Understanding the mechanisms by which nutritional signals impact upon root system architecture is a key facet in the drive for greater nutrient application efficiency in agricultural systems. Cereal plants reduce their rate of lateral root emergence under inorganic phosphate (Pi) shortage; this study uses molecular and pharmacological techniques to dissect this Pi response in Triticum aestivum. Plants were grown in coarse sand washed in high- or low-Pi nutrient solution before being assessed for their root branching density and expression of AUX/IAA and PIN genes. Seedlings were also grown on media containing [ 14C]indole acetic acid to measure basipetal auxin transport. Seedlings grown in low-Pi environments displayed less capacity to transport auxin basipetally from the seminal root apex, a reduction in root expression of PIN auxin transporter genes, and perturbed expression of a range of AUX/IAA auxin response genes. Given the known importance of basipetally transported auxin in stimulating lateral root initiation, it is proposed here that, in T. aestivum, Pi availability directly influences lateral root production through modulation of PIN expression. Understanding such processes is important in the drive for greater efficiency in crop use of Pi fertilizers in agricultural settings.