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Abstract

The chemical defence potential against herbivores of certain Dictyotalean brown algae increases after tissue disruption. This wound activated defence has been explored in bioassays, but the metabolic pathways behind it are unknown. Here we describe a metabolic profiling approach to identify the activated defence metabolites. Before and after tissue damage of Dictyota dichotoma modified diterpenes, non-volatile medium polar metabolites as well as volatile compounds were profiled. While comparison of extracted intact and mechanically wounded algae revealed no significant differences in structure and distribution of semi-volatile and reversed phase LC/MS detectable metabolites, a strong release of gaseous volatiles was observed. Solid phase micro extraction (SPME) and GC/MS were used for identification and quantification of these biogenic gases. This showed that D. dichotoma released elevated amounts of trimethylamine (TMA) and dimethylsulphide (DMS) after mechanical tissue damage. To study the ecological significance of compounds released post injury and of the biosynthetically connected non-volatile acrylate, choice assays were performed with the amphipod Amphithoe longimana. Behavioural assays on artificial diets did not reveal any repellent role for the single isolated metabolites. In strong contrast, a mixture of TMA, DMS and acrylate significantly reduced the association of the herbivores with the treated food pellets. This shows that mixtures of these biogenic gases and acrylate are recognized by the herbivores and influence food selection.