Background & aim This study tested the hypothesis that manure incorporation in the top of the B horizon of a texture-contrast duplex soil would increase root growth and water uptake by pasture grasses. Methods It involved soil columns consisting of a sandy loam topsoil over a dense clay. Four soil treatments included (i) a control profile, and modifications to the upper 10 cm of the clay subsoil involving (ii) a lower bulk density and (iii) the incorporation of poultry and (iv) dairy manure into this layer. Italian ryegrass and prairie grass were grown in separate columns, over 3 growth cycles. Results Manure incorporation decreased root biomass by up to 40% in both grasses, without any reduction in shoot growth. Root distribution varied with roots proliferating in the manured-layers compared with the same layer without the manures, resulting in a doubling of the root length density in the manured layer. Shoot yields in the 2nd growth cycle were similar between treatments, yet there was lower transpiration from the shoots in the manure treatments. This meant that manure incorporation increased transpiration efficiency, as it changed the distribution and architecture of the roots. Conclusion Manure incorporation into subsoil increased transpiration efficiency, and root surface area, rather than shoot biomass, was the controlling factor for transpiration from the shoots of the grasses.