Ijichi Nobuyuki, Kondo Natsuko, Matsumoto Rena, Shimada Masakazu, Ishikawa Hajime, Fukatsu Takema
National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8566, Japan.
Appl Environ Microbiol. 2002 Aug;68(8):4074-80. doi: 10.1128/AEM.68.8.4074-4080.2002.
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 10(8), 2 x 10(8), and 5 x 10(7) 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.
赤拟谷盗(Callosobruchus chinensis)感染了属于沃尔巴克氏体属(Wolbachia)的三种不同谱系的内共生细菌,分别命名为wBruCon、wBruOri和wBruAus。为了了解感染这三种微生物的潜在机制,采用定量PCR技术详细研究了这三种沃尔巴克氏体菌株的时空感染动态。在赤拟谷盗的发育过程中,wBruCon、wBruOri和wBruAus的感染水平持续上升,但增长模式有所不同。对于wBruCon、wBruOri和wBruAus,感染水平在蛹期达到平稳状态,每只昆虫的 wsp 拷贝当量分别约为3×10⁸、2×10⁸和5×10⁷。在全虫水平上,三种沃尔巴克氏体类型的种群密度在成年雄性和雌性之间没有显著差异。然而,在组织水平上,当比较不同组织和器官以及比较来自雄性和雌性的相同组织时,三种沃尔巴克氏体类型的总密度和相对水平存在显著差异。通过原位杂交和电子显微镜获得的组织学数据与定量PCR分析结果一致。基于组织学数据以及在滋养组织和卵母细胞中常见的特殊沃尔巴克氏体组成,我们认为沃尔巴克氏体菌株不是直接垂直传播到卵母细胞,而是通过滋养组织传播。根据我们的研究结果,我们讨论了三种共感染的沃尔巴克氏体类型之间的相互作用、沃尔巴克氏体的生殖策略以及涉及不同细胞质不亲和表型的因素。