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Biology and Diversity of Desmids

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Palacký University, Faculty of Science Department of Botany Biology and diversity of desmids Petra Mazalová Ph.D. thesis Olomouc, 2013 Supervised by: Prof. RNDr. Aloisie Poulíčková, CSc., Department of Botany, Palacký University I hereby declare that this thesis is my independent work, using the listed references, or in co- operation with co-authors of the papers. I did not submit any of the theses or their any part to acquire any other academic title. April 2013, Olomouc ................................. Mgr. Petra Mazalová Motto: „Only nature knows what it wants. It never makes jokes and never makes mistakes. Those make only men.” „In nature we never see anything isolated, but everything in connection with something else which is before it, beside it, under it and over it.” (Johann Wolfgang von Goethe) (Staurodesmus convergens; foto: P. Mazalová) Acknowledgements I would like to thank my supervisor, Aloisie Poulíčková, for her systematic guidance, collaboration, support and inspiration. I also express my appreciation to colleagues from our algological laboratory for their contribution to a good working atmosphere and for helpful advice whenever I needed it. Finally I kindly thank my family for their unwavering support throughout my life. Abstract Higher plants evolved from algae, and still share many common characteristics. However, studies on biology and diversity of algae lag behind those of higher plants. In this thesis, I have investigated the phylogenetic, morphological and geographical diversity of desmids and explored the DNA content variations for better understanding of the importance of polyploidy in their evolution. In addition, I propose there, a multi-step protocol that leads to the quantification of DNA content in desmids and some other microalgae using flow cytometry. In the course of the study of nine mires in Central and Northern Moravia, altogether 109 desmid taxa were found. Five of those taxa are considered to be new records for the Czech Republic, 42 of them are new records for Moravia. The findings were discussed with previous records from Moravia and Czech Republic. Moreover, the character and origin of the unique locality Slavkov mire were discussed. The results indicate a lack of knowledge of desmid diversity in the Moravia region. A pair of well-known desmid species, Micrasterias rotata and M. fimbriata, were tested for their phylogenetic and morphological homogeneity as well as their geographic distribution. The investigation was based on morphological and geometric morphometric studies, with genetic analyses of the trnGucc sequences of strains, isolated mainly from Europe. Micrasterias rotata proved to be phylogenetically homogenous across Europe, while M. fimbriata turned out to be composed of two firmly delimited lineages. The lineages differ genetically as well as in their morphology. There are also differences in their geographic distribution patterns and likely in their ecological preferences. It has been suggested to describe one lineage as a new species of the genus Micrasterias. Nuclear DNA content was measured in microalgae with a focus on desmids, using a new protocol based on flow cytometry measurements. To obtain the protoplasts, a mixture of enzymes (cellulase and macerozyme) was used. The nuclei suspension was prepared using LB01 nuclear isolation buffer, and nuclei were stained by propidium iodide. The leaves of four plant species were used as standards, and one new microalgal standard was suggested. The protocol proved to be useful for desmids and also worked for some Chlorophyta and Heterokontophyta. Optimization of enzyme mixture or nuclear isolation buffer for measuring other microalgae, as in those we did not succeed in measuring, deserves more attention. The nuclear DNA content of 34 Micrasterias strains (25 Micrasterias species) was measured. Intraspecific and interspecific variation was observed and estimated DNA contents were mapped along the phylogenetic tree, based on concatenated SSU rDNA, psaA, and coxIII alignment. The related species were mostly similar in their DNA content. Strong correlation between the absolute nuclear DNA content and chromosome numbers in species, for which the data were available, was found. Significant positive correlation between the DNA content and cell size of the investigated Micrasterias rotata strains was also found. In conclusion the evolutionary significance of genome size variation was confirmed in the genus Micrasterias and the importance of natural polyploidization in the evolution and speciation of it was verified. Abstrakt Vznik a vývoj vyšších rostlin úzce souvisí s řasami, vyšší rostliny s řasami doposud sdílejí mnoho společných znaků. Nicméně studie biologie a diverzity řas pokulhávají za rozsáhlostí studií vyšších rostlin. V této disertační práci byly zkoumány