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Research Article
Five new species of Graphidaceae (Ascomycota, Ostropales) from Thailand
expand article infoKhwanyuruan Naksuwankul, Ekaphan Kraichak§, Sittiporn Parnmen|, Robert Lücking, H. Thorsten Lumbsch|
‡ Mahasarakham University, Kantarawichai, Thailand
§ Kasetsart University, Bangkok, Thailand
| The Field Museum, Chicago, United States of America
¶ Botanical Museum Berlin, Berlin, Germany
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Abstract

Five new species of Graphidaceae are described from Thailand. Molecular evidence and phenotypical characters support their independent status from related and similar species. Glaucotrema thailandicum Naksuwankul, Lücking & Lumbsch is unique within the genus in having submuriform ascospores. Ocellularia klinhomii Naksuwankul, Lücking & Lumbsch is characterized by having a whitish gray, rimose thallus with ascomata in verrucae and surrounded by a black ring and lack of secondary metabolites. Ocellularia phatamensis Naksuwankul, Parnmen & Lumbsch has a grayish, thick and rimose thallus, differing from O. klinhomii in lacking a dark apothecial rim and having ascomata that are not immersed in verrucae. Ocellularia thailandica Naksuwankul, Kraichak & Lumbsch differs from O. albocincta in lacking a columella. Ocellularia rotundifumosa Naksuwankul, Lücking & Lumbsch differs from O. fumosa in having ascospores with rounded ends. An epitype for O. krathingensis is selected.

Key words

East Asia, lichens, taxonomy, thelotremoid lichens, tropical diversity

Introduction

Phenotypical characters, such as morphology of the thallus and ascomata and anatomy of the ascomata as well as secondary chemistry have traditionally guided species delimitation in lichenized ascomycetes. However, especially crustose lichens often exhibit only few traits and without independent markers, such as DNA sequence data, it is often difficult to assess whether variation is due to genetic differences or plasticity. Indeed, recent phylogenetic studies suggest high amounts of homoplasy in phenotypical characters used to delimit taxa in lichenized fungi (Grube et al. 2004; Tehler and Irestedt 2007; Schmitt et al. 2009; Rivas Plata and Lumbsch 2011; Lumbsch et al. 2014a). Hence, molecular data have greatly increased our ability to identify distinct lineages, including the detection of numerous cryptic lineages (Crespo and Lumbsch 2010; Lumbsch and Leavitt 2011; Leavitt et al. 2015). While numerous foliose and fruticose lichen groups have been studied in some detail, especially in the diverse Parmeliaceae, our knowledge on species delimitation in crustose lichens is still in its infancy. However, among predominantly crustose families, Graphidaceae is now relatively well known.

Graphidaceae constitutes the largest family of crustose tropical lichens with about 2100 accepted species (Rivas Plata et al. 2012; Lücking et al. 2013; Cáceres et al. 2014; Van den Broeck et al. 2014; Lumbsch et al. 2014b; Kraichak et al. 2014). The family has its center of distribution in the tropics, but also occurs in temperate regions with a smaller number of species, in some cases even extending towards the Sub-Antarctic region. The family is most common, however, in the tropics where its species occur often on bark, but can also be found on rocks, wood or soil and sometimes on leaves. Recently, the first author started a project on the diversity of thelotremoid Graphidaceae in East Asia (Papong et al. 2014). Thelotremoid Graphidaceae have rounded ascomata (formerly placed in Thelotremataceae), in contrast to species with lirellate ascomata. The group is still relatively poorly known in Thailand and generally in south-east Asia, but preliminary studies have provided important baseline data for the distribution of species and have indicated that numerous additional species can be expected in Thailand (Homchantara and Coppins 2002; Papong et al. 2010; Sutjaritturakan and Kalb 2015). Molecular data have been used to identify distinct lineages in this group of lichenized fungi and subsequent re-analysis of phenotypical characters often allowed identification of morphological or chemical traits to separate those species (Lumbsch et al. 2008; Mangold et al. 2014; Poengsungnoen et al. 2014; Medeiros et al. 2016). This paper employs molecular, morphological and chemical data to identify six distinct lineages of thelotremoid lichens from Thailand and to describe them as species new to science. Based on our limited sampling of thelotremoid Graphidaceae from other regions of southeast Asia, we expect the new species described here from Thailand to occur in other countries of the region.

