A growing number of studies have revealed the presence of closely related endosymbionts in phylogenetically distant arthropods, indicating horizontal transmission of these bacteria. Here we investigated the interspecific horizontal transmission of Rickettsia between two globally invasive whitefly species, Bemisia tabaci MEAM1 and B. tabaci MED, via cotton plants. We found both scattered and confined distribution patterns of Rickettsia in these whiteflies. After entering cotton leaves, Rickettsia was restricted to the leaf phloem vessels and could be taken up by both species of the Rickettsia-free whitefly adults, but only the scattered pattern was observed in the recipient whiteflies. Both the relative quantity of Rickettsia and the efficiency of transmitting Rickettsia into cotton leaves were significantly higher in MEAM1 females than in MED females. The retention time of Rickettsia transmitted from MEAM1 into cotton leaves was at least 5 days longer than that of MED. Phylogenetic analysis based on 16S rRNA and gltA genes confirmed that the Rickettsia extracted from the donor MEAM1, the cotton leaves, the recipient MEAM1 and the recipient MED were all identical. We conclude that cotton plants can mediate horizontal transmission of Rickettsia between different insect species, and that the transmission dynamics of Rickettsia vary with different host whitefly species.