fylogenetická, morfologická a geografická diverzita krásivek a rozdíly v množství jejich jaderné DNA, za účelem lepšího pochopení významu polyploidie v jejich evoluci. Navíc zde navrhuji protokol pro měření množství jaderné DNA u krásivek a několika dalších mikrořas, a to pomocí průtokové cytometrie. Během studia devíti mokřadních lokalit střední a severní Moravy bylo nalezeno celkem 109 taxonů krásivek. Pět z těchto taxonů je nových pro území ČR a 42 pro území Moravy. Byly diskutovány předchozí nálezy všech taxonů na území Moravy či celé České republiky. Zároveň byl zvážen charakter a původ unikátní lokality nacházející se poblíž vesnice Slavkov. Získaná data naznačila nízkou prozkoumanost krásivek na území Moravy. Dva dobře známé druhy krásivek, Micrasterias rotata a M. fimbriata, byly zkoumány z pohledu jejich fylogenetické a morfologické homogenity a geografického rozšíření. Výzkum byl proveden pomocí morfologických a morfometrických analýz, spolu s fylogenetickou analýzou trnGucc kmenů z různých lokalit Evropy. Micrasterias rotata se ukázala být fylogeneticky homogenní v rámci Evropy, zatímco M. fimbriata se rozdělila do dvou dobře podpořených linií. Populace těchto linií se lišily kromě fylogenetické analýzy také morfologickými a morfometrickými daty. Byly zjištěny rozdíly v jejich geografickém rozšíření a pravděpodobně rozdílné ekologické nároky. Na základě výsledků bylo navrženo popsat jednu z těchto linií jako nový druh rodu Micrasterias. Pomocí průtokové cytometrie bylo měřeno množství jaderné DNA u krásivek a několika dalších mikrořas. Byl použit nový protokol vzniklý na základě poznatků z měření u vyšších rostlin. Pro získání protoplastů byla použita směs enzymů celulázy a mecerozymu. Suspenze jader byla získána použitím LB01 lyzačního pufru a následně byla jádra obarvena propidium jodidem. Jako standard byly použity listy čtyř různých rostlin a byl navržen nový standard z řad mikrořas. Protokol se ukázal vhodný pro použití u krásivek a fungoval i u několika zástupců z řad Chlorophyta a Heterokontophyta. Optimalizace enzymatické směsi či lyzačního pufru pro jiné skupiny mikrořas, a pro ty, u kterých nebyla metoda úspěšná, si zaslouží další pozornost. U 34 kultur (25 druhů) rodu Micrasterias bylo změřeno množství jaderné DNA. Byla sledována mezidruhová a vnitrodruhová variabilita tohoto množství a zjištěné hodnoty DNA byly namodelovány na fylogenetický strom založený na propojení analýz SSU rDNA, psaA a coxIII. Příbuzné druhy vykázaly podobná množství DNA. U druhů, pro které byla známa data o počtu chromozomů, byla zjištěna silná pozitivní korelace mezi množstvím DNA a počtem chromozomů. Pozitivní korelace se projevila i mezi množstvím DNA a rozměry buněk u sledovaného druhu Micrasterias rotata. Experimenty poskytly důkaz významu změny velikosti genomu v evoluci rodu Micrasterias a potvrdily důležitost přirozené polyploidizace v jeho evoluci a speciaci. Contents 1 Introduction .................................................................................................................... 10 1.1 Systematic delimitation of desmids............................................................................. 10 1.1.1 Position of desmids within Plantae ......................................................................... 10 1.1.2 Desmid classification .............................................................................................. 11 1.2 Basic characteristics of desmids: an outline .............................................................. 12 1.2.1 Main morphological features of desmids................................................................ 12 1.2.2 Reproduction of desmids ......................................................................................... 13 1.2.3 Ecological preferences of desmids.......................................................................... 14 1.2.4 Genus Micrasterias ................................................................................................. 14 1.3 Polyphyletism and cryptic diversity within desmids................................................. 15 2 Aims of the thesis............................................................................................................ 17 3 Conclusions and future prospects................................................................................. 18 3.1 Desmid flora of mires in Central and Northern Moravia ........................................ 18 3.2 Delimitation of Micrasterias fimbriata and M. rotata ................................................ 19 3.3 Quantification of DNA content in microalgae using flow cytometry ...................... 20 3.4 DNA content in evolution of the genus Micrasterias ................................................
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  • Carbohydrate Release by a Subtropical Strain of Spondylosium Pygmaeum (Zygnematophyceae): Influence of Nitrate Availability and Culture Aging1

