The Caloplaca crenulatella species complex; Its intricate taxonomy and description of a new species

Abstract

The Caloplaca crenulatella species complex is monophyletic, based on analysis of ITS nrDNA sequences. It is characterized mainly by its ascospores and a simplified thallus. Caloplaca aquensis, C. borysthenica, C. interfulgens, C. lactea, C. marmorata and C. tominii represent well-defined monophyletic lineages within it. Caloplaca diffusa, described here as a new species, is a well-supported lineage with a distinctive phenotype. Caloplaca crenulatella s. lat. and C. ferrarii s. lat. are heterogeneous taxa appearing in four and two places respectively within our tree. Caloplaca ochracea also belongs to the group, although it has different ascospore characters. Caloplaca gyalolechiaeformis and C. pseudocitrina are later synonyms of C. tominii. Arctic and North American sorediate specimens morphologically very close to C. tominii represent a separate lineage recently recognized as C. erichansenii. The North American Caloplaca nashii has a C. crenulatella-like phenotype but does not belong to the C. crenulatella complex. The identities of Caloplaca lacteoides and Caloplaca epigaea are not clear, but the latter may be a terricolous ecotype of one of the entities within C. crenulatella s. lat. A key to the European species of the group is provided.

Full text

The Caloplaca crenulatella species complex; its intricate
taxonomy and description of a new species
Jan VONDRÁK, Pavel R
ˇÍHA, Olexii REDCHENKO,
Olga VONDRÁKOVÁ, Pavel HROUZEK and
Alexander KHODOSOVTSEV
Abstract: The Caloplaca crenulatella species complex is monophyletic, based on analysis of ITS
nrDNA sequences. It is characterized mainly by its ascospores and a simplified thallus. Caloplaca
aquensis,C. borysthenica,C. interfulgens,C. lactea,C. marmorata and C. tominii represent well-defined
monophyletic lineages within it. Caloplaca diffusa, described here as a new species, is a well-supported
lineage with a distinctive phenotype. Caloplaca crenulatella s. lat. and C. ferrarii s. lat. are heterogeneous
taxa appearing in four and two places respectively within our tree. Caloplaca ochracea also belongs to
the group, although it has different ascospore characters. Caloplaca gyalolechiaeformis and C. pseudo-
citrina are later synonyms of C. tominii. Arctic and North American sorediate specimens morphologi-
cally very close to C. tominii represent a separate lineage recently recognized as C. erichansenii. The
North American Caloplaca nashii has a C. crenulatella-like phenotype but does not belong to the
C. crenulatella complex. The identities of Caloplaca lacteoides and Caloplaca epigaea are not clear, but
the latter may be a terricolous ecotype of one of the entities within C. crenulatella s. lat. A key to the
European species of the group is provided.
Key words: Caloplaca lactea group, ITS nrDNA, lichens, paraphyletic taxa, phylogeny, Teloschistaceae
Introduction
Navarro-Rosinés & Hladun (1996) provided
a monograph of European species of the
Caloplaca lactea group, focused mainly on
the western Mediterranean. We deal with the
same group, but we prefer to name it after
C. crenulatella, a taxon that (though para-
phyletic) is common throughout Europe and
possesses a representative phenotype for the
group. In contrast, the name Caloplaca lactea
has been misapplied to various species in
Europe; C. lactea proper is restricted, in
Europe, to (sub)Mediterranean regions and
has a rather atypical phenotype within the
group (ascospore size and shape).
Our geographical focus is different from
that of Navarro-Rosinés & Hladun since we
have emphasized south-east Europe and
some parts of western Asia. The centre of our
field work was the Black Sea region.
Material and Methods
The main part of the material was collected by the first
author during field excursions in 2004 (Bulgaria), 2005
(Romania, Bulgaria, European part of Turkey), 2006
(Ukraine: Crimea), 2007 (Romania, Bulgaria, Turkey,
Georgia, Russia, Ukraine, Iran), 2008 (Ukraine: Steppe
zone and Crimea) and 2009 (Kazakhstan, Russia,
Ukraine). Additional material from the studied regions
was seen in KHER, LE and GZU. Comparative material
of the species from the western Mediterranean was
kindly provided by BCN.
J. Vondrák: Institut of Botany, Academy of Sciences,
Zámek 1, CZ-252 43 Pru˚honice, Czech Republic.
Email: j.vondrak@seznam.cz
P. R
ˇı´ha: Department of Zoology, Faculty of Science,
University of South Bohemia, Branišovská 31, C
ˇeské
Budeˇjovice.
O. Redchenko: Kholodny Institute of Botany, National
Academy of Sciences of Ukraine, 2 Tereshchenkivska
Street, 01601 Kiev, Ukraine.
O. Vondráková: Institute of Steppe (Urals Branch of
Russian Academy of Sciences), Pionerskaya st. 11,
Orenburg, RF–460000, Russia.
P. Hrouzek: Department of Autotrophic Microorgan-
isms, Institute of Microbiology, Academy of Sciences,
Opatovický mlýn, Trˇebonˇ, CZ-379 81, Czech Republic.
A. Khodosovtsev: Kherson State University, 40 Rokiv
Zhovtnya str. 27, 73000 Kherson, Ukraine.
The Lichenologist 43(5): 467–481 (2011) © British Lichen Society, 2011
doi:10.1017/S0024282911000466

T 1.Sample data and GenBank accession numbers of the ITS sequences used in the phylogenetic analysis
Taxon Voucher GenBank
accession nrs
Caloplaca aquensis Turkey: Black Sea coast: Kandıra, on coastal limestone rocks, 2007,
J. Vondrák (CBFS JV6923)
HQ699652
C. aquensis Ukraine: Crimean Peninsula: Sudak, coastal rocks at Cape Meganom,
2007, J. Vondrák (CBFS JV5970)
HQ699656
C. borysthenica Ukraine: Kherson region: Kherson, Stanislav, 2008, J. Vondrák (CBFS
JV7204)
HQ699649
C. borysthenica Ukraine: Kherson region: Kherson, Stanislav, 2008, J. Vondrák (CBFS
JV7205)
HQ699655
C. crenulatella s. lat. USA: California: Peninsular Range, San Bernardino NF, on carbonate
soil, 2006, K. Knudsen 5729 (UCR, dupl. CBFS)
HQ699635
C. crenulatella s. lat. Czech Republic: South Bohemia: Horažd’ovice, Rabı´, on limestone,
2008, J. Vondrák (CBFS JV6348)
HQ699642
C. crenulatella s. lat. Iran: Lake Urmia, Saraydeh, on siliceous rock, 2007, J. Vondrák
(CBFS JV5702)
HQ699628
C. crenulatella s. lat. Ukraine: Lugansk region: Rozkishne, protected area “Balka Polska”, on
shrub bark, 2007, O. Nadyeina (hb. Nadyeina)
HQ699645
C. crenulatella s. lat. Azerbaijan: Shemakhinskiy district: Sis, protected area “Pirkulinskiy
zap.”, on sandstone, 2004, O. Nadyeina (hb. Nadyeina)
HQ699654
C. crenulatella s. lat. Romania: Constanta, on concrete, 2007, J. Vondrák (CBFS JV5441) HQ699658
C. diffusa Spain: Catalonia: Girona, on siliceous block at stream, 2007, X.
Llimona (BCN)
HQ699659
C. diffusa Bulgaria: The Rhodopes: Madzharovo, on volcanic boulder, 2004,
J. Vondrák (CBFS JV2239)
HQ699632
C. diffusa Turkey: Black Sea coast: Demirköy, Limanköy, on coastal siliceous
rock, 2005, J. Vondrák (CBFS JV3041)
HQ699633
C. diffusa Bulgaria: Black Sea coast: Tsarevo, Rezovo, on coastal siliceous rock,
2005, J. Vondrák (CBFS JV3039)
HQ699646
C. diffusa Georgia: Black Sea coast: Batumi, coastal rocks near Buknari, 2007,
J. Vondrák (CBFS JV6536, holotypus)
HQ699660
C. diffusa Georgia: Black Sea coast: Batumi, coastal rocks near Sarpii, 2007,
J. Vondrák (CBFS JV6226)
HQ699647
C. diffusa Georgia: Black Sea coast: Batumi, coastal rocks near Buknari, 2007,
J. Vondrák (CBFS JV6227)
HQ699648
C. erichansenii USA: Montana:onsoil,T. Spribille (herb. Spribille) HQ699663
C. erichansenii Greenland: Søndre Strømfjord, on loess, 1998, E.S. Hansen (C,
isotypus)
HQ699638
C. ferrarii s. lat. Hungary: Mezo˝föld, Paks. Sánc-hegy, on loess cliff, T. Pócs 9768
(EGR 3814, dupl. in CBFS)
HQ699636
C. ferrarii s. lat. Iran; Lake Urmia, Shirin Bolagh, 2007, J. Vondrák (CBFS JV5715) HQ699643
C. ferrarii s. lat. Morocco: Atlas Mts: Agadir, Aurir, 2003, J. Vondrák (CBFS JV1432) HQ699653
C. ferrarii s. lat. Russia: Black Sea coast: Novorossiysk, coastal siliceous rocks near
Dyurso, 2007, J. Vondrák (CBFS JV6531)
HQ699662
C. ferrarii s. lat. Spain: Zaragoza (BCN 13509); details in Gaya et al. (2008) EU639622
C. ferrarii s. lat. Ukraine: Kherson region: Kherson, Stanislav, 2009, J. Vondrák (CBFS
JV7123)
HQ699651
C. ferrarii s. lat. Ukraine: Crimean Peninsula: Sudak, Morskoe, 2008, J. Vondrák
(CBFS JV7192)
HQ699657
C. ferrarii s. lat. Spain: on sandstone (BC13508) HQ699637
C. interfulgens Iran: Lake Urmia, Shirin Bolagh, 2007, J. Vondrák (CBFS JV5777) HQ699639
C. interfulgens Iran: Lake Urmia, Shirin Bolagh, 2007, J. Vondrák (CBFS JV5781) HQ699640
C. lactea Turkey: Black Sea coast:I
˙nebolu, Abana, on calcareous outcrop, 2007,
J. Vondrák (CBFS JV6910)
HQ699644
C. marmorata Czech Republic: Central Bohemia: Beroun, Srbsko, 2004, J. Vondrák
(CBFS JV2606)
HQ699629
468 THE LICHENOLOGIST Vol. 43

