Arbuscular mycorrhizal fungi (AMF) have been hypothesized to enhance plant adaptation and growth in petroleum-contaminated soils. Nevertheless, neither AMF-biodiversity under chronically petroleum-contaminated soils nor spore germination response to petroleum hydrocarbons has been well studied. Chronically petroleum-contaminated rhizosphere soil and roots from Echinochloa polystachya, Citrus aurantifolia and C. aurantium were collected from Activo Cinco Presidentes, Tabasco, Mexico. Root colonization and spore abundance were evaluated. Additionally, rhizosphere soil samples were propagated using Sorghum vulgare L. as a plant trap under greenhouse conditions; subsequently, AMF-spores were identified. AMF-colonization ranged from 63 to 77% while spore number ranged from 715 to 912 in 100 g soil, suggesting that AMF tolerate the presence of petroleum hydrocarbons in the rhizosphere. From grass species, four AMF-morphospecies were identified: Glomus ambisporum, G. sinuosum (previously described as Sclerocystis sinuosum), Acaulospora laevis, and Ambispora gerdermanni. From citrus trees, four AMF-species were also identified: Scutellospora heterogama, G. ambisporum, Acaulospora scrobiculata, and G. citricola. In a second study, it was observed that spore germination and hyphal length of G. mosseae, G. ambisporum, and S. heterogama were significantly reduced by either volatile compounds of crude oil or increased concentrations of benzo[a ]pyrene or phenanthrene in water-agar.