Ecological theory indicates that warmer and more stable climates should result in stronger biotic interactions. Therefore, plant species growing at lower elevations and experiencing greater herbivore pressure should invest in higher levels of defences than those at higher elevations. Nonetheless, there are a number of studies that have found no effect of elevational gradients on plant defensive traits. Several factors might explain the lack of consistency for the altitude-defence relationships, including (i) the reduction of all defensive traits into one measure of resistance; (ii) not considering plant defence as the simultaneous expression of several defensive traits; and (iii) not considering the relative influence of biotic (e.g. herbivory) and abiotic (e.g. climate and soil conditions) factors associated with the ecological gradient. Here, we present a comprehensive test of the effects of elevation and its associated biotic and abiotic factors on the individual and simultaneous expression of constitutive direct and indirect defences and their inducibility (i.e. expression of defences after herbivore attack). Specifically, we estimated climatic and soil variables and measured herbivore damage and constitutive and jasmonic acid-induced glucosinolate levels in the leaves as a proxy for direct defences, and volatile emission as a proxy for indirect defences in 16 Cardamine species naturally growing along the steep elevational gradient of the Alps. Within a phylogenetic comparative framework, we found that species growing at lower elevations invested more in the simultaneous inducibility of both direct and indirect defences, whereas species growing at higher elevations invested more in constitutive direct defences. Although we found strong elevational gradients in herbivory and climatic and soil variables, these biotic and abiotic factors only partially explained elevational patterns in plant defences.Synthesis. These results highlight that the complex regulation of multiple defence traits strongly vary across elevational gradients and build towards a better understanding of the multiple mechanisms underlying trait evolution and species interactions along ecological gradients.