Research Article |
Corresponding author: Beeyoung Gun Lee ( gitanoblue@koagi.or.kr ) Corresponding author: Jae-Seoun Hur ( jshur1@sunchon.ac.kr ) Academic editor: Xinli Wei
© 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 lecanoroid lichen species from the forested wetlands of South Korea, with a key for Korean Protoparmeliopsis species. MycoKeys 84: 163-183. https://doi.org/10.3897/mycokeys.84.70798
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Lecanora parasymmicta Lee & Hur and Protoparmeliopsis crystalliniformis Lee & Hur are described as new lichen species to science from the forested wetlands in southern South Korea. Molecular analyses employing internal transcribed spacer (ITS) and mitochondrial small subunit (mtSSU) sequences strongly support the two lecanoroid species to be distinct in their genera. Lecanora parasymmicta is included in the Lecanora symmicta group. It is morphologically distinguished from Lecanora symmicta (Ach.) Ach., its most similar species, by areolate-rimose thallus, blackish hypothallus, larger apothecia, absence of thalline excipulum from the beginning, narrower paraphyses, larger ascospores, smaller pycnoconidia, and the presence of placodiolic acid. The second new species Protoparmeliopsis crystalliniformis is included in a clade with Protoparmeliopsis bipruinosa (Fink) S.Y. Kondr. and P. nashii (B.D. Ryan) S.Y. Kondr., differs from Protoparmeliopsis ertzii Bungartz & Elix, its most morphologically similar species, by whitish thallus, flat to concave and paler disc, longer ascospores, thallus K+ yellow reaction, presence of atranorin and rhizocarpic acid, and the substrate preference to sandstone or basalt. A key is provided to assist in the identification of Protoparmeliopsis species in Korea.
Biodiversity, hygrophyte, Lecanoraceae, phylogeny, taxonomy
As the genus Lecanora has been considered one of the largest genera in lichens, several infrageneric groups have been specifically or comprehensively studied in diverse aspects in morphology, chemistry and molecular phylogeny (
Although there have been many groups classified as above, a few groups are proved more natural and homogenous and other groups are represented heterogenous without clarity in taxonomy (
This study describes two new lichen-forming fungi species to science in the genera Lecanora, i.e., the L. symmicta group, and Protoparmeliopsis. Field surveys for the lichen biodiversity in the forested wetlands of southern South Korea were accomplished during the summer of 2020, and a few dozen specimens were collected in the wetland forests nearby seashore or in islands (Fig.
Hand sections were prepared manually with a razor blade under a stereomicroscope (Olympus optical SZ51; Olympus, Tokyo, Japan), scrutinized under a compound microscope (Nikon Eclipse E400; Nikon, Tokyo, Japan) and pictured using a software program (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 examined 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 (SD), length-to-width ratio, 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 ten to twenty ascomata per collected specimen 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 with 16 μl of distilled water, 2 μl of DNA extracts and 2 μl of primers ITS5 and ITS4 (
All ITS and mtSSU sequences were aligned and edited manually using ClustalW in Bioedit V7.2.6.1 (
Four independent phylogenetic trees for the genera Lecanora and Protoparmeliopsis were produced from 117 sequences (71 for ITS, and 30 for mtSSU) from GenBank and, 16 new sequences (11 for ITS and 5 for mtSSU) from the new and compared species (Table
Species list and DNA sequence information employed for phylogenetic analysis.
