John Carroll University Carroll Collected 2017 Faculty Bibliography Faculty Bibliographies Community Homepage 2017 Cyanomargarita gen. nov. (Nostocales, Cyanobacteria): convergent evolution resulting in Jeffrey R. Johansen John Carroll University, [email protected] Sergei Shalygin Regina Shalygina Nicole Pietrasiak Follow this and additional works at: https://collected.jcu.edu/fac_bib_2017 Part of the Biology Commons, and the Ecology and Evolutionary Biology Commons Recommended Citation Johansen, Jeffrey R.; Shalygin, Sergei; Shalygina, Regina; and Pietrasiak, Nicole, "Cyanomargarita gen. nov. (Nostocales, Cyanobacteria): convergent evolution resulting in" (2017). 2017 Faculty Bibliography. 39. https://collected.jcu.edu/fac_bib_2017/39 This Article is brought to you for free and open access by the Faculty Bibliographies Community Homepage at Carroll Collected. It has been accepted for inclusion in 2017 Faculty Bibliography by an authorized administrator of Carroll Collected. For more information, please contact [email protected]. CYANOMARGARITA GEN. NOV. (NOSTOCALES, CYANOBACTERIA): CONVERGENT EVOLUTION RESULTING IN A CRYPTIC GENUS1 Sergei Shalygin 2 Department of Biology, John Carroll University, University Heights, Ohio 44118, USA Polar-Alpine Botanical Garden-Institute, Kola Science Center, Russian Academy of Science, Kirovsk-6 184230, Russia Regina Shalygina Institute of Industrial Ecology Problems of the North, Kola Science Center, Russian Academy of Science, Akademgorodok 14a, Apatity 184209, Russia Jeffrey R. Johansen Department of Biology, John Carroll University, University Heights, Ohio 44118, USA Department of Botany, Faculty of Science, University of South Bohemia, Branisovska 31, Ceske Budejovice 370 05, Czech Republic Nicole Pietrasiak Department of Plant and Environmental Sciences, New Mexico State University, 945 College Drive, Las Cruces, New Mexico 88003, USA Esther Berrendero Gomez Department of Botany, Faculty of Science, University of South Bohemia, Branisovska 31, Ceske Budejovice 370 05, Czech Republic Marketa Bohunicka Institute of Botany of the Academy of Sciences of the Czech Republic, Dukelska 135, Trebon 379 82, Czech Republic Research and Breeding Institute of Pomology, Holovousy 129, Horice 508 01, Czech Republic Jan Mares Department of Botany, Faculty of Science, University of South Bohemia, Branisovska 31, Ceske Budejovice 370 05, Czech Republic Institute of Botany of the Academy of Sciences of the Czech Republic, Dukelska 135, Trebon 379 82, Czech Republic Biology Centre of the Academy of Sciences of the Czech Republic, Institute of Hydrobiology, Na Sadkach 702/7, Ceske Budejovice 37005, Czech Republic and Christopher A. Sheil Department of Biology, John Carroll University, University Heights, Ohio 44118, USA Two populations of Rivularia-like cyanobacteria 16S rRNA and rbcLX phylogenies. We here name the were isolated from ecologically distinct and new cryptic genus Cyanomargarita gen. nov., with type biogeographically distant sites. One population was species C. melechinii sp. nov., and additional species from an unpolluted stream in the Kola Peninsula of C. calcarea sp. nov. We also name a new family for Russia, whereas the other was from a wet wall in the these taxa, the Cyanomargaritaceae. Grand Staircase-Escalante National Monument, a Key index words: 16S rRNA gene phylogeny; 16S-23S desert park-land in Utah. Though both were virtually ITS; cryptic genus; Cyanobacteria; Cyanomargarita; indistinguishable from Rivularia in field and cultured rbcLX phylogeny; Rivularia material, they were both phylogenetically distant from Rivularia and the Rivulariaceae based on both Abbreviations: BA, Bayesian Analysis; DIC, differen- tial interference contrast; ITS, internal transcribed spacer; ML, maximum likelihood; MP, maximum parsimony; RAS, russian academy of sciences 762 763 With the advent of molecular methods, many phy- hemispherical to spherical mucilage investment in a cologists, including those who study cyanobacteria, small, spring-fed, unpolluted stream near the town of began to recognize the existence of cryptic species Apatity in the Kola Peninsula, Russia. It was com- (Boyer et al. 2002, Casamatta et al. 2003, Erwin and pletely consistent with the description of Rivularia Thacker 2008, Joyner et al. 2008, Premanandh et al. C.Agardh ex Bornet & Flahault, the type genus of 2009, Ren~e et al. 2013, Johansen et al. 2014, Rivulariaceae, which contains tapering, heterocytous Muhlsteinov€ a et al. 2014a,b, Patzelt et al. 2014). taxa. This taxon fit no established species in Rivularia, However, while in these papers the existence of and upon sequencing was determined to be phyloge- cryptic species was suggested, the species were not netically distant from all members of that family. A