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Tamarixia radiata is one of the established biocontrol pests against the major Asian citrus psyllid, Diaphorina citri, a vector of Candidatus Liberibacter that is a causal agent of citrus Huanglongbing (HLB) disease. Updated information and regional exploration on biocontrol pests are important elements for effective disease management strategies. In this study, the diversity and parasitism rate of T. radiata populations were evaluated. Due to the importance of the host–parasitoid relationship, the presence of Wolbachia as an endosymbiont was also investigated. The parasitism rate of various T. radiata populations from Ecuador and China ranged between 57.27% and 66.32%, respectively, with a non-significant emergence rate and a statistically similar sex ratio. Sequence analysis of ITS and COI from T. radiata populations was consistent with the morphological hypothesis that the collections represent a single species, whereas phylogeny of the wsp gene confirmed the presence of Wolbachia pipientis as an endosymbiont within T. radiata populations. Based on partial COI sequences, the maximum genetic diversity such as total haplotype diversity (Hd = 0.788), nucleotide, diversity (π = 0.2439), and average nucleotide difference (k = 171.844) was also estimated for different T. radiata populations. Furthermore, neutrality tests based on COI sequences indicated an overall contraction in T. radiata populations, whereas an expansion trend was observed in associated W. pipientis strains. This study clearly demonstrated the presence of genetically diverse T. radiata populations that were able to parasitize D. citri effectively, and these can be further explored as promising biocontrol candidates in integrated pest management strategies to solve citriculture economic loss caused by D. citri. [ABSTRACT FROM AUTHOR]