No. | Species | ID (ITS) | ID (mtSSU) | Voucher |
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1 | Lecanora aitema | GU480092 | SPO1 | |
2 | Lecanora atrosulphurea | KY266931 | O-L-195558 | |
3 | Lecanora austrocalifornica | GU480103 | SPO2 | |
4 | Lecanora cinereofusca | KP224470 | KP224465 | Lendemer 34944 (NY) |
5 | Lecanora cinereofusca | KP224471 | KP224464 | Lendemer 35007 (NY) |
6 | Lecanora compallens | KY586043 | JM6948 | |
7 | Lecanora confusa | GU480093 | SPO10 | |
8 | Lecanora confusa | GU480120 | SPO9 | |
9 | Lecanora conizaeoides | AF189717 | U229 | |
10 | Lecanora conizaeoides | KJ766418 | AFTOL-ID 1858 | |
11 | Lecanora expallens | KY586040 | UGDA-L17316 | |
12 | Lecanora flavoleprosa | GU480101 | SPO18 | |
13 | Lecanora cf. fulvastra | GU480119 | SPO8 | |
14 | Lecanora helmutii | MG973240 | MA:Lichen:19506 | |
15 | Lecanora orosthea | AF070035 | U244 | |
16 | Lecanora parasymmicta | MW832793 | MW832799 | BDNA-L-0001218 |
17 | Lecanora parasymmicta | MW832794 | MW832800 | BDNA-L-0001220 |
18 | Lecanora parasymmicta | MW832795 | MW832801 | BDNA-L-0001235 |
19 | Lecanora perpruinosa | AF070025 | U176 | |
20 | Lecanora perpruinosa | DQ787344 | U506 | |
21 | Lecanora polytropa | DQ534470 | Hur ANT050752 | |
22 | Lecanora polytropa | HQ650643 | DQ986807 | AFTOL-ID 1798 |
23 | Lecanora polytropa | JN873881 | U.C. Riverside 47815UCR1 | |
24 | Lecanora polytropa | DQ787348 | U520 | |
25 | Lecanora saxigena | KP224467 | KP224460 | Lendemer 25832 (NY) |
26 | Lecanora saxigena | KP224468 | KP224461 | Lendemer 33186 (NY) |
27 | Lecanora solaris | MH512984 | LYF14–69 | |
28 | Lecanora solaris | MH520111 | ED (14336) & LY | |
29 | Lecanora stanislai | KY586041 | UGDA-L17244 | |
30 | Lecanora stanislai | MK778544 | J. Malicek 10367 | |
31 | Lecanora strobilina | MG973235 | MA:Lichen:19510 | |
32 | Lecanora strobilina | MG973236 | MA:Lichen:19511 | |
33 | Lecanora strobilina | MG973237 | MA:Lichen:19509 | |
34 | Lecanora strobilina | KJ766420 | DUKE:M. Kukwa 4761 | |
35 | Lecanora strobilinoides | MG973238 | MA:Lichen:19507 | |
36 | Lecanora subintricata | GU480112 | SPO28 | |
37 | Lecanora sulphurea | AF070030 | U212 | |
38 | Lecanora sulphurea | DQ787356 | U508 | |
39 | Lecanora symmicta | AF070024 | U205 | |
40 | Lecanora symmicta | GU480113 | SPO29 | |
41 | Lecanora symmicta | MH481912 | O-L-209831 | |
42 | Lecanora symmicta | MW832788 | BDNA-L-0000547 | |
43 | Lecanora symmicta | MW832789 | BDNA-L-0000548(br) | |
44 | Lecanora symmicta | MW832790 | BDNA-L-0000548(yel) | |
45 | Lecanora symmicta | MW832791 | BDNA-L-0000551 | |
46 | Lecanora symmicta | MW832792 | BDNA-L-0000642 | |
47 | Lecanora symmicta | KJ766421 | EGR:K. Molnar 23-08-2005/B | |
48 | Lecanora symmicta | KJ152466 | C. Printzen 9999a (FR) | |
49 | Lecanora varia | MK672852 | MK693694 | Kondratyuk S. 21325 (KW-L) |
50 | Polyozosia contractula | AF070032 | U236 | |
51 | Polyozosia contractula | HQ650604 | DQ986898 | AFTOL-ID 877 |
52 | Polyozosia poliophaea | MG925981 | MG925879 | O:L 200460 |
53 | Polyozosia sp. | MW832798 | BDNA-L-0001105 | |
54 | Protoparmeliopsis achariana | AF070019 | U155 | |
55 | Protoparmeliopsis achariana | DQ787342 | U525 | |
56 | Protoparmeliopsis bipruinosa | AF159932 | U354 | |
57 | Protoparmeliopsis bolcana | MK672838 | MK693686 | Kondratyuk S. 20309 (KW-L) |
58 | Protoparmeliopsis chejuensis | MK672839 | MK693687 | KoLRI 022622 |
59 | Protoparmeliopsis chejuensis | MK672840 | MK693688 | KoLRI 022618 |
60 | Protoparmeliopsis crystalliniformis | MW832796 | MW832802 | BDNA-L-0000298 |
61 | Protoparmeliopsis crystalliniformis | MW832797 | MW832803 | BDNA-L-0000349 |
62 | Protoparmeliopsis garovaglii | AF189718 | M107 | |
63 | Protoparmeliopsis garovaglii | KT453728 | KT453818 | Leavitt 089 (BRY-C) |
64 | Protoparmeliopsis garovaglii | KU934537 | Leavitt 199 (BRY-C) | |
65 | Protoparmeliopsis garovaglii | MK084624 | Szczepanska 1240 | |
66 | Protoparmeliopsis garovaglii | MK084626 | Flakus 21175 | |
67 | Protoparmeliopsis garovaglii | MK672841 | MK693689 | M. Haji Moniri (KW-L) |
