Skip to main content
SpringerLink
Log in

Microbial epibionts of the colonial ascidians Didemnum galacteum and Cystodytes sp.

  • Published:
Symbiosis Aims and scope Submit manuscript

Abstract

Symbiosis with microorganisms has been well documented for many marine invertebrate taxa. However, knowledge of the diversity of microorganisms associated with ascidians is still limited. This study assessed the microbial epibionts of Didemnum galacteum and Cystodytes sp., two ascidian species collected from the western coast of Ceará state (Brazil), at Dois Coqueiros beach and the port of Pecém, respectively. The microbiota were examined using optical microscopy, followed by subsequent analysis of fingerprinting profiles obtained by denaturing gradient gel electrophoresis (DGGE) and 16S rRNA clone libraries. The microscopy analysis showed for both ascidians a community comprising cyanobacteria, mainly Prochloron-like species, and diatoms. The DGGE results indicated that D. galacteum hosts a more diverse microbiota when compared to Cystodytes sp. The same analysis also suggested that the diversity of the seawater microbiota was higher than the diversity of the ascidian-associated microbiota. The analysis of the 16S rRNA clone libraries revealed the dominance of Proteobacteria symbionts associated with both ascidians, with Alphaproteobacteria as the major component in D. galacteum and Gammaproteobacteria the major component in Cystodytes sp. The analysis of the clone libraries also revealed the presence of other taxa such as Bacteroidetes, Planctomycetes, Actinobacteria, Cyanobacteria, and uncultured bacteria in D. galacteum, but not in Cystodytes sp. Among the bacteria found to be exclusively associated with the ascidians, none were shared by the two studied hosts. The combined results point to a diverse microbiota associated with the external surface of the ascidians, with a mixed composition including organisms typically found in the surrounding seawater, but also a more specific set of taxa.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Behrendt L, Larkum AWD, Trampe E, Norman A, Sørensen SJ, Kühl M (2012) Microbial diversity of biofilm communities in microniches associated with the didemnid ascidian Lissoclinum patella. ISME J 6:1222–1237

    Article  PubMed  CAS  Google Scholar 

  • Bright M, Bulgheresi S (2010) A complex journey: transmission of microbial symbionts. Nat Rev Microbiol 8:218–230

    Article  PubMed  CAS  Google Scholar 

  • Cox GC (1986) Comparison of prochloron from different hosts. i. structural and ultrastructural characteristics. New Phytol 104:429–445

    Article  Google Scholar 

  • Donia MS, Fricke WF, Partensky F, Cox J, Elshahawi SI, White JR, Phillippy AM, Schatz MC, Piel J, Haygood MG, Ravel J, Schmidt EW (2011) Complex microbiome underlying secondary and primary metabolism in the tunicate-Prochloron symbiosis. P Natl Acad Sci USA 108:E1423–E1432

    Article  CAS  Google Scholar 

  • Dunlap WC, Battershill CN, Liptrot CH, Cobb RE, Bourne DG, Jaspars M, Long PF, Newman DJ (2007) Biomedicinals from the photosymbionts of marine invertebrates: a molecular approach. Methods 42:358–376

    Article  PubMed  CAS  Google Scholar 

  • Groepler W, Schuett C (2003) Bacterial community in the tunic matrix of a colonial ascidian Diplosoma migrans. Helgoland Mar Res 57:139–143

    Article  Google Scholar 

  • Hirose E, Fukuda T (2006) Vertical transmission of photosymbionts in the colonial ascidian Didemnum molle: The larval tunic prevents symbionts from attaching to the anterior part of larvae. Zool Sci 23:669–674

    Article  PubMed  Google Scholar 

  • Hirose E, Hirose M, Neilan BA (2006) Localization of symbiotic cyanobacteria in the colonial ascidian Trididemnum miniatum (Didemnidae, Ascidiacea). Zool Sci 23:435–442

    Article  PubMed  Google Scholar 

  • Hirose E, Uchida H, Murakami A (2009) Ultrastructural and microspectrophotometric characterization of multiple species of cyanobacterial photosymbionts coexisting in the colonial ascidian Trididemnum clinides (Tunicata, Ascidiacea, Didemnidae). Eur J Phycol 44:365–375