Citations of specimens are abbreviated; full infor-
mation for a majority of the samples deposited in CBFS
(herbarium of the University of South Bohemia) is avail-
able in a database on the web page: http://botanika.
bf.jcu.cz/lichenology/data.php. For the common species
Caloplaca marmorata and C. ochracea, only selected
samples of particular geographical interest are cited (e.g.
new country records). For the taxonomically unresolved
species complexes C. crenulatella s. lat. and C. ferrarii
s. lat., only samples used in molecular analysis are
cited in Table 1. Simplified specimen data for other
sequenced lichens are also listed in Table 1.
Morphological investigations
Diagnostic characters for each species are provided in
the key and a detailed morphological description is only
given for the new species. Measurements are believed to
be accurate to 0·25 m (for cells, e.g. conidia and asco-
spores), 1 m (width of asci) or 10 m (larger structures,
e.g. hymenium and width of exciples). All measure-
ments of cells (ascospores, conidia, asci, paraphyses)
include their walls. Paraphyses tips were observed after
pre-treatment with c. 10% KOH. Only those ascospores
with well-developed septa were measured; in these asco-
spores loculi were connected with a thin cytoplasmatic
channel, never disconnected. Measurements are given
as (minimum–) x
¯
± SD (–maximum), where x
¯
= mean
value and SD = standard deviation. Total numbers
of measurements are given in square parentheses [n].
Morphological terminology mainly follows Smith et al.
(2009).
Secondary metabolites identification
HPLC chromatography was performed on samples
of Caloplaca crenulatella s. lat., C. diffusa,C. ferrarii,C.
marmorata and C. tominii according to Søchting (1997).
DNA extraction and amplification
Direct PCR was used for PCR–amplification of the
ITS regions, including the 5.8S gene of the nuclear
rDNA following Arup (2006). Primers for amplification
were ITS1F (Gardes & Bruns 1993) and ITS4 (White
et al. 1990). PCR cycling parameters follow Ekman
(2001).
Alignment and molecular analysis
The E-INS-i algorithm implemented in the online
version of MAFFT 6 (Katoh et al. 2002) was employed
to align sequence data sets. Further manual adjustments
were conducted in BioEdit (Hall 1999). Our final align-
ment included 45 terminals, of which 42 were ingroup
samples. A gap matrix was computed in SeqState 1.4
(Müller 2005) to encode indel-type mutations; as the
dedicated software computing the Bayesian inference
does not support cost matrices of MCIC, the simple
indel coding (SIC) algorithm was used. Bayesian analy-
sis coupled with Markov chain Monte Carlo (MCMC)
simulations was conducted in MrBayes (v3.12; Ronquist
& Huelsenbeck 2003). Two independent simultaneous
runs were run for 10 million generations using four
Markov chains heated incrementally by 0·25 and sam-
pled every 1000 generations, but the first 25% of trees
T 1.Continued
Taxon Voucher GenBank
accession nrs
C. marmorata France: Provence: Var, Massif de la Ste. Baume (BCN); details in
Gaya et al. (2008)
EU639621
C. marmorata Iran: Lake Urmia, Shirin Bolagh, on calcareous rock, 2007, J. Vondrák
(CBFS JV5713)
HQ699627
C. marmorata Czech Republic: Pavlovské vrchy hills: Mikulov, on limestone, 2004,
J. Vondrák (CBFS JV1800)
HQ699631
C. marmorata Bulgaria: The Rhodopes: Madzharovo, Silen, 2004, J. Vondrák (CBFS
JV2103)
HQ699630
C. nashii USA: California: Colorado desert, Anza Borrego State Park, on granite,
2006, K. Knudsen 5901 (UCR, dupl. CBFS)
HQ699624
C.aff.nashii USA: California: Channel Islands NP, on concrete, 2006, K. Knudsen
5335 (UCR, dupl. CBFS)
HQ699641
C. ochracea Spain: Tarragona (BCN 10053); details in Gaya et al. (2008) EU639620
C. tominii Kazakhstan: Western Kazakhstan region: lake Shalkar, on sandstone,
2009, J. Vondrák (CBFS JV7274)
HQ699625
C. tominii Kazakhstan: Western Kazakhstan region: lake Shalkar, on soil, 2009,
J. Vondrák (CBFS JV7273)
HQ699650
C. tominii Czech Republic: South Bohemia:C
ˇeské Budeˇjovice, Nové Hodeˇjovice,
on concrete and soil, 2009, J. Vondrák (CBFS JV6989)
HQ699626
C. trachyphylla Pakistan: North West Himalaya (GZU); details in Gaya et al. (2008) EU639651
Xanthoria elegans Spain: Catalonia (BCN): details in Gaya et al. (2008) EU639642
2011 Caloplaca crenulatella complex—Vondrák et al. 469

