High nitrogen loading impacts the temperature-size rule and heat tolerance in a nettle-feeding butterfly
Data files
Apr 16, 2026 version files 112.95 KB
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CaterpillarWeights_Alevana.csv
80.66 KB
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Determining_abdomenweight_Alevana.csv
5.05 KB
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Fresh_and_dryweigth_Alevana.csv
15.35 KB
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Heattolerance_file_ALevana.csv
4.59 KB
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N_P_mol_calculated.csv
1.15 KB
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README.md
4.35 KB
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survival_data_Alevana.csv
1.81 KB
Abstract
Host-plant quality and ambient temperature are key environmental drivers of herbivorous insect performance, affecting growth, development, and survival. While temperature accelerates physiological processes in ectothermic insects, nutrient limitation in host plants can constrain insect herbivore fitness. Yet, few studies have examined how these two factors interact to shape insect herbivore performance. We experimentally quantified the combined effects of host-plant stoichiometry and rearing temperature on the larval growth rate, development time, and adult heat tolerance of the Map butterfly (Araschnia levana L. 1758). Caterpillars were reared on Stinging nettle (Urtica dioica L.) cultivated under three nutrient addition regimes differing in N:P ratios—to alter the nutrient stoichiometry of their host plants—and under two diurnally fluctuating temperature regimes simulating ambient (day-night: 21 °C/12 °C) and elevated (day-night: 24 °C/16 °C) conditions. Both temperature and host-plant stoichiometry affected butterfly performance. Warming and a balanced stoichiometry enhanced growth and development; under these favourable conditions, caterpillars exhibited rapid growth, faster development, higher survival, and increased adult heat tolerance. A trade-off between size and age at maturity—individuals either matured rapidly at smaller sizes or more slowly at larger sizes—was observed only under conditions that were favourable for growth, suggesting that developmental time was the main limiting factor under these conditions for the final butterfly body size. Our results thus demonstrate that host-plant nutrient imbalances can mediate how insect herbivores prioritise between size and age at maturity under warming conditions, influencing thermal tolerance too. This suggests that eutrophication and climatic warming may jointly reshape the allocation strategies, performance, and resilience of insect herbivores in Anthropocene ecosystems.
Dataset DOI: 10.5061/dryad.jsxksn0r1
Description of the data and file structure
During the experiment, caterpillars of Araschnia levana were reared on stinging nettle (Urtica dioica) leaves, which received three different fertilization treatments, and reared under two different climate conditions. During the run of the experiment, caterpillars were weighed every couple of days, and pupation, as well as emergence day, were recorded. The fresh weight of both pupae and adults was determined.
Once adult butterflies had emerged, adult heat tolerance was determined. After adult butterflies were dried, their total dry weight and abdomen dry weight were determined.
For the leaves of U. dioica, the elemental composition was determined every couple of days and at the end of the experiment, especially focusing on C:N:P ratios.
Files and variables
File: CaterpillarWeights_Alevana.csv
Description:
Variables
- Date: Date of measurement
- Container#: ID number of container the caterpillar was reared in
- GroupID: Which treatment combination they belonged to
- Species: Which species was reared (we reared both A. levana and A. io in this experiment)
- Rearing_temperature: Which rearing temperature was used (°C)
- Plant_treatment: Which fertilization treatment was used
- Age (days): The age of the caterpillar in days at the moment of measurement
- stage: caterpillar/pupae/adult
- Weight: noted down the weight (g)
File: Determining_abdomenweight_Alevana.csv
Description:
Variables
- IND_ID: the individual ID a butterfly was given
- Species: Which species was reared (we reared both A. levana and A. io in this experiment)
- rearing_temperature: Which rearing temperature was used (°C)
- container: ID number of container the caterpillar was reared in
- treatment: Which fertilization treatment was used
- dryweight: the dryweight of the individual
- abdomen_weight: the abdomen weight of the individual
- sex_based on dryweight? -> not correct: (should be removed) male or female, based on a 2-split in the total abdomen weight, in which we did not look at the abdomen:total weight.
- concatenated: combination of rearing temperature x fertilization treatment combination
File: Fresh_and_dryweigth_Alevana.csv
Description:
Variables
- rearing_temperature: Which rearing temperature was used (°C)
- plant_treatment: which fertilization treatment was used
- type: whether it considers the fresh or the dryweight
- weight: Weight of the individual (g)
- treatment: treatment combination of rearing temperature and fertilization treatment
- stage: caterpillar, pupae, or butterfly
File: survival_data_Alevana.csv
Description:
Variables
- Container: which container the caterpillar was reared in
- GroupID: ID number of container the caterpillar was reared in.
- Species: Which species was reared (we reared both A. levana and A. io in this experiment)
- rearing_temperature:Which rearing temperature was used (°C)
- plant_treatment: which fertilization treatment was used
- survived: how many individuals survived
- survival_percentage: percentage of caterpillars that survived
- stage: caterpillar, pupae, or butterfly
File: Heattolerance_file_ALevana.csv
Description:
Variables
- tube: which tube was used in the waterbath (1 or 2)
- container: which container the caterpillar grew up in
- waterbath_temperature: the temperature of the waterbath the animal was tested in (°C)
- emergence date: the day a butterfly emerged
- measure_date: the day a butterfly was measured for heat tolerance
- duration_min_KO: the duration in minutes until the butterfly was KO
- duration_min_dead: the duration in minutes untill no more movement was observed
- test#: the file
- treatment: which fertilization treatment the caterpillar grew up with
- rearing_temperature: which rearing temperature the caterpiller grew up in (°C)
File: N_P_mol_calculated.csv
Description:
Variables
- ID: Id of sample
- N_P_mol: N:P ratio calculated using the mol weights
- treatment: Which treatment it considered (np, Np, or NP)
Code/software
Excel