    Article  Google Scholar 

  • Jimenez PC, Fortier SC, Lotufo TMC, Pessoa C, Moraes MEA, Moraes MO, Costa-Lotufo LV (2003) Biological activity in extracts of ascidians (Tunicata, Ascidiacea) from the northeastern Brazilian coast. J Exp Mar Biol Ecol 287:93–101

    Article  Google Scholar 

  • Kostka JE, Prakash O, Overholt WA, Green SJ, Freyer G, Canion A, Delgardio J, Norton N, Hazen TC, Huettel M (2011) Hydrocarbon-degrading bacteria and the bacterial community response in Gulf of Mexico beach sands impacted by the Deepwater Horizon oil spill. Appl Environ Microbiol 77:7962–7974

    Google Scholar 

  • Kott P (1984) Related species of Trididemnum in symbiosis with Cyanophyta. P Linn Soc NSW 107:515–520

    Google Scholar 

  • Kott P, Parry DL, Cox GC (1984) Prokaryotic symbionts with a range of ascidian hosts. B Mar Sci 34:308–312

    Google Scholar 

  • Lambert G (2005) Ecology and natural history of the Protochordates. Can J Zoolog 83:34–50

    Article  Google Scholar 

  • Lewin RA, Cheng L, Lafargue F (1980) Prochlorophytes in the Caribbean. B Mar Sci 30:744–745

    Google Scholar 

  • López-Legentil S, Song B, Bosch M, Pawlik JR, Turon X (2011) Cyanobacterial diversity and a new Acaryochloris-like symbiont from Bahamian sea-squirts. PLoS One 6:1–12

    Article  Google Scholar 

  • Lotufo TMC, Dias GM (2007) Didemnum galacteum, a new species of white Didemnid (Chordata: Ascidiacea: Didemnidae) from Brazil. Proc Biol Soc Wash 120:137–142

    Article  Google Scholar 

  • Lotufo TMC, Silva AMB (2006) Ascidiacea. In: Matthews-Cascon H, Lotufo TMC (eds) Biota Marinha da Costa Oeste do Ceará. Ministério do Meio Ambiente, Brasília, pp 221–248

    Google Scholar 

  • Martínez-García M, Díaz-Valdes M, Ramos-Esplá A, Salvador N, Lopez P, Larriba E, Antón J (2007a) Cytotoxicity of the ascidian Cystodytes dellechiajei against tumor cells and study of the involvement of associated microbiota in the production of cytotoxic compounds. Mar Drug 5:52–70

    Article  Google Scholar 

  • Martínez-García M, Díaz-Valdés M, Wanner G, Ramos-Esplá A, Antón J (2007b) Microbial community associated with the colonial ascidian Cystodytes dellechiajei. Environ Microbiol 9:521–534

    Article  PubMed  Google Scholar 

  • Martínez-García M, Díaz-Valdés M, Antón J (2010) Diversity of pufM genes, involved in aerobic anoxygenic phtosynthesis, in the bacterial communities associated with colonial ascidians. FEMS Microbiol Ecol 71:387–398

    Article  PubMed  Google Scholar 

  • Martínez-Garcia M, Koblĩzek M, López-Legentil S, Antón J (2011) Epibiosis of oxygenic phototrophs containing chlorophylls a, b, c and d on the colonial ascidian Cystodytes dellechiajei. Microbiol Ecol 61:13–19

    Article  Google Scholar 

  • Menezes CBA, Bonugli-Santos RC, Miqueletto PB, Passarini MRZ, Silva CHD, Justo MR, Leal RR, Fantinatti-Garboggini F, Oliveira VM, Berlinck RGS, Sette LD (2010) Microbial diversity associated with algae, ascidians and sponges from the north coast of São Paulo state, Brazil. Microbiol Res 165:466–482

    Article  PubMed  Google Scholar 

  • Millar RH (1971) The biology of ascidians. Adv Mar Biol 9:1–100

    Article  Google Scholar 

  • Monniot C, Monniot F, Laboute P (1991) Coral reef ascidians of New Caledonia. collections faune tropicale. ORSTOM Editions, Paris