Material and methods

This study is mainly based on new collections made by the first two authors deposited in F and MSUT. Sections of thalli and apothecia were cut using a razor blade and examined in water, a solution of KOH, and Lugol’s solution using a ZEISS Axioscope 2 plus compound microscope. Chromatography (HPTLC) was performed with standard solvent systems A and C (Culberson 1972; Arup et al. 1993).

We performed two different phylogenetic analyses: 1) sequences of six samples of the genus Glaucotrema were aligned with two outgroup taxa (Leptotrema wightii, Reimnitzia santensis) and 2) sequences of 35 samples of Ocellularia s. str. were aligned with O. cavata as outgroup. Selection of samples was done using Blast searches and included best hits to ensure that all similar sequences were included. In addition sequences of morphologically similar species were added to the data set. Sequences of mtSSU rDNA, nuLSU rDNA, and the protein-coding RPB2 gene were used for this study. Voucher information and GenBank numbers are listed in Table 1. DNA isolation, PCR, and direct cycle sequencing conditions were described previously (Kraichak et al. 2014).

GenBank numbers and voucher information of specimens used in this study. For author names see Index Fungorum (http://www.indexfungorum.org). Missing data are indicated by [–].

Species Country Collector Number mtSSU nuLSU RPB2
Glaucotrema glaucophaenum Philippines Rivas Plata 1099 JX421061 JX421501 JX420862
Glaucotrema glaucophaenum Thailand Lumbsch 19751g [—] JX421502 [—]
Glaucotrema glaucophaenum Australia Lumbsch 19127eA JX421060 [—] [—]
Glaucotrema stegoboloides Brazil Cáceres 11817 KJ435228 [—] [—]
Glaucotrema subcostaricense Tanzania Frisch 99Tz866 DQ384899 [—] [—]
Glaucotrema thailandicum Thailand Papong 8560 [—] KJ435152 [—]
Leptotrema wightii Costa Rica Nelsen 2034A JX421074 EU075622 [—]
Ocellularia albocincta Thailand Kalb 38891 JX421114 [—] [—]
Ocellularia albocincta Australia Mangold 43o EU075585 EU075633 [—]
Ocellularia albocincta Australia Mangold 34a JX421112 [—] [—]
Ocellularia ascidioidea New Caledonia Papong 7511 KJ435201 KJ435125 KJ435267
Ocellularia cavata Cameroon Frisch 99Ka403 DQ384879 DQ431935 [—]
Ocellularia diacida Australia Lumbsch 19120jB EU075583 EU075630 [—]
Ocellularia diacida Australia Lumbsch 19120jD JF828965 [—] [—]
Ocellularia exigua Thailand Papong 8434 KJ435244 [—] [—]
Ocellularia fumosa Thailand Lumbsch 19756n [—] JX421539 [—]
Ocellularia halei Brazil Cáceres 11071 KJ435218 [—] [—]
Ocellularia klinhomii Thailand Papong 8574 KJ435252 [—] [—]
Ocellularia krathingensis Thailand Papong 8478 KJ435248 KJ435153 [—]
Ocellularia krathingensis Thailand Papong 8479 KJ435246 [—] [—]
Ocellularia krathingensis Thailand Papong 8483 KJ435241 [—] [—]
Ocellularia krathingensis Thailand Papong 8496 KJ435232 KJ435143 [—]
Ocellularia mauritiana Peru Rivas Plata 803D JX421170 [—] [—]
Ocellularia natashae Peru Rivas Plata 1canopy JX421175 [—] JX420877
Ocellularia percolumellata Brazil Cáceres 6002a JX421180 [—] JX420888
Ocellularia polydiscus Brazil Lücking 27966 DQ384876 [—] [—]
Ocellularia portoricensis Puerto Rico Mercado F19 KJ435178 [—] KJ435256
Ocellularia phatamensis Thailand Papong 8541 KJ435239 KJ435150 [—]
Ocellularia phatamensis Thailand Papong 8542 KJ435249 KJ435154 [—]
Ocellularia phatamensis Thailand Papong 8552 KJ435236 KJ435147 [—]
Ocellularia phatamensis Thailand Papong 8557 KJ435238 KJ435149 [—]
Ocellularia phatamensis Thailand Papong 8566 KJ435233 KJ435144 [—]
Ocellularia phatamensis Thailand Papong 8567 KJ435245 [—] [—]
Ocellularia phatamensis Thailand Papong 8568 KJ435237 KJ435148 [—]
Ocellularia phatamensis Thailand Papong 8570 KJ435250 KJ435155 [—]
Ocellularia phatamensis Thailand Papong 8573 KJ435251 KJ435156 [—]
Ocellularia rhabdospora Puerto Rico Mercado F74 KJ435172 KJ435108 KJ435254
Ocellularia rotundifumosa Thailand Papong 8576 KJ435231 [—] [—]
Ocellularia thailandica Thailand Papong 8439 KJ435235 KJ435146 [—]
Ocellularia thailandica Thailand Papong 8458 KJ435247 [—] [—]
Ocellularia thryptica Peru Rivas Plata 103D JX421222 [—] [—]
Ocellularia violacea Brazil Cáceres sn JX421225 [—] [—]
Ocellularia xanthostromiza Peru Rivas Plata 809canopy JX421171 [—] [—]
Reimnitzia santensis El Salvador Lücking 28015 HQ639622 [—] JF828952

