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Catenarina (Teloschistaceae, Ascomycota), a New Southern Hemisphere Genus with 7-Chlorocatenarin

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Catenarina (Teloschistaceae, Ascomycota), a New Southern Hemisphere Genus with 7-Chlorocatenarin The Lichenologist 46(2): 175–187 (2014) 6 British Lichen Society, 2014 doi:10.1017/S002428291300087X Catenarina (Teloschistaceae, Ascomycota), a new Southern Hemisphere genus with 7-chlorocatenarin Ulrik SØCHTING, Majbrit Zeuthen SØGAARD, John A. ELIX, Ulf ARUP, Arve ELVEBAKK and Leopoldo G. SANCHO Abstract: A new genus, Catenarina (Teloschistaceae, Ascomycota), with three species is described from the Southern Hemisphere, supported by molecular data. All species contain the secondary metabolite 7-chlorocatenarin, previously unknown in lichens. Catenarina desolata is a non-littoral, lichenicolous species found on volcanic and soft sedimentary rock at 190–300 m in and near steppes in southernmost Chile and on the subantarctic island, Kerguelen. Catenarina vivasiana grows on maritime rocks and on rock outcrops in lowland Nothofagus forests, but has also been found at alti- tudes up to c. 580 m on moss and detritus on outcrops in Tierra del Fuego. The Antarctic species Caloplaca iomma is transferred to Catenarina based on chemical data; it grows on rocks near the coast in maritime Antarctica. Key words: Antarctica, Argentina, Caloplaca, Chile, HPLC, Kerguelen Island, lichen metabolites, molecular phylogeny Accepted for publication 18 November 2013 Introduction circumscribe with classical morphological, anatomical and secondary chemical char- The lichen family Teloschistaceae, with more acters. However, in some cases, secondary than 1000 species published worldwide metabolites provide synapomorphic charac- (Søchting & Lutzoni 2003; Gaya et al. ters, such as in the genus Shackletonia (Arup 2012), is currently receiving considerable et al. 2013). attention in order to subdivide the family Tierra del Fuego and southern Patagonia into molecularly-based genera (Arup et al. have recently been subjected to intensive 2013). The traditional genus Caloplaca has lichenological studies in connection with thus been divided into multiple genera based several Spanish research projects. In parti- on phylogenetic analyses of DNA data. In cular, the family Teloschistaceae has been the many cases, the monophyletic genera de- focus, resulting in the discovery of several fined by molecular characters are not easy to new species (Lumbsch et al. 2011; Søchting & Sancho 2012). U. Søchting and M. Z. Søgaard: Section for Ecology The present study describes a new genus and Evolution, Department of Biology, University of with three species, two of them new to Copenhagen, Universitetsparken 15, 2100 Copenhagen science, within Teloschistaceae. They form a K, Denmark. Email: [email protected] distinct molecular clade, have a charac- J. A. Elix: Research School of Chemistry, Building 33, Canberra, A.C.T. 0200, Australia. teristic reddish brown pigmentation, and the U. Arup: Botanical Museum, Lund University, Box thallus or disc contains 7-chlorocatenarin, a 117, SE-221 00 Lund, Sweden. secondary compound new to lichens. The A. Elvebakk: Tromsø University Museum, University of genus is so far known only from the south- Tromsø, N-9037 Tromsø, Norway. ernmost part of the Southern Hemisphere, L. G. Sancho: Universidad Complutense de Madrid, Departamento de Biologı´a Vegetal II, Plaza de Ramo´ny viz. Antarctica, southern Patagonia and Cajal s/n, S-28040 Madrid, Spain. Kerguelen Island. Downloaded from https://www.cambridge.org/core. University of Athens, on 30 Sep 2021 at 06:36:47, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S002428291300087X 176 THE LICHENOLOGIST Vol. 46 Material and Methods assumed to be significant if two different relationships (one monophyletic and one non-monophyletic) were The study includes material from southern Patagonia, both supported with posterior probabilities 0Á95 or collected by the fifth author in 1998 and 1999, and by higher (Buckley et al. 2002). A suitable model of molec- the first and the second authors in 1998, 2005 and 2008 ular evolution for each of the loci was selected, using within the framework of the Spanish project ‘‘Bio- BIC as implemented in the software jModeltest version complexity of Antarctic and Subantarctic ecosystems: 0.1.1 (Guindon & Gascuel 2003; Posada 2008; Darriba sensitivity to global change (ANT-COMPLEX)’’. The et al. 2012), evaluating only the 24 models available in collections are deposited at the Botanical Museum in MrBayes (Ronquist et al. 2012). The HKY+G+I was Copenhagen (C), unless stated otherwise. found to be optimal for the nrITS dataset for the com- bined analysis, SYM+G+I for the nrLSU dataset, and Morphology and anatomy K80+G for the mrSSU dataset. The ITS alignment was also analyzed