were discarded as burn-in. Model selection was made
in accordance with the Aikake information criterion
computed in MrModeltest 2.3 (Nylander et al. 2004).
In addition, the PAUP* software (Swofford 2002) was
necessary to get the ML scores. As suggested, the
nucleotide alignment was analyzed under the GTR+
model with the assumption of invariable sites. Distri-
bution of prior probability densities of the substitution
rates, along with the distribution of the stationary
nucleotide frequencies, was set to flat. The indel matrix
was treated as restriction data in MrBayes and analyzed
under the F81-like model. The stationarity of tree
topologies was assessed according to the average stan-
dard deviation of split frequencies between both runs (it
fell under 0·008). The resulting trees were used for
reconstruction of a 50% majority-rule consensus tree.
Results
Phylogeny
The Bayesian analysis of the ITS nrDNA
dataset with coded indels (656 nucleotide
positions and 101 indel characters) produced
a 50% majority rule consensus tree with an
average In L = −4020·305 (Fig.1). In gen-
eral, the Bayesian inference of our data failed
to reveal the relationship between species/
lineages, and several single terminals re-
mained ambiguously placed in the basal
polytomy. Nevertheless, it provided some
important information of interest about the
phylogeny of the group. Firstly, all ingroup
sequences studied formed a strongly sup-
ported monophyletic group (PP = 1·00) in
relation to the sequences of C. trachyphylla,
Xanthoria elegans and both C. nashii speci-
mens. Secondly, some phenotypically de-
fined species formed convincing ‘molecular
taxa’, such as C. diffusa and C. marmorata.
Thirdly, some phenotypically distinguish-
able groups of specimens, for this reason
formerly considered as good species (C.
crenulatella and C. ferrarii), appear as artificial
assemblages of two or more separate lineages.
These paraphyletic taxa will be discussed
below.
Phenotypic circumscription of the
group
Although morphologically variable, the
Caloplaca crenulatella group is diagnosed by
its long and narrow ellipsoid spores, c. 15–
20 × 6–8 m (except for C. lactea) with thin
septa, 1–4 m wide (except for young asco-
spores of C. ochracea with two small locules
and a very broad septum). The thallus is
usually reduced, inconspicuous (rarely areo-
late or exceptionally subsquamulose), yellow
or rarely pale grey. The apothecia are
zeorine, yellow to red with a thalline exciple
that may be enlarged and crenulate in some
species. Pycnidia were observed only in
C. borysthenica,C. crenulatella,C. diffusa and
C. ferrarii; conidia are ellipsoid or shortly
bacilliform c. 2·5–3·5 × 1·0–1·5 m.
The secondary metabolites are anthraqui-
nones of chemosyndrome A (sensu Søchting
1997). Apothecia always contain parietin
(main compound) with traces of emodin,
fallacinal, parietinic acid, and teloschistin.
The thallus, when present and yellow, has
the same anthraquinones.
Ecologically, the group shows a preference
for calcareous substrata (rock, soil, artificial
substrata), although some species tolerate
siliceous rocks; only C. diffusa grows mainly
on siliceous substrata. Epiphytic occurrences
are known in C. crenulatella s. lat., but are
exceptional. The biodiversity within the
group increases significantly from north to
south in Europe.
Another large group of European Calo-
placa species containing Caloplaca subpallida
H. Magn. and its relatives has a crustose
thallus, or no thallus, and ascospores with
thin septa. However, it differs from the C.
crenulatella group in having smaller and nar-
rower ascospores c. 12–17 × 4–6 m, and a
thallus which (if not entirely reduced) is grey
and lacks anthraquinones, but has the pig-
ment Sedifolia-grey. This phylogenetic group
is not closely related to the C. crenulatella
group (our unpublished data) and includes,
for example, C. arenaria (Pers.) Müll. Arg.,
C. scotoplaca (Nyl.) H. Magn., C. subpallida
and C. tristiuscula H. Magn. Species of this
group tend to occur on siliceous rocks.
Caloplaca luteoalba (Turner) Th. Fr. is also
similar to the C. crenulatella group in its nar-
row ascosopre septa and outer morphology,
but it is an epiphytic species and has smaller
ascospores, c. 8–12 × 3–6 m (Fletcher &
Laundon 2009).
470 THE LICHENOLOGIST Vol. 43

Taxonomy
The New Species
Caloplaca diffusa Vondrák & Llimona
sp. nov.
MycoBank No: MB561220
Caloplaca diffusa C. interfulgentis similis est, sed mar-
ginem thalli diffusum, et prothallus albus nonnumquam
adest. Habitat prope mare, in rupibus siliceis.
Typus: Georgia, Adjara, Batumi, coastal rocks near
Buknari, 41°45#58·31$N, 41°45#01·45$E, on siliceous
cliff in supralittoral zone, 27 April 2007, J. Vondrák-
(CBFS JV6536—holotypus; JV6227, JV6493—isotypi).
ITS sequence of the holotypus: HQ699660.
Caloplaca trachyphylla EU639651
Xanthoria elegans EU639642
Caloplaca nashii
HQ699624
Caloplaca cf. nashii HQ699641
Caloplaca ochracea EU639620
Caloplaca lactea HQ699644
Caloplaca crenulatella s. 536996QH.tal
Caloplaca crenulatella s. 246996QH.tal
Caloplaca aquensis HQ699652
Caloplaca aquensis HQ699656
Caloplaca interfulgens HQ699639
Caloplaca interfulgens HQ699640
Caloplaca tominii HQ699650
Caloplaca tominii HQ699625
Caloplaca tominii HQ699626
Caloplaca ferrarii s. 226936UE.tal
Caloplaca ferrarii s. 156996QH.tal
Caloplaca ferrarii s. 346996QH.tal
Caloplaca ferrarii s. 356996QH.tal
Caloplaca ferrarii s. lat. HQ699662
Caloplaca marmorata HQ699629
Caloplaca marmorata EU639621
Caloplaca marmorata HQ699627
Caloplaca marmorata HQ699630
Caloplaca marmorata HQ699631
Caloplaca diffusa HQ699659
Caloplaca diffusa HQ69963
2
Caloplaca diffusa HQ699633
Caloplaca diffusa HQ699646
Caloplaca diffusa HQ699661
Caloplaca diffusa HQ699660
Caloplaca diffusa HQ699647
Caloplaca diffusa HQ699648
Caloplaca crenulatella s. 826996QH.tal
Caloplaca erichansenii HQ699638
Caloplaca erichansenii HQ699663
Caloplaca crenulatella s. 546996QH.tal
Caloplaca borysthenica HQ699649
Caloplaca borysthenica HQ699655
Caloplaca crenulatella s. 456996QH.tal
Caloplaca crenulatella s. 856996QH.tal
Caloplaca ferrarii s. 756996QH.tal
Caloplaca ferrarii s. 636996QH.tal
Caloplaca ferrarii s. 736996QH.tal
0.0
0.05
1.001.00
1.001.00
1.001.00
1.001.00
1.001.00
1.001.00
1.001.00
1.001.00
1.001.00
1.001.00
1.001.00
1.001.00
1.001.00
1.001.00
1.001.00
1.001.00
1.001.00
1.001.00
0.650.65
0.650.65
0.840.84
0.840.84
1.001.00
1.001.00
1.001.00
1.001.00
0.990.99
0.990.99
1.001.00
1.001.00
1.001.00
1.001.00
0.930.93
0.930.93
0.990.99
0.990.99
1.001.00
1.001.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
0.65
0.91
1.00
1.00
0.99
1.00
0.65
A
1
0.85
0.99
1.00
1.00
0.80
0.84
0.59
0.99
1.00
0.66
0.99
0.93
0.65
1.00
B
C
C
1.00
1.00
D
1.00
2
F. 1.The 50% majority rule consensus phylogram of 15002 Bayesian MCMC trees (two simultaneous runs of
10 million generations sampled every 1000th generation, 25% burn-in) from gap-coded data set of ITS nrDNA
sequences from the Caloplaca crenulatella group together with four outgroup sequences. Support values are calculated
as Bayesian posterior probabilities. Encircled letters and numbers denote various lineages assigned to C. crenulatella
and C. ferrarii, respectively.
2011 Caloplaca crenulatella complex—Vondrák et al. 471

F. 2.A, Caloplaca diffusa (holotypus); B, C. borysthenica (CBFS JV7314); C, C. interfulgens (CBFS JV5777) together with C. marmorata in the lower part; D, C.
tominii (CBFS JV7274). Scales: A–D=1mm.
472 THE LICHENOLOGIST Vol. 43