For the phylogenetic analyses, the alignment of the nucleotide sequences for each dataset was performed separately using Geneious version 8.0.3 (Drummond et al. 2014) and manually inspected for removal of any ambiguous characters. We then performed a maximum likelihood analysis, using RAxML-HPC Blackbox version 8.2.8 (Stamatakis 2006) with the default rapid hill-climbing algorithm and the GTRGAMMA model of nucleotide substitution. The analysis was carried out on the online server CIPRES science Gateway version 3.3 (Miller et al. 2010) with a total of 1,000 pseudoreplicates to assess the rapid bootstrap value support. A bootstrap support value of 70 and above was considered a strong support for a clade. The resulting bipartitioned trees were visualized with the program FigTree version 1.4.2 (Rambaut 2012).

Results and discussion

Phylogenetic analysis

The final alignment of the combined data set for the Glaucotrema analysis consisted of 802 unambiguously aligned nucleotide positions for mtSSU, 865 for nuLSU, and for 985 RPB2. The final alignment of the dataset for the Ocellularia taxa consisted of 787 unambiguously aligned nucleotide positions for mtSSU, 879 for nuLSU, and for 913 RPB2. As the topologies of the single locus phylogenies for these two datasets did not show any conflicts, they were analyzed in a concatenated matrix.

