The distribution of free amino acids in different tissues of kiwifruit (Actinidia deliciosa) and changes in their concentration during maturation of the fruit were investigated. Amino acid concentrations in the fruit (and each fruit tissue except the skin) decreased as the fruit matured. Asparagine and glutamine were the main amino acids to decrease, allowing arginine and γ-aminobutyric acid to become the predominant amino acids in fruit harvested at the end of May. There were differences in individual and total amino acid concentrations between the core, inner and outer cortex fruit tissues, and with the fruit stalk. The core contained the highest concentration of amino acids and the outer cortex the least In February, asparagine was the major amino acid in the inner and outer cortex, whereas arginine and γ-aminobutyric acid were predominant in the core. By May, arginine and γ-aminobutyric acid were predominant in all tissues. Arginine and/or γ-aminobutyric acid always formed a longitudinal gradient within the fruit, regardless of tissue type. It was highest at the stem end and lowest at the blossom (distal) end. Concentrations of amino acids in the fruit stalk increased as the fruit matured. Arginine and/or γ-aminobutyric acid and asparagine were the major amino acids to increase, followed by glutamine.