(Fig. 2A)
Thallus yellow, ochre-yellow or grey-
yellow, areolate, forming spots up to several
cm diam.; thallus margin diffuse, often
rimmed by a white or pale grey prothallus.
Areoles angular, (170–) 438 ± 250 (–1100)
m diam. [20], up to 150 m thick. True
cortex absent but thin alveolate cortex (sensu
Vondrák et al. 2009) sometimes present, up
to 20 m thick. Medulla inconspicuous. Pro-
thallus white or pale grey, often present.
Apothecia zeorine, c. 0·4–0·7 mm (rarely to
1 mm) diam.; disc pale to dark orange. Proper
exciple covered by yellow-orange pigments,
(30–) 78 ± 38 (–150) m thick [14]; upper
part formed of isodiametric cells, c. 5–7 m
diam.; lower part formed of prosoplecten-
chyma. Thalline exciple covered by yellow
pigments, often crenulate, (60–) 104 ± 23
(–150) m thick [14]. Hymenium 70–90 m
thick. Hypothecium of variously shaped cells,
conically extended downwards. Asci clavate,
Teloschistes-type, (40–) 58 ± 8 (–70) × (12–)
17 ± 3 (–24) m [18]. Paraphyses branched,
thickened in upper part to (2·7–) 4·4 ± 0·7
(–5·7) m wide [35]. Ascospores (11·0–)
15·3 ± 1·4 (–17·7) × (5·0–) 7·0 ± 1·1 (–9·0)
m [45]; length / breadth ratio (1·5–)
2·2 ± 0·4 (–3·1). Ascospore septa thin, (2·0–)
2·7 ± 0·5 (–3·7) m thick [45]; ratio of
septum width / ascospore length (0·12–)
0·17 ± 0·03 (–0·26).
Pycnidia indistinct or forming orange
spots, up to c. 170 m diam. in section.
Conidia narrowly ellipsoid or shortly bacilli-
form, 2·7–3·2 × 1·0–1·7 m [10].
Chemistry. Thallus C−, K+ purple, P−,
UV+ orange; apothecia C−, K+ purple, P−,
UV+ orange. Compounds: parietin (major),
± traces of emodin, parietinic acid, fallacinal
and teloschistin.
Phylogeny. All eight sequences of C. diffusa
studied formed a well-supported monophyl-
etic group (PP = 1·00), but its precise phylo-
genetic position remains unclear due to the
basal polytomy.
Etymology. The name reflects the diffuse
thallus margin of the species.
Ecology and distribution. Caloplaca diffusa
prefers damp sites on coastal siliceous cliffs
or periodically inundated places where it
grows around seepage crevices. The only
known inland localities are in Bulgarian
Eastern Rhodopes, in NE Spain and in the
Peloponnese (Greece) where it grows in the
bottom of seasonal streams. It is well-
documented in the Black Sea region, but
there are two records only from Mediterra-
nean regions (Spain and Greece).
Remarks. Superficially similar species,
such as Caloplaca subsoluta (Nyl.) Zahlbr.,
C. maritima (B. de Lesd.) B. de Lesd. or C.
velana s. lat. differ considerably in ascospore
characters, mainly in their wider septum.
Caloplaca crenulatella s. lat. lacks a diffuse
thallus margin and its thallus is more re-
duced. Caloplaca interfulgens has a different
ecology and distribution (calcareous rocks in
semi-desert regions), does not have a diffuse
thallus margin and never has a distinct pro-
thallus.
Paratypes.Bulgaria: Black Sea coast: Burgas, Kiten,
2007, J. Vondrák (CBFS JV6592, 6593, 6595); Burgas,
Rezovo, 2005, J. Vondrák (CBFS JV3039, 3045, 6220);
Burgas, Sinemorets, 2007, J. Vondrák (CBFS JV6594,
dupl. in GZU). The Rhodopes: Kardzhali, Bregovo, on
volcanic rock, 2004, J. Vondrák (CBFS JV2215);
Haskovo, Madzharovo, on volcanic rock, 2004, J.
Vondrák (CBFS JV2239).—Georgia: Adjara: Batumi,
Sarpi, on coastal siliceous rock, 2007, J. Vondrák (CBFS
JV6226).—Greece: Peloponese: Argolis Peninsula
(Peiraias administrative district), Driopi, on serpentinite
in bottom of periodical stream, 2010, J. Vondrák &
O. Vondráková (CBFS JV8323).—Russia: Black Sea
coast: Tuapse, Gryaznova, on maritime schist, 2007, J.
Vondrák (CBFS JV6534).—Spain: Catalonia: Girona,
Port de la Selva, on siliceous block at brook, 2007, X.
Llimona (BCN).—Turkey: Black Sea coast: Demirköy,
Limanköy, on siliceous maritime rock, 2005, J. Vondrák
(CBFS JV3041); Istanbul, Kilyos, on siliceous maritime
rock, 2005, J. Vondrák (CBFS JV3352, dupl. in LD;
3362, 3372).
Other taxa recognized by the molecular
analysis
Caloplaca aquensis Houmeau & Cl. Roux
Bull. Soc. Bot. Centre-Ouest, nouv. sér.15: 143 (1984);
typus: France, Western Gallia, island Aquis (Aix: Le
Tridoux), on maritime limesone cliff, 1983, J. M.
Houmeau & C. Roux (hb. Houmeau—holotype;
MARSSJ— isotype; not seen).
2011 Caloplaca crenulatella complex—Vondrák et al. 473

This phenotypically well-circumscribed
species (Navarro-Rosinés & Hladun 1996)
forms a monophyletic group (PP = 1·0)
within the Caloplaca crenulatella group, but its
closer relationship is unclear as it fell into the
basal polytomy. It occurs on limestone cliffs
along the Atlantic coast of western Europe,
the western Mediterranean (Navarro-
Rosinés & Roux 1992), eastern Mediterra-
nean (e.g. Abbott 2009) and Black Sea coast
(Khodosovtsev 1999).
Specimens studied.Bulgaria: Black Sea coast: Kavarna,
2007, J. Vondrák (CBFS JV6210).—Croatia: Istria:
Porecˇ, Špadic´i, 2009, J. Vondrák (CBFS JV7248).—
Greece: Fokidas: Itea, Galaxidi, 2010, J. Vondrák & O.
Vondráková (CBFS JV8330).—Morroco: Mediterra-
nean Sea coast: Al-Hoceima, J. Šoun (CBFS JV6937).—
Russia: Black Sea coast: Novorossiysk, 2007, J. Vondrák
(CBFS JV6531).—Turkey: Black Sea coast: Kandıra,
2007, J. Vondrák (CBFS JV6120, 6923). Sea of
Marmara coast: Gallipoli peninsula, 2007, J. Vondrák
(CBFS JV7995).—Ukraine: Crimean Peninsula: Sudak,
2007, J. Vondrák (CBFS JV5970); Tarchankut penin-
sula, 1995, A. Khodosovtsev (KHER 176); ibid., 2008,
J. Vondrák (CBFS JV8189).
Caloplaca borysthenica Khodos. & S.Y.
Kondr.
in Kondratyuk, Khodosovtsev & Kärnefelt, Mycologia
Balcanica 3: 96 (2006); type: Ukraine, Kherson region,
Shirokaya balka, on loess, 2002, A. Khodosovtsev (KW—
holotype!; KHER—isotype!).
(Fig. 2B)
The description in Kondratyuk et al.
(2006) is based on a poorly developed speci-
men and the species was characterized by
asci with 2–4 (–6) spores that are often de-
formed. We examined the holotype, isotypes
and topotypes again and found some well-
developed asci with eight, regularly ellipsoid
spores, 14–20 × 4·5–8·5 m, with septa
1·5–2·5 m thick. The whole phenotype re-
sembles Caloplaca crenulatella s. lat., but C.
borysthenica is often sterile and forms patches
of diffuse yellow areoles. It is only known
from loess in the northern Black Sea region.
Based on ITS sequences, two topotype
specimens represent a well-supported mono-
phyletic group (PP = 1·00), which forms a
polytomy with one group of C. crenulatella
s. lat (clade D in Fig.1) and one group of C.
ferrarii s. lat (clade 2 in Fig.1). This clade is
weakly supported (PP = 0·65) and another
sequence considered as C. crenulatella s. lat is
placed as a sister taxon to this group (PP =
0·93). Whether C. borysthenica is a separate
species specialized in living on soil / loess, or
whether it is conspecific with the sister clade
of C. crenulatella s. lat. is at present unclear.
Specimens studied.Ukraine: Kherson region: Kherson,
Stanislav, 2007, J. Vondrák (CBFS JV6223); ibid., 2008
(CBFS JV6417, 7201, 7203, 7204, 7205); ibid., 2009
(CBFS JV7098, 7314, 7124, distributed in Sel. Exs.
Caloplaca, fasc. 2).
Caloplaca erichansenii S.Y. Kondr., A.
Thell, Kärnefelt & Elix
Bibliotheca Lichenologica 100: 239 (2009); type:
Greenland, SW Greenland, Head of Søndre Strømfjord
[Kangerlussuaq], Mt Hassel, south-facing slope, N
67°01 W 50°42, on loess, 1998, Lich. Groenl. Exs 722,
E.S. Hansen (C, GZU, Lich. Groenl. Exs. 722, sub C.
tominii—isotypes!). ITS sequence of the isotype (C):
HQ699638.
Collections from Greenland named Calo-
placa tominii were considered a separate
taxon by Kondratyuk et al. (2009), who de-
scribed it as C. erichansanii. According to our
observations, populations from Greenland (?
all arctic populations) and populations from
continental North America (e.g. Hansen
et al. 1987; Thomson 1997) are phenotypi-
cally indistinguishable from C. tominii s. str.
Kondratyuk et al. (2009) stated that asco-
spores of C. erichansenii are longer (18–23 m)
than in C. tominii (14–16 m), but we have
commonly observed ascospores longer than
20 minC. tominii s. str. Nevertheless, based
on two sequences from Greenland and
Montana, C. erichansenii forms a poorly sup-
ported clade that is not sister to C. tominii
s. str. (Fig.1). Instead, it forms a well sup-
ported clade (PP = 0·99) together with one
sequence of C. crenulatella s. lat. (terminal B
in Fig.1) and this group is sister to the C.
crenulatella s. lat. / C. borysthenica /C ferrarii
s. lat. clade (PP = 0·99).
Caloplaca interfulgens (Nyl.) J. Steiner
Verhandl. Zool.-Bot. Gesellsch. Wien 52: 479 (1902).—
Lecanora interfulgens Nyl., Flora 56: 340; type: Algeria,
474 THE LICHENOLOGIST Vol. 43

