Plant monocultures growing for extended periods face severe losses of productivity. This phenomenon, known as ‘yield decline', is often caused by the accumulation of above‐ and below‐ground plant antagonists. The effectiveness of plant defences against antagonists might help explain differences in yield decline among species. Using a trait‐based approach, we studied the role of 20 physical and chemical defence traits of leaves and fine roots on yield decline of 4‐ and 18‐year‐old monocultures of 27 grassland species. We hypothesized that yield decline is lower for species with high defences, that root defences are better predictors of yield decline than leaf defences, and that in roots, physical defences better predict yield decline than chemical defences, while the reverse is true for leaves. We additionally hypothesized that the relationship between defences and yield decline increases with time and that species increasing the expression of defence traits after long‐term monoculture growth would suffer less yield decline. We summarized leaf and fine root defence traits using principal component analyses and analysed the relationship between the most informative components along with their temporal changes and monoculture yield decline. The significant predictors of yield decline were traits related to the so‐called collaboration gradient of the root economics space (specific root length and root diameter) as well as their temporal changes and traits related to the leaf physical vs chemical defence tradeoff (leaf dry matter, silicon and cellulose content, toughness and phytochemical diversity). We were unable to unequivocally identify the mechanisms relating the effect of those traits to yield decline as they could mediate plant responses to several stressors such as antagonist accumulation, nutrient depletion or drought. Further studies are needed to differentiate between these alternative mechanisms and to gain a comprehensive understanding of the drivers of yield decline in relation to root and leaf defence traits.