Abstract
The cultivable bacterial communities associated with four South China Sea sponges—Stelletta tenuis, Halichondria rugosa, Dysidea avara, and Craniella australiensis in mixed cultures—were investigated by microbial community DNA-based DGGE fingerprinting and 16S rDNA phylogenetic analysis. Diverse bacteria such as α-, γ-, δ-Proteobacteria, Bacteroidetes, and Firmicutes were cultured, some of which were previously uncultivable bacteria, potential novel strains with less than 95% similarity to their closest relatives and sponge symbionts growing only in the medium with the addition of sponge extract. According to 16S rDNA BLAST analysis, most of the bacteria were cultured from sponge for the first time, although similar phyla of bacteria have been previously recognized. The selective pressure of sponge extract on the cultured bacterial species was suggested, although the effect of sponge extract on bacterial community in high nutrient medium is not significant. Although α- and γ-Proteobacteria appeared to form the majority of the dominant cultivable bacterial communities of the four sponges, the composition of the cultivable bacterial community in the mixed culture was different, depending on the medium and sponge species. Greater bacterial diversity was observed in media C and CS for Stelletta tenuis, in media F and FS for Halichondria rugosa and Craniella australiensis. S. tenuis was found to have the highest cultivable bacterial diversity including α-, γ-, δ-Proteobacteria, Bacteroidetes, and Firmicutes, followed by sponge Dysidea avara without δ-Proteobacteria, sponge Halichondria rugosa with only α-, γ-Proteobacteria and Bacteroidetes, and sponge C. australiensis with only α-, γ-Proteobacteria and Firmicutes. Based on this study, by the strategy of mixed cultivation integrated with microbial community DNA-based DGGE fingerprinting and phylogenetic analysis, the cultivable bacterial community of sponge could be revealed effectively.
Similar content being viewed by others
References
Cebron A, Coci M, Garnier J (2004) Denaturing gradient gel electrophoretic analysis of ammonia-oxidizing bacterial community structure in the lower Seine River: impact of Paris waster water effluents. Appl Environ Microbiol 70:6726–6737
Hentschel U, Usher KM, Taylor MW (2006) Marine sponges as microbial fermenters. FEMS Microbiol Ecol 55:167–177
Hentschel U, Schmid M, Wagner M, Fieseler L, Gernert C, Hacker J (2001) Isolation and phylogenetic analysis of bacteria with antimicrobial activities from the Mediterranean sponges Aplysina aerophoba and Aplysina cavernicola. FEMS Microbiol Ecol 35305–35312
Kaeberlein T, Lewis K, Epstein SS (2002) Isolating “uncultivable” microorganisms in pure culture in a simulated natural environment. Science 296:1127–1129
Kennedy J, Marchest J (2007) Metagenomic approaches to exploit the biotechnological potential of the microbial consortia of marine sponges. Appl Microbiol Biotechnol 75:11–20
Li Z, Liu Y (2006) Marine sponge Craniella austrialiensis-associated bacterial diversity revelation based on 16S rDNA library and biologically active Actinomycetes screening, phylogenetic analysis. Lett Appl Microbiol 43:410–416
Li Z, He L, Wu J, Jiang Q (2006) Bacterial community diversity associated with four marine sponges from the South China Sea based on 16S rDNA-DGGE fingerprinting. J Exp Marine Biol Ecol 329:75–85
Li Z, Hu Y, Liu Y, Huang Y, He L, Miao X (2007) 16S rDNA clone library based bacterial phylogenetic diversity associated with three South China Sea sponges. World J Microbiol Biotechnol 23:1265–1272
Li Z, Hu Y, Huang Y, Huang Y (2007) Three South China Sea sponges associated biologically active bacterial isolation and phylogenetic analysis. Microbiology 76:494–499
