Phosphorus (P) is an essential nutrient for plants and is acquired from the rhizosphere solution as inorganic phosphate. P is one of the least available mineral nutrients, particularly in highly weathered, tropical soils, and can substantially limit plant growth. The aim of this work was to study a possible effect of root morphology and the expression pattern of related candidate genes on P efficiency in maize. Our field phenotyping results under low and high P conditions enabled us to identify two contrasting genotypes for P acquisition efficiency that were used for the root traits studies. Root morphology was assessed in a paper pouch system to investigate root traits that could be involved in P acquisition efficiency. The genes, Rtcs, Bk2 and Rth3, which are known to be involved in root morphology, showed higher expression in the P efficient line relative to the P inefficient line. Overall, root traits showed high heritability and a low coefficient of variation. Principal component analysis revealed that out of the 24 root traits analysed, only four root traits were needed to adequately represent the diversity among genotypes. The information generated by this study will be useful for establishing early selection strategies for P efficiency in maize, which are needed to support subsequent molecular and physiological studies.