Our knowledge of the processes involved in speciation of microalgae remains highly limited. In the present study, we investigated a potential role of ecological speciation processes in diversification of the filamentous green alga Klebsormidium. We examined 12 strains representing four different genotypes. The strains were collected from sandstone and limestone rocks and were cultivated at five different pH levels ranging from pH 4 to pH 8. We determined the responses of the 12 strains to the experimental pH conditions by (1) measuring the effective quantum yield of photosystem II, and (2) determining the growth rates after cultivation at different pH levels. Strong differences were found between the results obtained by these two methods. Direct counting of cells revealed a strong ecological differentiation of strains of Klebsormidium isolated from different substrate types. Strains isolated from limestone showed the highest growth rates at higher pH levels; whereas, the strains isolated from sandstone exhibited two distinct growth responses with optima at pH 5 and 6, respectively. In contrast, the effective quantum yield of photosystem II was always down-regulated at lower pH values, probably due to dissolved inorganic carbon limitation. In general, we determined distinct ecophysiological differentiation among distantly and closely related lineages, thereby corroborating our hypothesis that the sympatric speciation of terrestrial algae is driven by ecological divergence. We clearly showed that pH is a critical ecological factor that influences the diversity of autotrophic protists in terrestrial habitats.

Charles University Prague Faculty of Science Department of Botany Benátská 2 12801 Prague 2 Czech Republic

University of Innsbruck Institute of Botany Functional Plant Biology Sternwartestraβe 15 A 6020 Innsbruck Austria

Zobrazit více v PubMed

Aguilera A, Zettler E, Gomez F, Amaral-Zettler L, Rodriguez N, Amils R. Distribution and seasonal variability in the benthic eukaryotic community of RioTinto (SW, Spain), an acidic, high metal extreme environment. Systematic and Applied Microbiology. 2007;30:531–546. PubMed

Allen ED, Spence DHN. The differential ability of aquatic plants to utilize the inorganic carbon supply in fresh waters. New Phytologist. 1981;87:269–283.

Amato A, Kooistra WHCF, Ghiron JHL, Mann DG, Pröschold T, Montresor M. Reproductive isolation among sympatric cryptic species in marine diatoms. Protist. 2007;158:193–207. PubMed

Baba M, Shiraiwa Y. High-CO2 response mechanisms in microalgae. In: Najafpour Mohammad., editor. Advances in photosynthesis-fundamental aspects. InTech; Rijeka, Croatia: 2012. pp. 299–320.

Bates ST, Clemente JC, Flores GE, Walters WA, Parfrey LW, Knight R, Fierer N. Global biogeography of highly diverse protistan communities in soil. The ISME Journal. 2012;7:1–8. PubMed PMC

Benton MJ, Pearson PN. Speciation in the fossil record. Trends in Ecology & Evolution. 2001;16:405–411. PubMed

Birky CW, Jr, Adams J, Gemmel M, Perry J. Using population genetic theory and DNA sequences for species detection and identification in asexual organisms. PLoS One. 2010;5(5):e10609. 101371/journal.pone.0010609. PubMed PMC

Brock TD. Lower pH limit for existence of blue-green algae: evolutionary and ecological implications. Science. 1973;179:480–483. PubMed

Caron DA. Past president’s address: protistan biogeography: why all the fuss? Journal of Eukaryotic Microbiology. 2009;56:105–112. PubMed

Casteleyn G, Leliaert F, Backeljau T, Debeer AE, Kotaki Y, Rhodes L, Lundholm N, Sabbe K, Vyverman W. Limits to gene flow in a cosmopolitan marine planktonic diatom. Proceedings of the National Academy of Sciences of the United States of America. 2010;107:12952–12957. PubMed PMC

De Meester L, Gomez A, Okamura B, Schwenk K. The monopolization hypothesis and the dispersal-gene flow paradox in aquatic organisms. Acta Oecologica. 2002;23:121–135.

De Vargas C, Norris R, Zaninetti L, Gibb SW, Pawlowski J. Molecular evidence of cryptic speciation in planktonic foraminifers and their relation to oceanic provinces. Proceedings of the National Academy of Sciences of the United States of America. 1999;96:2864–2868. PubMed PMC

Desrosiers C, Leflaive J, Eulin A, Ten-Hage L. Bioindicators in marine waters: benthic diatoms as a tool to assess water quality from eutrophic to oligotrophic coastal ecosystems. Ecological Indicators. 2013;32:25–34.

