Research Article |
Corresponding author: Beeyoung Gun Lee ( gitanoblue@koagi.or.kr ) Academic editor: Gerhard Rambold
© 2021 Beeyoung Gun Lee, Jae-Seoun Hur.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Lee BG, Hur J-S (2021) Two new calcicolous caloplacoid lichens from South Korea, with a taxonomic key to the species of Huriella and Squamulea. MycoKeys 84: 35-55. https://doi.org/10.3897/mycokeys.84.71227
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Pyrenodesmia rugosa Lee & Hur and Huriella aeruginosa Lee & Hur are described as new lichen-forming fungi from a calcareous mountain of South Korea. Pyrenodesmia rugosa is distinguishable from Pyrenodesmia micromontana (Frolov, Wilk & Vondrák) Hafellner & Türk, the most similar species, by thicker thallus, rugose areoles, larger apothecia, shorter hymenium, shorter hypothecium and narrower tip cells of paraphyses. Huriella aeruginosa, the second new species, differs from ‘Squamulea’ chelonia Bungartz & Søchting by dark greenish-grey to grey thallus without pruina, gold to yellow-brown epihymenium, larger ascospores and thallus K– and KC– reaction. Molecular analyses employing internal transcribed spacer (ITS), mitochondrial small subunit (mtSSU) and nuclear large subunit ribosomal RNA (LSU) sequences strongly support the two caloplacoid species to be distinct in their genera. A surrogate key is provided to assist in the identification of all 20 taxa in Huriella and Squamulea.
Biodiversity, phylogeny, saxicolous, taxonomy, Teloschistaceae
Many lichens are only detected in calcareous areas, particularly for crustaceous lichens, as many plants are never found, except on calcareous rocks and soils (
This study describes two new calcicolous caloplacoid lichens in the genera Pyrenodesmia and Huriella. Qualified field surveys for the lichen diversity on the Baekdudaegan Mountains, the main mountain range stretching across the entire Korean Peninsula, were accomplished during the summer of 2020 and a few dozen specimens of caloplacoid lichens were collected in Mt. Seokbyung, a calcareous mountain (Fig.
Hand-cut sections were prepared with a razor blade under a stereomicroscope (Olympus optical SZ51; Olympus, Tokyo, Japan), examined under a compound microscope (Nikon Eclipse E400; Nikon, Tokyo, Japan) and imaged using a software programme (NIS-Elements D; Nikon, Tokyo, Japan) and a DS-Fi3 camera (Nikon, Tokyo, Japan), mounted on a Nikon Eclipse Ni-U microscope (Nikon, Tokyo, Japan). The ascospores were investigated at 1000× magnification in water. The length and width of the ascospores were measured and the range of spore sizes was shown with average, standard deviation and number of measured spores. Thin-layer chromatography (TLC) was performed using solvent systems A and C according to standard methods (
Hand-cut sections of ascomata or thallus from all collected specimens were prepared for DNA isolation and DNA was extracted with a NucleoSpin Plant II Kit in line with the manufacturer’s instructions (Macherey-Nagel, Düren, Germany). PCR amplification for the internal transcribed spacer region (ITS1-5.8S-ITS2 rDNA), the mitochondrial small subunit and the nuclear large subunit ribosomal RNA genes was achieved using Bioneer’s AccuPower PCR Premix (Bioneer, Daejeon, Korea) in 20-μl tubes and primers ITS5 and ITS4 (
All ITS, mtSSU and LSU sequences were aligned and edited manually using ClustalW in Bioedit V.7.2.6.1 (
Three independent phylogenetic trees for Pyrenodesmia and one independent phylogenetic tree for Squamulea were produced from 165 sequences (96 for ITS, 37 for mtSSU and 32 for LSU) from GenBank and four new sequences (two for ITS, one for mtSSU and one for LSU) for the new species (Table
Species list and DNA sequence information employed for phylogenetic analysis.
