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Applied and Environmental Microbiology, August 2002, p. 4074-4080, Vol. 68, No. 8
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.8.4074-4080.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Internal Spatiotemporal Population Dynamics of Infection with Three Wolbachia Strains in the Adzuki Bean Beetle, Callosobruchus chinensis (Coleoptera: Bruchidae)

Nobuyuki Ijichi,1,2 Natsuko Kondo,3 Rena Matsumoto,1 Masakazu Shimada,3 Hajime Ishikawa,2,4 and Takema Fukatsu1*

National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8566,1 Department of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo 113-0033,2 Department of Systems Sciences (Biology), University of Tokyo, Tokyo 153-8902,3 University of the Air, Chiba 261-8586, Japan4

Received 19 February 2002/ Accepted 23 April 2002

The adzuki bean beetle, Callosobruchus chinensis, is infected with three distinct lineages of endosymbiotic bacteria belonging to the genus Wolbachia, which were designated wBruCon, wBruOri, and wBruAus. In an attempt to understand the mechanisms underlying the infection with these three organisms, the spatiotemporal infection dynamics of the three Wolbachia strains was investigated in detail by using a quantitative PCR technique. During the development of C. chinensis, the wBruCon, wBruOri, and wBruAus infection levels consistently increased but the growth patterns were different. The levels of infection plateaued at the pupal stage at approximately 3 x 108, 2 x 108, and 5 x 107 wsp copy equivalents per insect for wBruCon, wBruOri, and wBruAus, respectively. At the whole-insect level, the population densities of the three Wolbachia types did not show remarkable differences between adult males and females. At the tissue level, however, the total densities and relative levels of the three Wolbachia types varied significantly when different tissues and organs were compared and when the same tissues derived from males and females were compared. The histological data obtained by in situ hybridization and electron microscopy were concordant with the results of quantitative PCR analyses. Based on the histological data and the peculiar Wolbachia composition commonly found in nurse tissues and oocytes, we suggest that the Wolbachia strains are vertically transmitted to oocytes not directly, but by way of nurse tissue. On the basis of our results, we discuss interactions among the three coinfecting Wolbachia types, reproductive strategies of Wolbachia, and factors involved in the different cytoplasmic incompatibility phenotypes.

* Corresponding author. Mailing address: National Institute of Advanced Industrial Science and Technology (AIST), AIST Tsukuba Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan. Phone: 81-298-61-6087. Fax: 81-298-61-6080. E-mail: t-fukatsu{at}aist.go.jp.

Applied and Environmental Microbiology, August 2002, p. 4074-4080, Vol. 68, No. 8
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.8.4074-4080.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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