Taiwania 68(4): 417‒424, 2023
DOI: 10.6165/tai.2023.68.417

Highlighted Student Research

Racoleus japonicus sp. nov. (Teratosphaeriaceae, Ascomycota), a newsterile filamentous lichen collected from Japan
Kento MIYAZAWA1,*, Yoshihito OHMURA2,3
1. Degree Programs in Life and Earth Sciences, Graduate School of Science and Technology, University of Tsukuba, 1-1-1 Tennodai,
Tsukuba, Ibaraki, 305-8577, Japan. 2. Department of Botany, National Museum of Nature and Science, 4-1-1 Amakubo, Tsukuba,
305-0005, Japan. 3. School of Integrative and Global Majors, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8577,
Japan. *Corresponding author’s phone no.: +81298538335; email: miyazawa.kento.ss@alumni.tsukuba.ac.jp(Manuscript received 28 August 2023; Accepted 5 October 2023; Online published 12 October 2023)
ABSTRACT: Racoleus japonicus is described as a new species. It is characterized by a blackish brown minutely filamentous thallus
with lateral spines, vertical arrangement of hyphae with uneven and undulate to corrugated hyphal walls, and a Trentepohlia
photobiont. In this genus, only R. trichophorus was known before the present study. Racoleus japonicus is distinguished from thatspecies by having broader filaments (10–15 μm wide), larger hyphal cells (10–20 × 3–7 μm), and lateral sinuous spines. It wascollected on shady rocks from northern to southern Japan at elevations between 290 and 1700 m. A molecular phylogeny based on
nuSSU, nuLSU and mtSSU inferred that R. japonicus has a close relationship with unidentified fungi of Capnodiales, and that is
not related to Cystocoleus and Racodium which have similar filamentous thalli. Based on the molecular phylogenetic analyses, R.
japonicus is classified within Teratosphaeriaceae.
KEY WORDS: Capnodiales, Cystocoleus, lichenized fungi, mtSSU, nuLSU, nuSSU, Racodium, Trentepohlia.

INTRODUCTION
The genera Cystocoleus Thwaites, Racodium Fr. and
Racoleus R. Sant. & D. Hawksw. are known as sterilefilamentous lichens that are characterized by blackish
brown fungal hyphae surrounding a filamentous green
alga of the genus Trentepohlia. According to
Hawksworth et al. (2011), each genus is characterized by
the following morphology: Cystocoleus has a twisted
arrangement of hyphae with a corrugated hyphal wall;
Racodium has a vertical arrangement of hyphae with astraight hyphal wall; and Racoleus has a verticalarrangement of hyphae with a corrugated wall.
Furthermore, by scanning electronic microscopy, awarted ornamentation on the surface of hyphal wall isobserved only in Cystocoleus (vs. smooth surface in
Racodium and Racoleus). Emphasizing the lateral spines
as a feature of Racoleus by Hawksworth et al. (2011) will
be discussed later in this paper. These genera wereconsidered to belong to Capnodiales because of thesimilar appearance in morphology (Hawksworth et al.,2011). Abdollahzadeh et al. (2020) delimited
Capnodiales s. str. and transferred Cystocoleus to
Mycosphaerellales and Racodium to Racodiales, but
those authors did not mention Racoleus.
Cystocoleus and Racodium, but not Racoleus, wereanalyzed with other related fungi based on nuSSU,
nuLSU and mtSSU sequence data (Muggia et al., 2008)
or by using nuLSU, ITS rDNA, TEF-1α, and RPB2
sequences (Abdollahzadeh et al., 2020). The molecularphylogenetic analyses of those authors suggested that therelationship between Cystocoleus and Racodium is notsister and that the lichenization occurred independently ineach group within Capnodiales s. lat., a group of fungi inwhich other taxa are non-lichenized (i.e., saprophytes,
parasites, ectophytes, or epiphytes; Abdollahzadeh et al.,2020). While the taxonomic position of Cystocoleus andRacodium has been discussed based on molecularphylogeny and morphological data, the position of
Racoleus remains unclear due to a lack of DNA sequences.
While examining the Japanese sterile filamentous
lichens housed in the herbarium of the National Museumof Nature and Science (TNS), Tsukuba, Japan, several
specimens were recognized as an undescribed species of
the genus Racoleus. The aim of this study is to describe
and illustrate the new species, Racoleus japonicus, and toinfer its phylogenetic position based on molecular data.