Elster J, Degma P, Kováčík L, Valentová L, Šramková K, Pereira AB. Freezing and desiccation injury resistance in the filamentous green alga Klebsormidium from the Antarctic, Arctic and Slovakia. Biologia. 2008;63:843–851.

Evans KM, Chepurnov VA, Sluiman HJ, Thomas SJ, Spears BM, Mann DG. Highly differentiated populations of the fresh water diatom Sellaphora capitata suggest limited dispersal and opportunities for allopatric speciation. Protist. 2009;160:386–396. PubMed

Falkowski PG, Raven JA. Aquatic photosynthesis. Princeton University Press; Princeton, New Jersey: 2007. p. 488.

Fenchel T, Finlay BJ. Is microbial diversity fundamentally different from biodiversity of larger animals and plants? European Journal of Protistology. 2003;39:486–490.

Fenchel T, Finlay BJ. The ubiquity of small species: patterns of local and global diversity. Bioscience. 2004;54:777–784.

Finlay BJ. Global dispersal of free-living microbial eukaryote species. Science. 2002;296:1061–1063. PubMed

Finlay BJ, Fenchel T. Divergent perspective on protist species richness. Protist. 1999;150:229–233. PubMed

Finlay BJ, Monaghan EB, Maberly SC. Hypothesis: the rate and scale of dispersal of fresh water diatom species is a fiction of their global abundance. Protist. 2002;153:261–273. PubMed

Foissner W. Biogeography and dispersal of micro-organisms: a review emphasizing protists. Acta Protozoologica. 2006;45:111–136.

Foissner W, Chao A, Katz LA. Diversity and geographic distribution of ciliates (Protista: Ciliophora) Biodiversity and Conservation. 2008;17:235–242.

Fontaneto D, Herniou EA, Boschetti C, Caprioli M, Melone G, Ricci C, Barraclough TG. Independently evolving species in asexual bdelloid rotifers. PLoS Biology. 2007;5:e87. doi: 10.1371/journal.pbio.0050087. PubMed DOI PMC

Gächter E, Weisse T. Local adaptation among geographically distant clones of the cosmopolitan freshwater ciliate Meseres corlissi. I. Temperature response. Aquatic Microbial Ecology. 2006;45:291–300.

Gaysina LA, Purina ES, Safiullina LM, Bakieva GR. Resistance of Klebsormidium flaccidum (Streptophyta) to heavy metals. Plant Sciences. 2009;3:39–41.

Gross W. Ecophysiology of algae living in highly acidic environments. Hydrobiologia. 2000;433:31–37.

Hallmann C, Stannek L, Fritzlar D, Hause-Reitner D, Friedl T, Hoppert M. Molecular diversity of phototrophic biofilms on building stone. FEMS Microbiology Ecology. 2013;84:355–372. PubMed

Hammer O, Harper DAT, Ryan PD. PAST: Paleontological Statistics software package for education and data analysis. Palaeontologia Electronica. 2001;4(1):9.

Hepperle D. SeqAssem©. A sequence analysis tool, contig assembler and trace data visualisation tool for molecular sequences, version 09/2004. 2004 http://www.sequentix.de.

Herburger K, Holzinger A. Localization and quantification of callose in the streptophyte green algae Zygnema and Klebsormidium: Correlation with desiccation tolerance. Plant and Cell Physiology. 2015;56:2259–2270. PubMed PMC

Herburger K, Karsten U, Holzinger A. Entransia and Hormidiella, sister lineages of Klebsormidium (Streptophyta), respond differently to light, temperature and desiccation stress. Protoplasma. 2016 doi: 10.1007/s00709-015-0889-z. in press. PubMed DOI PMC

Hoffmann L. Algae of terrestrial habitats. The Botanical Review. 1989;55:77–105.

Holzinger A, Becker B. Desiccation tolerance in the streptophyte green alga Klebsormidium: the role of phytohormones. Communicative and Integrative Biology. 2015;8(4):e1059978. doi: 10.1080/19420889.2015.1059978. PubMed DOI PMC

Holzinger A, Kaplan F, Blaas K, Zechmann B, Komsic-Buchmann K, Becker B. Transcriptomics of desiccation tolerance in the streptophyte green alga Klebsormidium reveal a land plant-like defense. PLoS One. 2014;9(10):e110630. doi: 10.1371/journal.pone.0110630. PubMed DOI PMC

Hori K, Maruyama F, Fujisawa T, Togashi T, Yamamoto N, Seo M, Sato S, Yamada T, Mori H, Tajima N, Moriyama T, et al. Klebsormidium flaccidum genome reveals primary factors for plant terrestrial adaptation. Nature Communications. 2014;5:3978. PubMed PMC

Huelsenbeck JP, Ronquist F. MRBAYES: Bayesian inference of phylogeny. Bioinformatics. 2001;17:754–755. PubMed

Huss VAR, Frank C, Hartmann EC, Hirmer M, Kloboucek A, Seidel BM, Wenzeler P, Kessler E. Biochemical taxonomy and molecular phylogeny of the genus Chlorella sensu lato (Chlorophyta) Journal of Phycology. 1999;35:587–598.