“Biskra”, 1878, J. P. Norrlin (H-NYL 29579!
—holotype).
(Fig. 2C)
The holotype is broken into small pieces
but consists of a yellow areolate thallus and
matches the samples of Caloplaca interfulgens
that we examined. The species occurs in
arid regions (semi-deserts) of North Africa,
Central Asia and the Middle East. The two
sequences studied group together (PP =
1·00) with Caloplaca tominii as a sister clade.
This relation was strongly supported (PP =
1·00) and both species occur in dry regions
and have similar distributions.
Specimens examined.Iran: Lake Urmia, Shirin
Bolagh, 2007, J. Vondrák (CBFS JV5773, 5777,
5781).—Kazakhstan: Mangistau region: west chink
(slope) of Ustyurt plateau, Manashy, 2009, J. Vondrák
& A. Khodosovtsev (CBFS JV7953); Shetpe, Say-Utes,
2009, J. Vondrák & A. Khodosovtsev (CBFS JV8184).
Caloplaca lactea (A. Massal.) Zahlbr.
Österr. Botan. Zeitschrift 51: 347 (1901).—Callopisma
luteoalbum var. Lacteum A. Massal., Schedul. Critic.7:
133 (1856); type: Italy, Venetto, Verona, A. Massalongo
(A. Massal., Lich. Italici Exs. 236; M—lectotype,
selected by Navarro-Rosinés & Hladun 1996; LE—
isolectotype!).
The name was formerly used in many
European countries for specimens of Calo-
placa crenulatella or C. marmorata. However,
it is a well-characterized species restricted,
in Europe, to (sub)Mediterranean regions
(Navarro-Rosinés & Hladun 1996) with
the northernmost confirmed occurrence in
Hungary (Vondrák et al. 2010). Surprisingly,
the same species is considered common also
in the British Isles (Fletcher & Laundon
2009; material not seen by us and in need of
revision). The ITS sequence of C. lactea fell
into the C. crenulatella group, but its closer
relationships are still unresolved. In eastern
Europe, the species is restricted to the sub-
mediterranean zone of the Black Sea coast in
Crimea (Khodosovtsev & Redchenko 2002)
and southern Russia (our data, see below),
where it grows on limestone rocks and calcar-
eous schist.
Selected specimens studied.Azerbaijan: Murovdagh:
alt. 2900 m, 1912, Schelkovnikov (LE).—Greece:
Fokidas: Itea, Galaxidi, 2010, J. Vondrák & O. Von-
dráková (CBFS JV8331).—Russia.Black Sea coast:
Novorossiysk, 2007, J. Vondrák (CBFS JV7540);
Tuapse, 2007, J. Vondrák (CBFS JV7525).—Turkey:
Black Sea coast:I
˙nebolu, Abana, on calcareous outcrop,
2007, J. Vondrák (CBFS JV6910, dupl. in C, GZU).—
Ukraine: Black Sea coast: Crimean peninsula, Cape
Martian, 1999, A. Khodosovtsev (KHER 2713, 2717);
Kerch peninsula, Chokrak, 1996, A. Redchenko (KHER
2718).
Caloplaca marmorata (Bagl.) Jatta
Sylloge Lich. Ital.: 251 (1900). —Callopisma marmoratum
Bagl., Nuov. Giorn. Botan. Ital.11: 84 (1879); type:
Italy, Sardinia, Cagliari, on limestone, J. B. Canepa
(MOD?).
This widely distributed species in Europe
was formerly incorrectly named C. lactea but
the monograph of the group (Navarro-
Rosinés & Hladun 1996) clarified its tax-
onomy. In the eastern part of Europe, it is
common on inland, rarely coastal, calcareous
rocks (e.g. Khodosovtsev & Redchenko
2002; Khodosovtsev 2003; Vondrák et al.
2007). According to our phylogenetic analy-
sis, all sequences studied form one single
clade (PP = 0·91) with unresolved relation-
ships.
Selected specimens studied.Iran: Lake Urmia, Shirin
Bolagh, 2007, J. Vondrák (CBFS JV5713).—
Kazakhstan: Mangyshlag region: Usturt, 1982, M.
Andreev (LE, sub C. lactea).—Slovakia: West Carpathi-
ans: Spišské Podhradie, 1958, A. Veˇzda (Veˇzda: Lich.
Boh. Exs. 235, LE!, sub C. lactea).—Turkey: Sea of
Marmara coast: Gallipoli peninsula, Abide monument,
2007, J. Vondrák (CBFS JV6663).—Ukraine: Black
Sea coast: Kherson region, Tendrivska kosa, 1993,
A. Khodosovtsev (KHER 2963). Crimean peninsula:
Chatyrdag, 1999, A. Khodosovtsev (KW 2968);
Karabi-yaila, 2000, A. Khodosovtsev (KHER 2970);
Tyrke, 2000, A. Khodosovtsev (KHER 2964); Roman-
Kosh, 2001, A. Khodosovtsev (KHER 2090); Karadag,
Mt Besh-Tas, 2001, A. Khodosovtsev (KHER 2967);
Tarchankut peninsula, 1998, A. Khodosovtsev (KHER
2965); Kerch peninsula, Mt Opuk, 1994, A. Khodosovtsev
(KHER 2956); Chokrak, 1996, A. Redchenko (KHER
2959); Osoviny, 1994, A. Khodosovtsev (KHER 2957).
Caloplaca ochracea (Schaer.) Flagey
Mém. Soc. d’Emulat. Doubs: 257 (1886).—Lecidea ochra-
cea Schaer., Naturwiss. Anzeiger Allg. Schweizer. Ges.
Gesamten Naturwiss.2: 11 (1818); type: Alpes calcariae,
2011 Caloplaca crenulatella complex—Vondrák et al. 475

