SUMMARYNon‐mycorrhizal and vesicular‐arbuscular mycorrhizal Bouteloua gracilis infected with Glomus fasciculatus were grown in defined media containing different phosphate sources and compared with respect to phosphate content and form, biomass, chlorophyll concentration, and root phosphatase activity. The phosphate sources were sodium monobasic phosphate, a mixture of sodium monobasic phosphate and calcium phytate, and calcium phytate. Inositol and inositol plus calcium were added to the sodium phosphate medium as additional treatments. Mycorrhizal infection was highest in roots of plants grown in the presence of phytate (75%). Lower root infection levels were noted in plants from the sodium phosphate (19%) and mixed phosphate (22%) media. No penetration by fungi occurred in plants from the sodium phosphate plus inositol or inositol and calcium media. Dry wts of non‐mycorrhizal plants were highest when grown in media containing phytate and sodium phosphate plus inositol and calcium followed in decreasing order by sodium phosphate plus inositol, mixed phosphates, and sodium phosphate. Mycorrhizal infection increased leaf dry wt in plants from the sodium phosphate medium and root dry wt from the phytate medium. Phosphate concentrations in the plants were highest when grown in mixed phosphate medium followed by sodium phosphate and phytate. Mycorrhizal infection always increased significantly leaf phosphate concentrations but increased root phosphate concentrations only in the phytate medium. Phosphates were found predominantly as organically‐bound compounds in leaves of mycorrhizal plants whereas in leaves of non‐mycorrhizal plants, most of the phosphate was inorganic. Chlorophyll concentrations increased significantly with mycorrhizal infection with no change in a/b ratios. Mycorrhizal plants grown in the phytate medium had substantially higher alkaline phosphatase activity than did non‐mycorrhizal plants; acid phosphatase activity was not affected by mycorrhizal condition.These results suggest that form of the phosphate in the root environment influences mycorrhizal establishment and effect of mycorrhizae on plant growth.