raw data of the Papilio polytes experiments

Description

The file entitled "Raw data from the larval frass volatile compound test" presents the results of gas chromatography-mass spectrometry (GC-MS) analysis of volatile compounds emitted from the frass of larvae reared on lemon leaves, with three biological replicates conducted.The files titled "Raw data from the Y-tube olfactometer choice test" and "Raw data from the EAG response test" contain, respectively, the behavioral responses of adult Papilio polytes(common Mormon) to selected compounds assessed using a Y-tube olfactometer, and the corresponding electroantennographic (EAG) response data.

Steps to reproduce

Profiling the VOCs of larval frass： HS-SPME-GC-MS technique was used to identify the chemical composition and relative abundance of volatile compounds in P. polytes frass. Agilent 5975B gas chro-matograph-mass spectrometer (GC-MS) equipped with an HP-5MS capillary column (30 m × 0.25 mm × 0.25 μm film thickness) was used. The injector temperature was set to 250 °C, and the transfer line temperature to 280°C. High-purity helium gas (≥99.999% purity) was used as the carrier gas at a constant flow rate of 1.0 mL/min. Initial tem-perature of 50°C held for 2 min, ramped at 5°C /min to 180°C and held for 5 min, then increased at 10°C /min to 250°C and held for 5 min. Electron ionization source with an ion source temperature of 230°C and a quadrupole temperature of 150°C. Full-scan mode was used to acquire mass spectra over the range of 40–600 m/z.Frass samples were derived from 3rd–5th instar larvae of P. polytes reared under the aforementioned standardized conditions. Freshly excreted frass was collected at a fixed time each day, immediately sealed, and stored in a 4°C refrigerator until volatile compound extraction experiments were conducted. EAG Recordings： EAG recordings were performed using a Syntech (Germany) apparatus equipped with an IDAC-4 signal amplifier and EAG Pro software. Raw signals were amplified by the IDAC-4 unit and digitized by EAG Pro software. Behavioral ：Y-tube olfactometer choice A custom-built Y-tube olfactometer was used to assess the behavioral responses of P. polytes to various volatile compounds. The airflow system was constructed by se-quentially connecting the following components to generate a continuous, purified airstream: an atmospheric sampler, followed by an activated carbon scrubber bottle, a distilled water bubbler, a glass rotameter , and finally the Y-tube chamber (inner diameter: 12 mm; arm length: 30 cm; branching angle: 60°). An oil-free vacuum pump pulled air through the entire system. The airflow rate was regulated by the rotameter and maintained at a constant 800 mL/min. A single adult butterfly was introduced at the base of the olfactometer. A timer was initiated, and the insect's movement was observed. A choice was recorded if the butterfly entered an arm and remained there for at least 30 seconds. If no such com-mitment was made within 10 minutes, the trial was terminated and recorded as "no choice." Each compound was tested with 5 replicates per sex (totaling 10 butterflies per compound), ensuring a 1:1 sex ratio within each set of replicates. To minimize contamination and positional bias, the following procedures were implemented after every five trials: (1) the entire apparatus was thoroughly rinsed with anhydrous ethanol and air-dried; (2) fresh odor-source jars and filter paper discs were prepared; and (3) the left/right positions of the odor sources were switched.

Institutions

• Nanjing Forestry University

  Nanjing
• Jiangsu Academy of Agricultural Sciences

  Jiangsu, Nanjing

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