68 | Protoparmeliopsis kopachevskae | MK672845 | KoLRI 040224 | |
69 | Protoparmeliopsis kopachevskae | MK672846 | KoLRI 040267 | |
70 | Protoparmeliopsis kopachevskae | MK672847 | KoLRI 040276 | |
71 | Protoparmeliopsis laatokaensis | MN912366 | 20132508 | |
72 | Protoparmeliopsis macrocyclos | AF159933 | U273 | |
73 | Protoparmeliopsis muralis | KC791770 | BGK247 | |
74 | Protoparmeliopsis muralis | KP059048 | KP059054 | SK 765 |
75 | Protoparmeliopsis muralis | KT818623 | DNA 9890 (F) | |
76 | Protoparmeliopsis muralis | KU934555 | Leavitt 146 (BRY-C) | |
77 | Protoparmeliopsis muralis | KU934560 | Vondrak 106b (PRA) | |
78 | Protoparmeliopsis muralis | KY379232 | BGK257 | |
79 | Protoparmeliopsis muralis | LC547497 | CBM:FL-41434 | |
80 | Protoparmeliopsis muralis | KJ766466 | EGR:K. Molnar U0501/AO | |
81 | Protoparmeliopsis nashii | AF159931 | U253 | |
82 | Protoparmeliopsis peltata | KT453722 | KT453860 | |
83 | Protoparmeliopsis peltata | KT453723 | MS014622 | |
84 | Protoparmeliopsis peltata | KU934746 | Kaz 13085pelt | |
85 | Protoparmeliopsis peltata | KU934751 | Vondrak V127 (PRA) | |
86 | Protoparmeliopsis pseudogyrophorica | MK672851 | MK693693 | KoLRI 016651 |
87 | Protoparmeliopsis zareii | KP059049 | KP059055 | SK 480 |
88 | Protoparmeliopsis zareii | KP059056 | SK 481 | |
89 | Protoparmeliopsis sp. | KU934865 | Vondrak 9980 (PRA) | |
90 | Protoparmeliopsis sp. | KU934866 | Vondrak 10043 (PRA) | |
91 | Protoparmeliopsis sp. | KU934867 | Vondrak 10044 (PRA) | |
92 | Protoparmeliopsis sp. | KU934868 | Vondrak 10055 (PRA) | |
93 | Protoparmeliopsis sp. | KU934869 | Vondrak 9992 (PRA) | |
94 | Tephromela atra | HQ650608 | DQ986879 | AFTOL-ID 1373 |
Overall | 82 | 35 |
Phylogenetic relationships amongst available species in the Lecanora symmicta group based on a Maximum Likelihood analysis of the dataset of ITS sequences. The tree was rooted with five sequences of the Lecanora subfusca group and Tephromela. 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 sequences of Lecanora parasymmicta and Lecanora symmicta produced from this study are presented in bold, and all species names are followed by the GenBank accession numbers. Reference Table
Phylogenetic relationships amongst available species in the Lecanora symmicta group based on a Maximum Likelihood analysis of the dataset of the mitochondrial small subunit (mtSSU) sequences. The tree was rooted with four sequences of the Lecanora subfusca group. 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 Lecanora parasymmicta 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 Protoparmeliopsis based on a Maximum Likelihood analysis of the dataset of ITS sequences. The tree was rooted with five sequences of the genus Polyozosia. 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 Protoparmeliopsis crystalliniformis 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 Protoparmeliopsis based on a Maximum Likelihood analysis of the dataset of the mitochondrial small subunit (mtSSU) sequences. The tree was rooted with three sequences of the genus Polyozosia. 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 Protoparmeliopsis crystalliniformis is presented in bold, and all species names are followed by the GenBank accession numbers. Reference Table
Lecanora parasymmicta differs from L. symmicta, the most similar species, by its areolate-rimose thallus (vs. areolate to leprose thallus), blackish hypothallus (vs. hypothallus indistinct), larger apothecia (up to 1.7 mm diam. vs. up to 1 mm diam.), absence of thalline excipulum from the beginning (vs. presence of thalline excipulum when young at least), narrower paraphyses (1–1.5 μm vs. 2–2.5 μm), larger ascospores (11–18 × 4–7 μm vs. 9–15.5 × 4–5 μm), smaller pycnoconidia (12–21 × 0.5–0.8 μm vs. 18–25 × 0.5–1.0 μm), chemical reactions (thallus K± slightly yellow, C–, KC– and UV– vs. K–, C± orange, KC± slightly yellow, UV+ dull orange), and the presence of placodiolic acid (vs. presence of arthothelin and ±thiophanic acid).
