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Applied and Environmental Microbiology, May 2006, p. 3724-3732, Vol. 72, No. 5
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.5.3724-3732.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Characterization of a Culturable Alphaproteobacterial Symbiont Common to Many Marine Sponges and Evidence for Vertical Transmission via Sponge Larvae{dagger}

Julie J. Enticknap,1 Michelle Kelly,2 Olivier Peraud,1 and Russell T. Hill1*

Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore, Maryland 21202,1 National Centre for Aquatic Biodiversity and Biosecurity, National Institute of Water and Atmospheric Research (NIWA) Ltd., Auckland, New Zealand2

Received 28 June 2005/ Accepted 14 March 2006

A closely related group of alphaproteobacteria were found to be present in seven genera of marine sponges from several locations and were shown to be transferred between sponge generations through the larvae in one of these sponges. Isolates of the alphaproteobacterium were cultured from the sponges Axinella corrugata, Mycale laxissima, Monanchora unguifera, and Niphates digitalis from Key Largo, Florida; Didiscus oxeata and Monanchora unguifera from Discovery Bay, Jamaica; an Acanthostronglyophora sp. from Manado, Indonesia; and Microciona prolifera from the Cheasapeake Bay in Maryland. Isolates were very similar to each other on the basis of 16S rRNA gene sequence (>99% identity) and are closely related to Pseudovibrio denitrificans. The bacterium was never isolated from surrounding water samples and was cultured from larvae of M. laxissima, indicating that it is a vertically transmitted symbiont in this sponge. Denaturing gradient gel electrophoresis, 16S rRNA gene clone library analysis, and fluorescent in situ hybridization with probes specific to the alphaproteobacterium confirmed the presence of this bacterium in the M. laxissima larvae. The alphaproteobacterium was densely associated with the larvae rather than being evenly distributed throughout the mesohyl. This is the first report of the successful culture of a bacterial symbiont of a sponge that is transferred through the gametes.

* Corresponding author. Mailing address: Center of Marine Biotechnology, Columbus Center Suite 236, 701 E. Pratt Street, Baltimore, MD 21202. Phone: (410) 234 8883. Fax: (410) 234-8896. E-mail: hillr{at}umbi.umd.edu.

{dagger} Contribution no. 05-118 from the Center of Marine Biotechnology.

Applied and Environmental Microbiology, May 2006, p. 3724-3732, Vol. 72, No. 5
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.5.3724-3732.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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