In the Glaucotrema tree (Fig. 1), the Thai material formed an unsupported sister-group relationship with G. glaucophaenum, and G. stegoboloides. The latter two species were not separated in our analysis but were supported as different species in a broader analysis in Kraichak et al. (2014) with more samples, in which the Thai material was also included and supported as distinct species. In the Ocellularia tree (Fig. 2), O. aff. ascidioidea from Thailand did not form a monophyletic group with O. ascidioidea from New Caledonia but an unsupported sister-group relationship with Thai material of O. exigua, similar to the analysis by Kraichak et al. (2014). Subsequent morphological re-analysis revealed that the Thai samples previously identified as O. aff. ascidioidea are identical to O. krathingensis described from Thailand (Homchantara and Coppins 2002). As already indicated by Kraichak et al. (2014), Ocellularia aff. fumosa from Thailand did not cluster with O. fumosa but appeared closely related to O. natashae and O. thryptica. The latter differs in having a clear hymenium and containing protocetraric acid, whereas O. natashae has longer ascospores and contains the hirtifructic acid chemosyndrome (Hale 1973; Rivas Plata and Lücking 2013). The close phylogenetic relationship of these three taxa, which are not only phenotypically disparate but also have distinct geographic distributions, suggests that the loci here used may be of limited use for species delimitation in recently evolved complexes, which has already been discussed for mtSSU by Kraichak et al. (2014). Two samples, included as spec. nov. 8 in Kraichak et al. (2014), formed an unsupported sister-group relationship with O. albocincta, a species that differs morphologically (see below) and so the Thai material is described as a new species (O. siamensis) below. Nine samples included as spec. nov. 6 in Kraichak et al. (2014) from Thailand clustered together, related to O. diacida, which is readily distinguished by the presence of the hirtifructic acid chemosyndrome. The species is described new to science below as O. phatamensis. A single specimen, included as spec. nov. 7 in Kraichak et al. (2014), is also related to O. diacida but differs – among other characters – by the absence of secondary metabolites.

Figure 1.

Phenogram depicting phylogenetic relationships of Glaucotrema species. Only bootstrap support values above 70 are displayed on the nodes.

Figure 2.

Phenogram depicting phylogenetic relationships of Ocellularia species. Only bootstrap support values above 70 are displayed on the nodes.

Taxonomic novelties

Glaucotrema thailandicum Naksuwankul, Lücking & Lumbsch, sp. nov.

MycoBank No: 818194
Figure 3A–E

Type

THAILAND, Ubon Ratchathani Province, Pha Tam National Park, Sang Chan waterfall, 15°30'N, 105°35'E, 124 m, dry evergreen forest, on bark; 12 April 2013, K. Papong 8560 (holotype: MSUT; isotype: F).

Diagnosis

Characterized within the genus by having submuriform ascospores.

Etymology

The specific epithet refers to the country where the type specimen was collected.

Description

Thallus endophloeodal to epiphloeodal, up to c. 120 µm thick, pale green to yellowish green, smooth. True cortex ±continuous, to c. 25 µm thick. Algal layer poorly to well developed, ±continuous; calcium oxalate crystals sparse to abundant, large and clustered; medulla usually distinct. Vegetative propagules not seen. Ascomata conspicuous, to c. 0.8–1.2 mm diam., often larger when fused, ±rounded to irregular, apothecioid to somewhat chroodiscoid, solitary to more often fused, becoming slightly to distinctly emergent, mostly irregularly or regularly urceolate. Disc usually partly visible from above, rarely completely exposed, pale yellowish to whitish green. Pores broad to gaping, to c. 0.6–08 mm wide, ±rounded to irregular, entire to slightly ragged; thalline exciple often becoming apically visible, rarely completely visible from above, ±free, whitish. Thalline rim margin broad to gaping, ±rounded, more commonly irregular, thick, entire, concolorous to whitish. Thalline exciple fused to partly or entirely free, thick, hyaline internally, pale yellowish or greenish marginally, with calcium oxalate crystals. Hymenium to c. 120 µm thick, clear, strongly conglutinated; paraphyses thick, irregular and often distoseptate, ±interwoven, with thickened irregular tips; lateral paraphyses absent; columella whitish and reticulate. Epihymenium hyaline, with fine crystals. Asci 8-spored; tholus initially thick, thin when mature, 100–110 × 10–12 µm. Ascospores submuriform with 3 × 0–1 septa, hyaline, slightly amyloid, 15–20 × 7.5 µm. Pycnidia not seen.

Figure 3.