South Korea, Jeju Island, Aewol-eup, Gwangnyeongri/bongseongri, Mt. Halla, a forested wetland, 33°21.85'N, 126°26.91'E, 980 m alt., on bark of Maackia fauriei (H. Lév.) Takeda, 08 July 2020, B.G.Lee & H.J.Lee 2020-001020, with Graphis scripta (L.) Ach. (holotype: BDNA-L-0001220; GenBank MW832794 for ITS and MW832800 for mtSSU); same locality, on bark of Malus sieboldii (Regel) Rehder, 08 July 2020, B.G.Lee & H.J.Lee 2020-001018, (paratype: BDNA-L-0001218; GenBank MW832793 for ITS and MW832799 for mtSSU); same locality, on bark of Malus sieboldii, 08 July 2020, B.G.Lee & H.J.Lee 2020-001019, with Phaeographis aff. inusta (paratype: BDNA-L-0001219); same locality, on bark of Maackia fauriei, 08 July 2020, B.G.Lee & H.J.Lee 2020-001026, (paratype: BDNA-L-0001226); same locality, on bark of Maackia fauriei, 08 July 2020, B.G.Lee & H.J.Lee 2020-001035, with Lecanora megalocheila (Hue) H. Miyaw. (paratype: BDNA-L-0001235; GenBank MW832795 for ITS and MW832801 for mtSSU); same locality, on bark of Ligustrum obtusifolium Siebold & Zucc., 08 July 2020, B.G.Lee & H.J.Lee 2020-001036, with Graphis scripta (paratype: BDNA-L-0001236).
Thallus corticolous, crustose, areolate to rimose but not leprose, light olivish gray to light gray, margin determinate, not pruinose, 60–200 μm thick; cortex hyaline, 5–10 μm thick; medulla often intermixed with algae and even with bark layer, small crystals in cortex or between algae, dissolving in K; photobiont coccoid, cells globose to ellipsoid, 5–15 μm. Hypothallus blackish.
Apothecia abundant, rounded, often contiguous or even coalescent, emerging on the surface of thallus and sessile when mature but margin generally attached to thallus surface, constricted at the base, 0.3–1.7 mm diam. Disc flat in the beginning and soon convex, smooth or becoming rugose by apothecia adjoining, not pruinose or slightly pruinose, pale yellow in the beginning and slightly darker when mature, sometimes with dark spots (algae), 180–400 μm thick; biatorine. Thalline excipulum absent from the beginning, proper excipulum present and sometimes slightly paler than disc, more distinctive when young, hyaline but yellowish brown to pale brown at periphery with granules which dissolving in K, periphery color same to epihymenium, ca. 90 μm wide laterally and 70–80 μm wide at periphery, disappearing to the base. Epihymenium yellowish brown to pale brown, granular, dissolving in K, 10–20 μm high. Hymenium hyaline, 70–90μm high. Subhymenium hyaline, 30–50 μm high. Hypothecium hyaline, prosoplectenchymatous (irregular), 50–60 μm high. Crystals and oil droplets absent in apothecial section. Paraphyses septate, anastomosing, 1–1.5 μm wide, simple or branched at tips, tips not swollen or slightly swollen, not pigmented, epihymenium pigmented by granules, not by paraphysial tips, ca. 1.5 μm wide. Asci clavate, 8-spored, 50–60 × 13–21 μm (n = 7). Ascospores constantly simple but rarely 1-septate, coarsely biseriate or irregularly arranged, 11–18 × 4–7 μm (mean = 13.8 × 5.8 μm; SD = 1.62(L), 0.63(W); L/W ratio 1.8–4.0, ratio mean = 2.4, ratio SD = 0.3; n = 105). Pycnidia immersed, ostiolar region slightly projected with a thalline excipulum, round to irregularly asymmetric, brown to black, 220 × 180 μm. Pycnoconidia thread-like, generally curved, 12–21 × 0.5–0.8 μm.
