The aim of this study was to investigate the cause of variation in the digestibility of pea protein in poultry and to find a tool to select genotypes with high digestibility potential by using an in vitro hydrolysis assay. Eight pea genotypes were selected for their difference in seed protein content and composition. To reduce the variation due to tannins and particle size, seeds from these 8 genotypes were dehulled and micro-ground. They were incorporated as the only protein source in 8 different experimental isoproteinaceous diets with similar metabolisable energy content. The amino acid digestibility was studied in cecectomized chickens. A balance method was used to obtain apparent digestibility, and the isotope dilution technique was used to determine endogenous losses and true digestibility, after feeding a double labelled test meal containing chromic oxide and (15)N-labelled peas. The 8 diets showed differences in apparent amino acid digestibility. The average apparent digestibility for all amino acids varied between 79.5 and 86.3%, with the highest values for arginine (85.2 to 90.8%) and glutamic acid (85.2 to 90.5%), and the lowest values for cystine (63.3 to 69.7%) and tryptophan (69.1 to 80.3%). This variability of apparent amino acid digestibility was due to variations in endogenous losses and true digestibility among the 8 pea genotypes. The average endogenous losses as determined for 9 amino acids ranged from 3.6 to 5.4% of ingested amino acids, with the highest value for threonine (8.0 to 11.0%). The average true digestibility varied between 84.4 and 90.2%, with the highest values for lysine (89.0 to 95.0%), and the lowest for isoleucine (81.0 to 88.7%) and valine (82.4 to 88.7%). In vitro hydrolysis of protein from micro-ground seeds was performed for the 8 pea genotypes using three proteases (pepsin, trypsin and chymotrypsin). The quantity of small peptides (<3 kDa) that appeared after the combined hydrolysis with pepsin (3 h) followed by trypsin and chymotrypsin (15 min) was significantly correlated with the average true digestibility of the 8 genotypes (R = 0.74; P<0.05). (C) 2007 Elsevier B.V. All rights reserved.