Huss VAR, Ciniglia C, Cennamo P, Cozzolino S, Pinto G, Pollio A. Phylogenetic relationships and taxonomic position of Chlorella-like isolates from low pH environments (pH < 3.0) BMC Evolutionary Biology. 2002;26:2–13. PubMed PMC

Kaplan F, Lewis LA, Wastian J, Holzinger A. Plasmolysis effects and osmotic potential of two phylogenetically distinct alpine strains of Klebsormidium (Streptophyta) Protoplasma. 2012;249:789–804. PubMed

Karsten U, Holzinger A. Light, temperature, and desiccation effects on photosynthetic activity, and drought-induced ultrastructural changes in the green alga Klebsormidium dissectum (Streptophyta) from a high alpine soil crust. Microbial Ecology. 2012;63:51–63. PubMed

Karsten U, Holzinger A. Green algae in alpine biological soil crust communities: acclimation strategies against ultraviolet radiation and dehydration. Biodiversity and Conservation. 2014;23:1845–1858. PubMed PMC

Karsten U, Lütz C, Holzinger A. Ecophysiological performance of the aeroterrestrial green alga Klebsormidium crenulatum (Charophyceae, Streptophyta) isolated from an alpine soil crust with an emphasis on desiccation stress. Journal of Phycology. 2010;46:1187–1197. PubMed

Karsten U, Pröschold T, Mikhailyuk T, Holzinger A. Photosynthetic performance of different genotypes of the green alga Klebsormidium sp. (Streptophyta) isolated from biological soil crusts of the Alps. Algological Studies. 2013;142:45–62.

Karsten U, Herburger K, Holzinger A. Dehydration, temperature and light tolerance in members of the aeroterrestrial green algal genus Interfilum (Streptophyta) from biogeographically different temperate soils. Journal of Phycology. 2014;50:804–816. PubMed PMC

Karsten U, Herburger K, Holzinger A. Living in biological soil crust communities of African deserts – physiological traits of green algal Klebsormidium species (Streptophyta) to cope with desiccation, light and temperature gradients. Journal of Plant Physiology. 2016 doi: 10.1016/j.jplph.2015.09.002. in press. PubMed DOI PMC

Kitzing C, Karsten U. UV-induced effects on growth, photosynthetic performance and sunscreen contents in members of the genera Interfilum, Klebsormidium, Hormidiella and Entransia (Klebsormidiophyceae, Streptophyta) European Journal of Phycology. 2015;50:279–287.

Kitzing C, Pröschold T, Karsten U. UV-induced effects on growth, photosynthetic performance and sunscreen contents in different populations of the green alga Klebsormidium fluitans (Streptophyta) from alpine soil crusts. Microbial Ecology. 2014;67:327–340. PubMed

Lane AE, Burris JE. Effects of environmental pH on the internal pH of Chlorella pyrenoidosa, Scenedesmus quadricauda and Euglena mutabilis. Plant Physiology. 1981;68:439–442. PubMed PMC

Logares R, Rengefors K, Kremp A, Shalchian-Tabrizi K, Boltovskoy A, Tengs T, Shurtleff A, Klaveness D. Phenotypically different microalgal morphospecies with identical ribosomal DNA: a case of rapid adaptive evolution? Microbial Ecology. 2007;53:549–561. PubMed

Lomin SN, Krivosheev DM, Steklov MY, Arkhipov DV, Osolodkin DI, Schmülling T, Romanov GA. Plant membrane assays with cytokinin receptors underpin the unique role of free cytokinin bases as biologically active ligands. Journal of Experimental Botany. 2015;66:1851–1863. PubMed PMC

Lowe RL, Furey PC, Ress JA, Johansen JR. Diatom biodiversity and distribution on wet walls in Great Smoky Mountains National Park. Southeastern Naturalist. 2007;6:135–152.

Lukešová A. Soil algae in brown coal and lignite post-mining areas in central Europe (Czech Republic and Germany) Restoration Ecology. 2001;9:341–350.