MATERIALS AND METHODSMorphology and chemistry
All voucher specimens examined in this study arehoused in the herbarium of the National Museum of
Nature and Science (TNS), Tsukuba, Japan.
Morphological observations and photography were
performed using a dissecting microscope (SZX16;
Olympus) and a differential interference contrast
microscope (BX53; Olympus) with a digital camera (EOS
Kiss X10i; Canon). Anatomical examinations were
performed using hand-cut sections mounted in GAW(glycerin: ethanol: water = 1: 1: 1) solution (Asahina, 1936).
Color spot tests for K, C, KC, and Pd were performed
according to Orange et al. (2001). Secondary substanceswere examined using high-performance thin layer
417

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Vol. 68, No. 4

Table 1. Collections/strains and their GenBank accession numbers. New sequences obtained in this study are in bold.
Taxa
Capnodiales sp.

Collection/strain No. nuSSU
A557
KT263481
A571
KT263492
A577
KT263494
A863
KT263485
A886
KT263488
A913
KT263484
A951
KT263482
A959
KT263487
A960
KT263486
A1043
KT263480
Capnodium coffeae
CBS 147.52
DQ247808
Cladosporium cladosporioides CBS 170.54
DQ678004
Cystocoleus ebeneus
L161
EU048571
L348
EU048573
Friedmanniomyces endolithicus CCFEE 522 & 524 DQ066715
Mycosphaerella fijiensis
OSC 100622
DQ767652
Myriangium duriaei
CBS 260.36
AY016347

nuLSU
KT263447
KT263457
KT263459
KT263451
KT263453
KT263450
KT263448
KT263460
KT263452
KT263446
DQ247800
DQ678057
EU048578
EU048580
GU250364
DQ678098
AY016365

mtSSU
KT263516
KT263527
KT263529
KT263520
KT263523
KT263519
KT263517
KT263522
KT263521
KT263515
FJ190609
FJ190628
EU048584
EU048586
GU250409
FJ190656
AY571389

Neodevriesia strelitziae
Petrophila incerta

GU301810
GU323961
GU323963
EU048583
EU048582
LC779821
LC779822
LC779825
LC779823
LC779824
LC779826
DQ470968
KC315876
DQ678075

GU561845
GU324022
GU324024
EU048588
EU048589

Racodium rupestre
Racoleus japonicus

Ramularia punctiformis
Recurvomyces mirabilis
Scorias spongiosa

CBS 122379
TRN 62TRN 77
L346
L424
YO9473YO9651 (Holotype)
KeM1561
KeM1700
KeM1701
KeM1702
CBS 113265
CCFEE 5475
CBS 325.33

GU296146
GU323991
GU323993
EU048575
EU048577
LC779815
LC779816
LC779817
LC779818
LC779819
LC779820
DQ471017
KC315865
DQ678024

chromatography (HPTLC) following Schumm and Elix
(2015). Solvent system B′ (n-hexane: methyl tert-butyl
ether: formic acid, 140: 72: 18) (Culberson and Johnson,1982) was used for HPTLC. The spot color was checked
under 254 and 366 nm UV light and visible light, before
and after spraying with 10% sulfuric acid on the HPTLC
plate and charring at 90°C for 20 minutes.
DNA extraction, PCR amplification and sequencing
DNA was extracted from fresh material collected
within one year according to a modified method of
Izumitsu et al. (2012) (see also Miyazawa et al., 2022) ora modified CTAB protocol (Hosaka, 2009). The voucher
specimens for DNA extractions are housed in TNS.
For PCR amplification, 10 μL of PCR mix contained
1 μL genomic DNA extraction, 0.25 μL of each primer(10 pmol/μL) and 5 μL EmeraldAmp® MAX PCR
Master Mix (TaKaRa Bio Inc.). The partial sequences ofthe small subunit and the large subunit of the nuclear
ribosomal RNA gene (nuSSU and nuLSU) and of thesmall subunit of the mitochondrial ribosomal RNA gene
(mtSSU) were amplified with the primer sets nuSSU0021(Gargas and DePriest, 1996) and nuSSU0852 (Gargas
418