Lecanora parasymmicta morphology (BDNA-L-0001235, paratype in A BDNA-L-0001220, holotype in B–M) A–C habitus and apothecia, thalline margin of apothecia consistently absent from the beginning D blackish hypothallus (red arrows) E apothecia in vertical section F biatorine apothecia without thalline margin G–J clavate asci with eight spores K ascospores constantly simple but rarely 1-septate L immersed pycnidia M thread-like, curved pycnoconidia. Scale bars: 1 mm (A–D); 200 μm (E); 50 μm (F); 10 μm (G–K); 100 μm (L); 10 μm (M).
Thallus K– or K+ slightly yellowish, KC–, C–, Pd–. Hymenium, epihymenium and ascus tholus I+ blue. UV–. Usnic acid, zeorin, and placodiolic acid were detected by TLC.
Comparison of the new species with close species in the Lecanora symmicta group.
Species | L. parasymmicta | L. aitema | L. confusa | L. strobilina | L. symmicta |
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Thallus growth form | areolate-rimose | granular-areolate | granular-areolate | granular-subareolate | areolate-reprose |
Thallus color | olive-gray to gray | cream-white | green gray to yellow gray | white to pale yellow-green | variable (pale yellow-green, white or green-gray) |
Prunia | absent or slightly pruinose on disc | absent | absent | present | absent |
Hypothallus | blackish | indistinct or pale brown | absent or indistinct | indistinct | indistinct |
Apothecia (mm diam.) | 0.3–1.7 | 0.2–0.5 | 0.4–0.7 | 0.4–1.0 | 0.3–1.0 |
Thalline excipulum | absent from beginning | present when young | present when young | present when young | present when young |
Epihymenium | yellow-brown | yellow-brown | brown | colorless | colorless, yellow-brown to olive |
Paraphyses (μm) | 1–1.5 | 2–2.5 | 1–2 | 1–1.5 | 2–2.5 |
Asci (μm) | 50–60 × 13–21 | 35–45 × 10–15 | 32–45 × 11–15 | 35–45 × 10–17 | 30–47 × 8–12* |
Ascospores (μm) | 11–18 × 4–7 | 12–17 × 4.5–5.5 | 10–14 × 4–5 | 10–15 × 4–6 | 9–15.5 × 4–5 8–12 × 4–6* |
Pycnoconidia (μm) | 12–21 × 0.5–0.8 | not observed | not observed | 25 × 1.0 | 18–25 × 0.5–1.0 |
Spot test | thallus K± slightly yellow, C–, KC– | thallus K–, KC± slightly yellow | thallus K–, C+ orange, KC+ orange | thallus K+ yellow to brown, KC± yellow | thallus K–, C± orange, KC± slightly yellow |
UV | negative | pale orange | bright orange | pale orange | dull orange |
Substance | usnic acid, zeorin, placodiolic acid | ±usnic acid, ±zeorin | usnic acid, ±zeorin, thiophanic acid, ±arthothelin | usnic acid, zeorin | usnic acid, zeorin, arthothelin, ±thiophanic acid |
Reference | BDNA-L-0001218 (paratype), BDNA-L-0001220 (holotype), and BDNA-L-0001235 (paratype) |
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The species occurs on the bark of Ligustrum obtusifolium, Maackia fauriei, and Malus sieboldii. The species is currently known from the type collections.
The species epithet indicates the lichen’s morphological similarity to the close species Lecanora symmicta.