No | Species | ID (ITS) | ID (mtSSU) | ID (LSU) | Voucher |
---|---|---|---|---|---|
1 | Amundsenia approximata | KJ789965 | L08179 (LD) | ||
2 | Amundsenia austrocontinentalis | KJ789962 | 21966 (HO) | ||
3 | Athallia holocarpa | MG954144 | Vondrak 18072 | ||
4 | Athallia vitellinula | FJ346556 | Arup L03052 | ||
5 | Caloplaca monacensis | MG773668 | MG773679 | Malicek 8255 | |
6 | Caloplaca sp. | KC611244 | CBFS:JV6943 | ||
7 | Erichansenia sauronii | KC179120 | Sochting 7654 | ||
8 | Huriella aeruginosa | MW832829 | BDNA-L-0001072 | ||
9 | Huriella flakusii | MT967442 | Bungartz 4131 (CDS 28162) | ||
10 | Huriella flakusii | MT967443 | Bungartz 4157 (CDS 28188) | ||
11 | Huriella flakusii | MT967444 | Aptroot 65261 (CDS 31847) | ||
12 | Huriella loekoesiana | KY614406 | KoLRI 15423 | ||
13 | Huriella loekoesiana | KY614407 | KoLRI 19017 | ||
14 | Huriella loekoesiana | KY614408 | KoLRI 40141 | ||
15 | Huriella loekoesiana | KY614409 | KoLRI 40236 | ||
16 | Huriella loekoesiana | KY614410 | KoLRI 40238 | ||
17 | Huriella loekoesiana | MK499351 | HKAS 102112 | ||
18 | Huriella sp. | MN108089 | KRAM-L-70242 | ||
19 | Kuettlingeria albolutescens | KC179423 | KC179502 | MT952898 | Arup L09030 (LD) |
20 | Kuettlingeria areolata | MN305805 | MN305825 | MN305847 | Vondrak 10843 |
21 | Kuettlingeria atroflava | MH104921 | MH100775 | Vondrak 8723 (PRA) | |
22 | Kuettlingeria cretensis | MH104925 | MH100783 | MH100751 | Frolov s.n. |
23 | Kuettlingeria diphyodes | MH104926 | MH100785 | MH100753 | Frolov 1430 |
24 | Kuettlingeria emilii | KC416102 | MH100787 | MH100754 | JV9358 |
25 | Kuettlingeria erythrocarpa | KC179427 | KC179506 | KC179173 | Arup L07109 (LD) |
26 | Kuettlingeria neotaurica | MN305807 | MN305829 | MN305849 | Vondrak 7213 |
27 | Kuettlingeria percrocata | MH104931 | MH100794 | Vondrak 4634 (PRA) | |
28 | Kuettlingeria soralifera | MN305808 | MN305830 | MN305850 | Vondrak 10813 |
29 | Kuettlingeria aff. soralifera | JN641781 | CBFS:JV8325 | ||
30 | Kuettlingeria teicholyta | MH104935 | MH100797 | MH100767 | Vondrak 6943 (PRA) |
31 | Kuettlingeria xerica | MN305809 | MN305831 | MN305851 | Vondrak 14544 |
32 | Kuettlingeria aff. xerica | HQ611275 | CBFS:JV7618 | ||
33 | Lendemeriella borealis | MG954129 | Vondrak 11073 | ||
34 | Lendemeriella exsecuta | MG954227 | Spribille 24441 | ||
35 | Lendemeriella nivalis | MG954222 | Spribille 29306 | ||
36 | Lendemeriella reptans | MH104934 | MH100796 | MH100766 | Lendemer 48186 (NY) |
37 | Lendemeriella sorocarpa | MG954132 | Vondrak12695 | ||
38 | Lendemeriella tornoensis | MG954221 | Spribille 29473 | ||
39 | Olegblumia demissa | KT220203 | KT220221 | KT220212 | SK C65 |
40 | Pyrenodesmia aetnensis | EU639590 | KT291476 | X. Llimona (BCN) | |
41 | Pyrenodesmia albopruinosa | EF093577 | MH100770 | TSB 37658 | |
42 | Pyrenodesmia albopustulata | MH104918 | MH100771 | MH100741 | Vondrak 10463 (PRA) |
43 | Pyrenodesmia alociza | EF090931 | MH100772 | MH100742 | TSB 37735 |
44 | Pyrenodesmia aractina | GU723415 | Bornholm 5907 | ||
45 | Pyrenodesmia aractina | GU723418 | Bornholm 6911 | ||
46 | Pyrenodesmia aractina | MH104919 | MH100773 | Vondrak 6702 (PRA) | |
47 | Pyrenodesmia atroalba | MH104920 | MH100774 | Spribille s.n. | |
48 | Pyrenodesmia badioreagens | EF081035 | MH100776 | MH100745 | TSB 36422 |
49 | Pyrenodesmia bicolor | MH104922 | MH100777 | MH100746 | Vondrak 10373 (PRA) |
50 | Pyrenodesmia ceracea | HQ234603 | BM-6656 | ||
51 | Pyrenodesmia chalybaea | KC884498 | MH100779 | MH100747 | CBFS:JV4059 |
52 | Pyrenodesmia circumalbata | MH104923 | MH100780 | MH100748 | Halici s.n. |
53 | Pyrenodesmia concreticola | KC884506 | MH100781 | MH100749 | CBFS:JV9443 |
54 | Pyrenodesmia duplicata | HQ611272 | TUR-V-7513 | ||
55 | Pyrenodesmia erodens | MH104927 | MH100788 | MH100755 | Vondrak 12733 (PRA) |
56 | Pyrenodesmia haematites | GU723420 | MH100789 | MH100756 | Vondrak 7278 (PRA) |