Schaerer (G—lectotype designated by Hafellner & Poelt
1979).
Caloplaca ochracea is safely placed in the
ingroup (as in the case of C. lactea), but it is
an atypical member of the C. crenulatella
group for it possesses spores with three thin
septa when mature. Two to three-septate
spores are not rare among tropical Caloplaca
species (Hafellner & Poelt 1979), but they
are probably unrelated to C. ochracea. This
species is widely distributed on calcareous
rocks in Europe and is easily recognizable.
In Eastern Europe, it occurs on inland cal-
careous rocks and also on coastal cliffs (e.g.
Khodosovtsev & Redchenko 2002; Vondrák
& Slavı´ková-Bayerová 2006).
Selected specimens studied.Romania: Munt¸ii Banatu-
lui Mts:Ba˘ile Herculane, slope of Mt Domogled, 2005,
J. Vondrák (CBFS JV3628).—Ukraine: Crimean Penin-
sula: Perevalnoye, 1999, A. Khodosovtsev (KHER 2832);
Chiginitra, 2000, A. Khodosovtsev (KHER 2834); Cape
Martian, 2000, A. Khodosovtsev (KHER 2831, 2833).
Caloplaca tominii Savicz
Isswest. Glawn. Botan. Sada 29: 194 (1930); type:
Russia, Astrakhan region, Baskunchak, on soil in
steppe, 1926, Savicz (Lich. Ross. 24; LE—holotype!).—
Caloplaca gyalolechiaeformis Szatala, Ann. Hist.-Nat.
Mus. Natl. Hungarici, s.n.7: 276 (1956); type: Hungary,
Várhegy, Nógrád, on siliceous rock, 1937, V. Ko˝faragó-
Gyelnik (BP 27571—holotype!; BP 27569, 27570—
isotypes).—Caloplaca pseudocitrina Khodos. &
Kudratov, in Kondratyuk, Kärnefelt, Kudratov &
Khodosovtsev, Nordic Journal of Botany 22: 633 (2002);
type: Tajikistan, Southern Tajikistan, Chormagzak pass,
1968, I. Kudratov (KW—isotype!).
(Fig. 2D)
The name C. tominii was previously used
for sorediate terricolous populations, com-
mon in Central Asian steppes. We discovered
that sorediate saxicolous populations named
C. gyalolechiaeformis or C. pseudocitrina are
conspecific with terricolous C. tominii. The
species occurs mainly in the steppe and semi-
desert zones of Eurasia with the westernmost
records being from Hungary (Vondrák et al.
2010) and the Czech Republic (this paper).
The identity of populations from continental
Scandinavia (Nordin 1972), Pakistan (Poelt
& Hinteregger 1993) and the Alps (Hafellner
& Türk 2001) needs further investigation.
Sequences of specimens from the temperate
to arid zone of Eurasia formed a well sup-
ported clade (PP = 1·00), closely related to
C. interfulgens (PP = 1·00 for their mutual
sisterhood).
Caloplaca tominii often occurs sterile and
cannot then be separated morphologically
from some other yellow sorediate species,
mainly C. flavocitrina (Nyl.) H. Olivier.
However, sterile terricolous samples from
continental Eurasia should be identified as
C. tominii on ecological grounds.
Selected specimens studied.Czech Republic: C
ˇeské
Budeˇjovice, Nové Hodeˇjovice, on concrete and soil,
2009, J. Vondrák (CBFS JV6989).—Iran: Lake Urmia,
Bandar-e-Rahmanlu, on soil and over mosses, 2007,
J. Vondrák (CBFS JV5591).—Kazakhstan: “Naur-
zumskiy zapovednik” protected area, on soil, 1939, S. S.
Levitskiy (LE). Western Kazakhstan region: lake Shalkar,
on soil, 2009, J. Vondrák (CBFS JV7273); ibid., on
sandstone (CBFS JV7274).—Kyrgysztan: Kyrgyzskiy
khrebet Mts: Kochkorki, alt. 2700, on soil, 1972, L.
Bredkina (LE); between Frundze and Osh, alt. 2100 m,
on soil, 1870, L. Bredkina (LE).—Russia: Astrakhan
region: Baskunchak, on soil, 2009, J. Vondrák (CBFS
JV7946, topotype). Orenburg region: Svetliy, locality
Aschisayskaya step, on soil, 2004, O. Merkulova (LE,
ORIS); Aytuarskaya step, on conglomerate, 2009,
J. Vondrák (CBFS JV8234).—Tajikistan: Eastern
Pamir: Murghab, alt. 3860 m, 1966, N. S. Golubkova &
L. Bredkina (LE); ibid., alt. c. 4000 m (LE).—Turkey:
Kurdistan: Lake Van, lava stream near NW shore of Lake
Sodalı, on soil among mosses, 2007, J. Vondrák (CBFS
JV6629); Kars, Aygir Lake, on siliceous stone, 2007,
J. Vondrák (CBFS JV6084).—Ukraine: Kherson region:
Stanislav, Shiroka Balka, on loess, 2003, A. Khodosovtsev
(KHER 2878); Sofiyevka, on loess, 2008, J. Vondrák
(CBFS JV7200).
Unresolved Groups
Caloplaca crenulatella (Nyl.) H. Olivier
s. lat.
According to our phylogenetic study, this
taxon is paraphyletic and contains at least
four lineages, which we cannot currently
distinguish using conventional characters.
However, this outcome may be caused par-
tially by the poor resolution of our clado-
gram. One of the lineages (clade of two
sequences, PP = 0·85, marked as A in Fig.1)
fell within the basal polytomy and can there-
fore not be taken into phylogenetic assess-
ment. Another sequence of C. crenulatella
476 THE LICHENOLOGIST Vol. 43

(terminal B) forms a clade together with two
sequences of C. erichansenii sequences, with
strong support (PP = 0·99), and the remain-
ing three sequences (clades C and D) showed
clear affinity to C. borysthenica and the ‘clade
2’ of C. ferrarii (PP = 0·93). All studied
specimens share characters described in
Navarro-Rosinés & Hladun (1996) for C.
crenulatella (Nyl.) H. Olivier. The variability
of some characters (e.g. ascospore size and
width of septa) within individual samples is
often large; this makes it difficult to find
diagnostic characters of the lineages (poss-
ible phylospecies). The degree of thallus
development also varies greatly, from absent
to subsquamulose and this depends, at least
in part, on the age of the lichen. To under-
stand this species complex we need molecu-
lar data from more samples to see whether
the phenotypic, geographical and ecological
characteristics of each clade can be better
interpreted. Caloplaca crenulatella s. lat. is
common on various base-rich siliceous or
calcareous rocks and on artificial substrata,
often concrete (e.g. Khodosovtsev 2001;
Vondrák & Slavı´ková-Bayerová 2006).
Caloplaca ferrarii (Bagl.) Jatta s. lat.
Navarro-Rosinés & Hladun (1996) char-
acterized Caloplaca ferrarii by apothecia be-
tween 0·7–1·5 mm diam. and ascospores
with septa 1–2 m. Based on our observa-
tions, apothecia in young specimens may be
much smaller, usually 0·3–0·7 mm diam.
and the ascospore septa are 1–4 m wide.
Caloplaca ferrarii s. lat. (in our sense) is
a heterogeneous group appearing in two
different clades in our tree (Fig. 1). One of
the lineages (clade 1 in Fig.1) contains a
sequence of Spanish saxicolous material col-
lected by Navarro-Rosinés (cf. Gaya et al.
2008) together with saxicolous and terricol-
ous material from a wide geographical range.
It is a well supported clade (PP = 0·99), but
its position with the sister clade (C. interful-
gens +C. tominii) is not strongly supported.
The second lineage (clade 2, PP = 1·00)
contains a sequence of material identified by
Navarro-Rosinés (HQ699637), a sequence
of a terricolous sample from Hungary and a
sequence of a Ukrainian specimen from con-
crete. It clusters together with sequences of
C. borysthenica and the C and D lineages of
C. crenulatella s. lat. (PP = 0·93). Although
both clades of C. ferrarii clearly represent
separate phylogenetic lineages, we cannot
currently distinguish them by their pheno-
types.
Species omitted from the molecular
analysis
Two more European species have been
described from the Caloplaca crenulatella
group, but were omitted from our analysis as
source material was unavailable. We have
some doubts about their delimitation, but
include them here for completeness.
Caloplaca epigaea Søchting, Huneck &
Etayo
Bibliotheca Lichenologica 96: 282 (2007); type: Spain,
Aragon, Zaragoza, on gypsic soil, 2004, A. & J. Etayo
21453 (JACA—holotype; C—isotype!).
The description of the species (Søchting
et al. 2007) corresponds with Caloplaca borys-
thenica or C. crenulatella s. lat. The authors
use well-developed thallus areoles and terri-
colous occurrence as diagnostic characters,
but other taxa may have the same morpho-
logy and terricolous occurrences are not
exceptional among them (Lo˝kös 2003;
Vondrák et al. 2007).
Caloplaca lacteoides Nav.-Ros. &
Hladun
Bull. Soc. Linn. Provence,47: 156 (1996); type: España,
Cataluña, Prov. Barcelona, Vallès Oriental, el Figaró,
al lado del camino hacia Montmany, U.T.M.
31TDG3718 600 m alt., 22 October 1992, P. Navarro-
Rosinés (BCC—holotype).
According to the description (Navarro-
Rosinés & Hladun 1996), this species is char-
acterized by isodiametric cells in the outer
part of the proper exciple and by unbranched
paraphyses with strongly widened tips.
Branching and anastomosing of paraphyses
is, however, a variable character within and
among samples of the same species and in
our opinion it is very unlikely that some
2011 Caloplaca crenulatella complex—Vondrák et al. 477

