Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a major pest of the tomato (Solanum lycopersicum L.) crop worldwide, mostly because of resistance to insecticide that has been associated with endosymbionts. Although knowledge of biotypes in the B. tabaci complex and its endosymbionts is critical for developing pest management strategies, limited information is available on the pest in the Culiacan Province of Mexico. In total, 249 samples of whiteflies were collected from tomato plants of four growers in the Culiacan Valley to analyze the genetic identity of the B. tabaci complex and its endosymbionts. PCR and phylogenetic analyses on the mitochondrial cytochrome oxidase I gene (mtCO1) revealed MEAM1, MED, and indigenous genetic groups in 53, 19, and 28%, respectively, of B. tabaci on tomatoes. Individual whiteflies were examined for endosymbionts through specific primers and phylogenetic analyses on the 16S rRNA and the 23S rRNA genes. Incidence of Portiera, Hamiltonella, Rickettsia, Wolbachia, and Cardinium was 81, 46, 28, 9, and 13%, respectively, in the B. tabaci samples analyzed. A new bacterium-endosymbiont in the genus Orientia was found and described in 63% of the samples of B. tabaci. A mixture of the endosymbionts Rickettsia+UAS_511+Cardinium was observed in 10% of samples of B. tabaci MED, while in the MEAM1 biotype, a combination of Hamiltonella+UAS_511 was in 32% of the samples analyzed. The study highlights the diversity and distribution of B. tabaci and its endosymbionts throughout the Culiacan Province and provides evidence for control of the insect pest.