57 | Pyrenodesmia haematites | GU723421 | JS280 | ||
58 | Pyrenodesmia haematites | MH104928 | Vondrak 7278 (PRA) | ||
59 | Pyrenodesmia helygeoides | MH104929 | MH100790 | MH100757 | Frolov 1414 |
60 | Pyrenodesmia micromarina | NR_156257 | CBFS:JV8199 | ||
61 | Pyrenodesmia micromarina | MH100791 | MH100758 | Vondrak 7236 (PRA) | |
62 | Pyrenodesmia micromontana | NR_158297 | MH100792 | MH100759 | CBFS:JV9467 |
63 | Pyrenodesmia microstepposa | NR_156260 | MH100760 | CBFS:JV9141 | |
64 | Pyrenodesmia molariformis | KC416145 | MH100793 | MH100761 | Nadyeina 132 (KW) |
65 | Pyrenodesmia obscurella | MH104938 | MH100762 | Vondrak 7641 (PRA) | |
66 | Pyrenodesmia peliophylla | MG733135 | Jason Hollinger:16476 | ||
67 | Pyrenodesmia pratensis | MH104933 | MH100795 | MH100765 | MIN 891605 |
68 | Pyrenodesmia rugosa | MW832828 | MW832825 | MW832904 | BDNA-L-0001099 |
69 | Pyrenodesmia transcaspica | MH104936 | MH100799 | MH100768 | Vondrak 9430 (PRA) |
70 | Pyrenodesmia variabilis | KT291466 | KT291514 | KT291561 | Ulf Arup L07196 (LD) |
71 | Shackletonia buelliae | KC179117 | Sochting 7583 | ||
72 | Shackletonia siphonospora | KC179121 | Sochting 7883 | ||
73 | Squamulea galactophylla | KC179122 | Morse 10997 (LD) | ||
74 | Squamulea kiamae | KC179123 | Kondratyuk 20480 (LD) | ||
75 | Squamulea parviloba | KC179124 | Wetmore 87830 (LD) | ||
76 | Squamulea squamosa | MT967462 | Moberg 8782 (UPS) | ||
77 | Squamulea squamosa | KC179125 | Karnefelt AM960105 (LD) | ||
78 | Squamulea ‘squamosa’ | MT967465 | Bungartz 7428 (CDS 37915) | ||
79 | Squamulea subsoluta | AF353954 | Arup L97072 | ||
80 | Squamulea subsoluta | DQ173238 | Arup L97829 | ||
81 | Squamulea subsoluta | KJ133480 | KoLRI 011067 | ||
82 | ‘Squamulea’ chelonia | MT967448 | Bungartz 4521 (CDS 28607) | ||
83 | ‘Squamulea’ chelonia | MT967451 | Bungartz 9251 (CDS 46069) | ||
84 | ‘Squamulea’ chelonia | MT967452 | Bungartz 6146 (CDS 34358) | ||
85 | ‘Squamulea’ humboldtiana | MT967439 | Buck 29560 (MIN) | ||
86 | ‘Squamulea’ humboldtiana | MT967440 | Bungartz 4711B (CDS 56235) | ||
87 | ‘Squamulea’ humboldtiana | MT967441 | Bungartz 9985 (CDS 47354) | ||
88 | ‘Squamulea’ oceanica | MT967445 | Yánez-Ayabaca 2023 (CDS 48373) | ||
89 | ‘Squamulea’ oceanica | MT967446 | Bungartz 10152 (CDS 47571) | ||
90 | ‘Squamulea’ oceanica | MT967447 | Bungartz 9857 (CDS 47195) | ||
91 | ‘Squamulea’ osseophila | MT967455 | Aptroot 65489 (CDS 32078) | ||
92 | ‘Squamulea’ phyllidizans | MT967456 | Aptroot 65468 (CDS 32057) | ||
93 | ‘Squamulea’ phyllidizans | MT967457 | Bungartz 4710 (CDS 28808) | ||
94 | ‘Squamulea’ phyllidizans | MT967458 | Bungartz 4158 (CDS 28189) | ||
95 | ‘Squamulea’ subsoluta | KJ133481 | KoLRI 012491 | ||
96 | ‘Squamulea’ sp. | MG954160 | Vondrak 18682 | ||
97 | Usnochroma carphineum | KC179468 | KC179598 | KC179259 | Roux s.n. |
98 | Usnochroma scoriophilum | JQ301664 | JQ301496 | JQ301560 | P. & B. v.d. Boom 38386 |
Overall | 98 | 38 | 33 |
Phylogenetic relationships amongst available species in the genus Pyrenodesmia, based on a Maximum Likelihood analysis of the dataset of ITS sequences. The tree was rooted with the sequences of the genera Caloplaca, Lendemeriella, Olegblumia and Usnochroma. Maximum Likelihood bootstrap values ≥ 70% and posterior probabilities ≥ 95% are shown above internal branches. Branches with bootstrap values ≥ 90% are shown in bold. The new species Pyrenodesmia rugosa is presented in bold and all species names are followed by the GenBank accession numbers. Reference Table
Phylogenetic relationships amongst available species in the genus Pyrenodesmia, based on a Maximum Likelihood analysis of the dataset of the mitochondrial small subunit (mtSSU) sequences. The tree was rooted with five sequences of the genera Caloplaca, Lendemeriella, Olegblumia and Usnochroma. Maximum Likelihood bootstrap values ≥ 70% and posterior probabilities ≥ 95% are shown above internal branches. Branches with bootstrap values ≥ 90% are shown in bold. The new species Pyrenodesmia rugosa is presented in bold and all species names are followed by the GenBank accession numbers. Reference Table