Morphology and anatomy of Glaucotrema thailandicum (A–E) A–B habitat of ascoma C cross-section of ascoma show whitish and reticulate columella D asci with spores and E submuriform ascospores (holotype), Ocellularia klinhomii (F–K) F–G ascomata immersed in verrucae and surrounded by a black ring H cross-section of ascoma with carbonized columella and apically carbonized exiple I–K ascus and ascospores (holotype). Scale bar A–B, F–G = 1 mm, H = 100 µm, C–D, I = 50 µm, E, J–K = 20 µm.

Secondary chemistry

Thallus K+ yellowish, C–, P+ yellow; containing psoromic acid.

Distribution and ecology

The new species was found in northeastern Thailand, growing on bark in a dry evergreen forest. It is known only from the type locality.

Remarks

This new species is unique within the genus in having submuriform ascospores, whereas all other described species have transversely septate ascospores. In addition, the ascospores in G. bahianum, G. costaricense and G. stegoboloides are smaller than in the new species. Molecular data support the distinction of the new taxon. In morphology it resembles G. bahianum and G. stegoboloides.

Ocellularia klinhomii Naksuwankul, Lücking & Lumbsch, sp. nov.

MycoBank No: 818195
Figure 3F–K

Type

THAILAND, Ubon Ratchathani Province, Pha Tam National Park, Sang Chan waterfall, 15°30'N, 105°35'E, 124 m, dry evergreen forest, on bark; 12 April 2013, K. Papong 8566 (holotype: MSUT; isotype: F).

Diagnosis

Differing from the similar O. krathingensis in having a whitish grey, rimose thallus.

Etymology

The specific epithet refers to the collector Mr. Winia Klinhom, mycologist from Thailand.

Description

Thallus corticolous, epiperidermal, up to c. 5 cm diam., continuous; surface rimose, whitish grey, medulla white; prothallus absent. Thallus in section 30–40 µm thick, with prosoplectenchymatous cortex, 5–10 µm thick, photobiont layer 15–20 µm thick, and medulla 20–25 µm thick, with scattered clusters of calcium oxalate crystals. Photobiont Trentepohlia; cells rounded to irregular in outline, in irregular groups, green, 7–9 × 6–8 µm. Ascomata rounded, verrucae and surrounded by a black ring, erumpent to immersed, with complete thalline margin, 0.4–0.7 mm diam., 0.15–0.2 mm high; disc covered by 0.05–0.1 mm wide pore more or less filled with black-tipped columella but columella often immersed; proper margin indistinct, entire to slightly fissured, visible as whitish rim around the pore; thalline margin entire to slightly fissured, smooth, yellowish green. Excipulum entire, prosoplectenchymatous, brown with apically carbonized, 15–20 µm wide, fused with thalline margin and difficult to separate from the bordering periderm; laterally covered by algiferous, corticate thallus containing periderm layers; columella present, finger-like, carbonized, up to 100 µm broad and 120–140 µm high; hypothecium prosoplectenchymatous, 5–10 µm high, light brown; hymenium 125–150 µm high, hyaline, clear; epithecium indistinct, 5–7 µm high, hyaline. Paraphyses unbranched, apically smooth; periphysoids absent; asci cylindrical to narrowly clavate, 110–115 × 12–15 µm. Ascospores 8 per ascus, ellipsoid, 6–9-septate, 25–38 × 7–8 µm, hyaline, distoseptate with lens-shaped lumina, I+ violet-blue. Pycnidia not seen.

Secondary chemistry

No substances detected by TLC.

Distribution and ecology

The new species was collected in northeastern Thailand, growing on bark in a dry evergreen forest. It is known only from the type locality.