LC779827

LC779828
LC779829
FJ190611
KC315887
FJ190643

Reference
Muggia et al., 2016
Muggia et al., 2016
Muggia et al., 2016
Muggia et al., 2016
Muggia et al., 2016
Muggia et al., 2016
Muggia et al., 2016
Muggia et al., 2016
Muggia et al., 2016
Muggia et al., 2016
Schoch et al., 2006a, 2009
Schoch et al., 2006b, 2009
Muggia et al., 2008
Muggia et al., 2008
Selbmann et al., 2005
Schoch et al., 2006a, 2009
Lumbsch and Lindemuth, 2001;
Lumbsch et al., 2005
Ruibal et al., 2009; Schoch et al., 2009
Ruibal et al., 2009
Ruibal et al., 2009
Muggia et al., 2016
Muggia et al., 2016
This study
This study
This study
This study
This study
This study
Spatafora et al., 2006; Schoch et al., 2009
Unpublished
Schoch et al., 2006a, 2009

and Taylor, 1992) for nuSSU, LIC24R (Miadlikowska
and Lutzoni, 2000) and LR7 or LR3 (Vilgalys and Hester,1990) for nuLSU, and mrSSU1 and mrSSU3R (Zoller etal., 1999) for mtSSU, according to the modified PCR
conditions of Frisch et al. (2014) (45 cycles to 35 cycles)
using an Applied Biosystems Veriti® 96-Well Thermal
Cycler (Thermo Fisher Scientific). The PCR products
were purified with illustra™ ExoProStar™ (GEHealthCare). 1.2 μL of PCR products were incubated with
0.2 μL ExoProStar™ and 0.5 μL ddH2Oat 37°C for 30
minutes and then at 80°C for 15 minutes.
DNA sequencing was performed on an Applied
Biosystems™ 3500xL Genetic Analyzer (Thermo Fisher
Scientific) using the BigDye® Terminator v3.1 Cycle
Sequencing Kit (Thermo Fisher Scientific) following the
manufacturer’s instructions. The taxon name and the
GenBank accession numbers for the obtained sequences
are shown in Table 1.
Molecular phylogenetic analyses
The newly obtained nuSSU, nuLSU and mtSSU
sequences of Racoleus japonicus from the Japanesematerial were aligned with sequences of selected taxa in2023

Miyazawa & Ohmura: Racoleus japonicus, a new species from Japan

Fig. 1. Maximum likelihood tree of selected taxa in Capnodiales s. lat. showing the phylogenetic position of Racoleus japonicus
collected from Japan (in bold). Myriangium duriei is used as the outgroup. NJ and ML support values are presented for each node.
Branches highly supported (≥70) by both analyses are indicated with bold black lines.

Capnodiales s. lat. from GenBank (Table 1) in MAFFTver. 7 (Katoh et al., 2019) using the default settings. Taxa
included in the alignment, including Myriangium duriaei(Myrangiales) as the outgroup (see Table 1), wereselected according to the BLAST results in GenBank andprevious phylogenetic studies (Muggia et al., 2008, 2016;
Abdollahzadeh et al., 2020). Each single locus data set(nuSSU, nuLSU and mtSSU) was separately aligned.
After removing sites with gaps, missing or ambiguous
data, the initial molecular phylogenetic trees werereconstructed based on each single locus, then the data
sets were concatenated. The final alignment of 1153 sites
was used for the molecular phylogenetic analyses.
Neighbor-joining (NJ) and maximum likelihood (ML)analyses were performed using the Tamura-Nei model(Tamura and Nei, 1993) plus gamma distribution withinvariant sites (G + I), which was selected as the best fitting
model based on the lowest Bayesian information criterion
score. The bootstrap values (≥50%) of 1,000 replicates for
NJ and ML are shown on each branch. All calculations
were conducted in MEGA Ⅹ (Kumar et al., 2018).

