Trichomes play a key role in both heavy metal tolerance and herbivory defense, and both stressors have been shown to induce increased trichome density. However, the combined effect of these stressors on trichome density in general, and specifically on metal-hyperaccumulating plants, has yet to be examined. The aim of this study was to test the effect of cadmium availability and herbivory on leaf trichome density and herbivore deterrence in the metal hyperaccumulator Helianthus annuus. To test this, H. Annuus plants were grown in control pots or pots inoculated with 10 mg/kg cadmium and were subjected to either no herbivory or simulated herbivory using mechanical damage and foliar jasmonic acid application. Herbivore deterrence was tested in a feeding assay using Spodoptera littoralis caterpillars. Interestingly, while the trichome density of H. annuus increased by 79% or 53.5% under high cadmium availability or simulated herbivory, respectively, it decreased by 26% when the stressors were combined. Furthermore, regardless of cadmium availability, simulated herbivory induced a 40% increase in deterrence of S. littoralis. These findings suggest that the combination of metal availability and herbivory might present excessive stress to hyperaccumulators. Moreover, they suggest that the risk of metal bioaccumulation in phytoremediation can be reduced by simulated herbivory.

Soil pollution by metals and metalloids as a consequence of anthropogenic industrialisation exerts a seriously damaging impact on ecosystems. However, certain plant species, termed hyperaccumulators, are able to accumulate extraordinarily high concentrations of these metal(loid)s in their aboveground tissues. Such hyperaccumulation of metal(loid)s is known to act as a defence against various antagonists, such as herbivores and pathogens. We investigated the influences of metal(loid)s on potential defence traits, such as foliar elemental, organic and mechanical defences, in the hyperaccumulator plant species Arabidopsis halleri (Brassicaceae) by artificially amending the soil with common metallic pollutants, namely cadmium (Cd) and zinc (Zn). Additionally, unamended and metal-amended soils were supplemented with the metalloid silicon (Si) to study whether Si could alleviate metal excess. Individuals originating from one non-/low- and two moderately to highly metal-contaminated sites with different metal concentrations (hereafter called accessions) were grown for eight weeks in a full-factorial design under standardised conditions. There were significant interactive effects of metal amendment and Si supplementation on foliar concentrations of certain elements (Zn, Si, aluminium (Al), iron (Fe), potassium (K) and sulfur (S), but these were accession-specific. Profiles of glucosinolates, characteristic organic defences of Brassicaceae, were distinct among accessions, and the composition was affected by soil metal amendment. Moreover, plants grown on metal-amended soil contained lower concentrations of total glucosinolates in one of the accessions, which suggests a potential trade-off between inorganic defence acquisition and biosynthesis of organic defence. The density of foliar trichomes, as a proxy for the first layer of mechanical defence, was also influenced by metal amendment and/or Si supplementation in an accession-dependent manner. Our study highlights the importance of examining the effects of co-occurring metal(loid)s in soil on various foliar defence traits in different accessions of a hyperaccumulating species.