Phylogenetic Position of Endosymbiotic Green Algae in Paramecium bursaria Ehrenberg from Japan

Abstract

Endosymbiotic green algae of Japanese Paramecium bursaria were phylogenetically analyzed based on DNA sequences from the ribosomal DNA operon (18S rDNA, ITS1, 5.8S rDNA, and ITS2). Phylogenetic trees constructed using 18S rDNA sequences showed that the symbionts belong to the Chlorella sensu stricto (Trebouxiophyceae) group. They are genetically closer to the C. vulgaris Beijerinck group than to C. kessleri Fott et Nováková as proposed previously. Branching order in C. vulgaris group was unresolved in 18S rDNA trees. Compared heterogeneities of 18S rDNA, ITS1, 5.8S r, and ITS2 among symbionts and two Chlorella species, indicated that the ITS2 region (and probably also ITS1) is better able to resolve phylogenetic problems in such closely related taxa. All six symbiotic sequences obtained here (approximately 4000-bp sequences of 18S rDNA, ITS1, 5.8S rDNA, and ITS2) were completely identical in each, strongly suggesting a common origin.

Key words

18S rDNA - Chlorella - endosymbiont - ITS - Paramecium bursaria - phylogeny

References

• 1 Bhattacharya D., Friedl T., Damberger S.. Nuclear-encoded rDNA group I introns, origin and phylogenetic relationships of insertion site lineages in the green algae.  Mol. Biol. Evol.. (1996);  13 978-989
• 2 Cozzolino S., Campo I., Moretti M. A., Pollio A.. The use of nuclear ribosomal ITS1 DNA sequences for the identification of Chlorella strains.  Algological Studies. (1999);  95 31-42
• 3 Douglas A. E., Huss V. A. R.. On the characteristics and taxonomic position of symbiotic Chlorella. .  Arch. Microbiol.. (1986);  145 80-84
• 4 Dahlkild Å., Källersjö M., Lohtander K., Tehler A.. Photobiont diversity in the Physciaceae (Lecanorales).  Bryologist. (2001);  104 527-536
• 5 Felsenstein J.. Evolutionary trees from DNA sequences: a maximum-likelihood approach.  J. Mol. Evol.. (1981);  17 368-376
• 6 Felsenstein J.. Confidence intervals on phylogenies: an approach using the bootstrap.  Evolution. (1985);  39 783-791
• 7 Friedl T., Zeltner C.. Assessing the relationships of some coccoid green lichen algae and the Microthamniales (Chlorophyta) with 18S ribosomal RNA gene sequence comparisons.  J. Phycol.. (1994);  30 500-506
• 8 Friedl T., Rokitta C.. Species relationships in the lichen alga Trebouxia (Chlorophyta, Trebouxiophyceae): molecular phylogenetic analyses of nuclear-encoded large subunit rRNA gene sequences.  Symbiosis. (1997);  23 125-148
• 9 Hasegawa M., Kishino H., Yano T. A.. Dating of human-ape splitting by a molecular clock of mitochondrial DNA.  J. Mol. Evol.. (1985);  22 160-174
• 10 Helms G., Friedl T., Rambold G., Mayrhofer H.. Identification of photobionts from the lichen family Physciaceae using algal-specific ITS rDNA sequencing.  Lichenologist. (2001);  33 73-86
• 11 Hershkovitz M. A., Lewis L. A.. Deep-level diagnostic value of the rDNA-ITS region.  Mol. Biol. Evol.. (1996);  13 1276-1295
• 12 Hoshina R., Hasegawa K., Tanaka J., Hara Y.. Molecular phylogeny of the Dictyotaceae (Phaeophyceae) with emphasis on their morphology and its taxonomic implication.  Jpn. J. Phycol.. (2004);  52 (Suppl.) 189-194
• 13 Huss V. A. R., Sogin M. L.. Primary structure of the Chlorella vulgaris small subunit ribosomal RNA coding region.  Nucl. Acids Res.. (1989);  17 1255