separately using the evolutionary model Macroscopic descriptions are based on observations GTR+G+I. Bayesian tree inference was carried out made with a Wild Heerbrugg M5-53204 dissecting using Markov chain Monte Carlo (MCMC) as imple- microscope. Measurements were made using a mounted mented in MrBayes ver. 3.2 (Ronquist et al. 2012). In Nikon DS-Fi1 camera combined with the software NIS- the combined analysis, the three genes included were Elements. Sections were made by hand or using a treated as separate partitions. Parameters used in the Reichert-Jung Cryostat 2800 Frigocut E microtome. analyses followed those of Arup et al. (2013), except for Measurements were performed using an Olympus the branch length prior that was set to an exponential BX60 microscope. All measurements were made on with mean 1/10. Three parallel runs of Markov chain material mounted in water. Spores were measured out- Monte Carlo were performed, each with 8 chains, 7 of side the asci. which were incrementally heated with a temperature of 0Á10. Analyses were diagnosed every 100 000 genera- PCR amplification and alignment tions and automatically halted when convergence was reached. Convergence was defined as a standard de- Nine specimens of the genus Catenarina were used viation of splits (with frequency b0Á1) between runs to produce nine new nuclear rDNA sequences of the below 0Á01. Every 1000th tree was sampled and the first internal transcribed spacer regions (nrITS), and two 50% of runs were removed as burn-in. Majority-rule sequences of the small subunit of the mitochondrial consensus trees were constructed from the post burn-in ribosomal RNA gene (mrSSU). The PCR amplifica- tree samples, using FigTree 1.4 (http://tree.bio.ed.ac. tions were produced using direct PCR following Arup uk/software/figtree/). (2006). The primers used were ITS1F (Gardes & Bruns 1993), ITS4 (White et al. 1990), mrSSU1 (Zoller et al. 1999) and mrSSU7 (Zhou & Stanosz 2001). The PCR Secondary chemistry settings followed Ekman (2001) for the ITS and Arup et al. (2013) for mrSSU. The primers used for the PCR The secondary metabolite patterns were identified were also used in the sequencing reaction, which was using HPLC, and analyzed separately for the thallus and carried out by Macrogen Inc., Korea. apothecia. The relative composition of the secondary Two alignments were produced, one combined align- compounds was calculated based on absorbance at ment with 41 species including three loci, nrITS, nrLSU 270 nm according to Søchting (1997). and mrSSU, representing the three subfamilies in Telo- schistaceae, and one alignment with 48 ITS sequences of the subfamily Teloschistoideae (Arup et al. 2013). In Catenarin identification addition to the newly produced sequences, additional Catenarin was first isolated from cultures of Helmin- sequences were downloaded from GenBank. Voucher thosporium catenarium Drechsler, and subsequently also information and GenBank accession numbers are indi- isolated from various other microfungi including species cated in Table 1. For the combined analysis, Amandinea of Aspergillus and Penicillium (Raistrick et al. 1934). The punctata (Hoffm.) Coppins & Scheid. was used as an structure of this anthraquinone was ultimately estab- outgroup, with Physcia aipolia (Ehrh. ex Humb.) Fu¨rnr. lished by synthesis (Anslow & Raistrick 1941) (Fig. 1). as internal outgroup. For the ITS analysis, Caloplaca In the present work, the occurrence of catenarin in the cerina (Hedw.) Fr. of the subfamily Caloplacoideae was Catenarina species was established by HPLC com- used as outgroup and Leproplaca xantholyta (Nyl.) Nyl. parisons of the lichen extracts with authentic material. and Variospora velana (A. Massal.) Arup et al. as internal The HPLC was coupled to a photodiode array detector outgroup. Ambiguously aligned regions were removed for ultraviolet spectroscopic comparisons. By this means from all alignments before analyses. the ultraviolet spectra observed for natural catenarin eluting in the HPLC chromatogram was recorded, and Phylogenetic analysis computer-matched against the ultraviolet spectrum recorded for authentic catenarin under identical con- The alignments of the three different genes were first ditions. The correlation of ultraviolet spectra of the analyzed separately to check for incongruence between synthetic and natural material was >99Á9%. genes, but no incongruences were found. A conflict was Downloaded from https://www.cambridge.org/core. University of Athens, on 30 Sep 2021 at 06:36:47, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S002428291300087X https://www.cambridge.org/core/terms Downloaded from Table 1. Sequences used in either of the two analyses; those newly produced are in bold and the others were downloaded from Genbank. 2014 https://www.cambridge.org/core Species Country, collector,
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