The new species is morphologically similar to Lecanora symmicta in its areolate and gray thallus, yellowish apothecia without developed thalline excipulum, yellowish brown epihymenium filled with pigmented granules which dissolving in K, and the presence of conidia. However, the new species differs from L. symmicta by its areolate-rimose thallus, blackish hypothallus, larger apothecia, absence of thalline excipulum from the beginning, narrower paraphyses, larger asci, larger ascospores, smaller pycnoconidia, chemical reaction, and the presence of placodiolic acid (
The new species is comparable to Lecanora aitema (Ach.) Hepp, L. confusa, and L. strobilina in the L. symmicta group as all those are corticolous without soredia or leprose thallus. However, the new species differs from L. aitema by olive-gray to gray thallus, blackish hypothallus, larger and paler apothecia, absence of thalline excipulum from the beginning, larger asci, wider ascospores, chemical reaction, presence of placodiolic acid, and the substrate preference to deciduous trees/shrubs (vs. conifers) (
The new species is different from Lecanora confusa by the absence of thalline excipulum from the beginning, larger asci, larger ascospores, chemical reaction, and the presence of placodiolic acid (
The new species is distinguished from Lecanora strobilina by olive-gray to gray thallus without pruina, presence of black hypothallus, absence of thalline excipulum from the beginning, yellow-brown epihymenium, absence of crystals in apothecial section, larger asci, larger ascospores, smaller pycnoconidia, chemical reaction, and the presence of placodiolic acid (
All above compared species do not contain placodiolic acid and Lecanora species with placodiolic acid, such as L. placodiolica Lumbsch & Elix, L. cinereofusca H. Magn., L. sarcopidoides (A. Massal.) Hedl., L. subravida Nyl., L. semitensis (Tuck.) Zahlbr. and L. opiniconensis Brodo, are considered for discriminating the new species. Lecanora placodiolica differs from the new species by yellowish thallus, absence of hypothallus, presence of thalline excipulum, and darker (red-brown) discs (
South Korea, Gangwon Province, Gangneung, Seongsan-myeon, Eoheul-ri, a forested wetland, 37°43.61'N, 128°48.13'E, 212 m alt., on bark of Alnus sibirica Fisch. ex Turcz., 02 June 2020, B.G.Lee & H.J.Lee 2020-000347, with Lecanora strobilina, Lecidella euphorea (Flörke) Kremp., Traponora varians (Ach.) J. Kalb & Kalb (BDNA-L-0000547; GenBank MW832788 for ITS); same locality, on bark of Alnus sibirica, 02 June 2020, B.G.Lee & H.J.Lee 2020-000348, two variants (one with pale brown discs and the other with yellow discs) of Lecanora symmicta with Lecidella euphorea, Rinodina sp., Traponora varians (BDNA-L-0000548; GenBank MW832789 for ITS of the former variant and GenBank MW832790 for ITS of the latter); same locality, on bark of Alnus sibirica, 02 June 2020, B.G.Lee & H.J.Lee 2020-000351, two above variants of Lecanora symmicta with Traponora varians (BDNA-L-0000551; GenBank MW832791 for ITS); Pyeongchang-gun, Daegwallyeong-myeon, Hoenggye-ri, a forested wetland, 37°46.00'N, 128°42.33'E, 1,047 m alt., on bark of Maackia amurensis Rupr. & Maxim., 03 June 2020, B.G.Lee & H.J.Lee 2020-000442, with Buellia disciformis (Fr.) Mudd, Buellia sp., Catillaria nigroclavata (Nyl.) J. Steiner, Lecanora megalocheila, Lecidella euphorea, Rimularia cf. caeca, Rinodina sp. (BDNA-L-0000642; GenBank MW832792for ITS).
Protoparmeliopsis crystalliniformis differs from P. ertzii by thallus color (grayish white to white vs. pale beige to ochraceous), flat to concave disc (vs. flat to convex disc), paler disc color (pale brown to dark brown vs. deep reddish brown), longer ascospores (8.5–17 × 4.2–7 μm vs. 9.4–11.3 × 5.3–6.6 μm), chemistry (thallus K+ yellow, and the presence of atranorin and rhizocarpic acid vs. all spot tests negative and no substance), and the substrate preference (sandstone or basalt vs. exposed lava).
South Korea, South Jeolla Province, Sinan, Ja-Eun Island, a wetland just nearby coast, 34°55.96'N, 126°04.30'E, 5 m alt., on rock (sandstone), 16 April 2020, B.G.Lee & D.Y.Kim 2020-000149, with Ramalina yasudae Räsänen, Xanthoparmelia coreana (Gyeln.) Hale (holotype: BDNA-L-0000349; GenBank MW832797 for ITS, MW832803 for mtSSU, and MW832822 for LSU); same locality, on rock (sandstone, not calcareous), 16 April 2020, B.G.Lee & D.Y.Kim 2020-000151, with Buellia spuria (Schaer.) Anzi, Ramalina yasudae, Xanthoparmelia coreana (paratype: BDNA-L-0000351).
Thallus saxicolous, areolate to squamulose, linearly or web-like dispersed following furrows of substrate, not forming a rosette, pale grayish white to white, margin indeterminate, not pruinose, 100–450 μm thick; cortex pale brown, 10–20 μm thick; medulla below algal layer, 30–50 μm (sometimes 150–200 μm) thick; algal layer 50–80 μm thick, small crystals in cortex or between algal cells, dissolving but remaining in K; photobiont coccoid, cells globose to ellipsoid, 5–15 μm. Hypothallus absent.