Phylogenetic relationships amongst available species in the genus Pyrenodesmia , based on a Maximum Likelihood analysis of the dataset of the nuclear large subunit ribosomal RNA (LSU) sequences. The tree was rooted with three sequences of the genera Lendemeriella and Usnochroma. Maximum Likelihood bootstrap values ≥ 70% and posterior probabilities ≥ 95% are shown above internal branches. Branches with bootstrap values ≥ 90% are shown in bold. The new species Pyrenodesmia rugosa is presented in bold and all species names are followed by the GenBank accession numbers. Reference Table
Phylogenetic relationships amongst available species in the genera Huriella and Squamulea, based on a Maximum Likelihood analysis of the dataset of ITS sequences. The tree was rooted with the sequences of the genera Amundsenia, Erichansenia and Shackletonia. Maximum Likelihood bootstrap values ≥ 70% and posterior probabilities ≥ 95% are shown above internal branches. Branches with bootstrap values ≥ 90% are shown in bold. The new species Huriella aeruginosa is presented in bold and all species names are followed by the GenBank accession numbers. Reference Table
Pyrenodesmia rugosa differs from P. micromontana by thicker thallus (125–200 μm vs. 95–125 μm), rugose areoles (vs. flat areoles), larger apothecia (0.2–0.7 mm diam. vs. 0.2–0.4 mm diam.), shorter hymenium (60–70 μm vs. 80–100 μm), shorter hypothecium (50–55 μm vs. 80–100 μm) and narrower tip cells of paraphyses (3–4.5 μm vs. 5–6 μm).
Pyrenodesmia rugosa (BDNA-L-0001102, holotype) in morphology A–C habitus and apothecia. Rugose thallus brown with orange spots and without pruina, but black apothecia often white pruinose D–E zeorine apothecia with well-developed parathecium. Algal layers continue to the base and underlying the hypothecium F epihymenium K+ purple and tiny granules not dissolving in K G–K asci oblong to narrowly clavate with eight spores K in the lactophenol cotton blue L ascospores simple in the beginning and developed polarilocular at maturity M paraphyses anastomosing in lactophenol cotton blue. Paraphysis tips slightly swollen. Scale bars: 1 mm (A–C); 100 μm (D); 50 μm (E, F); 10 μm (G–M).
South Korea, Gangwon Province, Gangneung, Okgye-myeon, Mt. Seokbyung (summit), 37°35.21'N, 128°53.87'E, 1,072 m alt., on calcareous rock, 17 June 2020, B.G.Lee & H.J.Lee 2020-000902, with Athallia cf. vitellinula (Nyl.) Arup, Frödén & Søchting, Bagliettoa baldensis (A. Massal.) Vězda, Catillaria lenticularis (Ach.) Th. Fr. and Staurothele aff. succedens (Rehm) Arnold (holotype: BDNA-L-0001102!); same locality, on calcareous rock, 17 June 2020, B.G.Lee & H.J.Lee 2020-000899, with Athallia cf. holocarpa (Hoffm.) Arup, Frödén & Søchting and Staurothele cf. rupifraga (A. Massal.) Arnold (paratype: BDNA-L-0001099; GenBank MW832828 for ITS, MW832825 for mtSSU and MW832804 for LSU).
Thallus saxicolous (calcicolous), crustose, mainly areolate or slightly rimose, rugose, greyish-brown to pale brown, often with orange spots, margin indeterminate or determinate when placodioid areoles are arranged around edge, vegetative propagules absent, areoles 0.4–1.0 mm diam., 125–200 μm thick; cortex hyaline with pale brown pigment layer, pale brown pigment K+ purple, 10–40 μm thick, cortical cells granular, 5–10 μm diam., with epinecral layer, 5–7 μm thick; medulla 60–110 μm thick below algal layer or inconspicuous and algal layer shown just above substrate; photobiont coccoid, cells globose to oval, 5–15 μm diam., algal layer 50–70 μm thick. Small crystals present between algal cells, not dissolving in K. Prothallus absent.
