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Phototrophic Microorganisms in Extreme Environments

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Phototrophic Microorganisms in Extreme Environments University of South Bohemia in České Budějovice Faculty of Science PHOTOTROPHIC MICROORGANISMS IN EXTREME ENVIRONMENTS Ph.D. Thesis Hana Medová supervisor: Mgr. Michal Koblížek, Ph.D. Institute of Microbiology CAS, Třeboň scientific advisor: Doc. Libor Pechar, CSc. University of South Bohemia. České Budějovice Třeboň, Czech Republic 2013 This thesis should be cited as: Medová H, 2013: Phototrophic microorganisms in extreme environments. Ph.D. Thesis Series, No. 1. University of South Bohemia, Faculty of Science, České Budějovice, Czech Republic, 123 pp. Annotation This thesis focuses on photosynthetic prokaryotes living in extreme saline, thermal, or acidic conditions. Their distribution and physiology was assesed by a range of physical and chemical techniques such as kinetic fluorometry, epifluorescence and light microscopy, emission spectroscopy, or high performance liquid chromatography. Chapter 1 provides a brief overview of phototrophic organisms inhabiting various extreme environmens and serves as a general introduction. Chapter 2 provides a field study of phototrophic communities inhabiting saline and soda lakes in the Great Steppe region. A small survey of bacterial phototrophic community in freshwater polar lakes on Svalbard (Spitsbergen) and James Ross Island, Antarctica is presented in Chapter 3. Further, a laboratory based study focusing on thermal stability of the anoxygenic reaction centers is presented in Chapter 4. The last Chapter 5 describes experiments studying the response of plaktonic cyanobacteria to acidic post-mining waters. Declaration (in Czech) Prohlašuji, že svoji disertační práci jsem vypracoval/a samostatně pouze s použitím pramenů a literatury uvedených v seznamu citované literatury. Prohlašuji, že v souladu s § 47b zákona č. 111/1998 Sb. v platném znění souhlasím se zveřejněním své disertační práce, a to elektronickou cestou ve veřejně přístupné části databáze STAG provozované Jihočeskou univerzitou v Českých Budějovicích na jejích internetových stránkách, a to se zachováním mého autorského práva k odevzdanému textu této kvalifikační práce. Souhlasím dále s tím, aby toutéž elektronickou cestou byly v souladu s uvedeným ustanovením zákona č. 111/1998 Sb. zveřejněny posudky školitele a oponentů práce i záznam o průběhu a výsledku obhajoby kvalifikační práce. Rovněž souhlasím s porovnáním textu mé kvalifikační práce s databází kvalifikačních prací Theses.cz provozovanou Národním registrem vysokoškolských kvalifikačních prací a systémem na odhalování plagiátů. V Třeboni dne 21.3. 2013 ......................................... Hana Medová This thesis originated from a partnership of the Faculty of Science, the University of South Bohemia, and the Institute of Microbiology CAS, Třeboň, supporting doctoral studies in the biophysics study programme. Financial support Grant Agency of CR No. P501/10/0221 Bilateral Czech-Russian Academy of Science exchange program No. 22 Research concepts of the Institute of Physical Biology, South Bohemia University No. MSM6007665808 Research concept of the Institute of Microbiology ASCR No. AV0Z50200510 Project ‘Czech Polar’ of the Ministry of Education, Sports and Youth of CR No. LM2010009 Grant Agency of ASCR No. IAA608170901 Project ‘Algatech’ of the Research, Development and Innovation program of the Ministry of Education, Sports and Youth of CR No. CZ.1.05/2.1.00/03.0110 Research program of the Ministry of Education, Youth and Sport of CR No. 2B08006 Research, Development and Innovation program of the Ministry of Agriculture of CR No. Qh82078 Acknowledgment (in Czech) Ráda bych vyjádřila své poděkování všem lidem, kteří mě podporovali a provázeli po celou dobu mého studia. Mé nekonečné díky patří školiteli Michalu Koblížkovi, jenž mi umožnil studovat a pobývat v Třeboni. Vážím si jeho trpělivosti, přísnosti i vlídnosti. Cením si jeho rad, postřehů a doporučení, která se vždy ukázala jako přínosná a pravdivá. Skládám hold Ondřeji Prášilovi za vtisknutí přátelské, otevřené a podnětné atmosféry celému areálu Opatovického mlýna. Děkuji svým kolegům (Zuzce Bláhové, Jasonu Deanovi, Evě Kolářů) za veškeré jejich laskavé rady i pomoc, byli mi podporou, hromosvodem i vrbou. Ráda bych vyjádřila svůj vděk kolektivu laboratoře ENKI (Janě Šulcové, Lence Kröpfelové, Evě Chmelové, Zdeňce Benedové, Liborovi Pecharovi, Ivu Přikrylovi, Mirku Kosíkovi) za načerpání cenných a zajímavých zkušeností z třeboňského i sokolovského terénu. Děkuji svým rodičům za nekonečnou starost, lásku a pochopení a v neposlední řadě za pomoc s opatrováním mých dvou miláčků. Vážím si laskavosti Mišky Knížové, Helči Handrkové, Zuzičky Schwabové, Ondry Palkovského, Petry Hesslerové, Venduly Křinické, kteří mě slovy i beze