Remarks

Similar in ascospore size, lack of secondary metabolites and only apically carbonized exciple to O. krathingensis but differing in having a whitish gray, rimose thallus with ascomata in verrucae and surrounded by a black ring, reminiscent of O. wirthii (Mangold et al. 2008). The latter species is readily distinguished by having a broader, carbonized columella and the presence of the psoromic acid chemosyndrome. The species would key out at alternative 60 in the Ocellularia key for Thailand (Sutjaritturakan and Kalb 2015).

Additional specimen examined

THAILAND, Ubon Ratchathani Province, Pha Tam National Park, Sang Chan waterfall, 15°30'N, 105°35'E, 124 m, dry evergreen forest, on bark; 12 April 2013, K. Papong 8568, 8552, 8567, 8570, 8542, 8541, 8573, 8574 (MSUT), K. Papong 8557 (RAMK).

Ocellularia phatamensis Naksuwankul, Parnmen & Lumbsch, sp. nov.

MycoBank No: 818196
Figure 4A–B

Type

THAILAND, Ubon Ratchathani Province, Pha Tam National Park, Sang Chan waterfall, 15°30'N, 105°35'E, 124 m, dry evergreen forest, on bark; 12 April 2013, K. Papong 8574 (holotype: MSUT; isotype: F).

Diagnosis

Differing from the similar O. krathingensis in having an a grayish, thick and rimose thallus.

Etymology

The specific epithet refers to the name of the Pha Tam National Park in Ubon Ratchathani Province, Thailand.

Description

Thallus corticolous, epiperidermal, up to c. 5 cm diam., continuous; surface uneven-verrucose to rimose, grayish, medulla white; prothallus absent. Thallus in section 60–75 µm thick, with prosoplectenchymatous cortex, 5–8 µm thick, photobiont layer 20–25 µm thick, and medulla 35–40 µm thick, with scattered clusters of calcium oxalate crystals. Photobiont Trentepohlia; cells rounded to irregular in outline, in irregular groups, green, 8–10 × 6–7 µm. Ascomata rounded, erumpent, with complete thalline margin, 0.4–0.7 mm diam., 0.15–0.2 mm high; disc covered by 0.07–0.1 mm wide pore more or less filled with black-tipped columella but columella often immersed; proper margin indistinct; thalline margin entire to slightly fissured, smooth, light yellowish green. Excipulum entire, prosoplectenchymatous, apically carbonized, 15–20 µm wide, fused with thalline margin and difficult to separate from the bordering periderm; laterally covered by algiferous, corticate thallus containing periderm layers; columella present, finger-like, carbonized, up to 110 µm broad and 120–135 µm high; hypothecium prosoplectenchymatous, 5–10 µm high, light brown; hymenium 120–150 µm high, hyaline, clear; epithecium indistinct, 5–10 µm high, hyaline. Paraphyses unbranched, apically smooth; periphysoids absent; asci cylindrical to narrowly clavate, 100–110 × 12–15 µm. Ascospores 8 per ascus, ellipsoid, 7–8-septate, 25–30 × 7.5–8 µm, hyaline, distoseptate with lens-shaped lumina, I+ violet-blue. Pycnidia not seen.

Figure 4.

Morphology and anatomy of Ocellularia phatamensis (A–B) A habitat of ascomata B ascospores (K. Papong 8574, holotype MSUT!), O. rotundifumosa (C–E) C ascomata D hymenium with ascus and E ascospores (holotype MSUT), O. thailandica (F–H) F habitat of ascomata G ascus with ascospores and H ascospores (holotype). Scale bar A, C, F = 1 mm, D, G = 50 µm, B, E, H = 20 µm.

Secondary chemistry

No substances detected by TLC.

Distribution and ecology

The new species was collected in northeastern Thailand, growing on bark in a dry evergreen forest. It is known only from the type locality.