Apothecia abundant, scattered or concentrated in centre, rounded, often contiguous or even coalescent when mature, emerging on the surface of thallus, immersed or adnate, slightly constricted at the base, 0.2–0.7 mm diam. Disc flat when young and flat or concave when mature, often white pruinose, black, 200–300 μm thick; zeorine, margin persistent, slightly prominent, generally entire or rarely slightly crenulate, thalline margin paler to disc and showing brown colour, often inconspicuous due to locating below proper margin, proper margin concolorous to disc. Amphithecium present, with small crystals between algal cells, not dissolving in K, 80–130 μm wide laterally, algal layers continuous to the base and underlying the hypothecium, algal cells 5–15 μm diam., cortical layer hyaline with pale brownish pigment at periphery, 10–40 μm thick. Parathecium well-developed, hyaline, but grey with slightly brown pigment concolorous to epihymenium at periphery, 20–40 μm wide laterally and 50–90 μm wide at periphery. Epihymenium grey with slightly brown pigment, K+ purple, tiny granules abundant on surface, not dissolving in K, 5–10 μm high. Hymenium hyaline, 60–70 μm high. Hypothecium hyaline, base open and extending downwards, 50–55 μm high. Oil droplets present in upper hypothecium, but absent in hymenium. Paraphyses septate, often anastomosing, 2–2.5 μm wide, generally simple, but occasionally branched at tips, tips slightly swollen, not pigmented, 3.0–4.5 μm wide. Asci oblong to narrowly clavate, 8-spored, 52–60 × 14–18 μm (n = 5). Ascospores ellipsoid, 1-septate, polarilocular when mature or narrow septum remaining, hyaline permanently, 11–18 × 5.5–11 μm (mean = 14.1 × 7.6 μm; SD = 1.6(L), 1.0(W); L/W ratio 1.5–2.5, ratio mean = 1.9, ratio SD = 0.3; n = 105), septum 1.5–3.0 μm. Pycnidia not detected.
Thallus K–, KC–, C–, Pd–. Epihymenium K+ purple. Hymenium I+ blue. UV–. No lichen substance was detected by TLC.
The species occurs on the calcareous rock. The species is currently known from the type collections.
The species epithet indicates the lichen’s thallus texture, rugose or wrinkled, which is the key characteristic distinguished from closely-related calcicolous species in the genus Pyrenodesmia.
The new speices is similar to P. micromontana, P. microstepposa and Caloplaca micromarina Frolov, Khodos. & Vondrák in having epilithic thallus without vegetative propagules, small apothecia generally less than 0.5 mm diameter and the substrate preference to calcareous rocks. The new species differs from P. micromontana by thicker thallus (125–200 μm vs. 95–125 μm), rugose areoles (vs. flat areoles), larger apothecia (0.2–0.7 mm diam. vs. 0.2–0.4 mm diam.), shorter hymenium (60–70 μm vs. 80–100 μm), shorter hypothecium (50–55 μm vs. 80–100 μm) and narrower tip cells of paraphyses (3–4.5 μm vs. 5–6 μm) (
The new species is different from P. microstepposa by darker thallus (greyish-brown to pale brown vs. ochre, grey or grey-white), rugose thallus (vs. flat thallus), thinner thallus (125–200 μm vs. 85–370 μm), smaller algal cells (5–15 μm diam. vs. 13.5–20.5 μm diam.), presence of pruina on disc (vs. absence of it), absence of oil droplets in hymenium (vs. presence of it), greyish epihymenium (vs. brownish epihymenium), wider ascospores (11–18 × 5.5–11 μm with the L/W ratio of 1.5–2.5 vs. 13.6–18.4 × 6–7.9 μm with the ratio of 1.9–2.9) (
The new species is distinguished from C. micromarina by darker thallus (greyish-brown to pale brown vs. ochre to grey), rugose thallus (vs. flat thallus), absence of pruina on thallus (vs. presence of it), shorter hymenium (60–70 μm vs. 90–100 μm), shorter septum (1.5–3 μm vs. 2.6–3.4 μm) and the habitat preference to mountain rocks (vs. coastal rocks) (
Additional specimens examined: South Korea, Gangwon Province, Okgye-myeon, Mt. Seokbyung (summit), 37°35.21'N, 128°53.87'E, 1,072 m alt., on calcareous rock, 17 June 2020, B.G.Lee & H.J.Lee 2020-000889, with Bagliettoa baldensis, Catillaria lenticularis, Fulgogasparrea decipioides (Arup) S.Y. Kondr., M.H. Jeong, Kärnefelt, Elix, A. Thell & Hur and Laundonia flavovirescens (Wulfen) S.Y. Kondr., Lőkös & Hur (BDNA-L-0001089); same locality, on calcareous rock, 17 June 2020, B.G.Lee & H.J.Lee 2020-000909, with Bagliettoa baldensis, Rusavskia elegans (Link) S.Y. Kondr. & Kärnefelt and Verrucaria nigrescens Pers. (BDNA-L-0001109); same locality, on calcareous rock, 17 June 2020, B.G.Lee & H.J.Lee 2020-000910, with Bagliettoa baldensis, Catillaria lenticularis and Laundonia flavovirescens (BDNA-L-0001110); same locality, on calcareous rock, 17 June 2020, B.G.Lee & H.J.Lee 2020-000911, with Athallia cf. vitellinula, Bagliettoa baldensis, Lichenella sp. and Rusavskia elegans (BDNA-L-0001111); same locality, on calcareous rock, 17 June 2020, B.G.Lee & H.J.Lee 2020-000913, with Athallia cf. vitellinula, Bagliettoa baldensis, Endocarpon sp., Laundonia flavovirescens, Lichenella sp. and Rusavskia elegans (BDNA-L-0001113); same locality, on calcareous rock, 17 June 2020, B.G.Lee & H.J.Lee 2020-000916, with Caloplaca sp., Endocarpon sp., Lichenella sp. and Rusavskia elegans (BDNA-L-0001116).