    Google Scholar 

  • Moss C, Green DH, Pérez B, Velasco A, Henríquez R, Mckenzie JD (2003) Intracellular bacteria associated with the ascidian Esteinascidia turbinata: phylogenetic and in situ hydridization analysis. Mar Biol 14:99–110

    Article  Google Scholar 

  • Nogales B, Aguiló-Ferretjans MM, Martín-Cardona C, Latucat J, Bosch R (2007) Bacterial diversity, composition and dynamics in and around recreational coastal areas. Environ Microbiol 9:1913–1929

    Article  PubMed  CAS  Google Scholar 

  • Oka AT, Suetsugu M, Hirose E (2005) Two new species of Diplosoma (Ascidiacea: Didemnidae) bearing prokaryotic algae Prochloron from Okinawajima (Ryukyu Archipelago, Japan). Zool Sci 22:367–374

    Article  PubMed  Google Scholar 

  • Øvreås L, Forney L, Daae FL, Torsvik V (1997) Distribuition of bacterioplankton in meromictic Lake Saelenvannet as determined by denaturing gradient gel electrophoresis of PCR amplified gene fragments coding for 16S rRNA. Appl Environ Microbiol 63:3367–3373

    PubMed  Google Scholar 

  • Pérez-Matos AE, Rosado W, Govind NS (2007) Bacterial diversity associated with the Caribbean tunicate Ecteinascidia turbinata. Anton Leeuw Int J G 92:155–164

    Article  Google Scholar 

  • Roger SO, Bendich AJ (1985) Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant-tissues. Plant Mol Biol 5:69–76

    Article  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Schmidt EW, Sudek S, Haygood MG (2004) Genetic evidence supports secondary metabolic diversity in Prochloron spp., the cyanobacterial symbiont of a tropical ascidian. J Nat Prod 67:1341–1345

    Article  PubMed  CAS  Google Scholar 

  • Schmidt EW, Donia MS, McIntosh JA, Fricke WF, Ravel J (2012) Origin and variation of tunicate secondary metabolites. J Nat Prod 75:295–304

    Article  PubMed  CAS  Google Scholar 

  • Schuett C, Doepke H, Groepler W, Wichels A (2005) Diversity of intratunical bacteria in the tunic matrix of the colonial ascidian Diplosoma migrans. Helgoland Mar Res 59:136–140

    Article  Google Scholar 

  • Tait E, Carman M, Sievert SM (2007) Phylogenetic diversity of bacteria associated with ascidians in Eel Pond (Woods Hole, Massachusetts, USA). J Exp Mar Biol Ecol 342:138–146

    Article  Google Scholar 

Download references

Acknowledgment

This research project was funded through a scholarship for F.A.S.O. from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior.

Author information

Authors and Affiliations

  1. Programa de Pós-Graduação em Engenharia de Pesca, Departamento de Engenharia de Pesca, Universidade Federal do Ceará, 60021-970, Fortaleza, Brazil

    Francisca Andréa da Silva Oliveira & Tito M. C. Lotufo

  2. Laboratório de Ecologia Microbiana e Biotecnologia, Departamento de Biologia, Universidade Federal do Ceará, 60455-760, Fortaleza, Brazil

    Geórgia B. Colares, Denise C. Hissa, Alysson L. Angelim & Vânia M. M. Melo

  3. Laboratório de Ecologia Animal, Instituto de Ciências do Mar, Universidade Federal do Ceará, 60165-081, Fortaleza, Brazil

    Francisca Andréa da Silva Oliveira & Tito M. C. Lotufo

Authors
  1. Francisca Andréa da Silva Oliveira
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Geórgia B. Colares
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Denise C. Hissa
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Alysson L. Angelim
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Vânia M. M. Melo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tito M. C. Lotufo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tito M. C. Lotufo.

Rights and permissions

Reprints and permissions

About this article

Cite this article

da Silva Oliveira, F.A., Colares, G.B., Hissa, D.C. et al. Microbial epibionts of the colonial ascidians Didemnum galacteum and Cystodytes sp.. Symbiosis 59, 57–63 (2013). https://doi.org/10.1007/s13199-012-0210-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13199-012-0210-2

Keywords

Search

Navigation