Phosphate (P) uptake is critical for plant growth, but to date little is known about P uptake and transport in the pasture grass Lolium perenne L. We have identified a putative P transporter (PT) from L. perenne mycorrhizal roots (LpPT1) and assessed its transcriptional regulation by soil P availability and mycorrhizal colonisation. We also investigated transcript levels of fungal PTs from the two arbuscular mycorrhizal species Rhizophagus intraradices and Funneliformis mosseae. Our analyses indicated that LpPT1 codes for a high affinity PT most likely responsible for direct P uptake from the soil. LpPT1 is highly expressed in roots of plants grown at low P, whereas high P repressed its expression. LpPT1 was not expressed in above-ground plant tissues. Colonisation with R. intraradices did not affect expression of LpPT1 significantly. Transcript levels of the R. intraradices PT were not affected by P availability but the F. mosseae PT was repressed by high P supply, particularly in intraradical hyphae. Our study could assist in deciphering the molecular mechanisms of P uptake in the pasture grass L. perenne.