species of this group exists with entirely un-
branched paraphyses. We have observed
swollen cell lumina in the outer part of
proper exciples in most species in the group,
but this character is variable among samples
and we do not consider it to be a good taxo-
nomic character. We have not seen the type
of Caloplaca lacteoides, but we are sceptical
about the delimitation of this species. Some
East European Caloplaca marmorata-like
specimens seen by us (KHER 2845, 2916,
from Crimea) with large cell lumina in para-
physes tips and in the upper exciple corre-
spond to C. lacteoides, but their identity
remains unclear.
Key to the species
1 Thallus sorediate / blastidiate; vegetative diaspores c. 20–80 m diam. (when sterile,
it may be indistinguishable from some other sorediate Caloplaca species)....2
Thallus not sorediate / blastidiate ..........................3
2(1) Specimens from continental Eurasia .................. C. tominii
Specimens from the Arctic and continental N. America ..... C. erichansenii
3(1) Ascospores 3-septate when mature .................. C. ochracea
Ascospores polarilocular with septum 1–4 m thick ................4
4(3) Thallus yellow, well-developed ...........................5
Thallus not yellow, or if yellow then poorly developed ...............8
5(4) Thallus of granules, squamules or diffuse convex areoles in apothecial surroundings
or forming small sterile spots ...........................6
Thallus areolate, effuse, not restricted to apothecial surroundings.........7
6(5) Often sterile but pycnidia frequent; thallus forming spots a few mm diam.; on
loess; often infected by Llimoniella caloplacae S. Y. Kondr. & Khodos.......
..................................C. borysthenica
Fertile; pycnidia rare or absent; variable in thallus extension; on various sub-
strata ............................C. crenulatella s. lat.
7(5) Ascospores c. 14–17 × 6–8 m; thallus diffuse at margin, often with distinct grey-
white prothallus; on siliceous (maritime) rocks.............C. diffusa
Ascospores c. 15–19 × 5–7·5 m; thallus sharply delimited by marginal areoles;
prothallus indistinct; on calcareous (inland) rocks; in N Africa, Near East and
Central Asia (not confirmed from Europe)............C. interfulgens
8(4) Apothecia small, up to 0·5 mm ...........................9
Apothecia larger, over 0·5 mm (except for young or grazed thalli)........ 11
9(8) Ascospores broadly ellipsoid, c. 11–16 × 6–9 m, apothecia in shades of yellow to
orange ..................................C. lactea
Ascospores ellipsoid, more than 15 m long, apothecia in shades of red, rarely
orange ......................................10
10(9) Uppermost cells of proper exciple c. 5–9 m wide ..........C. lacteoides
Uppermost cells of proper exciple c. 3–5 m wide .........C. marmorata
11(8) Ascospores c. 13–18 × 6–8 m; thallus endolithic; on calcareous maritime cliffs . .
.....................................C. aquensis
Ascospores c. 15–20 × 6–8 m; thallus usually partly epilithic; on calcareous or
siliceous, inland or maritime sites........................12
478 THE LICHENOLOGIST Vol. 43

12(11)Proper exciple swollen, c. 120–150 m thick, thalline exciple not enlarged and not
distinctly crenulate; thallus pale grey, devoid of anthraquinones .........
..................................C. ferrarii s. lat.
Proper exciple thinner, c. 80–100 m thick, thalline exciple (at least in older
apothecia) somewhat enlarged, paler than proper exciple (yellow) and often
crenulate; thallus (if present) yellow ...........C. crenulatella s. lat.
Discussion
The biodiversity of the group is very un-
equally distributed in Europe. In Scandina-
via, only two species occur (Santesson et al.
2004), C. crenulatella (in southern territories)
and C. tominii (isolated occurrences in
central Norway). From the British Isles
only three species are reported (Fletcher &
Laundon 2009). The situation is similar in
most of central Europe, where only C. crenu-
latella,C. marmorata and C. ochracea are
regularly collected. The European centre of
biodiversity is the Mediterranean, from
where most of the species have been de-
scribed. This has caused misunderstandings
of some names by several European authors;
for example, the name C. lactea was used for
various central European species and C.
interfulgens for C. maritima. The monograph
of the group by Navarro-Rosinés & Hladun
(1996), which focused on Mediterranean
regions, greatly clarified the taxonomy of this
group, which was previously poorly under-
stood.
Outside Europe, we have reliable distri-
bution data for species of the C. crenulatella
group only from North Africa, Near East,
Middle East and Central Asia (Navarro-
Rosinés & Hladun 1996; this study); records
from other regions may be unreliable. Some
European species have been reported from
North America, but we consider these re-
ports to be doubtful; for example, Wetmore
(2007) listed Caloplaca crenulatella,C.
marmorata and C. tominii from the Sonoran
Desert. We obtained one sequence from a
Californian collection called C. crenulatella;it
is placed within the group and, surprisingly,
groups with a collection of C. crenulatella
s. lat. from the Czech Republic (clade A, PP
= 0·85). The sequence from a collection
from Montana, called C. tominii, falls into the
group with the arctic C. erichansenii and is
unrelated to the Eurasian C. tominii s. str. We
do not have molecular data for the Sonoran
lichen called C. marmorata, but its ascospores
are only 14–15·5 m long, considerably
shorter than the 14–19 m in the European
C. marmorata s. str. The North American C.
nashii Nav.-Ros., Gaya & Hladun was con-
sidered a member of this group on morpho-
logical grounds by Navarro-Rosinés et al.
(2001), but we obtained ITS sequences of C.
nashii and C. cf. nashii (second sample differs
from the description of C. nashii in its spore
characters) from California; they fall outside
the group and the BLAST search showed
that their closest relationship is to C. bolacina
(Tuck.) Herre.
We are grateful to Ester Gaya, Laszlo Lo˝kös, Olga
Nadyeina, Toby Spribille and Kerry Knudsen for pro-
viding their herbarium material. Ulf Arup and Ulrik
Søchting commented on the manuscript and Ulf Arup
also provided one sequence of Caloplaca diffusa. Linda
in Arcadia kindly revised the English and Latin. This
research received support from the Visegrad Fund
(grant 51000067) and the SYNTHESYS Project http://
www.synthesys.info/ which is financed by European
Community Research Infrastructure Action under the
FP7 “Capacities” Programme.
R
Abbott, B. F. M. (2009) Checklist of the lichens and
lichenicolous fungi of Greece. Bibliotheca Licheno-
logica 103: 1–368.
Arup, U. (2006) A new taxonomy of the Caloplaca citrina
group in the Nordic countries, except Iceland.
Lichenologist 38: 1–20.
Ekman, S. (2001) Molecular phylogeny of the Bacidi-
aceae (Lecanorales, lichenized Ascomycota). Myco-
logical Research 105: 783–797.
Fletcher, A. & Laundon, J. R. (2009) Caloplaca.InThe
Lichens of Great Britain and Ireland. (C. W. Smith, A.
Aptroot, B. J. Coppins, A. Fletcher, O. L. Gilbert,
P. W. James & P. A. Wolseley, eds): 245–273.
London: British Lichen Society.
2011 Caloplaca crenulatella complex—Vondrák et al. 479

