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Bacterial diversity associated with the Caribbean tunicate Ecteinascidia turbinata

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Abstract

The Caribbean tunicate, Ecteinascidia turbinata produces the anti-cancer agent ET-743 that could well be a metabolite of an associated bacterial strain. This current study aims at the analysis of bacteria that are persistently and specifically associated with this invertebrate. Utilizing techniques such as denaturing gradient gel electrophoresis, DNA sequencing and phylogenetic analysis of bacteria from E. turbinata collected from different locations in the Caribbean Sea, we report here the identification of five possible persistently associated bacteria. Of these, only one organism, Candidatus Endoecteinascidia frumentensis, was found specifically associated to E. turbinata from the Caribbean and has also been found to be associated with E. turbinata from the Mediterranean. These experiments suggest that assessment of bacterial diversity associated with invertebrates from different geographical sites might be an effective way of identifying persistently and specifically associated bacteria.

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References

  • Altschul SF, Lipman DJ (1990) Protein database searches for multiple alignments. Proc Natl Acad Sci USA 87(14):5509–5513

    Article  PubMed  CAS  Google Scholar 

  • Bernan VS, Greenstein M, Maiese WM (1997) Marine microorganisms as a source of new natural products. Adv Appl Microbiol 43:57–89

    Article  PubMed  CAS  Google Scholar 

  • Brennan Y, Callen WN, Christoffersen L, Dupree P, Goubet F, Healey S, Hernandez M, Keller M, Li K, Palackal N, Sittenfeld A, Tamayo G, Wells S, Hazlewood GP, Mathur EJ, Short JM, Robertson DE, Steer BA (2004) Unusual microbial xylanases from insect guts. Appl Environ Microbiol 70(6):3609–3617

    Article  PubMed  CAS  Google Scholar 

  • Colwell RR, Grimes DJ (2000) Semantic and strategies, In: Colwell RR, Grimes DJ (eds) Nonculturable microorganisms in the environment, ASM Press, Washington, D.C pp 1–6

    Google Scholar 

  • Davidson SK, Allen SW, Lim GE, Anderson CM, Haygood MG (2001) Evidence for the biosynthesis of bryostatins by the bacterial symbiont “Candidatus Endobugula sertula” of the bryozoan Bugula neritina. Appl Environ Microbiol 67(10):4531–4537

    Article  PubMed  CAS  Google Scholar 

  • Diez B, Pedros-Alio C, Marsh TL, Massana R (2001) Application of denaturing gradient gel electrophoresis (DGGE) to study the diversity of marine picoeukaryotic assemlages and comparison of DGGE with other molecular techniques. Appl Environ Microbiol 67(7):2942–2951

    Article  PubMed  CAS  Google Scholar 

  • Farrelly V, Rainey FA, Stackebrandt E (1995) Effect of genome size and rrn gene copy number on PCR amplification of 16S rRNA genes from a mixture of bacterial species. Appl Environ Microbiol 61(7):2798–2801

    PubMed  CAS  Google Scholar 

  • Flam F (1994) Chemical prospectors scour the seas for promising drugs. Science 266(5189):1324–1325

    Article  PubMed  CAS  Google Scholar 

  • Friedrich AB, Fischer I, Proksch P, Hacker J, Hentschel U (2001) Temporal variation of the microbial community associated with the Mediterranean sponge Aplysina aerophoba. FEMS Microbiol Ecol 1301:1–9

    Google Scholar 

  • Fukatsu T (2001) Secondary intracellular symbiotic bacteria in aphids of the genus Yamatocallis (Homoptera: Aphididae: Drepanosiphinae). Appl Environ Microbiol 67(11):5315–5320

    Article  PubMed  CAS  Google Scholar 

  • Goto S, Anbutsu H, Fukatsu T (2006) Asymmetrical interactions between Wolbachia and Spiroplasma endosymbionts coexisting in the same insect host. Appl Environ Microbiol 72(7):4805–4810

    Article  PubMed  CAS  Google Scholar 

  • Grobkopf R, Janssen PH, Liesack W (1998) Diversity and structure of the methanogenic community in anoxic rice paddy soil microcosms as examined by cultivation and direct 16S rRNA sequence retrieval. Appl Environ Microbiol 64(3):960–969

    Google Scholar 

  • Hentschel U, Hopke J, Horn M, Friedrich AB, Wagner M, Hacker J, Moore BS (2002) Molecular evidence for a uniform microbial community in sponges from different oceans. Appl Environ Microbiol 68:4431–4440

    Article  PubMed  CAS  Google Scholar 

  • Hildebrand M, Waggoner LE, Lim GE, Sharp KH, Ridley CP, Haygood MG (2004) Approaches to identify, clone, and express symbiont bioactive metabolite genes. Nat Prod Rep 21(1):122–142

    Article  PubMed  CAS  Google Scholar 

  • Hirose E, Maruyuama T, Cheng L, Lewin RA (1996) Intracellular symbiosis of a photosynthetic prokaryote, Prochloron sp., in a colonial ascidian. Invert Biol 115:343–348

    Google Scholar 

  • König GM, Kehraus S, Seibert SF, Abdel-Lateff A, Müller D (2005) Natural products from marine organisms and their associated microbes. Chem BioChem 6:1–10

    Google Scholar 

  • Kumar S, Tamura K, Nei M (2004) MEGA3: Integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163