Lukešová A, Hoffmann L. Soil algae from acid rain impacted forest areas of Krušné hory mountains (Czech Republic). 2. Effects of pH on growth. Algological Studies. 1995;78:39–51.

Machová-Černá K, Neustupa J. Spatial distribution of algal assemblages in a temperate lowland peat bog. International Review of Hydrobiology. 2009;94:40–56.

Mikhailyuk T, Holzinger A, Mlassalski A, Karsten U. Morphology and ultrastructure of Interfilum and Klebsormidium (Klebsormidiales, Streptophyta) with special reference to cell division and thallus formation. European Journal of Phycology. 2014;49:395–412. PubMed PMC

Mikhailyuk T, Glaser K, Holzinger A, Karsten U. Biodiversity of Klebsormidium (Streptophyta) from alpine biological soil crusts (Alps, Tyrol, Austria and Italy) Journal of Phycology. 2015;51:750–767. PubMed PMC

Moniz MB, Rindi F, Novis PM, Broady PA, Guiry MD. Molecular phylogeny of antarctic Prasiola (Prasiolales, Trebouxiophyceae) reveals extensive cryptic diversity. Journal of Phycology. 2012;48:940–955. PubMed

Nagao M, Matsui K, Uemura M. Klebsormidium flaccidum, a charophycean green alga, exhibits cold acclimation that is closely associated with compatible solute accumulation and ultrastructural changes. Plant, Cell & Environment. 2008;31:872–885. PubMed

Novis PM. Taxonomy of Klebsormidium (Klebsormidiales, Charophyceae) in New Zealand streams and the significance of low-pH habitats. Phycologia. 2006;45:293–301.

Novis PM, Harding JS. Extreme acidophiles: freshwater algae associated with acid mine drainage. In: Seckbach J, editor. Extremophilic algae, cyanobacteria and non-photosynthetic protists: from prokaryotes to astrobiology. Springer Verlag; Dordrecht, the Netherlands: 2007. pp. 445–463.

Nylander JAA. MrModeltest, version 2. 2004 http://www.abc.se/~nylander.

Peksa O, Škaloud P. Do photobionts influence the ecology of lichens? A case study of environmental preferences in symbiotic green alga Asterochloris (Trebouxiophyceae) Molecular Ecology. 2011;20:3936–3948. PubMed

Pietrasiak N, Johansen JR, Drenovsky RE. Geologic composition influences distribution of microbiotic crusts in the Mojave and Colorado Deserts at the regional scale. Soil Biology and Biochemistry. 2011;43:967–974.

R Core Team. R: a language and environment for statistical computing. R Foundation for Statistical Computing. Vienna, Austria: 2016. version R 2.15.1 http://www.R-project.org/

Reynolds CS. The ecology of freshwater phytoplankton. Cambridge University Press; Cambridge, UK: 1984. p. 384.

Rindi F, Guiry MD. Diversity, life history and ecology of Trentepohlia and Printzina (Trentepohliales, Chlorophyta) in urban habitats in western Ireland. Journal of Phycology. 2002;38:39–54.

Rindi F, Guiry MD. Composition and spatial variability of terrestrial algal assemblages occurring at the bases of urban walls in Europe. Phycologia. 2004;43:225–235.

Rindi F, Mcivor L, Sherwood AR, Friedl T, Guiry MD, Sheath RG. Molecular phylogeny of the green algal order Prasiolales (Trebouxiophyceae, Chlorophyta) Journal of Phycology. 2007;43:811–822.

Rindi F, Guiry MD, López-Bautista JM. Distribution, morphology, and phylogeny of Klebsormidium (Klebsormidiales, Charophyceae) in urban environment in Europe. Journal of Phycology. 2008;44:1529–1540. PubMed

Rindi F, Mikhailyuk TI, Sluiman HJ, Friedl T, LÓpez-Bautista JM. Phylogenetic relationships in Interfilum and Klebsormidium (Klebsormidiophyceae, Streptophyta) Molecular Phylogenetics and Evolution. 2011;58:218–231. PubMed

Ryšánek D, Hrčková K, Škaloud P. Global ubiquity and local endemism of free-living terrestrial protists: phylogeographic assessment of the streptophyte alga Klebsormidium. Environmental Microbiology. 2015;17:689–698. PubMed

Shelp BJ, Canvin DT. Utilization of exogenous inorganic carbon species in photosynthesis by Chlorella pyrenoidosa. Plant Physiology. 1980;65:774–779. PubMed PMC

Shields LM, Durell LW. Algae in relation to soil fertility. The Botanical Review. 1964;30:92–128.

