Abstract
Plants have evolved a diverse arsenal of defensive secondary metabolites in their evolutionary arms race with insect herbivores. In addition to the bottom-up forces created by plant chemicals, herbivores face top-down pressure from natural enemies, such as predators, parasitoids and parasites. This has led to the evolution of specialist herbivores that do not only tolerate plant secondary metabolites but even use them to fight natural enemies. Monarch butterflies (Danaus plexippus) are known for their use of milkweed chemicals (cardenolides) as protection against vertebrate predators. Recent studies have shown that milkweeds with high cardenolide concentrations can also provide protection against a virulent protozoan parasite. However, whether cardenolides are directly responsible for these effects, and whether individual cardenolides or mixtures of these chemicals are needed to reduce infection, remains unknown. We fed monarch larvae the four most abundant cardenolides found in the anti-parasitic-milkweed Asclepias curassavica at varying concentrations and compositions to determine which provided the highest resistance to parasite infection. Measuring infection rates and infection intensities, we found that resistance is dependent on both concentration and composition of cardenolides, with mixtures of cardenolides performing significantly better than individual compounds, even when mixtures included lower concentrations of individual compounds. These results suggest that cardenolides function synergistically to provide resistance against parasite infection and help explain why only milkweed species that produce diverse cardenolide compounds provide measurable parasite resistance. More broadly, our results suggest that herbivores can benefit from consuming plants with diverse defensive chemical compounds through release from parasitism.
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Data Availability
All data presented in this manuscript is publicly available in GitHub repository: https://github.com/mhoogshagen/cardenolide-mixtures.
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Acknowledgements
We thank Chris Catano, Gabe DuBose, Mitchell Kendzel, and two anonymous reviewers for helpful comments on the manuscript. We thank Erik Edwards for growing the plants used in these experiments. Ron White and Christophe Duplais helped with cardenolide isolation, purification, and identification.
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This research was supported by NSF grant IOS-2202255 to JCdR and IOS-2209762 to AAA; MH was supported by NSF GRFP 2022324290.
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All authors contributed to study design and data collection. MH and JCdR wrote the main manuscript and prepared figures 1 and 2. All authors reviewed and edited the manuscript.
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Hoogshagen, M., Hastings, A.P., Chavez, J. et al. Mixtures of Milkweed Cardenolides Protect Monarch Butterflies against Parasites. J Chem Ecol 50, 52–62 (2024). https://doi.org/10.1007/s10886-023-01461-y
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DOI: https://doi.org/10.1007/s10886-023-01461-y