| Summary: | <i>Wolbachia</i> and <i>Rickettsia</i> are bacterial endosymbionts that can induce a number of reproductive abnormalities in their arthropod hosts. We screened and established the co-infection of <i>Wolbachia</i> and <i>Rickettsia</i> in <i>Bemisia tabaci</i> and compared the spatial and temporal distribution of <i>Wolbachia</i> and <i>Rickettsia</i> in eggs (3–120 h after spawning), nymphs, and adults of <i>B. tabaci</i> by qPCR quantification and fluorescent in situ hybridization (FISH). The results show that the titer of <i>Wolbachia</i> and <i>Rickettsia</i> in the 3–120 h old eggs showed a “w” patterned fluctuation, while the titers of <i>Wolbachia</i> and <i>Rickettsia</i> had a “descending–ascending descending–ascending” change process. The titers of <i>Rickettsia</i> and <i>Wolbachia</i> nymphal and the adult life stages of Asia II1 <i>B. tabaci</i> generally increased with the development of whiteflies. However, the location of <i>Wolbachia</i> and <i>Rickettsia</i> in the egg changed from egg stalk to egg base, and then from egg base to egg posterior, and finally back to the middle of the egg. These results will provide basic information on the quantity and localization of <i>Wolbachia</i> and <i>Rickettsia</i> within different life stages of <i>B. tabaci.</i> These findings help to understand the dynamics of the vertical transmission of symbiotic bacteria.
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