Gardes, M. & Bruns, T. D. (1993) ITS primers with
enhanced specificity for basidiomycetes. Applica-
tion for the identification of mycorrhizae and rusts.
Molecular Ecology 2: 113–118.
Gaya, E., Navarro-Rosinés, P., Llimona, X., Hladun, N.
& Lutzoni, F. (2008) Phylogenetic reassessment
of the Teloschistaceae (lichen-forming Ascomycota,
Lecanoromycetes). Mycological Research 112: 528–
546.
Hafellner, J. & Poelt, J. (1979) Die Arten der Gattung
Caloplaca mit plurilocularen Sporen (Meroplacis,
Triophthalmidium, Xanthocarpia). Journal of the
Hattori Botanical Laboratory 46: 1–41.
Hafellner, J. & Türk, R. (2001) Die lichenisierten Pilze
Österreichs – eine Checkliste der bisher nachgew-
iesenen Arten mit verbreitungsangaben. Stapfia 76:
1–167.
Hall, T. A. (1999) BioEdit: a user-friendly biological
sequence alignment editor and analysis program for
Windows 95/98/NT. Nucleic Acids Symposium Series
41: 95–98.
Hansen, E. S., Poelt, J. & Søchting, U. (1987) Die
Flechtengattung Caloplaca in Grönland. Meddelelser
om Gronland, Bioscience 25: 1–52.
Katoh, K., Kuma, K., Toh, H. & Miyata, T. (2002)
MAFFT: a novel method for rapid multiple se-
quence alignment based on fast Fourier transform.
Nucleic Acids Research 30: 3059–3066.
Khodosovtsev, A. Y. (1999)
The Lichens of the Black Sea’s
Steppes
. Kiev: Phytosociocentre. [in Ukrainian].
Khodosovtsev, A. Y. (2001) Species of the genus Calo-
placa Th. Fr. (Teloschistaceae) new to Ukraine.
Ukrainian Botanical Journal 58: 460–465 [in
Ukrainian].
Khodosovtsev, A. Y. (2003) An annotated list of the
lichen forming fungi of the Karadag Natural
Reserve. Visti Biosphernogo zapovednika “Askania-
Nova”5: 31–43 [in Ukrainian].
Khodosovtsev, A. Y. & Redchenko, O. O. (2002) An
annotated list of the lichen forming fungi of the
natural reserve “Mis Martian”. Ukrainian Botanical
Journal 59: 64–71 [in Ukrainian].
Kondratyuk, S., Khodosovtsev, A. & Kärnefelt, I.
(2006) Llimoniella caloplacae sp. nova (Leothiales), a
new lichenicolous fungus on Caloplaca borysthenica
sp. nova (Lecanorales, Ascomycota). Mycologia
Balcanica 3: 95–98.
Kondratyuk, S. Y., Kärnefelt, I., Elix, J. A. & Thell, A.
(2009) Contribution to the Teloschistaceae, with
particular reference to the Southern Hemisphere.
Bibliotheca Lichenologica 100: 207–282.
Lo˝ kös, L. (2003) [Loess colonizing lichens in Hungary.]
Acta Academiae Paedagogicae Agriensis, Sectio Bio-
logia 24: 159–188.
Müller, K. F. (2005) SeqState – primer design and
sequence statistics for phylogenetic DNA data sets.
Applied Bioinformatics 4: 65–69.
Navarro-Rosinés, P. & Hladun, N. L. (1996) Las espe-
cies saxı´colo-calcı´ colas del grupo de Caloplaca lactea
(Teloschistaceae,lı´quenes), en las regiones mediter-
ránea y medioeuropea. Bulletin de la Société linéenne
de Provence 47: 139–166.
Navarro-Rosinés, P. & Roux, C. (1992) Présence de
Caloplaca aquensis sur le littoral Méditerranéen.
Cryptogamie, Bryologie-Lichénologie 13: 355–358.
Navarro-Rosinés, P., Gaya, E. & Hladun, N. L. (2001)
Caloplaca nashii sp. nov. (Teloschistaceae, Lichenes),
a North American species of the C. lactea-group
growing in caliche. Mycotaxon 79: 29–41.
Nordin, I. (1972) Caloplaca sect. Gasparrinia in
Nordeuropa. Taxonomiska och Ekologiska Studier.
Uppsala: Skriv Service AB.
Nylander, J. A. A., Ronquist, F., Huelsenbeck, J. P. &
Nieves-Aldrey, J. L. (2004) Bayesian phylogenetic
analysis of combined data. Systematic Biology 53:
47–67.
Poelt, J. & Hinteregger, E. (1993) Beiträge zur Kenntnis
der Flechtenflora des Himalaya. VII. Die Gattun-
gen Caloplaca, Fulgensia und Ioplaca.Bibliotheca
Lichenologica 50: 1–247.
Ronquist, F. & Huelsenbeck, J. P. (2003) MRBAYES 3:
Bayesian phylogenetic inference under mixed mod-
els. Bioinformatics 19: 1572–1574.
Santesson, R., Moberg, R., Nordin, A., Tønsberg, T. &
Vitikainen, O. (2004) Lichen-forming and Lichen-
icolous Fungi of Fennoscandia. Uppsala: Museum of
Evolution, Uppsala University.
Smith, C. W., Aptroot, A., Coppins, B. J., Fletcher, A.,
Gilbert, O. L., James, P. W. & Wolseley, P. A. (eds)
(2009) The Lichens of Great Britain and Ireland.
London: British Lichen Society.
Søchting, U. (1997) Two major anthraquinone chem-
osyndromes in Teloschistaceae.Bibliotheca Licheno-
logica 68: 135–144.
Søchting, U., Huneck, S. & Etayo, J. (2007) Caloplaca
epigaea sp. nova from arid soil in Spain and dead
grass in Germany. Bibliotheca Lichenologica 96: 279–
286.
Swofford, D. L. (2002) PAUP*: Phylogenetic Analysis
Using Parsimony (*and Other Methods), Version 4.
Sunderland, Massachusetts: Sinauer Associates.
Thomson, J. W. (1997) American Arctic Lichens. 2. The
Microlichens. The University of Madison: Wisconsin
Press.
Vondrák, J. & Slavı´ková-Bayerová, Š. (2006): Contribu-
tion to the lichenized and lichenicolous fungi in
Bulgaria. II, the genus Caloplaca.Mycologia Bal-
canica 3: 61–69.
Vondrák, J., Kocourková, J., Palice, Z. & Liška, J.
(2007) New and noteworthy lichens in the Czech
Republic – genus Caloplaca.Preslia 79: 163–184.
Vondrák, J., R
ˇı´ha, P., Arup, U. & Søchting, U. (2009)
The taxonomy of the Caloplaca citrina group (Tel-
oschistaceae) in the Black Sea region; with contribu-
tions to the cryptic species concept in lichenology.
Lichenologist 41: 571–604.
Vondrák, J., Khodosovtsev, A., Lo˝kös, L. & Merkulova,
O. (2010) The identity of type specimens in BP of
some names in Caloplaca.Mycotaxon 111: 241–250.
Wetmore, C. M. (2007) Caloplaca.InLichen Flora of the
Greater Sonoran Desert Region. Vol. 3. (T. H. Nash
III, C. Gries & F. Bungartz, eds): 179–220. Tempe:
Lichens Unlimited, Arizona State University.
480 THE LICHENOLOGIST Vol. 43

White, T. J., Bruns, T. D., Lee, S. & Taylor, J. (1990)
Amplification and direct sequencing of fungal ribo-
somal RNA genes for phylogenies. In PCR Protocols:
a Guide to Methods and Applications (M. A. Innis,
D. H. Gelfand, J. J. Sninsky, & T. J. White, eds):
315–322. San Diego: Academic Press.
Accepted for publication 28 May 2011
2011 Caloplaca crenulatella complex—Vondrák et al. 481