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_dp.xlsx * OUTdata_dp.xlsx * workingdataGH.xlsx * workingdataOUT.xlsx * GH_Sinigrin_Behaviour_dp.R * OUT_Sinigrin_Behaviour_dp.R **These files are related to the electrophysiological taste recordings performed:** * pierislegtaste.xlsx * pierislegtaste script.R **Pieris_ovi.mp4**: Mp4 video of oviposition instance by a female Pieris brassicae on green paper disc impregnated with sinigrin. ##### MEASUREMENTS IN \"GHdata\_DP\" and \"OUTdata\_DP\": * Consolidation: time (expressed in minutes) used to consolidate memory formation after conditioning. * Testing time: length (expressed in minutes) of the behavioural observation. * Flight: binary expression (1 or 0) to express whether flight (i.e. behavioural respon...# 菜白蝶（*Pieris brassicae*）的嗅觉学习依赖于植物来源产卵刺激的强度 与多数昆虫类似，大菜粉蝶（*Pieris brassicae*，Linnaeus）天生依靠味觉与嗅觉线索评估产卵位点的适宜性，且能够将颜色、叶片形状与产卵奖励建立关联。已有针对其他昆虫的研究证实，奖励质量是形成关联记忆的关键影响因素。本研究旨在探究大菜粉蝶是否能够将产卵经历与中性嗅觉线索建立关联；此外，我们还验证了此类关联的强度是否取决于对黑芥子苷（sinigrin）的味觉反应——黑芥子苷是已知的大菜粉蝶产卵刺激物，属于硫代葡萄糖苷（glucosinolate）类化合物。 实验结果显示，无论是在温室还是半自然室外环境中，雌性大菜粉蝶仅需一次产卵经历，即可将中性气味与产卵经历建立关联。不仅如此，当使用能够引发最强特异性反应的黑芥子苷浓度开展训练时，菜粉蝶的表现最佳[原文此处存在截断]。 本研究通过将1毫升黑芥子苷溶液喷洒于直径9.5厘米的绿色纸碟上作为非条件刺激（unconditioned stimulus），同时搭配20微升香草气味作为条件刺激（conditioned stimulus），对菜粉蝶进行条件化训练。实验共设置3种不同浓度的黑芥子苷溶液处理组，外加一组未经过条件化的空白对照组，总计4种实验处理。 测试环节具体为：将单头蝴蝶释放至测试环境中，该环境内放置4个与训练阶段完全一致的绿色纸碟，但未涂抹黑芥子苷。其中2个纸碟涂抹香草气味，剩余2个则涂抹30%乙醇水溶液。每项测试持续30分钟。本次测试记录的各项参数如下并将逐一说明，请注意并非所有记录参数均用于后续数据分析。 ### 电生理学实验 我们采用单感器记录（single sensillum recordings）技术，评估了毛形味觉感器对黑芥子苷的反应。黑芥子苷溶解于1 mM氯化钾（KCl）溶液中[原文此处存在截断]。 --- ## 文件夹结构与内容 ### 室内外行为观测相关文件 * GHdata_dp.xlsx * OUTdata_dp.xlsx * workingdataGH.xlsx * workingdataOUT.xlsx * GH_Sinigrin_Behaviour_dp.R * OUT_Sinigrin_Behaviour_dp.R ### 味觉电生理记录相关文件 * pierislegtaste.xlsx * pierislegtaste script.R ### 实验视频 * Pieris_ovi.mp4：雌性大菜粉蝶在涂抹黑芥子苷的绿色纸碟上产卵的MP4视频。 ##### GHdata_DP与OUTdata_DP中的测量指标 * 记忆巩固时长：指条件化训练后记忆形成巩固所耗费的时间，单位为分钟。 * 测试时长：行为观测的持续时长，单位为分钟。 * 飞行行为：二进制变量（1或0），用于表征是否发生飞行行为（即行为反应……[原文此处存在截断]）