Huriella aeruginosa differs from ‘Squamulea’ chelonia by dark greenish-grey to grey thallus without pruina (vs. yellow orange to deep orange thallus with white pruina), gold to yellow-brown epihymenium (vs. orange epihymenium), larger ascospores (7.5–12 × 4.5–7.5 μm vs. 8–10.4 × 4.7–6.0 μm) and the chemistry (thallus K–, KC– and no substance vs. thallus K+ purple, KC± purplish and the presence of parietin, teloschistin, fallacinal, parietinic acid and emodin).
Huriella aeruginosa (BDNA-L-0001072, holotype) in morphology A–C habitus and apothecia. Thallus dark greenish-grey to grey with no pruina. Thalline margin of apothecia concolorous to disc D apothecia adnate or rarely sessile. Amphithecium well-developed, but parathecium inconspicuous. E thallus with dark green pigment layer under cortex F–G clavate asci containing 8-spores H ascospores generally ellipsoid, but occasionally globose, developing polarilocular in both types. Two blue coloured spores in lactophenol cotton blue. Scale bars: 1 mm (A–C); 100 μm (D);10 μm (E–H).
South Korea, Gangwon Province, Gangneung, Okgye-myeon, Mt. Seokbyung (summit), 37°35.21'N, 128°53.87'E, 1,072 m alt., on calcareous rock, 17 June 2020, B.G.Lee & H.J.Lee 2020-000872, with Bagliettoa baldensis, Catillaria lenticularis, Endocarpon subramulosum Y. Joshi & Hur, Laundonia flavovirescens, Rusavskia elegans and Verrucaria nigrescens (holotype: BDNA-L-0001072!; GenBank MW832829 for ITS).
Thallus saxicolous (calcicolous), crustose, mainly areolate or slightly rimose, placodioid around edge, but without distinct lobes, thin, dark greenish-grey to grey, occasionally pale yellowish-grey when young, margin indeterminate or determinate when placodioid areoles are arranged around edge, vegetative propagules absent, areoles 0.3–0.7 mm diam., 150–200 μm thick; cortex hyaline with dark green pigment layer, 15–25 μm thick, cortical cells granular, coarsely anticlinally arranged, 5–10 μm diam., with epinecral layer, up to 5 μm thick; medulla 80–100 μm thick, below algal layer, with large crystals (materials of substrate possibly) and brown cells (dead algal cells possibly); photobiont coccoid, cells globose to oval, 5–25 μm. Small crystals in cortex, medulla and between algal cells, dissolving in K. Prothallus absent.
Apothecia abundant, scattered and not concentrated in centre, rounded, often contiguous when mature, emerging on the surface of thallus, immersed, adnate or rarely sessile, constricted at the base, 0.2–0.4 mm diam. Disc flat when young and flat or slightly convex when mature, not pruinose, orange from the beginning, 110–230 μm thick; margin persistent, even to disc or slightly prominent, generally entire or slightly crenulate, thalline margin concolorous to disc, proper margin inconspicuous. Amphithecium well-developed, with small crystals between algal cells, dissolving in K, 50–55 μm wide laterally, algal layers continuous to the base or solitarily remaining in amphithecium, algal cells 5–25 μm diam., cortical layer hyaline with gold to yellow-brown pigment concolorous to epihymenium at periphery, 15–20 μm thick. Parathecium inconspicuous, hyaline but gold to yellow-brown at periphery, ca. 10 μm wide laterally and ca. 20 μm wide at periphery. Epihymenium gold to yellow-brown, granular, pigment K+ wine red and dissolved, 10–20 μm high. Hymenium hyaline, 45–55 μm high. Hypothecium hyaline, 35–45 μm high. Oil droplets present, small, along paraphyses and more in the base of hymenium and hypothecium. Paraphyses septate, anastomosing, 2–3 μm wide, simple or branched at tips, tips swollen or slightly swollen, not pigmented, 3.5–5.5 μm wide. Asci clavate, 8-spored, 35–48 × 14–17 μm (n = 5). Ascospores generally ellipsoid, occasionally globose, 1-septate, polarilocular or narrow septum remaining, hyaline permanently, 7.5–12 × 4.5–7.5 μm (mean = 9.9 × 5.7 μm; SD = 0.9(L), 0.6(W); L/W ratio 1.2–2.3, ratio mean = 1.8, ratio SD = 0.2; n = 104), globose spores 7.5–9 × 7.0–9.2 μm (mean = 8.0 × 7.7 μm; SD = 0.8(L), 0.9(W); L/W ratio 1.0–1.1, ratio mean = 1.0, ratio SD = 0.1; n = 11). Pycnidia not detected.