    Carbohydrate Release by a Subtropical Strain of Spondylosium Pygmaeum (Zygnematophyceae): Influence of Nitrate Availability and Culture Aging1

    J. Phycol. 46, 477–483 (2010) Ó 2010 Phycological Society of America DOI: 10.1111/j.1529-8817.2010.00823.x CARBOHYDRATE RELEASE BY A SUBTROPICAL STRAIN OF SPONDYLOSIUM PYGMAEUM (ZYGNEMATOPHYCEAE): INFLUENCE OF NITRATE AVAILABILITY AND CULTURE AGING1 Fernanda Reinhardt Piedras Po´s-graduac¸a˜o em Oceanografia Biolo´gica, Instituto de Oceanografia, Universidade Federal de Rio Grande- FURG, Av. Italia, Km 8, Rio Grande, RS 96201-900, Brasil Paulo Roberto Martins Baisch, Maria Isabel Correˆa da Silva Machado Laborato´rio de Oceanografia Geolo´gica Instituto de Oceanografia, Universidade Federal de Rio Grande- FURG, Av. Italia, Km 8, Rio Grande, RS 96201-900, Brasil Armando Augusto Henriques Vieira Departamento de Botanica, Unversidade Federal de Sao Carlos, Via Washington Luis, Km 235, Sao Carlos, SP 13565-905, Brasil and Danilo Giroldo2 Laborato´rio de Botaˆnica Criptogaˆmica, Instituto de Cieˆncias Biolo´gicas, Universidade Federal de Rio Grande – FURG, Av. Italia, Km 8, Rio Grande, RS 96201-900, Brasil This paper describes the influence of nitrate avail- availability. EPS molecules >12 kDa were composed ability on growth and release of dissolved free and mainly of xylose, fucose, and galactose, as for other combined carbohydrates (DFCHOs and DCCHOs) desmids. However, a high N-acetyl-glucosamine con- produced by Spondylosium pygmaeum (Cooke) W. tent was found, uniquely among desmid EPSs. West (Zygnematophyceae). This strain was isolated Key index words: carbohydrate; desmid; growth; from a subtropical shallow pond, located at the nitrate; Spondylosium extreme south of Brazil (Rio Grande, RS). Experi- ments were carried out in batch culture, comparing Abbreviations: Ara, arabinose; DCCHO, dissolved two initial nitrate levels (10 ⁄ 100 lM) in the medium.
  • New Desmid Records from High Mountain Lakes in Artabel Lakes Nature Park, Gümüşhane, Turkey

    New Desmid Records from High Mountain Lakes in Artabel Lakes Nature Park, Gümüşhane, Turkey