Shiraiwa Y, Goyal A, Tolbert NE. Alkalization of the medium by unicellular green algae during uptake dissolved inorganic carbon. Plant and Cell Physiology. 1993;34:649–657.

Silva PC, Mattox KR, Blackwell WH. The generic name Hormidium as applied to green algae. Taxon. 1972;21:639–645.

Škaloud P, Peksa O. Evolutionary inferences based on ITS rDNA and actin sequences reveal extensive diversity of the common lichen alga Asterochloris (Trebouxiophyceae, Chlorophyta) Molecular Phylogenetics and Evolution. 2010;54:36–46. PubMed

Škaloud P, Rindi F. Ecological differentiation of cryptic species within an asexual protest morphospecies: a case study of filamentous green alga Klebsormidium (Streptophyta) Journal of Eukaryotic Microbiology. 2013;60:350–362. PubMed

Škaloud P, Kynčlová A, Benada O, Kofroňová O, Škaloudová M. Toward a revision of the genus Synura, section Petersenianae (Synurophyceae, Heterokontophyta): morphological characterization of six pseudo-cryptic species. Phycologia. 2012;51:303–329.

Škaloud P, Lukešová A, Malavasi V, Ryšánek D, HrčKová K, Rindi F. Molecular evidence for the polyphyletic origin of low pH adaptation in the genus Klebsormidium (Klebsormidiophyceae, Streptophyta) Plant Ecology and Evolution. 2014;147:333–345.

Sluiman HJ, Guihal C, Mudimu O. Assessing phylogenetic affinities and species deliminations in Klebsormidiales (Streptophyta): nuclear-encoded rDNA phylogenies and ITS secondary structure models in Klebsormidium, Hormidiella, and Entransia. Journal of Phycology. 2008;44:183–195. PubMed

Starks TL, Shubert LE, Trainor FR. Ecology of soil algae: a review. Phycologia. 1981;20:65–80.

Starr RC, Zeikus JA. UTEX – the Culture Collection of Algae at the University of Texas at Austin 1993 List of Cultures. Journal of Phycology. 1993;29:1–106.

Svoboda P, Kulichová J, Šťastiný J. Spatial and temporal community structure of desmids on a small spatial scale. Hydrobiologia. 2014;722:291–303.

Swofford DL. PAUP* Phylogenetic Analysis Using Parsimony (*and other methods) Sinauer Associates; Sunderland, Massachusetts: 2002. Version 4.

Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution. 2011;28:2731–2739. PubMed PMC

Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: improving sensitivity of progressive multiple sequence alignment through sequence weighting, position 86 specific gap penalties, and weight matrix choice. Nucleic Acids Research. 1994;22:4673–4680. PubMed PMC

Vanelslander B, Creach V, Vanormelingen P, Ernst A, Chepurnov VA, Sahan E, Sabbe K. Ecological differentiation between sympatric pseudocryptic species in the estuarine benthic diatom Navicula phyllepta (Bacillariophyceae) Journal of Phycology. 2009;45:1278–1289. PubMed

Vanormelingen P, Vyverman W, De Bock D, Van der Gucht K, De Meester L. Local genetic adaptation to grazing pressure of the green alga Desmodesmus armatus in a strongly connected pond system. Limnology and Oceanography. 2009;54:503–511.

Von der Heyden S, Chao E, Cavalier-Smith T. Genetic diversity of goniomonads: an ancient divergence between freshwater and marine species. European Journal of Phycology. 2004;39:343–350.

Weisse T. Distribution and diversity of aquatic protists: an evolutionary and ecological perspective. Biodiversity and Conservation. 2008;17:243–259.

Zwickl DJ. Dissertation. The University of Texas at Austin; Texas: 2006. Genetic algorithm approaches for the phylogenetic analysis of large biological sequence datasets under the maximum likelihood criterion; p. 115. Available from Digital Repository, University of Texas Libraries. http://hdl.Handle.net/2152/2666.

Seasonal Dynamics of Zygnema (Zygnematophyceae) Mats from the Austrian Alps

Terrestrial adaptation of green algae Klebsormidium and Zygnema (Charophyta) involves diversity in photosynthetic traits but not in CO2 acquisition

DNA-Based Taxonomy in Ecologically Versatile Microalgae: A Re-Evaluation of the Species Concept within the Coccoid Green Algal Genus Coccomyxa (Trebouxiophyceae, Chlorophyta)