slov dávali sílu vytrvat v mém třeboňském studentském životě. List of publications relevant to the thesis Prášil O, Bína D, Medová H, Řeháková K, Zapomělová E, Veselá J, Oren A (2009) Emission spectroscopy and kinetic fluorometry studies of phototrophic microbial communities along a salinity gradient in solar saltern evaporation ponds of Eilat, Israel. Aquat Microbial Ecology, 56: 285-296 HM conducted kinetic fluorometry measurements and analyzed pigment composition of phototrophic community by HPLC, processed the data and revised the manuscript. Řeháková K, Zapomělová E, Prášil O, Veselá J, Medová H, Oren A (2009) Composition changes of phototrophic microbial communities along the salinity gradient in the solar saltern evaporation ponds of Eilat, Israel. Hydrobiologia, 636: 77- 88 HM conducted kinetic fluorometry measurements and analyzed pigment composition of phototrophic community by HPLC and analyzed recorded data. Medová H, Boldareva EN, Hrouzek P, Borzenko SV, Namsaraev ZB, Gorlenko VM, Namsaraev BB, Koblížek M (2011) High abundance of aerobic anoxygenic phototrophs in saline steppe lakes. FEMS Microbiol Ecol., 76: 393-400 HM performed epifluorescence microscopy and HPLC analyses of collected samples, processed the data and wrote the manuscript. Medová H, Kaftan D, Koblížek M. Structural basis of thermostability in purple bacterial reaction centers. in preparation HM cultivated the studied bacterial strains, performed fluorometric measurements, processed,and analyzed the data, and wrote the manuscript. Medová H, Přikryl I, Zapomělová E, Pechar L (2013) Effect of post-mining waters on cyanobacterial photosynthesis. Water Environ Res, submitted HM sampled the studied localities, conducted all the field and laboratory measurements, processed and analysed the fluorometric data and wrote the manuscript. List of abbreviations AAPs aerobic anoxygenic phototrophs AMD acid mine drainage APB anoxygenic phototrophic bacteria BChl bacteriochlorophyll Chl chlorophyll rETR relative electron transport rate FETR forward electron transport rate FV/FM maximum photochemical yield LH1 light-harvesting complex 1 (inner antenna) LH2 light harvesting complex 2 (outer antenna) PCA principal canonical analysis pHopt optimal growth pH value RC reaction center RDA redundancy analysis RMSD root mean square deviation topt optimal growth temperature Content 1. Introduction to Extremophily ......................................................................................1 1.1 Phototrophic organisms.........................................................................................1 1.2 Definition of extremes and extremophiles ............................................................3 1.3 Main extremophilic groups ...................................................................................5 1.4 References ...........................................................................................................15 2. Anoxygenic Bacteria in Saline and Soda Lakes........................................................21 2.1 Summary .............................................................................................................21 2.1 Introduction .........................................................................................................22 2.2 Methods...............................................................................................................24 2.3 Results .................................................................................................................28 2.4 Discussion ...........................................................................................................33 2.5 References ...........................................................................................................37 2.6 Supplements ........................................................................................................42 3. Anoxygenic Phototrophic Bacteria in Polar Regions................................................45 3.1 Summary .............................................................................................................45 3.2 Introduction .........................................................................................................45 3.3 Methods...............................................................................................................46 3.4 Results and discussion.........................................................................................52 3.5 Conclusions .........................................................................................................56 3.6 References ...........................................................................................................57 4. Thermostability of Anoxygenic Reaction Centers ....................................................59
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