    Turkish Journal of Botany Turk J Bot (2019) 43: 570-583 http://journals.tubitak.gov.tr/botany/ © TÜBİTAK Research Article doi:10.3906/bot-1810-71 New desmid records from high mountain lakes in Artabel Lakes Nature Park, Gümüşhane, Turkey 1, 2 Bülent ŞAHİN *, Bülent AKAR 1 Department of Biology Education, Fatih Education Faculty, Trabzon University, Trabzon, Turkey 2 Department of Food Engineering, Faculty of Engineering and Natural Sciences, Gümüşhane University, Gümüşhane, Turkey Received: 30.10.2018 Accepted/Published Online: 15.04.2019 Final Version: 08.07.2019 Abstract: The algal flora of 17 lakes and 1 pond in the Artabel Lakes Nature Park were investigated during two summer seasons (2013 and 2016). In total, 26 desmid taxa were found and identified as new records for the desmid flora of Turkey based on their morphotaxonomic characteristics and ecological preferences. The taxa identified belong to the genera Actinotaenium (1), Closterium (1), Cosmarium (15), Micrasterias (1), Spondylosium (1), Staurastrum (5), Teilingia (1), and Tetmemorus (1). Morphotaxonomy, ecology, and distribution of each species were discussed in detail. Key words: Desmids, new records, high mountain lakes, Artabel Lakes Nature Park, Turkey 1. Introduction Desmids are an integral part of benthic habitats of Desmid habitats are exclusively freshwater (Coesel and high mountain lakes; in particular, those of the Northern Meesters, 2007; Kouwets, 2008). Desmids usually prefer Hemisphere (Medvedeva, 2001; Sterlyagova, 2008). In acidic or pH-circumneutral, nutrient-poor, and clear the period from 1998 to 2014, 43 new records of desmid waters (Lenzenweger, 1996; Coesel and Meesters, 2007). species from high mountain lakes in the eastern Black It is well known that members of order Desmidiales Sea Region were identified and published (Şahin, 1998, exhibit great diversity in their external morphology and 2000, 2002, 2007, 2008, 2009; Şahin and Akar, 2007; Akar also have remarkably complex cell symmetry (Lee, 2015).
  • Some Freshwater Green Algae of Raja-Rani Wetland, Letang, Morang: New for Nepal

    Some Freshwater Green Algae of Raja-Rani Wetland, Letang, Morang: New for Nepal

    2020J. Pl. Res. Vol. 18, No. 1, pp 6-26, 2020 Journal of Plant Resources Vol.18, No. 1 Some Freshwater Green Algae of Raja-Rani Wetland, Letang, Morang: New for Nepal Shiva Kumar Rai1*, Kalpana Godar1and Sajita Dhakal2 1Phycology Research Lab, Department of Botany, Post Graduate Campus, Tribhuvan University, Biratnagar, Nepal 2National Herbarium and Plant Laboratories, Department of Plant Resources, Godawari, Lalitpur, Nepal *E-mail: [email protected] Abstract Freshwater green alga of Raja-Rani wetland has been studied. A total 36 algal samples were collected from 12 sites by squeezing the submerged aquatic plants. Present paper describes 35 green algae under 18 genera from Raja-Rani wetland as new record for Nepal.Genus Euastrum consists 5 species; genera Cosmarium, Staurodesmus, and Staurastrum consist 4 species each; genera Scenedesmus, Closterium, Pleurotaeniuum and Xanthidium consist 2 species each; and rest genera consist only single taxa each. Water parameters of the wetland of winter, summer and rainy seasons were also recorded. Keywords: Chlorophyceae, Cosmarium, New report, Staurodesmus, Triploceras,Xanthidium Introduction climatic condition and rich aquatic habitats for algae, extensive exploration is lacking in the history.Suxena Algae are the simplest photosynthetic thalloid plants, & Venkateswarlu (1968), Hickel (1973), Joshi usually inhabited in water and moist environment (1979), Subba Raju & Suxena (1979), Shrestha & throughout the world. Green algae are the largest Manandhar (1983), Hirano (1984), Ishida (1986), and most diverse group of algae, with about 8000 Watanabe & Komarek (1988), Haga & Legahri species known (Guiry, 2012). They have wide range (1993), Watanabe (1995), Baral (1996, 1999), Das of habitats as they grow in freshwater, marine, & Verma (1996), Prasad (1996), Komarek & subaerial, terrestrial, epiphytic, endophytic, parasitic, Watanabe (1998), Simkhada et al.