• 14 Huss V. A. R., Sogin M. L.. Phylogenetic position of some Chlorella species within the chlorococcales based upon complete small-subunit ribosomal RNA sequences.  J. Mol. Evol.. (1990);  31 432-442
• 15 Huss V. A. R., Holweg C., Seidel B., Reich V., Rahat M., Kessler E.. There is an ecological basis for host/symbiont specificity in Chlorella/Hydra symbioses.  Endocyt. Cell Res.. 1993/94;  10 35-46
• 16 Huss V. A. R., Frank C., Hartmann E. C., Hirmer M., Kloboucek A., Seidel B. M., Wenzeler P., Kessler E.. Biochemical taxonomy and molecular phylogeny of the genus Chlorella sensu lato (Chlorophyta).  J. Phycol.. (1999);  35 587-598
• 17 Huss V. A. R., 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 Evol Biol.. (2002);  2 13
• 18 Ichimura T.. Sexual cell division and conjugation-papilla formation in sexual reproduction of Closterium strigosum. .  Proc. Int. Seaweed Symp.. (1971);  7 208-214
• 19 Jukes T. H., Cantor C. H.. Evolution of protein molecules. Munro, H. M., ed. Mammalian Protein Metabolism. New York; Academic Press (1969): 21-123
• 20 Kessler E., Huss V. A. R.. Biochemical taxonomy of symbiotic Chlorella strains from Paramecium and Acanthocystis. .  Bot. Acta. (1990);  103 140-142
• 21 Kessler E., Schäfer M., Hümmer C., Kloboucek A., Huss V. A. R.. Physiological, biochemical, and molecular characters for the taxonomy of the subgenera of Scenedesmus (Chlorococcales, Chlorophyta).  Bot. Acta. (1997);  110 244-250
• 22 Kimura M.. A simple method for estimating rate of base substitutions through comparative studies of nucleotide sequences.  J. Mol. Evol.. (1980);  16 111-120
• 23 Kroken S., Taylor J. W.. Phylogenetic species, reproductive mode, and specificity of the green alga Trebouxia forming lichens with the fungal genus Letharia. .  Bryologist. (2000);  103 645-660
• 24 Kvitko K. V., Migunova A. V., Karelov D. V., Prokosheva M.  Ju.. Molecular taxonomy of virus-sensitive Chlorella sp. - symbionts of Paramecium bursaria. .  Protistology. (2001);  2 96-104
• 25 Lanave C., Preparata G., Saccone C., Serio G.. A new method for calculating evolutionary substitution rates.  J. Mol. Evol.. (1984);  20 86-93
• 26 Linz B., Linz A., Migunova A. V., Kvitko K. V.. Correlation between virus-sensitivity and isoenzyme spectrum in symbiotic Chlorella-like algae.  Protistology. (1999);  1 76-81
• 27 Litaker R. W., Tester P. A., Colorni A., Levy M. G., Noga E. J.. The phylogenetic relationship of Pfiesteria piscicida, Cryptoperidiniopsoid sp., Amyloodinium ocellatum and a Pfiesteria-like dinoflagellate to other dinoflagellates and apicomplexans.  J. Phycol.. (1999);  35 1379-1389
• 28 Marin B., Melkonian M.. Mesostigmatophyceae, a new class of Streptophyte green algae revealed by SSU rRNA sequence comparisons.  Protist. (1999);  150 399-417
• 29 Moncalvo J. M., Lutzoni F. M., Rehner L. S., Johnson J., Vilgalys R.. Phylogenetic relationships of agaric fungi based on nuclear large subunit ribosomal DNA sequences.  Syst. Biol.. (2000);  49 278-305
• 30 Nakayama T., Watanabe S., Mitsui K., Uchida H., Inouye I.. The phylogenetic relationship between the Chlamydomonadales and Chlorococcales inferred from 18S rDNA sequence data.  Phycol. Res.. (1996);  44 47-55
• 31 Nishihara N., Horiike S., Takahashi T., Kosaka T., Shigenaka Y., Hosoya H.. Cloning and characterization of endosymbiotic algae isolated from Paramecium bursaria. .  Protoplasma. (1998);  203 91-99
