We report the results of a 2-yr survey that determined some of the host plant and parasitoid associations of Anastrepha fruit flies (Diptera: Tephritidae) in the “Montes Azules” tropical rainforest biosphere reserve (State of Chiapas, Mexico). We collected a total of 57.38 kg of fruit representing 47 native species from 23 plant families. Of these, 13 plant species from eight plant families were found to be native hosts of 9 species of Anastrepha. The following Anastrepha host associations were observed: Bellucia pentamera Naudin (Melastomataceae) with A. coronilli Carrejo y González; Malmea gaumeri (Greenm.) Lundell (Annonaceae) with A. bahiensis Lima; Tabernamontana alba Mill. (Apocynaceae) with A. cordata Aldrich; Quararibea yunckeri Standl. (Bombacaceae) with A. crebra Stone; Ampelocera hottlei (Standl.) Standl. (Ulmaceae) with A. obliqua (Macquart) and A. fraterculus (Wiedemann); Zuelania guidonia Britton and Millsp. and Casearia tremula (Griseb.) Griseb. ex C. Wright (Flacourtaceae) with A. zuelaniae Stone; Psidium sartorianum (O. Berg.) Nied (Myrtaceae) with A. fraterculus; Psidium guajava L. and P. sartorianum (Myrtaceae) with A. striata Schiner; and Manilkara zapota (L.) Van Royen, Pouteria sp., Bumelia sebolana Lundell, and Calocarpum mammosum (L.) Pierre (Sapotaceae) with A. serpentina (Wiedemann). The following are new host plant records: Malmea gaumeri for A. bahiensis; Quararibea yunckeri for A. crebra; Ampelocera hottlei for A. fraterculus and A. obliqua; Bumelia sebolana for A. serpentina; and Casearia tremula for A. zuelaniae. A. coronilli is reported for the first time in Mexico. Infestation levels were variable and ranged between 0 and 1.63 larvae/g of fruit depending on host species. Larvae of eight species of Anastrepha on nine plant species from six plant families were found to be parasitized by Doryctobracon areolatus Szepligeti, D. crawfordi Viereck, D. zeteki Musebeck (new report for Mexico and northernmost record for the species), Opius hirtus Fisher, Utetes anastrephae Viereck (all Hymenoptera: Braconidae), and Aganaspis pelleranoi Brethes (Hymenoptera: Figitidae). Percent parasitism ranged from 0 to 76.5%. We discuss our findings in light of their practical (e.g., biological control) and theoretical (e.g., species radiation) implications and highlight the importance of these types of studies given the rampant deforestation of tropical forests in Latin America and the risk of extinction of rare fruit fly species that could shed light on the evolution of host plant and parasitoid associations within the genus Anastrepha.