Apothecia abundant, rounded, often contiguous or even coalescent, emerging on the surface of thallus and sessile when mature, constricted at the base, 0.3–1.7 mm diam. Disc flat or slightly concave, crenulate or entire, smooth or becoming rugose by apothecia adjoining, not pruinose, pale brown to dark brown from the beginning, 250–350 μm thick; lecanorine. Thalline excipulum persistent or rarely excluded, concolorous to thallus, 125–160 μm laterally, 80–150 μm at periphery, cortex inconspicuous, concolorous to epihymenium or slightly paler, up to 5 μm, with small and large crystals, small crystals dissolving but remaining in K, large crystals not dissolving in K. Proper excipulum inconspicuous. Epihymenium brown to pale brown, with tiny granules, granules and pigments dissolving in K, 10–20 μm high. Hymenium hyaline, 80–100μm high. Subhymenium hyaline, 30–50 μm high. Hypothecium hyaline, prosoplectenchymatous (irregular), 100–150 μm high. Oil droplets present in hymenium to upper hypothecium. Paraphyses septate, anastomosing, 1–1.5 μm wide, generally simple or occasionally branched at tips, tips not swollen or slightly swollen, not pigmented, 1.5–2 μm wide. Asci narrowly clavate, 8-spored, 40–65 × 10–12 μm (n = 6). Ascospores simple and often biguttulate in the beginning then having an oval-shaped oil drop by assembly of guttules when mature, ellipsoid to narrowly ellipsoid, rarely globose, 8.5–17 × 4.2–7 μm (mean = 11.8 × 5.5 μm; SD = 1.9(L), 0.6(W); L/W ratio 1.4–3.1, ratio mean = 2.2, ratio SD = 0.4; n = 102). Pycnidia not detected.
Protoparmeliopsis crystalliniformis morphology (BDNA-L-0000349, holotype) A–C habitus and apothecia, areolate to squamulose thallus in white to whitish gray color D–E apothecia in vertical section F–G well-developed thalline margin H large crystals present in the thalline margin, not dissolving in KOH I clavate ascus J ascospores constantly simple and ellipsoid, often biguttulate in the beginning. Scale bars: 1 mm (A–C); 200 μm (D–E); 100 μm (F–G); 50 μm (H); 10 μm (I–J).
Thallus K+ yellow, KC–, C–, Pd–. Hymenium I+ blue. UV–. Atranorin and rhizocarpic acid were detected by TLC.
Comparison of the new species with close species in the genus Protoparmeliopsis.
Species | P. crystalliniformis | P. bipruinosa | P. ertzii | P. nashii |
---|---|---|---|---|
Thallus color | gray-white to white | pale yellow-green to gray-green-yellow, finally pale brown | pale beige to ochraceous | various shades of yellow to orange-brown cast |
pruina | not present | pruina on both thallus and disc | not present | not on thallus, but present on disc |
Disc evenness | flat to slightly concave | flat to slightly convex | flat to convex | flat to slightly concave |
Disc color | pale brown to dark brown | yellow-brown to pale orange or green | deep reddish brown | weakly yellow to strongly yellow |
Crystals | large crystals, insoluble | not observed | large crystals, insoluble | not observed |
Ascospores (μm) | 8.5–17 × 4.2–7 | 10–14 × 4–7.5 | 8.8–12.7 × 4.9–6.9 | 8–14 × 4–9 |
Spot test | thallus K+ yellow, KC–, C–, Pd– | thallus K–, C–; cortex KC+ yellow, P–; medulla KC–, P+ yellow or P– | all negative | thallus K– or occasionally K+, C–; cortex KC+ yellow, P–; medulla KC–, P+ yellow or less often P– |
Substance | atranorin,rhizocarpic acid | usnic acid, psoromic acid, or fatty acids | no substance | usnic acid, psoromic acid, or fatty acids |
Substrate | sandstone or basalt on seashore | volcanic tuff, basalt, rhyolite, or sedimentary rocks from desert scrub to woodlands | exposed lava on island | siliceous rocks (conglomerate to volcanic rocks), rarely on limestone in woodlands, desert scrub or grassland |
Reference | BDNA-L-0000298, BDNA-L-0000349 (holotype), and BDNA-L-0000351 (paratype) |
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The species occurs on the rock (sandstone or basalt) nearby coast. The species is currently known from two localities in the southern coast of South Korea.