    Article  PubMed  CAS  Google Scholar 

  • Lane DJ, Pace B, Olsen GJ, Stahl DA, Sogin ML, Pace NR (1985) Rapid determination of 16S ribosomal RNA sequences for phylogenetic analyses. Proc Natl Acad Sci USA 82(20):6955–6959

    Article  PubMed  CAS  Google Scholar 

  • Lim GE, Haygood MG (2004) “Candidatus Endobugula glebosa,” a specific bacterial symbiont of the marine bryozoan Bugula simplex. Appl Environ Microbiol 70(8):4921–4929

    Article  PubMed  CAS  Google Scholar 

  • Lindhal V, Bakken LR (1995) Evaluation of methods for extraction of bacteria from soil. FEMS Microbiol Ecol 16:135–142

    Article  Google Scholar 

  • Mendola D (2000) Aquacultural production of bryostatin 1 and ecteinacidin 743., In: Fusetani N (ed) Drugs from the sea, Karger Baser, pp 120–133

  • Moss C, Green DH, Perez B, Velasco A, Henriquez R, McKenzie JD (2003) Intracellular bacteria associated with the ascidian Ecteinascidia turbinata: phylogenetic and in situ hybridization analysis. Mar Biol 143:99–110

    Article  CAS  Google Scholar 

  • Muyzer G, De Waal EC, Uitterlinden AG (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol 59(3):695–700

    PubMed  CAS  Google Scholar 

  • Proksch P, Edrada RA, Ebel R (2002) Drugs from the seas-current status and microbiological implications. Appl Environ Microbiol 59:125–134

    CAS  Google Scholar 

  • Rinehart KL, Holt TG, Fregeau NL, Stroh JG, Keifer PA, Sun F, Li LH, Martin DJ (1990) Ecteinascidins 729, 743, 745, 759A, 759B, and 770: Potent antitumor agents from the Caribbean tunicate Ecteinascidia turbinata. J Org Chem 55(15):4512–4515

    Article  CAS  Google Scholar 

  • Rosado W, Govind NS (2003) Identification of carbohydrate degrading bacteria in sub-tropical regions. Rev Biol Trop 51:205–210

    PubMed  Google Scholar 

  • Schirmer A, Gadkari R, Reeves CD, Ibrahim F, DeLong EF, Hutchinson CR (2005) Metagenomic analysis reveals diverse polyketide synthase gene clusters in microorganisms associated with the marine sponge Discodermia dissoluta. Appl Environ Microbiol 71(8):4840–4849

    Article  PubMed  CAS  Google Scholar 

  • Schloss PD, Handelsman J (2005) Introducing DOTUR, a computer program for defining operational taxonomic units and estimating species richness. Appl Environ Microbiol 71(3):1501–1506

    Article  PubMed  CAS  Google Scholar 

  • Schmidt EW, Nelson JT, Rasko DA, Sudek S, Eisen JA, Haygood MG, Ravel J (2005) Patellamide A and C biosynthesis by a microcin-like pathway in Prochloron didemni, the cyanobacterial symbiont of Lissoclinum patella. Proc Natl Acad Sci USA 102(20):7315–7320

    Article  PubMed  CAS  Google Scholar 

  • Schmidt EW, Obraztsova AY, Davidson SK, Faulkner DJ, Haygood MG (2000) Identification of the antifungal peptide-containing symbiont of the marine sponge Theonella swinhoei as a novel d-proteobacterium “Candidatus Entothenonella palauensis”. Mar Biol 136:969–977

    Article  CAS  Google Scholar 

  • Suzuki MT, Taylor LT, DeLong EF (2000) Quantitative analysis of small-subunit rRNA genes in mixed microbial populations via 5′-nuclease assays. Appl Environ Microbiol 66(11):4605–4614

    Article  PubMed  CAS  Google Scholar 

  • Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progresive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

    Article  PubMed  CAS  Google Scholar 

  • Velasco A, Acebo P, Gomez A, Schleissner C, Rodriguez P, Aparicio T, Conde S, Munoz R, de la Calle F, Garcia JL, Sanchez-Puelles JM (2005) Molecular characterization of the safracin biosynthetic pathway from Pseudomonas fluorescens A2-2: designing new cytotoxic compounds. Mol Microbiol 56(1):144–154

    Article  PubMed  CAS  Google Scholar 

  • Wang Y, Stingl U, Anton-Erxleben F, Zimmer M, Brune A (2004) ‘ Candidatus Hepatincola porcellionum’ gen. nov., sp. nov., a new, stalk-forming lineage of Rickettsiales colonizing the midgut glands of a terrestrial isopod. Arch Microbiol 181(4):299–304

    Article  PubMed  CAS  Google Scholar 

  • Webster NS, Wilson KJ, Blackall LL, Hill RT (2001) Phylogenetic diversity of bacteria associated with the marine sponge Rhopaloeides odorabile. Appl Environ Microbiol 67(1):434–444

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

This study has been supported, in part by a seed money grant from the Sea Grant College program of the University of Puerto Rico (to AEP and WR) and the N.I.H MBRS-SCORE program (SO6-GM 08103 to NSG).

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Correspondence to Nadathur S. Govind.

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Pérez-Matos, A.E., Rosado, W. & Govind, N.S. Bacterial diversity associated with the Caribbean tunicate Ecteinascidia turbinata . Antonie van Leeuwenhoek 92, 155–164 (2007). https://doi.org/10.1007/s10482-007-9143-9

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