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Phenotypic plasticity in chemical defence allows butterflies to diversify host use strategies


de Castro, Erika et al. (2021), Phenotypic plasticity in chemical defence allows butterflies to diversify host use strategies, Dryad, Dataset,


Hostplant specialization is a major force driving ecological niche partitioning and diversification in insect herbivores. The cyanogenic defences of Passiflora plants keeps most herbivores at bay, but not larvae of Heliconius butterflies, which can both sequester and biosynthesize cyanogenic compounds. Here, we demonstrate that both Heliconius cydno chioneus, a host plant generalist, and H. melpomene rosina, a specialist, have remarkable plasticity in their chemical defence. When feeding on Passiflora species with cyanogenic compounds they can readily sequester, both species downregulate the biosynthesis of these compounds. In contrast, when fed on Passiflora plants that do not contain cyanogenic glucosides that can be sequestered, both species increase biosynthesis. This biochemical plasticity comes at a significant fitness cost for specialist like H. m. rosina, as growth rates for this species negatively correlate with biosynthesis levels, but not for a generalist like H. c. chioneus. In exchange, H. m rosina has increased performance when sequestration is possible as on its specialised hostplant. In summary, phenotypic plasticity in biochemical responses to different host plants offers these butterflies the ability to widen their range of potential host within the Passiflora genus, while maintaining their chemical defences.


This dataset correspond to body measurements (forewing length, body length, adult weight, and pupal weight) and cyanogen concentrations of H. cydno chioneus and H. melpomene rosina raised on P. biflora, P. menispermifolia, P. vitifolia and P. platyloba. The common garden experiment was conducted at the Smithsonian Tropical Research Institute (Panama).

Usage Notes

LC-MS analyses were carried out as described in Pinheiro de Castro, É. C., Zagrobelny, M., Zurano, J. P., Zikan Cardoso, M., Feyereisen, R., & Bak, S. (2019). Sequestration and biosynthesis of cyanogenic glucosides in passion vine butterflies and consequences for the diversification of their host plants. Ecology and evolution9(9), 5079-5093.

Each whole butterfly was  homogeinized in 1.5 mL methanol 80%. All samples were centrifuged at 10,000 x g for 5 min and the supernatant filtered (Anapore 0.45 µm, Whatman) to remove insoluble components. Samples were diluted 50X and injected (3 uL) in a Agilent 1100 Series LC (Agilent Technologies, Germany) hyphenated to a Bruker HCT-Ultra ion trap mass spectrometer (Bruker Daltonics, Bremen, Germany).

Deidaclin/Tetraphyllin A (RT 5.5 min, [M+Na]+ at m/z 294), linamarin (RT 2.4 min, [M+Na]+ at m/z 310), lotaustralin (RT 5.5 min, [M+Na]+ at m/z 284), prunasin (RT 7.0 min, [M+Na]+ at m/z 317) and amygdalin (RT 6.6 min, [M+Na]+ at m/z 480)  were detected and their RTs compared to authentic standards (Engler et al. 2000; Jaroszewski et al. 2002; Møller et al. 2016).  Tetraphyllin B-sulfate (RT 1.3 min, [M+Na]+ at m/z 390) and passibiflorin (RT 5.8 min, [M+Na]+ at m/z 456) were identified in the plant samples by their m/z, fragmentation pattern (MS/MS) and comparison with data reported in the literature regarding these compounds.

Please utilize the software  Compass Dataanalysis from Bruker to open the raw chemical data (samples, washed and standards). In the excel file "Cyanogen Datasheet.xlsx", you will find information about the species/diet treatment corresponded to each sample (1-105) as well as the  cyanogenic glucoside concentrations in the standard curve (ST1-ST7).  README.txt contains a description of all the information in the file "Cyanogen Datasheet.xlsx".  A file with all the data Analysed in this srudy is also available as .csv (("alldata.csv").


European Research Council, Award: Grant number: 339873 (Acronym: SpeciationGenetics)

Horizon 2020, Award: Marie Curie Actions, grant award: 841230 (Acronym: CyanideEvolution)

Danmarks Frie Forskningsfond, Award: FNU - 1323-00088