The Andean Puna is an arid, high-elevation plateau in which plants such as grasses experience high abiotic stress and distinctive environmental conditions. We assessed colonization by arbuscular mycorrhizal fungi (AMF) and dark septate endophytes (DSE) in the roots of 20 native grass species and examined the relationship between root-associated fungi (AMF and DSE) as a function of the elevation of study sites, the photosynthetic pathways of the grass hosts, and the hosts' life cycles. In general, grasses were co-colonized by AMF and DSE and the colonization by AMF and DSE was not extensive. The extension of colonization of AMF and that of DSE were positively correlated, as were number of arbuscules and DSE colonization extension. The extension of AMF colonization differed among sites with different elevations, but DSE colonization was similar across sites. Overall, AMF and DSE patterns shifted as a function of elevation in most grass species, with no general trends observed with respect to host photosynthetic pathway or life cycle. In general, our observations differ from previous studies in the Northern Hemisphere. Variation among sites in AMF and DSE colonization was greater than variation that could be explained by the other factors considered here, suggesting a strong influence of environmental factors. We predict that both AMF and DSE may have established synergistic and beneficial associations with grasses in these distinctive and harsh ecosystems.