Remarks

The new species is similar to O. krathingensis in having an apically carbonized exciple and columella, transversely septate, amyloid ascospores, and lacking secondary metabolites, but differs in having a grayish and thicker thallus (Homchantara and Coppins 2002). Another similar species is O. klinhomii, but differs in lacking a dark apothecial rim and the ascomata are not immersed in verrucae. Molecular data support the distinction of these two species (Fig. 2). Another similar and related species is O. diacida, which is readily distinguished by the presence of the hirtifructic acid chemosyndrome. The species would key out at alternative 60 in the Ocellularia key for Thailand (Sutjaritturakan and Kalb 2015).

Ocellularia rotundifumosa Naksukankul, Lücking & Lumbsch, sp. nov.

MycoBank No: 818197
Figure 4C–E

Type

THAILAND, Ubon Ratchathani Province, Pha Tam National Park, Sang Chan waterfall, 15°30'N, 105°35'E, 124 m, dry evergreen forest, on bark; 12 April 2013, K. Papong 8576 (holotype: MSUT; isotype: F).

Diagnosis

Differing from O. fumosa in having ascospores with rounded ends.

Etymology

The specific epithet refers to the ascospore shape with rounded ends and to the similarity with O. fumosa.

Description

Thallus corticolous, endophloeodal to epiphloeodal, up to c. 200 µm thick, greenish gray to olive, slightly glossy, smooth, rarely continuous to usually ±verrucose. True cortex discontinuous, to c. 15 µm thick, formed by irregular hyphae. Algal layer well developed, continuous; calcium oxalate crystals moderately large, scattered. Photobiont Trentepohlia; cells rounded to irregular in outline, in irregular groups, green, 7–9 × 6–9 µm. Vegetative propagules not seen. Ascomata rounded with complete thalline margin, 0.4–0.9 mm diam., solitary to marginally fused, immersed to rather emergent, then verrucose-hemispherical to urceolate. Disc with the columella visible from above, entire, free, slightly pruinose, dark gray. Pores formed by the thalline rim margin, c. 0.5 mm diam., the apex of the proper exciple becoming visible from above as a brownish to dark gray line, moderately thick, concolorous with the thallus or brighter; thalline rim incurved. Proper exciple fused, dark brown to carbonized marginally and towards the tips, usually distinctly amyloid at the base. Hymenium to c. 150 µm thick, densely inspersed, distinctly conglutinated; paraphyses slightly bent, ±interwoven, unbranched, with moderately thickened tips; columellar structures moderately well developed, to 150 µm wide, entire, the upper parts brownish to carbonized. Epihymenium brownish, with grayish or brownish granules. Asci 8-spored; tholus initially thick, thin when mature. Ascospores 7–9-septate, fusiform to oblong-fusiform, rarely clavate, with rounded ends, 24–35 × 7–10 µm, hyaline, distoseptate with lens-shaped lumina, I+ violet-blue. Pycnidia not seen.

Secondary chemistry

No compounds detectable by TLC.

Distribution and ecology

The new species was collected in northeastern Thailand, growing on bark in a dry evergreen forest. It is known only from the type locality.

Remarks

Similar to O. fumosa, but differing in having rounded ends of the ascospores instead of acute ones in O. fumosa. Molecular data support the distinction of the species (Fig. 2). Characters to separate the related O. natashae and O. thryptica are discussed above. The species would key out at alternative 23 in the Ocellularia key for Thailand (Sutjaritturakan and Kalb 2015).

Ocellularia thailandica Naksuwankul, Kraichak & Lumbsch, sp. nov.

MycoBank No: 818198
Figure 4F–H

Type

THAILAND, Ubon Ratchathani Province, Pha Tam National Park, trail to Huai Sanom, 15°27'N, 105°34'E, 245 m, dry evergreen forest, on bark; 12 April 2013, K. Papong 8458 (holotype: MSUT; isotype: F).

Diagnosis

Differing from the similar O. viridipallens in having broader ascospores with up to 7 septa.

Etymology

The specific epithet refers to the country where the type specimen was collected.

