The Lichenologist 50(4): 411–424 (2018) © British Lichen Society, 2018 doi:10.1017/S0024282918000142 Caloplaca sol (Teloschistaceae), a new coastal lichen from Great Britain Alan ORANGE Abstract: Caloplaca sol is described as a new species from limestone and basic siliceous rocks on the southern and western coasts of Great Britain. It is characterized by a well-developed, crustose, non- placodioid, epilithic, cracked, orange-yellow thallus, almost concolorous apothecia up to 0·66 mm diameter, and ascospores c.11·0–12·2–13·0 µm long with a septum c.0·4 × the ascospore length. Caloplaca dalmatica is related but differs in the endolithic or only thinly epilithic thallus. Caloplaca marina is darker orange in colour, with more convex areoles, and is mostly confined to the splash zone of the seashore. Caloplaca maritima differs in the typically more convex, sometimes isolated areoles, and often in the presence of a crenulate thalline margin in young apothecia. Caloplaca itiana is newly reported from Great Britain from coastal limestone; it differs from C. sol in the thallus being endolithic or almost so, and from C. dalmatica in the more completely endolithic thallus and the larger ascospores. Key words: Flavoplaca, France, key, maritime, taxonomy, Wales Accepted for publication 16 November 2017 Introduction Caloplaca is restricted to 12 species related to the type of the genus, C. cerina, and the Species of Caloplaca in the broad sense often remaining species are distributed throughout form a conspicuous part of the lichen numerous resurrected or newly described communities in which they occur due to the genera (Arup et al. 2013). yellow to orange colour of the thallus or A conspicuous unidentified species of apothecia in many taxa. The species-level Caloplaca in the broad sense has been known taxonomy in Europe has changed sig- on coastal rocks in Great Britain for many nificantly in recent years due to more detailed years and is described below. It is related to, studies, supported by DNA sequencing, among others, the species currently known as which have revealed additional taxa within C. dalmatica in Great Britain. The relationship formerly more broadly defined species or of these species to the recently described species-groups, including C. cerina (Šoun C. itiana was investigated, with the result that et al. 2011), C. citrina (Arup 2006; Vondrák C. itiana is newly recorded from Great Britain. et al. 2009), C. crenulatella (Vondrák et al. 2011), C. herbidella (Arup & Åkelius 2009) and C. holocarpa (Arup 2009; Vondrák et al. Materials and Methods 2012, 2016). Traditionally, growth habit Thallus and apothecial sections were mounted in water was used as a major generic character in and 5% KOH (K). Ascospores were measured in water. Teloschistaceae and the genus Caloplaca was The spores of recently collected specimens were first restricted to species with crustose or placo- killed by warming apothecial sections in a drop of water, then allowing them to dry, and then remounting in dioid thalli. Caloplaca in this sense is now water. Ascospore measurements for each species are known to be non-monophyletic (e.g. Gaya given in the following format (after Frolov et al. 2016): et al. 2012). Recently, a new arrangement of ðminimumÞ x1 À x2 À x3ðmaximumÞ,whereminimum the family has been proposed in which and maximum are extreme values, x1 is the smallest mean value measured for each individual collection, x2 is the mean of all values measured in that species, and x3 is the A. Orange: Natural Sciences, National Museum of largest mean value measured for each individual Wales, Cardiff CF10 3NP, U.K. Email: alan.orange@ collection. The measurements are followed by the museumwales.ac.uk standard deviation (SD) of x2,N= the total number of Downloaded from https://www.cambridge.org/core. IP address: 170.106.40.219, on 25 Sep 2021 at 06:44:00, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0024282918000142 412 THE LICHENOLOGIST Vol. 50 measurements for all samples of that species and et al. 2007). Support values of ≥70% maximum likelihood n = the number of samples from which measurements were bootstrapping were regarded as significant. made. Sequences used in the analysis are shown in Table 1. Recently collected or frozen material was used to generate DNA sequences. BLAST searches (http://blast. Additional figured specimens. Caloplaca marina: ncbi.nlm.nih.gov/Blast.cgi) were used to find sequences Great Britain: Wales: V.C. 41, Glamorgan, Gower, similar to those of the new species. Additional possible Port Eynon, grid ref. 21/4649.8463, 51·539638°N, related taxa, and potential outgroup taxa, were suggested −04·214855°W, on limestone in splash zone of seashore, by the analyses in Arup et al. (2013). A sequence of 