RESULTS AND DISCUSSION
Molecular analysis
Among the aligned sites of each locus within theJapanese material of Racoleus japonicus, those of nuSSU(857 sites) had ten variable sites including singleton
and/or parsimony-informative sites (98.9–100% identity
among six specimens), those of nuLSU (1285 sites) had
twelve variable sites and one gap site (99.4–99.9%
identity among six specimens), and those of mtSSU (826sites) had no variable sites or gaps (100% identity among
three specimens).
Before conducting the phylogenetic analysis withcombined sequences of three loci, each locus was
separately analyzed. The phylogenetic tree based on
nuSSU could not resolve the relationships among thefamilies within Capnodiales s. lat. However, each tree
based on nuLSU or mtSSU shows supportable bootstrap
values on some branches in the trees.
The concatenated phylogenetic tree based on threeloci for Racoleus japonicus within Capnodiales s. lat. is419

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shown in Fig. 1. The topology of our phylogenetic ML
tree fundamentally shows no conflict with those of
Muggia et al. (2008, 2016) and Abdollahzadeh et al.
(2020), and the bootstrap values were higher than thosein the separate analysis of each locus.
The monophyly of R. japonicus is confirmed withhigh support values (NJ/ML = 100/100). This new species
shows a close phylogenetic relationship with unidentified
fungi of Capnodiales s. lat. (A557 and A951 in Fig. 1)that were collected from alpine rock lichen communities(Muggia et al. 2016) which is supported by relatively highsupport values (NJ/ML = 92/87, Fig. 1). These lichenassociated fungi of Capnodiales s. lat. were treated under
Teratosphaeriaceae (Cometto et al., 2023). This clade and
other
fungi
of
Teratosphaeriaceae
includingFriedmanniomyces endolithicus and Recurvomyces
mirabilis form a common clade that is not well-supported
in our analyses (NJ/ML = 62/68, Fig. 1). Regardless,based on the currently available sequence data andphylogenetic analyses, we consider it is appropriate toplace R. japonicus in Teratosphaeriaceae. Furthermore,
our phylogenetic analyses revealed that Racoleus is not
closely related to Cystocoleus (Cystocoleaceae) and
Racodium (Racodiaceae) despite having extremely
similar appearance in morphology with their unusual,sterile, filamentous thalli. Although they all belong to abroadly defined Capnodiales s. lat., each genus appears to
represent an independent lichenization event.
Teratosphaeriaceae has been delimited based on
phylogenetic analysis of nuLSU sequence data (Crous etal., 2007), and presently does not have well defined
phenotypic synapomorphies. However, within this family,
there are taxa with similar traits to R. japonicus such as
the brown fungal filaments (Crous et al., 2007; Si et al.,2023) and living in harsh environments like rock surfaces(Ruibal et al., 2008, 2009).
Because DNA sequences of the type species ofRacoleus, R. trichophorus R. Santt. & D. Hawksw., havenot been obtained, further study is needed to confirm themonophyly including both R. trichophorus and R.
japonicus. However, based on the morphological featuresdescribed and discussed below, we consider the new
species belongs to Racoleus.

TAXONOMIC TREATMENT
Racoleus japonicus K. Miyaz. & Y. Ohmura, sp. nov.
Figs. 2 & 3C,F
MycoBank No.: MB 850089
Japanese common name: Iwa-goke.
Similar to Racoleus trichophorus but differs by wider
filaments (10–15 μm vs. 7–9 μm wide in R. trichophorus),larger hyphal cells (3–7 μm vs. 2–3 μm wide in R.
trichophorus), and irregularly sinuous lateral spines.
Type: JAPAN. Honshu. Shinano Prov. (Nagano
Pref.): Azusayama, Kawakami-mura, Minamisaku-gun
420

