Potent Nitrogen-containing Milkweed Toxins are Differentially Regulated by Soil Nitrogen and Herbivore-induced Defense.

Theories have been widely proposed and tested for impacts of soil nitrogen (N) on phytochemical defenses. Among the hundreds of distinct cardenolide toxins produced by milkweeds (Asclepias spp.), few contain N, yet these appear to be the most toxic against specialist herbivores. Because N- and non-N-cardenolides coexist in milkweed leaves and likely have distinct biosynthesis, they present an opportunity to address hypotheses about drivers of toxin expression. We tested effects of soil N and herbivore-damage on cardenolide profiles of two milkweed species differing in life-history strategies (Asclepias syriaca and A. curassavica), and the toxicity of their leaves. In particular leaf extracts were tested against the target enzymes (Na ⁺ /K ⁺ -ATPase extracted from neural tissue) from both monarch butterflies (Danaus plexippus) as well as less cardenolide-resistant queen butterflies, D. gilippus. Increasing soil N enhanced biomass of Asclepias syriaca but had weak effects on cardenolides, including causing a significant reduction in the N-cardenolide labriformin; feeding by monarch caterpillars strongly induced N-cardenolides (labriformin), its precursors, and total cardenolides. Conversely, soil N had little impact on A. curassavica biomass, but was the primary driver of increasing N-cardenolides (voruscharin, uscharin and their precursors); caterpillar induction was weak. Butterfly enzyme assays revealed damage-induced cardenolides substantially increased toxicity of both milkweeds to both butterflies, swamping out effects of soil N on cardenolide concentration and composition. Although these two milkweed species differentially responded to soil N with allocation to growth and specific cardenolides, leaf toxicity to butterfly Na ⁺ /K ⁺ -ATPases was primarily driven by herbivore-induced defense. Thus, both biotic and abiotic factors shape the composition of phytochemical defense expression, and their relative importance may be dictated by plant life-history differences.
(© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)