2014, Orange 21907 (NMW–C.2014.008.60). England: Caloplaca oasis was used as outgroup in the ITS analysis. V.C. 9, Dorset, Isle of Portland, below Grove Cliff, Specimens used in the analyses are shown in Table 1. grid ref. 30/7047.7166, 50·543955°N, −02·418112°W, DNA was extracted using the Qiagen DNeasy Plant on limestone on rocks on slope above seashore, 2012, Mini Kit; the manufacturer’s instructions were followed Orange 21071 (NMW–C.2012.002.70). except that warm water was used for the final elution. Caloplaca maritima: Great Britain: Wales: V.C. 52, PCR amplification was carried out using Bioneer Accu- Anglesey, Amlwch, Parys Mountain, grid ref. 23/ Power PCR Premix in 50 µl tubes. The two internal 4433.9052, 53·388457°N, 04·342421°W, on rock in mor- transcribed spacer regions and the 5.8S region (ITS1- tared wall of disused windmill, 2011, Orange 20529 5.8S-ITS2) of the nuclear ribosomal genes were ampli- (NMW–C.2011.014.34); Benllech, Trwyn Dwlban, fied, using the primers ITS1F and ITS4. The PCR grid ref. 23/5317.8203, 53·314746°N, 04·206073°W, on thermal cycling parameters were: initial denaturation for limestone rocks above seashore, 2012, Orange 21047 5 min at 94 °C, followed by 5 cycles of 30 s at 94 °C, 30 s (NMW–C.2012.002.66). at 55 °C and 1 min at 72 °C, then 30 cycles of 30 s at 94 °C, 30 s at 52 °C and 1 min at 72 °C. PCR products were visualized on agarose gels stained with ethidium Results bromide and purified using the Sigma GenElute PCR Clean-Up Kit. DNA sequencing was performed by DNA ITS analysis Sequencing & Services (MRC I PPU, School of Life Sciences, University of Dundee, Scotland, www.dnaseq. The tree resulting from analysis of the co.uk) using Applied Biosystems BigDye v3.1 chemistry ITS1-5.8S-ITS2 region is shown in Fig. 1. on an Applied Biosystems 3730 automated capillary All species included in the tree, with the DNA sequencer. Sequences were assembled and edited using GeneStudio software (http://www.genestudio. exception of the outgroup, belong to the com). Alignment was carried out using BioEdit (http:// genus Flavoplaca,asdefined by Arup et al. www.mbio.ncsu.edu/BioEdit/bioedit.html); ClustalW (2013). The lower nodes of the tree are was used to create an initial alignment which was edited poorly supported. Specimens from Great manually. Ambiguously aligned regions were deleted Britain described below as the new species before further analysis of the alignment. ITS sequences were newly prepared for 40 specimens. Caloplaca sol form a well-supported clade of The ITS alignment was 574 bp in length. A total of 66 bp seven sequences showing little variation. were excluded from the analysis as they could not be A well-supported clade with Caloplaca unambiguously aligned (ITS1: 2 regions with a total of dalmatica and a single sequence named as 41 bp, ITS2: 3 regions with a total of 25 bp). After deletion of ambiguous regions, the alignment was 509 C. navasiana is a sister clade to the C. sol characters in length. clade but with very low support. A number of Maximum likelihood (ML) analyses were used to species are not resolved in the tree, being investigate phylogenetic relationships and support values. recovered as non-monophyletic. This is likely Analysis was performed using RAxML v8.2.10 (Stama- to be due to a combination of factors takis et al. 2008; Stamatakis 2014), as hosted on the fi CIPRES Science Gateway (Miller et al.2010). including insuf cient resolving power of the PartitionFinder v2.1.1 (Guindon et al. 2010; Lanfear et al. ITS region between certain closely related 2012, 2017) was used to optimize partitions and sub- species, misidentification of specimens, and stitution models. Three regions of the ITS (ITS1, 5.8S, insufficient taxon and specimen sampling. ITS2) were used as input. Parameters included linked fi branch lengths and greedy search, and the ‘–raxml’ For example, ve sequences of C. maritima command was used to limit the number of models to form a well-supported clade, but it is nested those used in RAxML. PartitionFinder returned GTR + within a poorly supported clade including G for ITS1 and ITS2, and GTR + G + I for the 5.8S C. communis and C. havaasii. Another region. Analyses with RAxML used rapid bootstrapping well-supported clade includes sequences with 1000 iterations and the GTRGAMMAI substitution model; a search for the best-scoring ML tree was carried named as C. calcitrapa, C. itiana and out with the bootstrap analysis in a single run. The C. navasiana, but which are not resolved into resulting tree was visualized using MEGA v4 (Tamura individual taxa. It is possible that these three Downloaded from https://www.cambridge.org/core. IP address: 170.106.40.219, on 25 Sep 2021 at 06:44:00, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms.