Hypogymnia Phylogeny, Including Cavernularia, Reveals Biogeographic Structure Author(S): Jolanta Miadlikowska, Conrad L

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Hypogymnia Phylogeny, Including Cavernularia, Reveals Biogeographic Structure Author(S): Jolanta Miadlikowska, Conrad L Hypogymnia phylogeny, including Cavernularia, reveals biogeographic structure Author(s): Jolanta Miadlikowska, Conrad L. Schoch, Stacie A. Kageyama, Katalin Molnar, François Lutzoni, and Bruce McCune Source: The Bryologist, 114(2):392-400. 2011. Published By: The American Bryological and Lichenological Society, Inc. DOI: 10.1639/0007-2745-114.2.392 URL: http://www.bioone.org/doi/full/10.1639/0007-2745-114.2.392 BioOne (www.bioone.org) is an electronic aggregator of bioscience research content, and the online home to over 160 journals and books published by not-for-profit societies, associations, museums, institutions, and presses. Your use of this PDF, the BioOne Web site, and all posted and associated content indicates your acceptance of BioOne’s Terms of Use, available at www.bioone.org/page/ terms_of_use. Usage of BioOne content is strictly limited to personal, educational, and non-commercial use. Commercial inquiries or rights and permissions requests should be directed to the individual publisher as copyright holder. BioOne sees sustainable scholarly publishing as an inherently collaborative enterprise connecting authors, nonprofit publishers, academic institutions, research libraries, and research funders in the common goal of maximizing access to critical research. Hypogymnia phylogeny, including Cavernularia, reveals biogeographic structure Jolanta Miadlikowska1,6, Conrad L. Schoch2,3, Stacie A. Kageyama2,4, Katalin Molnar1,5, Franc¸ois Lutzoni1, and Bruce McCune2 1 Department of Biology, Duke University, Durham, NC 27708-0338, U.S.A.; 2 Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR 97331-2902, U.S.A.; 3 Current address: National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, 45 Center Drive, MSC 6510, Bethesda, MD 20892-6510, U.S.A.; 4 Current address: USDA Agricultural Research Service, Fort Keogh Livestock and Range Research Laboratory, Miles City, MT 59301-4016, U.S.A.; 5 Current address: Institute of Ecology and Botany, Hungarian Academy of Sciences, 2163 Va´cra´to´t, Alkotma´ny u. 2-4, Hungary ABSTRACT. We inferred phylogenetic relationships using Bayesian and maximum likelihood approaches for two genera of lichenized fungi, Hypogymnia and Cavernularia (Parmeliaceae). Based on the combined ITS and GPD1 dataset from 23 species (49 specimens) of Hypogymnia and two species (8 specimens) of Cavernularia, we conclude that Hypogymnia is paraphyletic, and that it should include Cavernularia to retain its monophyly. Hypogymnia hultenii (5 Cavernularia hultenii) and H. lophyrea (5 C. lophyrea) are accepted here. Five species of Hypogymnia represented by more than a single individual were found to be monophyletic and significantly supported. The phylogeny reflects a statistically significant biogeographic pattern where continental-scale endemic taxa tend to occur within the same phylogenetic group. Sorediate taxa, which have worldwide or broader geographical ranges than affiliated species lacking soredia, are spread across the phylogenetic tree. Hypogymnia contains three species pairs: H. krogiae and the sorediate counterpart H. incurvoides, H. minilobata and the sorediate H. mollis, and H. lophyrea and the sorediate H. hultenii. In the case of H. minilobata, both members of the pair are restricted to a small area in southern California. In the other two cases, the fertile counterpart occurs only in North America, while the sorediate species occurs in both North America and Fennoscandia. This suggests but not proves an origin of each species pair in North America, with migration of the sorediate member to Fennoscandia following the prevailing wind direction. KEYWORDS. Biogeography, Cavernularia, DNA sequences, GPD1, Hypogymnia, ITS, lichenized ascomycetes, Parmeliaceae, species pair. 6 Corresponding author’s e-mail: [email protected] DOI: 10.1639/0007-2745-114.2.392 The Bryologist 114(2), pp. 392–400 0007-2745/11/$1.05/0 Copyright E2011 by The American Bryological and Lichenological Society, Inc. Miadlikowska et al.: Phylogeny of Hypogymnia 393 ¤¤¤ The Parmeliaceae is a large, conspicuous family of Hypogymnia in apothecial anatomy, lobe lichenized fungi, and one of the best studied based on morphology, and chemistry. Both Ra¨sa¨nen (1943) both morphological and molecular data (Blanco et al. and Krog (1951) merged Cavernularia into 2006; Crespo et al. 2007). This family, as defined by Hypogymnia, but this concept was not adopted by Crespo et al. (2007), appears to be monophyletic. The later authors, including later works by Krog. cupulate exciple is currently the only phenotypic The species pair concept (Du Rietz 1924; synapomorphy that unites the Parmeliaceae. In this Mattson & Lumbsch 1989; Poelt 1970) has not been apothecial anatomy, the proper exciple isolates the applied within Hypogymnia. McCune and Schoch hypothecium from the medulla. The ‘‘parmelioid’’ (2009) suggested that the sorediate H. mollis was group within the family appears also to be sister to the otherwise similar but esorediate monophyletic, but Hypogymnia (Nyl.) Nyl. and H. minilobata, both endemic to southern California. Cavernularia Degel. are placed outside that group In Cavernularia, however, C. hultenii and C. lophyrea (Crespo et al. 2007). are an obvious species pair. In this study we examine Hypogymnia occurs in temperate to subpolar the phylogenetic and geographic relationships of environments with the greatest diversity in oceanic to other potential species pair within Hypogymnia (e.g., suboceanic climates. It is found on all continents H. incurvoides and H. krogiae). except Antarctica. In tropical to subtropical latitudes The phylogeny of Hypogymnia has not been the genus occurs only at high elevations. studied. In this paper we focus on selected species of Approximately 100 species are recognized at present Hypogymnia and Cavernularia, based on (McCune unpubl.). All Hypogymnia species lack reconstructed phylogeny using the combined entire rhizines and have thickened lobes (either solid or nuclear ribosomal internal transcribed spacer region hollow), bifusiform spermatia, substipitate apothecia, (ITS: ITS1, 5.8S, and ITS2) and partial and asci with eight simple, hyaline, ellipsoid to glyceraldehyde-3-phosphate dehydrogenase gene subspherical spores. Most have hollow lobes, a black (GPD1). We address the following questions: 1) are lower cortex, contain atranorin, physodic acid and species represented in our data by at least two related compounds, and have small spores (,9 mm individuals (12 out of 25) monophyletic? 2) what are long). A few have solid lobes or a brown to dark the phylogenetic relationships among species, brown lower cortex or larger spores or contain usnic including potential species pairs? 3) does acid instead of atranorin. Cavernularia represent a separate genus or should it The related genus Cavernularia occurs in be included in Hypogymnia as suggested by Ra¨sa¨nen oceanic temperate regions of North America and (1943) and Krog (1951)? 4) can we detect a Fennoscandia (Ahti & Henssen 1965; Degelius 1952; biogeographic pattern based on their inferred Printzen & Ekman 2002). Cavernularia is a genus of phylogenetic history? only two species, appearing like a tiny Hypogymnia but with a dense array of small pits in the lower MATERIAL AND METHODS surface. In contrast to the holes in the lower surface, Sampling and alignments. Fungal genomic lobe tips, and axils of some species of Hypogymnia, DNA was isolated from approximately 5 mm long the pits in Cavernularia do not open into a lobe sections of lichen thalli using the FastDNAH kit and cavity, but instead open into a small, broadened The FastPrep instrument from MPI Biochemicals chamber that is completely lined by the lower cortex. (Irvine, CA). DNA amplifications were completed These minute cavities (ca. 0.1 mm diameter) were using PCR Master Mix from Promega Corporation termed ‘‘cavernulae’’ by Degelius (1937), and (Madison, WI) under the following PCR conditions: differentiate Cavernularia from all other parmelioid 94u C for 2 min; five cycles at 94u C for 40 s, 55u C for lichens. Otherwise Cavernularia resembles 45 s lowering by 0.8u C per cycle and 72u C for 90 s; 394 The Bryologist 114(2): 2011 30 cycles at 94u C for 30 s, 52u C for 45 s and 65u C supported at $ 70% as monophyletic in one for 120 s and a final cycle for 10 min at 72u C. The majority rule bootstrap tree, but supported as ITS region was amplified with primers ITS4 and ITS5 non-monophyletic in another. No conflict was (White et al. 1990). Primers Gpd1LM and Gpd2LM detected, therefore, the two single locus datasets were were used to obtain a partial sequence (1 kb) of the concatenated in an ITS+GPD1 59-OTU combined GPD1 gene (Myllys et al. 2002; Thell et al. 2004). One dataset (TreeBASE; http://purl.org/phylo/treebase/ ITS PCR product (for H. macrospora 2) had to be phylows/study/TB2:S11110). A maximum likelihood cloned (due to the presence of multiple peaks in the search using RAxML, 1000 replicates, GTR model chromatograms) using the Topo-TA 5-minute PCR with gamma distribution and four discrete rate cloning kit (Invitrogen, Carlsbad, CA, USA) categories (GTRGAMMA) was performed on the following the manufacturer’s protocol. We sampled 59-OTU combined dataset. Phylogenetic confidence 23 species of Hypogymnia represented by 49 was estimated for the combined dataset with ML individuals and two species of Cavernularia bootstrapping (RAxML), using the same settings as represented by eight individuals. Brodoa for the single-locus
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