. O. Sharma & Khadilkar which has smaller, 50–80×12–20 µm ascospores (Sharma & Khadilkar Reference Sharma and Khadilkar2011). It also resembles P. chlorocarpoides (Nyl.) Staiger et al. and P. chrysenteron (Mont.) Staiger et al., but P. chlorocarpoides is distinguished by its 2–4-spored asci and presence of stictic and constictic acids, while P. chrysenteron differs by having 6–8-spored asci and smaller (33–65×10–15 µm; vide Staiger Reference Staiger2002) ascospores, and also by the production of stictic and constictic acids. Pallidogramme awasthii is endemic to India and distributed in the states of Karnataka, Kerala and West Bengal.
Pallidogramme divaricoides (Räsänen) Pushpi Singh & Kr. P. Singh comb. nov.
MycoBank No.: MB 820017
Phaeographis divaricoides Räsänen, Arch. Soc. Zool. Bot. Fenn. Vanamo 5: 31 (1951); type: India, E. Himalayas, Sikkim State, Gangtok, alt. 2100 m, D. D. Awasthi 219 (LWG-AWAS!—isotype).
Acanthothecis subconsocians Pooja Gupta & G. P. Sinha, J. New Biol. Rep. 4(2): 98 (2015); type: India, Sikkim, East District, Pangthang-Rokshe, alt. c. 1900 m, 2006, G. P. Sinha 3621 (BSA!—holotype).
(Fig. 1D)
Thallus corticolous, brownish, glossy, smooth, corticate.
Apothecia lirelliform, emergent to prominent, dull brownish, divaricately branched; disc closed to narrow, brownish, epruinose; exciple uncarbonized, brownish, striate; hymenium inspersed; ascospores hyaline to brown, (5–)7–9-septate, 20–35×7–10 µm, I+ reddish brown; no lichen substances present.
Notes. Examination of the type material of Phaeographis divaricoides, from LWG-LWU, revealed that it belongs to the genus Pallidogramme. Only two other Pallidogramme species with transversely septate ascospores, viz. P. indica A. Dube & Makhija and P. undulatolirellata A. Dube & Makhija, occur in India. Both species can be distinguished from P. divaricoides by their clear hymenia, larger ascospores and presence of secondary metabolites (Chitale et al. Reference Chitale, Makhija and Sharma2009). The type of Acanthothecis subconsocians Pooja Gupta & G. P. Sinha, preserved at BSA, agrees with that of Pallidogramme divaricoides in morphology, anatomy, ascospore size and chemistry. Thus, the two taxa are conspecific and P. divaricoides is the correct name. In India, P. divaricoides is widely distributed in Arunachal Pradesh, Nagaland, Sikkim and hilly areas of West Bengal.
Additional specimens examined. India: Arunachal Pradesh: West Kameng District, Simipam forest, alt. 1500–1250 m, K. P. Singh 9904C (BSA). Sikkim: North Sikkim District, Tangla Br., alt. 1689 m, G. Swarnlatha 5903C (BSA). West Bengal: Darjeeling District, Takdah Tea Estate, near Char mile, V. K. Singh 2313 (BSA).
Phaeographis firmula (Stirt.) Pushpi Singh & Kr. P. Singh comb. nov.
MycoBank No.: MB 820019
Graphis firmula Stirt., Proc. Roy. Soc. Glasgow 13: 186 (1881).—Phaeographina firmula (Stirt.) Zahlbr., Cat. Lich. Univ. 2: 438 (1923); type: Assam, A. Watt (BM 001106502!—lectotype).
(Fig. 1E)
Thallus corticolous, smooth, shiny, brownish, corticate.
Apothecia lirelliform, immersed, simple to trifurcate; disc exposed, epruinose; labia entire; exciple apically pale brown to brown; hymenium clear; asci 8-spored; ascospores brown, muriform, 18–26×8–12 µm; no lichen substances present.
Notes. Examination of the type material of Graphis firmula, from BM, revealed that it belongs to the genus Phaeographis. It closely resembles P. noralboradians (Patw. & C. R. Kulk.) Makhija & Chitale which has a densely pruinose disc and norstictic acid as a secondary substance (Makhija et al. Reference Makhija, Chitale and Dube2014). In ascospore characters it resembles Phaeographis schizolomoides Poengs. & Kalb, but the latter species differs in the inspersed hymenium and the norstictic acid chemistry. The species is endemic to India.
Nitidochapsa leprieurii (Mont.) Parnmen, Lücking & Lumbsch
Bryologist 116(2): 129 (2013).—Stictis leprieurii Mont., Ann. Sci. Nat., Bot., sér. 4 3: 97 (1855).—Chapsa leprieurii (Mont.) Frisch, Biblioth. Lichenol. 92: 105 (2006); type: French Guiana, Cayenne, Leprieur 804 (PC— lectotype; G—isolectotype).
Phaeographis albolabiata Patw. & C. R. Kulk., Norw. J. Bot. 26(1): 48 (1979); type: India, Kerala, Anamalai Hills, Sholayar Forest, C. R. Kulkarni 76.229 (AMH!—holotype).
(Fig. 1F)
Notes. Patwardhan & Kulkarni (Reference Patwardhan and Kulkarni1979) described Phaeographis albolabiata Patw. & C. R. Kulk. as a new species from India. We have examined the type (AMH!) and confirmed that it agrees with Nitidochapsa leprieurii in morphology, anatomy and chemistry. The former is therefore synonymized here.
The authors thank the Director of the Botanical Survey of India, Kolkata, for encouragement, and Dr G. P. Sinha, Head of Office, Botanical Lichen Society Survey of India, Central Regional Centre, Allahabad, for providing facilities. The authors are also grateful to the curators of herbaria AMH, BM, LWG and M for the loan of type specimens. We thank Dr Robert Lücking for valuable suggestions and two anonymous reviewers for useful comments and suggestions on the manuscript. We also thank Dr D. K. Singh, Lucknow, for revising the manuscript. One of the authors (PS) is grateful to the authorities at the Botanical Survey of India for financial assistance under the ‘Flora of India Project’.