Vol. 68, No. 4(N35°57′, E138°40′), 1370 m elev., on rock, 26 May 2013,
Y. Ohmura 9651, K. Yoshida & A. Frisch [TNS-L-132529,
holotype: TNS, isotypes: B, BG, BM, BRY, C, CANB,CANL, DUKE, F, FH, G, GZU, H, KRAM, LIV, M, MIN,
MVM, NY, O, S, TAI, TSB, TUR, UPS, US, W, and hb.
Kalb, distributed to each herbarium as an exsiccata
specimen of Y. Ohmura: Lich. Minus Cogn. Exs. 494 as“Racodium rurestre Dill.” in Ohmura (2014)].
THALLUS minutely filamentous, blackish brown,
forming a dense fluffy colony reaching up to ca. 30 mm
diam. Filaments suberect to decumbent or spreading on
the surface of substrate, occasionally sympodiallybranched, 10–15(–20) μm wide, with lateral spines; outerwall undulating and irregularly corrugated, reflecting themorphology of the fungal hyphae. HYPHAE surrounding
the algal filaments in a single layer, orientated verticallyalong and parallel to the axis of the filament, brown, 3–7
μm wide, septate; the septa generally 10–20 μm apart,thick-walled; walls uneven and undulate to corrugated,
corrugations tending to interlink with those to adjacenthyphae. SPINES arising at broadly acute to almost rightangles from the vertical axis, brown, septate, stiff,
irregularly sinuous, up to 150 μm in length and 2–3 μmwide, the base expanded into a foot like cell (12–15 × 4–
5 μm). CONIDIOGENOUS CELLS and CONIDIA not
observed. PHOTOBIONT Trentepohlia sp., in single
filaments surrounded by fungal hyphae; cells roundedrectangular, 25–50 × 8–10 μm.
Chemistry: C–, K–, KC–, Pd–. No secondary
substances were detected by HPTLC.
Etymology: The epithet ‘japonicus’ refers to Japan
where the new species was collected.
Habitat and distribution: This species grows on
shady rocks or rock walls (Fig. 2A) in subboreal totemperate regions of Japan (i.e., from Hokkaido to
Kyushu) at elevations between 290 and 1700 m.
Remarks: Racoleus japonicus closely resembles R.
trichophorus by its dense, minutely filamentous, blackish
brown fluffy colonies (Fig. 2B), that are formed bysympodially branching filaments (Fig. 2C) withcorrugated hyphal walls (Fig. 2C). However, R. japonicus
differs from R. trichophorus in wider filaments (10–15
μm vs. 7–9 μm wide in R. trichophorus), larger hyphal
cells (3–7 μm vs. 2–3 μm wide in R. trichophorus), andirregularly sinuous lateral spines (Figs. 2D, 3F; vs.
straight in R. trichophorus). Furthermore, R. japonicus isdistributed in temperate to subboreal regions, whereas R.
trichophorus is found in tropical regions of Africa (Ivory
Coast), Asia (southern China), and South America (Peru)(Hawksworth et al., 2011).
The lateral spines of Racoleus were considered as animportant feature for the genus distinguishing it from
Cystocoleus and Racodium (Hawksworth et al., 2011).
Indeed, there is a difference that the spines are straight in
R. trichophorus (Hawksworth et al., 2011), sinuous in R.
japonicus (Figs. 2D, 3F), Cystocoleus (Sukuja and Ore,

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Miyazawa & Ohmura: Racoleus japonicus, a new species from Japan

Fig. 2. Racoleus japonicas. A. Habitat of type locality. B. Thallus (holotype, TNS). C. Sympodially branched filamentous thallus withcorrugated hyphal walls (K. Miyazawa 1700, TNS). D. Filamentous thallus with spines (lateral hyphae) (holotype, TNS). Scale bars: B= 0.5 mm; C = 50 μm; D = 10 μm.

1934; Fig. 3D) and Racodium (Fig. 3E), but the following
morphological features are common in these genera. The
spines are composed of a single hypha extending outside
the lichenized part of the thallus. The hyphae are 1.5–4
μm wide and up to 150 μm in length with some septa [thepresence of septa in R. trichophorus was confirmed by thephoto in Hawksworth et al., (2011, Fig. 1D)]. There is nolarge morphological difference except being straight orsinuous in the lateral spine among these taxa, and thisdifference serves as taxonomic character at species level

rather than for genus level. The arrangement of thehyphae and the presence/absence of corrugated walls in
the filamentous thallus should be an important taxonomic
character to recognize each genus.
Racoleus japonicus may be confused with
Cystocoleus ebeneus (Dillwyn) Thwaites and Racodium
rupestre Pers. because of the similar blackish brown
filamentous thalli in which the fungal hyphae surround
Trentepohlia filaments. However, Racoleus japonicus
differs in the vertical arrangement of hyphae with a421