Thallus K–, KC–, C–, Pd–. Apothecia K+ wine red. Epihymenium K+ wine red. Epihymenium and hymenium I+ blue. UV–. No lichen substance was detected by TLC.
The species occurs on the calcareous rock. The species is currently known from the type collection.
The species epithet indicates the lichen’s thallus colour, dark green, which is the key characteristic distinguished from all the species in the genus Huriella.
The morphological classification of the new species is not clear between Huriella and Squamulea because the new species has some characteristics for the former genus and others for the latter, i.e. the new species represents mainly areolate thallus without lobed margin and smaller apothecia for the former, whilst showing some squamulose thallus and wider ascospores for the latter (Table
Comparison of the new species with two type species in Huriella and Squamulea.
Species | Huriella aeruginosa | Huriella loekoesiana | Squamulea subsoluta |
---|---|---|---|
Thallus | mainly areolate, rimose or placodioid around edge, but without lobes | areolate (not squamulose) | squamulose, areolate or subsquamulose, margin slightly lobed |
Apothecia (mm in diam.) | 0.2–0.4 | 0.2–0.4(–0.5) | 0.1–0.6 |
Ascospores (μm) | 7.5–12 × 4.5–7.5 | (8.5–)9–11(–12) × (4.5)5–6 | 9.5–12.5 × 5.5–7 |
Molecular phylogeny | Huriella | Huriella | Squamulea |
Reference | – |
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The new species is unique with the key characteristics of green pigmented thallus (with a distinct green layer in a section) and the substrate preference to calcareous rocks amongst all Huriella species.
The new species is similar to ‘Squamulea’ chelonia, Squamulea galactophylla,‘Squamulea’ humboldtiana, S. parviloba and S. subsoluta in the substrate preference to calcareous rocks. However, the new species is different from ‘Squamulea’ chelonia by dark greenish-grey to grey thallus without pruina (vs. yellow orange to deep orange thallus with white pruina), gold to yellow-brown epihymenium (vs. orange epihymenium), larger ascospores (7.5–12 × 4.5–7.5 μm vs. 8–10.4 × 4.7–6.0 μm) and the chemistry (thallus K–, KC– and no substance vs. thallus K+ purple, KC± purplish and the presence of parietin, teloschistin, fallacinal, parietinic acid and emodin) (
The new species differs from S. galactophylla by thallus colour (dark greenish-grey to grey vs. dirty white to yellowish-brown), flat to convex disc (vs. flat disc only), yellowish-orange apothecia (vs. cinnamon-brown apothecia), smaller ascospores (7.5–12 × 4.5–7.5 μm vs. 10–15 × 5–7 μm) (
The new species is distinguished from‘Squamulea’ humboldtiana by dark greenish-grey to grey thallus without pruina (vs. yellow-orange to deep orange thallus with pruina), absence of prothallus (vs. presence of prothallus), larger ascospores (7.5–12 × 4.5–7.5 μm vs. 8.1–9.9 × 4.8–5.9 μm) and the chemistry (thallus K–, KC– and no substance vs. thallus K+ purple, KC± purplish and the presence of parietin, teloschistin, fallacinal, parietinic acid and emodin) (
The new species differs from S. parviloba by dark greenish-grey to grey thallus (vs. yellow-orange to orange thallus), absence of lobes (vs. short narrow elongated lobes around edge), convex and yellow-orange disc (vs. flat and deep orange disc), smaller ascospores (7.5–12 × 4.5–7.5 μm vs. 11–14 × 5.5–7 μm) and the chemistry (thallus K– vs. thallus K+ red) (
The new species is different from S. subsoluta by dark greenish-grey to grey thallus (vs. yellow-orange, orange to reddish-orange thallus), absence of prothallus (vs. black prothallus), flat to convex, yellow-orange apothecia (vs. flat to concave, deep orange apothecia) and the chemistry (thallus K– and no substance vs. thallus K+ red, the presence of parietin, fallacinal, emodin and teloschistin) (
The most distinctive characteristic of the new species is the thallus colour, i.e. dark greenish-grey to grey, which is different from all comparable calcicolous species in the genus Squamulea.