The species epithet indicates the insoluble large crystals present in the thalline excipulum of the lichen.
The new species is morphologically similar to Protoparmeliopsis ertzii in having insoluble large crystals in the thalline excipulum and the absence of usnic acid, which are the key characteristics distinguishing them from all other species in the genus Protoparmeliopsis. However, the new species differs from P. ertzii by whitish thallus, flat to concave disc, paler disc color, longer ascospores, chemical reaction, presence of atranorin and rhizocarpic acid, and the substrate preference (
The new species is compared with P. bipruinosa and P. nashii as those are closest to the new species in molecular results (Figs
The new species is different from P. nashii by whitish thallus, absence of pruina, presence of large crystals, and the presence of atranorin and rhizocarpic acid (
South Korea, South Jeolla Province, Goheung, Yeongnam-myeon, Ucheon-ri, a coastal area, 34°37.02'N, 127°29.82'E, 31 m alt., on rock (basalt), 14 April 2020, B.G.Lee 2020-000098, with Caloplaca bogilana Y. Joshi & Hur, Circinaria caesiocinerea (Nyl. ex Malbr.) A. Nordin, Savić & Tibell, Pertusaria flavicans Lamy (BDNA-L-0000298; GenBank MW832796 for ITS, MW832802 for mtSSU, and MW832821 for LSU); same locality, on rock (basalt), 14 April 2020, B.G.Lee 2020-000099, with Buellia sp., Circinaria caesiocinerea (BDNA-L-0000299); same locality, on rock (basalt), 14 April 2020, B.G.Lee 2020-000100, with Buellia aff. nashii (BDNA-L-0000300); same locality, on rock (basalt), 14 April 2020, B.G.Lee 2020-000102, with Buellia sp., Caloplaca bogilana, Circinaria caesiocinerea, Endocarpon maritimum Y. Joshi & Hur, Parmotrema grayanum (Hue) Hale (BDNA-L-0000302); same locality, on rock (basalt), 14 April 2020, B.G.Lee 2020-000103, with Circinaria caesiocinerea, Endocarpon maritimum, Pertusaria flavicans (BDNA-L-0000303); same locality, on rock (basalt), 14 April 2020, B.G.Lee 2020-000105, with Buellia aff. nashii, Circinaria caesiocinerea, Pertusaria flavicans (BDNA-L-0000305); same locality, on rock (basalt), 14 April 2020, B.G.Lee 2020-000107, with Xanthoparmelia mexicana (Gyeln.) Hale (BDNA-L-0000307); same locality, on rock (basalt), 14 April 2020, B.G.Lee 2020-000108, with Caloplaca bogilana, Endocarpon maritimum, Pertusaria flavicans (BDNA-L-0000308); same locality, on rock (basalt), 14 April 2020, B.G.Lee 2020-000110, with Buellia aff. nashii, Buellia sp., Lecanora oreinoides (Körb.) Hertel & Rambold (BDNA-L-0000310).
1 | Thalline margin with large crystals, containing atranorin and rhizocarpic acid | P. crystalliniformis |
– | Thalline margin without large crystals | 2 |
2 | Thallus whitish | 3 |
– | Thallus yellowish, brownish, or greenish | 4 |
3 | Apothecia 0.4–0.7 mm diam., disc with white pruina, epihymenium brownish, hymenium 30–40 μm high, hypothecium 70–100 μm high, ascospores wider 6–7 μm | P. chejuensis |
– | Apothecia 0.5–1.5 mm diam., disc without pruina but thalline margin with pruina, epihymenium dull yellowish, hymenium 45–55 μm high, hypothecium 60–70 μm high, ascospores narrower 4.5–5.5 μm | P. kopachevskae |
4 | Soralia developed on thallus, apothecia absent | P. zerovii |
– | Soralia absent, apothecia present | 5 |
5 | Thallus greenish gray, disc light yellow to pale brown, ascospores 8–13 × 4.5–7 μm, medulla KC– (not containing gyrophoric acid) | P. muralis |
– | Thallus yellowish, green to grayish yellow, disc dull brown to dark brown, ascospores 17–21 × 5.5–6.5 μm, medulla KC+ yellow (containing gyrophoric acid) | Sedelnikovaea pseudogyrophorica (P. pseudogyrophorica) |
This work was supported by a grant from the Korean National Research Resource Center Program (NRF-2017M3A9B8069471) and the Korean Forest Service Program through the Korea National Arboretum (KNA-202003127AF-00) for the forested wetland conservation of Korea.