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Vol. 68, No. 4

Fig. 3. Hyphal arrangements of Cystocoleus, Racodium and Racoleus. (A–C) and their lateral spines (hyphae) (D–F). A.
Cystocoleus ebeneus. The hyphae with corrugated walls are arranged twisted to the algal filament (Follmann s.n. [Follmann & Werner:
Lich. Exs. Sel. Colon. 505], TNS). B. Racodium rupestre. The hyphae with straight walls are arranged parallel to the algal filament(R. Santesson 22567 [Moberg: Lich. Sel. Exs. Upsal. 45], TNS). C. Racoleus japonicus. The hyphae with corrugated walls arearranged parallel to the algal filament (K. Miyazawa 1700, TNS). D. A lateral hypha of Cystocoleus ebeneus (W. A. Weber s.n. [Weber:Lich. Exs. 491], TNS)]. E. A lateral hypha of Racodium rupestre (R. Santesson 22567 [Moberg: Lich. Sel. Exs. Upsal. 45], TNS). F.
A lateral hypha of Racoleus japonicus (K. Miyazawa 1700, TNS). Scale bars: A–F = 10 μm.

corrugated wall (Fig. 3C). On the other hand, Cystocoleus
has a weakly twisted arrangement of hyphae with acorrugated wall (Fig. 3A), and Racodium has a verticalarrangement of hyphae without a corrugated wall (Fig.
3B). Racoleus japonicus might be confused with taxa thatbelong to Spilonema Bornet (Coccocarpiaceae) and
Thermutis Fr. (Lichinaceae), which have blackish
filamentous thalli. However, Spilonema and Thermutis
have stiffer thalli composed of wider filaments (at least
20 μm vs. up to 20 μm wide in R. japonicus), blackish
apothecia, and cyanobacteria photobionts (observed
materials in TNS are cited below).
Some Japanese specimens previously identified as
422‘Racodium rupestre’ housed in TNS were found to be R.
japonicus by our study, while R. rupestre was confirmed tooccur in Japan (at about 2480 m elevation in Mt. Kinpu).
The habitat of R. rupestre in Japan seems to be alpine tosubalpine areas. The collections of ‘Racodium rupestre’
reported from Mt. Ryokami by Yoshimura (1964) wereconfirmed as Racoleus japonicus in the present study byexamining materials collected from the same locality. The
taxon reported by Kato and Harada (2011) from Mt.
Haruna, central Honshu in Japan, should be Racoleus
japonicus according to their illustration and description.
Additional specimens examined. JAPAN. Hokkaido.
Kitami Prov.: Ikutahara-Kiyosato, Engaru-cho, Monbetsu-gun (N43°51′,

