A study was performed to determine the effect of arbuscular mycorrhizal (AM) fungal inoculation, using a locally isolated Claroideoglomus claroideum (Gc) ecotype, on the seedling development of chilean pepper plants, and to select an appropriate growth substrate. The first experiment consisted of two stages: (i) a seedling production stage of pepper plants were germinated and grown, inoculated or not with C. claroideum; and (ii) a seedling transplanting stage, in which seedlings were transplanted to a wider capacity containers, using the same substrates as in the seedling production stage. These were inoculated or not with C. claroideum and/or Penicillium albidum (Pa). A soil mixture was selected as the target substrate for further studies. A second experiment was then carried out and three new phosphate-solubilizing Penicillium that is, Penicillium frequentans, Penicillium jensenii and Penicillium restrictum, were incorporated as inoculation variables, in addition to the already tested P. albidum, inoculated either alone or in co-inoculation with C. claroideum. After 28 weeks, plants were harvested and fruit number, weight, and length were recorded. A synergistic interaction between C. claroideum and P. albidum to improve fruit weight and phosphorous (P) concentration was evidenced, suggesting a sustainable alternative for chilean pepper production.