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Applied and Environmental Microbiology, September 2009, p. 5676-5686, Vol. 75, No. 17
0099-2240/09/$08.00+0     doi:10.1128/AEM.01172-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Complete WO Phage Sequences Reveal Their Dynamic Evolutionary Trajectories and Putative Functional Elements Required for Integration into the Wolbachia Genome{triangledown} ,{dagger}

Kohjiro Tanaka,1 Seiichi Furukawa,2 Naruo Nikoh,3 Tetsuhiko Sasaki,2 and Takema Fukatsu1*

National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8566, Japan,1 Honeybee Science Research Center, Tamagawa University, Tokyo 194-8610, Japan,2 Division of Natural Sciences, The Open University of Japan, Chiba 261-8586, Japan3

Received 20 May 2009/ Accepted 1 July 2009

Wolbachia endosymbionts are ubiquitously found in diverse insects including many medical and hygienic pests, causing a variety of reproductive phenotypes, such as cytoplasmic incompatibility, and thereby efficiently spreading in host insect populations. Recently, Wolbachia-mediated approaches to pest control and management have been proposed, but the application of these approaches has been hindered by the lack of genetic transformation techniques for symbiotic bacteria. Here, we report the genome and structure of active bacteriophages from a Wolbachia endosymbiont. From the Wolbachia strain wCauB infecting the moth Ephestia kuehniella two closely related WO prophages, WOcauB2 of 43,016 bp with 47 open reading frames (ORFs) and WOcauB3 of 45,078 bp with 46 ORFs, were characterized. In each of the prophage genomes, an integrase gene and an attachment site core sequence were identified, which are putatively involved in integration and excision of the mobile genetic elements. The 3' region of the prophages encoded genes with sequence motifs related to bacterial virulence and protein-protein interactions, which might represent effector molecules that affect cellular processes and functions of their host bacterium and/or insect. Database searches and phylogenetic analyses revealed that the prophage genes have experienced dynamic evolutionary trajectories. Genes similar to the prophage genes were found across divergent bacterial phyla, highlighting the active and mobile nature of the genetic elements. We suggest that the active WO prophage genomes and their constituent sequence elements would provide a clue to development of a genetic transformation vector for Wolbachia endosymbionts.

* Corresponding author. Mailing address: National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba 305-8566, Japan. Phone: 81 29 861 6087. Fax: 81 29 861 6080. E-mail: t-fukatsu{at}aist.go.jp

{triangledown} Published ahead of print on 10 July 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, September 2009, p. 5676-5686, Vol. 75, No. 17
0099-2240/09/$08.00+0     doi:10.1128/AEM.01172-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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