2023

Miyazawa & Ohmura: Racoleus japonicus, a new species from Japan

E143°29′), 290 m elev., on shady rock wall in riverine forest, 29 May
2012, A. Frisch 12/Jp152 & Y. Ohmura (TNS). Honshu. Musashi Prov.
(Saitama Pref.): mountain path between Hinataooyaguchi and thesummit of the Mt. Ryokami, Ogano-machi, Chichibu-gun (N36°01′,
E138°50′), about 1500 m elev., on shady rock, 17 August 2023, K.
Miyazawa 1700 (TNS); ditto, about 1700 m elev., K. Miyazawa 1701
(TNS), K. Miyazawa 1702 (TNS). Aki Prov. (Hiroshima Pref.): Mt.
Misen, Miyajima Island, Hatsukaichi-city (N34°16′, E132°19′), 340 m
elev., on rock, 8 November 2012, Y. Ohmura 9473, A. Frisch & K.
Fedrowitz (TNS). Kyushu. Higo Prov. (Kumamoto Pref.):
Hakusuishizenshinrin Park, Mizukami-mura, Kuma-gun (N32°22′,
E131°02′), 780 m elev., on rock, 29 May 2023, K. Miyazawa 1561 (TNS).
Exsiccati of other species examined. [Cystocoleus niger(Huds.) Har.]: AUSTRIA. Westösterreich, Stubaier Alpen: flockenrasiganschattig-trockenen,aber
luftfeuchtenüberhängenden
Urgesteinswänden im Cystocoleo-Racodietum rupestris SCHADE,
1900 m elev., NW, pH 6.2, Osthang des Lisenser Tales bei Jufenau unterdem Windegg, October 1975, G. Follmann s.n. (Follmann: Lich. Exs.
Sel. Cassel. 167, TNS). [Cystocoleus ebeneus (Dillwyn) Thwaites]:
GERMANY: Rheinlasnd-Pfalz, southern Eifel Mountains, GermanLuxemburgian Nature Park, Gutland District, on vertical or slightly
overhanging faces of red sandstone cliffs in open woodland not far fromthe Prümerburg north of Irrel, occasionally growing intermixed with
Racodium rupestre Pers. which is, nevertheless, always of secondary
importance in comparison with the above species, about 300 m elev.,
W-S exposition, forming extensive mats, October 1989, Follmann s.n.
(Follmann & Werner: Lich. Exs. Sel. Colon. 505, TNS). COLOMBIA:
Depart. Cundinamarca, Municipio Supatá, Alto El Tablazo, along trail
from radar station to Suptatá, subparamo scrub, 3400 m elev., on
perpendicular sandstone rockface, 11 September 1984. J. Aguirre C. &H. Sipman 5228 (Sipman: Lichenoth. Latinoamer. 17, TNS). U.S.A.
Colorado. Rocky Mountain National Park: Odessa Gorge, on trail from
Odessa to Fern Lake; vertical faces of granite cliffs, locally humid site,
3000–3100 m elev., 28 September 1975, W. A. Weber s.n. (Weber: Lich.
Exs. 491, TNS). [Racodium rupestre Pers.]: SWEDEN, Härjedalen
Prov.: Tännäs par., southern slope of Mt. Gruvvålen (SW of Mt.
Mittåkläppen) (N62°43′, E12°25′), about 900 m elev., on exposed rockin the upper part of the subalpine (birch) region, 1 September 1970, R.
Santesson 22567 (Moberg: Lich. Sel. Exs. Upsal. 45, TNS). Czech
Republic, Sudeti occident., Teplice nad Metují: loco dicto ‘Skalníměsto’, 620 m elev., ad parietes altos umbrosos rupium arenacearum,
18 June 1962, A. Vězda s.n. (Vězda: Lich. Sel. Exs. 450, TNS).
[Spilonema paradoxum Bornet]: U. S. A., Alabama, Lee Co., Waverly,
auf zeitweise überrieseltem Glimmerscheifer am Bachufer, 23 February
1963, A. Henssen & H. McCullough 15166 (Henssen: Lich. Cyanoph. 15,
TNS). [Spilonema revertens Nyl.] U. S. A., California, Humboldt Co.,
Coast Range, Pine Ridge Summit bei Arcata, in Sickerwasserstreifen auf
vulkanischem Gestein, ca. 1000 m elev., 31 October 1961, A. Henssen
13627a (Henssen: Lich. Cyanoph. 14, TNS). [Thermutis velutina (Ach.)
Flot.]: SWEDEN, Bohuslän: Ödsmål, Kolhättan, on irrigated, sunny rocks
near the see, fertile, 13 August 1932, A. H. Magnusson s. n. (Magnusson:
Lich. Sel. Scand. Exs. 296, TNS). CROATIA, Dalmatia: Duba prope
vicum Drvenik, 30 m elev., ad saxa calcarean in litore maris, A. Vězda s.n.
(Vězda: Lich. Sel. Exs. 509, TNS).
Specimens examined of other species. [Racodium
rupestre Pres.: Japanese common name “Iwa-goke-modoki”]: JAPAN.
Honshu. Kai Prov. (Yamanashi Pref.): Mt. Kinpu, Hokuto-city (N35°52′
E138°37′), about 2480 m elev., on shady rock-wall in forest, 4 July 2012,
A. Frisch 12/Jp464, Y. Ohmura & G. Thor (TNS).

ACKNOWLEDGMENTS
We thank two anonymous reviewers for carefully reading of
our manuscript and providing valuable comments. This study
was partly supported by JSPS KAKENHI 22J20567 for the firstauthor. Permissions for the collections were kindly granted by
the Ministry of the Environment Government of Japan (Nos.
091105003, 110913004).

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