recognized taxonomically. Subsequently, cryptic spe- second species belonging to the same clade as the cies have been named in several algal groups, Russian material was found, and sequenced several including euglenids (Marin et al. 2003, Kosmala years earlier from a wet wall in the Grand Staircase- et al. 2007, Kosmala et al. 2009, Karnkowska-Ishi- Escalante National Monument in Utah, USA. These kawa et al. 2010, Kim et al. 2010, 2013, Linton et al. two populations differ morphologically and ecologi- 2010), eustigmatophytes (Fawley and Fawley 2007, cally, and are described herein as two new species Fawley et al. 2015), chlorophytes (Fawley et al. 2005, based on a modified phylogenetic species concept 2011, Fucıkova et al. 2014), and cyanobacteria (Oso- (Mishler and Theriot 2000, Johansen and Casamatta rio-Santos et al. 2014, Pietrasiak et al. 2014). 2005) in a newly proposed genus, Cyanomargarita gen. Some cyanobacterial systematists have suggested nov. This genus cannot be placed in any family-level the existence of cryptic genera as well (Komarek grouping of taxa based on the phylogenetic analyses et al. 2014, Dvorak et al. 2015a), although very few performed – we place these taxa in a new family to cryptic genera have actually been described. Pinoc- science, the Cyanomargaritaceae fam. nov. chia, which is morphologically identical to Pseudan- abaena, has a phylogenetic position distant from that genus, and therefore was described as a cryptic MATERIALS AND METHODS genus (Dvorak et al. 2015b). Kovacikia, which is Isolation and strain characterization. Both strains of morphologically similar to Phormidesmis but molecu- Cyanomargarita were isolated from natural populations into larly distinct, would also fit the definition of a cryp- unialgal cultures using standard microbiological methods, tic genus, although the authors did not label it as including enrichment plates and direct isolation from the orig- such (Miscoe et al. 2016). There are also a number inal samples, in Z8 medium (Kotai 1972, Carmichael 1986). Cultures were observed under a Zeiss Axioskop photomicro- of pseudocryptic genera (genera defined by mor- scope with both bright field and DIC optics. All morphological phological traits that are minor or phenotypically measurements were obtained using AxioVision 4.8 software plastic, and therefore not always expressed in the provided by Zeiss (Oberkochen, Germany). Living cultures population) as well, such as Nodosilinea, Oculatella, were deposited into the Cyanobacterial Culture Collection at Limnolyngbya, Pantanalinema, and Alkalinema. These John Carroll University, Cleveland, OH, USA. Natural popula- genera belong to the Synechococcales, an order tions of material from which the strain C. melechinii APA-RS9 containing taxa with few morphological characteris- was derived were dried and deposited as an isotype in the Her- barium of the Polar-Alpine Botanical Garden-Institute, Kola tics (simple filamentous forms, variations in tri- Science Centre, RAS, Kirovsk-6, Murmansk Region, Russia, and chome width and sheath characteristics). information about habitat, coordinates and locality can be Outside of the Synechococcales, few cryptic gen- found in the online database Cyanopro (Melechin et al. 2013). era have been recognized. In the Chroococcales, The dried holotype material of this strain was deposited in the Chalicogloea is similar to Gloeocapsa and could be con- Herbarium for Nonvascular Cryptogams in the Monte L. Bean sidered a cryptic genus (Roldan et al. 2013). There Museum, Provo, UT, USA. Liquid materials of both species fixed in 4% formaldehyde, as well as dried materials of C. cal- are likely more cryptic genera in this order, but carea, were also deposited in the Herbarium for Nonvascular identifying them is more problematic because mem- Cryptogams in the Monte L. Bean Museum, Provo, UT, USA. bers of the genus are difficult to grow in culture, Molecular methods. Genomic DNA was extracted following and consequently, fewer sequences are available. In techniques described in Pietrasiak et al. (2014). PCR amplifi- the Oscillatoriales, Ammassolinea is the one of the cation of the 16S rRNA gene was accomplished following described cryptic genera (Hasler et al. 2014), being Osorio-Santos et al. (2014), with the exception that forward morphologically inseparable from Phormidium,asit primer 8F was used instead of forward primer VRF2, for amplification of a longer sequence, starting near the begin- is presently defined, as well as Moorea, Okeania and ning of the 16S rRNA gene (Perkerson et al. 2011). The 16S Microseira (Engene et al. 2012, 2013, McGregor and rRNA amplicons were cloned to avoid problems in sequenc- Sendall 2015). Within the Nostocales, there is much ing of multiple operons (Siegesmund et al. 2008).