• 32 Pearson W. R., Lipman D. J.. Improved tools for biological sequence analysis.  PNAS. (1988);  85 2444-2448
• 33 Reisser W.. The taxonomy of green algae endosymbiotic in Ciliates and Sponge.  Br. Phycol. J.. (1984);  19 309-318
• 34 Reisser W.. Naturally occurring and artificially established associations of ciliates and algae. Models for different steps in the evolution of stable endosymbioses.  Ann. N.Y. Acad. Sci.. (1987);  503 316-329
• 35 Reisser W., Vietze S., Widowski M.. Taxonomic studies on endocytobiotic Chlorophycean algae isolated from different American and European strains of Paramecium bursaria. .  Symbiosis. (1988);  6 253-270
• 36 Romeike J., Friedl T., Helms G., Ott S.. Genetic diversity of algal and fungal partners in four species of Umbilicaria (Lichenized A scomycetes) along a transect of the Antarctic peninsula.  Mol. Biol. Evol.. (2002);  19 1209-1217
• 37 Saitou N., Nei M.. The neighbor-joining method: A new method for reconstructing phylogenetics trees.  Mol. Biol. Evol.. (1987);  4 406-425
• 38 Sonneborn T. M.. Breeding systems, reproductive methods and species problems in Protozoa. Mayr, E., ed. The Species Problem. Washington DC; American Assoc. Adv. Science (1957): 155-324
• 39 Swofford D. L.. PAUP. Phylogenetic Analysis Using Parsimony (and other methods), Version 4. Sunderland, Massachusetts; Sinauer Associates (2000)
• 40 Takeda H.. Cell wall composition and taxonomy of symbiotic Chlorella from Paramecium and Acanthocystis. .  Phytochemistry. (1995);  40 457-459
• 41 Takeda H., Sekiguchi T., Nunokawa S., Usuki I.. Species-specificity of Chlorella for establishment of symbiotic association with Paramecium bursaria - Does infectivity depend upon sugar components of the cell wall?.  Europ. J. Protistol.. (1998);  34 133-137
• 42 Thompson J. D., Gibson T. J., Plewniak F., Jeanmougin F., Higgins D. G.. The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.  Nucleic Acids Res.. (1997);  24 4876-4882
• 43 Ustinova I., Krienitz L., Huss V. A. R.. Closteriopsis acicularis (G. M. Smith) Belcher et Swale is a fusiform alga closely related to Chlorella kessleri Fott et Nováková (Chlorophyta, Trebouxiophyceae).  Eur. J. Phycol.. (2001);  36 341-351
• 44 Van de Peer Y., De Wacher R.. Evolutionary relationships among the eukaryotic crown taxa taking into account site-to-site rate variation in 18S rRNA.  J. Mol. Evol.. (1997);  45 619-630
• 45 Waddle J. A., Schuster A. M., Lee K. W., Meints R. H.. The mitochondrial genome of an exsymbiotic Chlorella-like green alga.  Plant Mol. Biol.. (1990);  14 187-195
• 46 White T. J., Bruns T., Lee S., Taylor J.. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. Innis, M., Gelfand, D., Sninsky, J., and White, T., eds. PCR Protocols: A Guide to Methods and Application. San Diego; Academic Press (1990): 315-322
• 47 Wolf M., Krienitz L., Hepperle D.. Phylogenetic Position of Actinastrum hantzschii Lagerheim 1882 (Chlorophyta, Trebouxiophyceae).  Arch. Hydrobiol. Suppl. Algol. Stud.. (2002);  104 59-67
• 48 Yang Z.. Estimating the pattern of nucleotide substitution.  J. Mol. Evol.. (1994);  39 105-111
• 49 Zuker M.. Mfold web server for nucleic acid folding and hybridization prediction.  Nucleic Acids Res.. (2003);  31 1-10

Section Editor: F. Salamini