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Journal of Bacteriology, October 2004, p. 6626-6633, Vol. 186, No. 19
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.19.6626-6633.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Coexistence of Wolbachia with Buchnera aphidicola and a Secondary Symbiont in the Aphid Cinara cedri

Laura Gómez-Valero,1 Mario Soriano-Navarro,1 Vicente Pérez-Brocal,1 Abdelaziz Heddi,2 Andrés Moya,1 José Manuel García-Verdugo,1 and Amparo Latorre1*

Institut Cavanilles de Biodiversitat i Biologia Evolutiva, Universitat de València, València, Spain,1 Laboratoire de Biologie Fonctionnelle Insectes et Interactions, UMR INRA/INSA de Lyon, Villeurbanne Cedex, France2

Received 19 May 2004/ Accepted 7 July 2004

Intracellular symbiosis is very common in the insect world. For the aphid Cinara cedri, we have identified by electron microscopy three symbiotic bacteria that can be characterized by their different sizes, morphologies, and electrodensities. PCR amplification and sequencing of the 16S ribosomal DNA (rDNA) genes showed that, in addition to harboring Buchnera aphidicola, the primary endosymbiont of aphids, C. cedri harbors a secondary symbiont (S symbiont) that was previously found to be associated with aphids (PASS, or R type) and an {alpha}-proteobacterium that belongs to the Wolbachia genus. Using in situ hybridization with specific bacterial probes designed for symbiont 16S rDNA sequences, we have shown that Wolbachia was represented by only a few minute bacteria surrounding the S symbionts. Moreover, the observed B. aphidicola and the S symbionts had similar sizes and were housed in separate specific bacterial cells, the bacteriocytes. Interestingly, in contrast to the case for all aphids examined thus far, the S symbionts were shown to occupy a similarly sized or even larger bacteriocyte space than B. aphidicola. These findings, along with the facts that C. cedri harbors the B. aphidicola strain with the smallest bacterial genome and that the S symbionts infect all Cinara spp. analyzed so far, suggest the possibility of bacterial replacement in these species.

* Corresponding author. Mailing address: Institut Cavanilles de Biodiversitat i Biologia Evolutiva, Universitat de València, Apartat 2085, 46071 València, Spain. Phone: (34) 963543649. Fax: (34) 963543670. E-mail: amparo.latorre{at}uv.es.

Journal of Bacteriology, October 2004, p. 6626-6633, Vol. 186, No. 19
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.19.6626-6633.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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