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Applied and Environmental Microbiology, August 2005, p. 4833-4839, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4833-4839.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Aphid-Symbiotic Bacteria Cultured in Insect Cell Lines

A. C. Darby,^1* S. M. Chandler,² S. C. Welburn,¹ and A. E. Douglas²

Centre for Infectious Diseases, College of Medicine and Veterinary Medicine, The University of Edinburgh, Easter Bush, Edinburgh EH25 9RG, United Kingdom,¹ Department of Biology, University of York, P.O. Box No. 373, York YO10 5YW, United Kingdom²

Received 7 December 2004/ Accepted 10 March 2005

The cells and tissues of many aphids contain bacteria known as "secondary symbionts," which under specific environmental circumstances may be beneficial to the host insect. Such symbiotic bacteria are traditionally described as intractable to cultivation in vitro. Here we show that two types of aphid secondary symbionts, known informally as T type and U type, can be cultured and maintained in three insect cell lines. The identities of the cultured bacteria were confirmed by PCR with sequencing of 16S rRNA gene fragments and fluorescence in situ hybridization. In cell lines infected with bacteria derived from aphids harboring both T type and U type, the U type persisted, while the T type was lost. We suggest that the two bacteria persist in aphids because competition between them is limited by differences in tropism for insect tissues or cell types. The culture of these bacteria in insect cell lines provides a new and unique research opportunity, offering a source of unibacterial material for genomic studies and a model system to investigate the interactions between animal cells and bacteria. We propose the provisional taxon names "Candidatus Consessoris aphidicola" for T type and "Candidatus Adiaceo aphidicola" for U type.

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* Corresponding author. Mailing address: Centre for Infectious Diseases, College of Medicine and Veterinary Medicine, The University of Edinburgh, Easter Bush, Edinburgh EH25 9RG, United Kingdom. Phone: 44 (0)131 6506261. Fax: 44 (0)131 445 5099. E-mail: Alistair.Darby{at}ed.ac.uk.

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Applied and Environmental Microbiology, August 2005, p. 4833-4839, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4833-4839.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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