Li Z, Qin E, Jiang Q (2005) Distribution characteristics of sponge Stelletta tenuis associated microorganisms by TEM. J Fisheries China 29:38–42
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 59:695–700
Müller WEG, Brümmer F, Batel R, Müller IZ, Schröder HC (2003) Molecular biodiversity. Case study: Porifera (Sponges). Naturwissensehaften 90:103–120
Olson JB, Lord CC, McCarthy PJ (2000) Improved recoverability of microbial colonies from marine sponge samples. Microbial Ecol 40:139–147
Piel J, Hui D, Wen G, Butzke D, Platzer M, Fusetani N, Matsunaga S (2004) Antitumor polyketide biosynthesis by an uncultivated bacterial symbiont of the marine sponge Theonella swinhoei. Proc Natl Acad Sci USA 101:16222–16227
Pimentel-Elardo S, Wehrl M, Friedrich AB, Jensen PR, Hentschel U (2003) Isolation of planctomycetes from Aplysina sponges. Aquatic Microb Ecol 33:239–245
Ramm W, Schantton W, Wagner-Döbler I, Wray V, Nimtz M, Tokuda H, Enjyo F, Nishino H, Bell W, Heckmann R, Lurtz V, Lang S (2004) Diglucosyl-glycerolipids from the marine sponge-associated Bacillus pumilus strain AAS3: their production, enzymatic modification and properties. Appl Microbiol Biotechnol 64:497–504
Rath J, Wu KY, Herndl GJ, DeLong EF (1998) High phylogenetic diversity in a marine-snow-associated bacterial assemblage. Aquatic Microb Ecol 14:262–269
Rochelle PA, Will JAK, Fry JC, Jenkins GJS, Parkers RJ, Turley CM, Weightman AJ (1995) Extraction and amplification of 16S rRNA genes from deep marine sediments and seawater to assess bacterial community diversity. In: Trevors JT, Elsas JD (eds) Nucleic acids in the environment. Berlin: Springer-Verlag, pp 219–239
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 δ-Proteobacterium,”Candidatus Entotheonella palauensis”. Marine Biol 136:969–977
Thakur NL, Anil AC (2000) Antibacterial activity of the sponge Ircinia ramose: importance of its surface-associated bacteria. J Chem Ecol 26:57–71
Thiel V, Neulinger SC, Staufenberger T, Schmaljohann R, Imhoff JF (2007) Spatial distribution of sponge-associated bacteria in the Mediterranean sponge Tethya aurantium. FEMS Microbiol Ecol 59:47–63
Thiel V, Leininger S, Schmaljohann R, Bruemmer F, Imhoff J (2007) Sponge-specific bacterial associations of the Mediterranean sponge Chondrilla nucula (Demospongiae, Tetractinomorpha). Microbial Ecol 54:101–111
Thiel V, Imhoff JF (2003) Phylogenetic identification of bacteria with antimicrobial activities isolated from Mediterranean sponges. Biomolec Eng 20:421–423
Webster NS, Wilson KJ, Blackall LL, Hill RT (2001) Phylogenetic diversity of bacteria associated with the marine sponge Rhopaloeides odorabile. Appl Environ Microbiol 67:434–444
Webster NS, Hill RT (2001) The culturable microbial community of the Great Barrier Reef sponge Rhopaloeides odorabile is dominated by an alpha-Proteobacterium. Marine Biol 138:843–851
Wichels A, Würtz S, Döpke H, Schütt C, Gerdts G (2006) Bacterial diversity in the breadcrumb sponge Halichondria rugosa panacea (pallas). FEMS Microbiol Ecol 56:102–118
Acknowledgments
Financial supports to Zhiyong Li from “High-Tech Research and Development Program of China” (2007AA09Z447), Rising-star Program of Science & Technology, Shanghai, China (04QMX1411) are gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Li, Z., He, L. & Miao, X. Cultivable Bacterial Community from South China Sea Sponge as Revealed by DGGE Fingerprinting and 16S rDNA Phylogenetic Analysis. Curr Microbiol 55, 465–472 (2007). https://doi.org/10.1007/s00284-007-9035-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00284-007-9035-2