Although some species of Huriella have distinct characteristics, different from Squamulea, such as mainly areolate and non-squamulose thallus without lobes at margin, smaller apothecia and narrower ascospores (
1 | Not directly on rock, but on lichen or bone | 2 |
– | On rock | 4 |
2 | On lichen (Aspicilia) living on rock | Squamulea nesodes |
– | On bone | 3 |
3 | Thallus generally areolate, without blastidia, not pruinose | ‘ Squamulea’ osseophila |
– | Thallus generally (sub)squamulose, blastidia abundant, not pruinose or faintly orange pruinose on thallus | ‘ Squamulea’ phyllidizans |
4 | On calcareous rocks | 5 |
– | On siliceous rocks | 10 |
5 | Thallus pruinose | 6 |
– | Thallus not pruinose | 7 |
6 | Thallus angular, areolate to subsquamulose, prothallus absent | ‘ Squamulea’ chelonia |
– | Thallus areolate or bullate, prothallus black when present | ‘ Squamulea’ humboldtiana |
7 | Thallus whitish, greyish or greenish | 8 |
– | Thallus yellow-orange to orange | 9 |
8 | Thallus dirty whitish, disc cinnamon-brown | Squamulea galactophylla |
– | Thallus dark greenish-grey to grey, disc orange | Huriella aeruginosa |
9 | Areole margins with small lobules | Squamulea parviloba |
– | Areole margins without lobules | Squamulea squamosa (S. subsoluta) |
10 | With blastidia or soredia | 11 |
– | Without blastidia or soredia | 13 |
11 | Thallus brownish-orange, apothecia rare, disc reddish to reddish-brown, ascospores 11–16 × 6–8 μm, isthmus 1–3 μm | Squamulea kiamae |
– | Thallus yellowish-orange to deep orange, apothecia common, disc concolorous to thallus or slightly deeper, ascospores 8.4–13.3 × 5–7.1 μm, isthmus 2.5–4.6 μm | 12 |
12 | Blastidia abundant, sometimes faintly orange pruinose on thallus, but not pruinose on disc | ‘ Squamulea’ phyllidizans |
– | Soredia rarely present, rarely white pruinose on disc, but not pruinose on thallus | Squamulea squamosa (S. subsoluta) |
13 | Thallus areolate to (sub)squamulose | 14 |
– | Thallus areolate or bullate, but not squamulose | 21 |
14 | Prothallus distinctly blackened | ‘ Squamulea’ oceanica |
– | Prothallus absent | 15 |
15 | Disc brownish to reddish or blackish | 16 |
– | Disc orangish | 19 |
16 | Thallus orange, disc reddish, ascospores 11–14.2 × 5.9–7.5 μm | Huriella flakusii |
– | Thallus brownish, disc pale brown, brownish-orange to blackish-brown | 17 |
17 | Disc dark brown-orange to black-brown, hypothecium 20–30 μm high, ascospores 7–9 × 4.5–6.5 μm | Huriella salyangiana |
– | Disc pale brown to brownish-orange, hypothecium 50–150 μm high, ascospores 9–13 × 4.5–6 μm | 18 |
18 | Disc 0.4–0.9 mm diam., hypothecium 50–100 μm high, ascospores 9–13 × 5–6 μm | Squamulea coreana |
– | Disc 0.2–0.4 mm diam., hypothecium 100–150 μm high, ascospores 10–10.5 × 4.5–6 μm | Squamulea uttarkashiana |
19 | Areole margins with small lobules | Squamulea parviloba |
– | Areole margins without lobules | 20 |
20 | Ascospores 8–10.4 × 4.7–6 μm, isthmus 2.1–3.3, not pruinose on disc | ‘ Squamulea’ chelonia |
– | Ascospores 8.4–13.3 × 5.2–7 μm, isthmus 2.5–4 μm, rarely pruinose on disc | Squamulea squamosa (S. subsoluta) |
21 | Thallus yellow-orange to deep orange, prothallus black when present, ascospores 8.1–9.9 × 4.8–5.9 μm, isthmus 2.7–3.2 μm | ‘ Squamulea’ humboldtiana |
– | Thallus yellow-brownish or yellow-greenish, prothallus absent, ascospores 9–15 × 5–8 μm, isthmus 2–5 μm | 22 |
22 | Apothecia 0.2–0.3 mm diam., disc dull brown, dull yellow to bright yellow | 23 |
– | Apothecia 0.3–1 mm diam., disc orange, brownish-yellow to reddish-orange | 24 |
23 | Disc dull yellow to bright yellow, hymenium 50–60 μm high, hypothecium 20–30 μm high, ascospores 9–11 × 5–6 μm, isthmus 4–5 μm | Huriella loekoesiana |
– | Disc dull brown, hymenium 80–100 μm high, hypothecium 80–110 μm high, ascospores 13–14.5 × 7–8 μm, isthmus 3–4 μm | Huriella upretiana |
24 | On mountain, thallus yellow-brown, disc orange, isthmus 3–4 μm | Squmulea micromera |
– | On coast, thallus dull green-yellow to yellow-brown, disc orange to red-orange, isthmus 2–3 μm | Huriella pohangensis |
This work was supported by a grant from the Korean National Research Resource Center Program (NRF-2017M3A9B8069471).