Olfactory learning in Pieris brassicae butterflies is dependent on the intensity of a plant-derived oviposition cue

Butterflies, like many insects, use gustatory and olfactory cues innately to assess the suitability of an oviposition site and are able to associate colours and leaf shapes with an oviposition reward. Studies on other insects have demonstrated that the quality of the reward is a crucial factor in forming associative memory. We set out to investigate whether the large cabbage white Pieris brassicae (Linnaeus) has the ability to associate an oviposition experience with a neutral olfactory cue. In addition, we tested whether the strength of this association is dependent on the gustatory response to the glucosinolate sinigrin, which is a known oviposition stimulus for P. brassicae. Female butterflies were able to associate a neutral odour with an oviposition experience after a single oviposition experience, both in a greenhouse and in a semi-natural outdoor setting. Moreover, butterflies performed best when trained with concentrations of sinigrin that showed the strongest response by specif..., Butterflies were conditioned by spraying 1 ml sinigrin solution on a green paper disc (9.5 cm in diameter) as an unconditioned stimulus, coupled with 20 microliters of vanilla scent as a conditioned stimulus. Three different concentrations of sinigrin solution were used. 3 different conditioning treatments plus one group of non-conditioned naive animals --> four different treatments. Tests consisted of releasing one animal in a testing environment provided with four green paper discs, identical to the ones used during training, but without sinigrin. Two of these were scented with vanilla scent, while the other two were scented with 30% alcohol-water solution. Each test lasted 30 minutes. Parameters recorded for each test are listed and explained below. Please note that not all of the recorded parameters have been used for analysis.
Electrophysiology
Response of the trichoid taste sensilla to sinigrin was assessed with single sensillum recordings. Sinigrin was dissolved in 1 mM KCl, 4..., , # Olfactory learning in *Pieris brassicae* butterflies is dependent on the intensity of a plant-derived oviposition cue

## Folder structure and contents:

**These files are related to the behavioural observations in both indoor and outdoor settings:**

* GHdata.xlsx
* OUTdata.xlsx
* GH_Sinigrin_Behaviour.R
* OUT_Sinigrin_Behaviour.R
* Peftuloglu_et_al_Electronic_Supplementary_Material.pdf

**These files are related to the electrophysiological taste recordings performed:**

* pierislegtaste.csv
* Pierislegtaste script.R

**Pieris_ovi.mp4**: Mp4 video of oviposition instance by a female Pieris brassicae on green paper disc impregnated with sinigrin.

**Peftuloglu_et_al_Electronic_Supplementary_Material.pdf**: This file contains a picture of the indoor set-up, as well as graphs showing the proportion of time spent on vanilla-scented vs. unscented discs for control butterflies as well as butterflies trained with different concentrations of sinigrin in the indoor trails (figure S1). The fil...