Description

Thallus corticolous, epiperidermal, up to c. 5 cm diam., continuous; surface uneven-verrucose to rimose, light yellowish green, medulla white; prothallus absent. Thallus in section 40–60 µm thick, with prosoplectenchymatous cortex, 5–7 µm thick, photobiont layer 15–25 µm thick, and medulla 20–30 µm thick, with scattered clusters of calcium oxalate crystals. Photobiont Trentepohlia; cells rounded to irregular in outline, in irregular groups, green, 7–8 × 5–9 µm. Ascomata rounded, erumpent, with complete thalline margin, 0.3–0.5 mm diam., 0.12–0.2 mm high; disc covered by 0.05–0.1 mm wide pore; proper margin indistinct, entire to slightly fissured, visible as whitish rim around the pore; thalline margin entire to slightly fissured, smooth, light yellowish green. Excipulum entire, prosoplectenchymatous, brown to dark brown, 15–20 µm wide, fused with thalline margin and difficult to separate from the bordering periderm; laterally covered by algiferous, corticate thallus containing periderm layers; columella present, finger-like, carbonized, up to 100 µm broad and 120–135 µm high; hypothecium prosoplectenchymatous, 5–10 µm high, hyaline; hymenium 125–140 µm high, hyaline, clear; epithecium indistinct, 5–10 µm high, hyaline. Paraphyses unbranched, apically smooth; periphysoids absent; asci cylindrical to narrowly clavate, 87–100 × 12–15 µm. Ascospores 8 per ascus, ellipsoid, 5–7-septate, 20–23 × 7–8 µm, hyaline, distoseptate with lens-shaped lumina, I+ violet-blue. Pycnidia not seen.

Secondary chemistry

No substances detected by TLC.

Distribution and ecology

The new species was collected in northeastern Thailand, growing on bark in a dry evergreen forest. It is known only from the type locality.

Remarks

This new species is closely related to O. albocincta (Fig. 2). However, this species differs in lacking a columella (Papong et al. 2010). Morphologically it resembles O. viridipallens, which differs in having narrower ascospores. The species would key out at alternative 60 in the Ocellularia key for Thailand (Sutjaritturakan and Kalb 2015).

Additional specimen examined

THAILAND, Ubon Ratchathani Province, Pha Tam National Park, trail to Huai Sanom, 15°27'N, 105°34'E, 245 m, dry evergreen forest, on bark; 12 April 2013, K. Papong 8439 (MSUT).

Ocellularia krathingensis Homchant. & Coppins

Figure 5D–F

Epitype

THAILAND. Ubon Ratchathani Province: Pha Tam National Park, trail to Huai Sanom, 15° 27’ 620” N, 105° 34’ 615” E, 245 m, dry evergreen forest, on bark; 12 Apr. 2013, K. Papong 8479 (epitype MSUT!).

In order to clarify the application of the name Ocellularia krathingensis, we propose an epitype for this species that agrees morphologically well with the holotype (RAMK!) and has been sequenced.

Additional specimens examined

THAILAND, Ubon Ratchathani Province, Pha Tam National Park, trail to Huai Sanom, 15°27'N, 105°34'E, 245 m, dry evergreen forest, on bark; 12 April 2013, K. Papong 8496, 8478, 8483 (F, MSUT) (Figure 5A–C).

Figure 5.

Morphology and anatomy of O. krathingensis (A–F); A habitat of ascomata B hymenium with ascus and C ascospores. (D–F) D erumpent ascomata E ascus and F ascospores A–C K. Papong 8483 D–F K. Papong 8479 (epitype). Scale bar A, D = 1 mm, B, E = 50 µm, C, F = 20 µm.

Acknowledgments

This study was financially supported by grants of the Mahasarakham University and Thai Research Fund to the first author (K. Papong RSA 5580045) and the grant ATM – Assembling a taxonomic monograph: The lichen family Graphidaceae (DEB-1025861 to The Field Museum; PI T. Lumbsch, CoPI R. Lücking) by the National Science Foundation.

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