DOI: 10.5061/dryad.4f4qrfjnt
title: Data for: Synchronous seasonal plasticity in colouration, behaviour, and visual gene expression in a wild butterfly population

description: This data corresponds to the paper: "Synchronous seasonal plasticity
 in colouration, behaviour, and visual gene expression in a wild butterfly
 population." It contains multiple spreadsheets with behavioral data,
 wing reflectance measurements, qualitative wing characteristics, real-time
 quantitative PCR results, and primer effiency data. We investigate if
 there are covarying seasonal changes to morphology, behaviour, and the
 visual system in the seasonally plastic butterfly Junonia coenia.
 Wild, native butterflies in tallgrass prairies were collected and observed
 during their flight period (May to November) from 2018 to 2021. We found
 at our field sites in northwest Arkansas, more J. coenia exhibit seasonal
 dark wing patterns in September and October compared to butterflies
 collected in previous months, independent of sex. This change in wing
 pattern correlates to an increase in observed basking behaviour in the
 field. Basking changed in both focal watches and point count surveys.
 RNA-Seq analysis revealed eye tissues of dark fall animals and lighter
 summer animals exhibit overall different patterns of gene expression,
 including in visual system development genes, pigmentation genes, and
 clock genes. Subsequent qPCR analysis of monthly variation in opsin gene
 expression in eye tissues confirmed that opsin genes are not
 differentially expressed throughout the year, though females have higher
 expression of blue sensitive opsin than males. Subsequent analysis also
 confirmed that eye tissue from fall caught animals has higher expression
 levels of the clock gene period, a gene implicated in seasonal physiology
 and circadian activity in other species. This concurrent seasonal
 shift in colouration, behaviour, and underlying visual physiology
 indicates that natural populations of  J. coenia undergoes a
 complex shift in phenotype encompasses more than simple changes to
 thermoregulation.
description: Behavioral data includes counted instances and durations of
 behaviors made during point counts and focal watches. Qualitative wing
 score data was scored blind in regards to sex and collection date by
 single individual in a single session.  Reflectance measurements were
 recorded with a spectrophotometer (Jaz; Ocean Optics) and proccessed in R
 to extract total area under reflectance curves. Temperature data in
 spreadsheets is from wunderground.com and daylength from NOAA (https://gml.noaa.gov/grad/solcalc/calcdetails.html). Real-time quantitative PCR data are results from reactions run on a CFX96 machine (Bio-Rad). 
description: # Data for: Synchronous seasonal plasticity in colouration, behaviour, and
 visual gene expression in a wild butterfly population
 [https://doi.org/10.5061/dryad.4f4qrfjnt](https://doi.org/10.5061/dryad.4f4qrfjnt) ## Description of the data and file structure This dataset (WildJunonia_Data_Dryad.xlsx) contains wing measurements, behavioral observations, and gene expression levels taken from wild *Junonia coenia* observed and captured from wild populations in northwest Arkansas from May to November in 2018-2021. Wing measurements are both quantitative (reflectance measurements) and qualitative. Behavioral observations are from point counts and focal watches. Gene expression values are the results of real-time quantitative PCR examining opsin (*BRh*, *LWRh*, and *UVRh*) and *period* genes in eye tissue. Dataset also includes environmental conditions gathered from online sources. ### Files and variables #### File: WildJunonia_Data_Dryad.xlsx #### Description of the data and file structure **Sheet 1: Quantitative Wing Scores** Contains qualitative wing scores with associated metadata from time of collections (including collection date, average daylength of collection week, and average temperature of collection week) Wings were scored based on criteria outlined by Smith(1). Details about columns: Year: Collection Year Julian.Week: Week of the year animal was collected Julian.WeekPlus: Week after animal was collected Julian.WeekMinus: Week prior to collection week PWeek_Start: Start of the week prior to collection (Month/Day/Year) All_Daylengths: Number of minutes of daylength from each day of previous week, starting with "PWeek_Start" P_Avg_DL: Average daylength (minutes) for the week prior to collection (Mean of All_Daylengths) Wing.Score: Qualitative ventral wing score ranging from 1 (summer-like) to 5 (fall-like) with intermediate forms (2, 3, 4). Week.AvgF: Average Weekly Temperature in Fahrenheit Wool.AvgFP: Average Weekly Temperature in Fahrenheit of week prior to collection Week.AvgC: Average Weekly Temperature in Celsius of (converted from Fahrenheit) Wool.AvgPC: Average Weekly Temperature in Celsius of week prior to collection (converted from Fahrenheit) **Sheet 2: Ventral Wing Reflectance** Contains wing reflectance values of the ventral wing of *J. coenia* caught throughout the year, with associated metadata Details about columns: ID: Wing ID Date: Collection date (Month/Date/Year, or for animals where only Week & Year are known: Month'Year or MonthWeek) Year: Collection year Week: Collection week of the year WeekPlus: Week after collection week Sex: Sex of animal Notes: If the wing was faded or worn (Blanks indicate a wing was undamaged) Columns H-J: Area under the % reflection curve for a ventral part of the wing. Each individual was measured three times. Mean.auc: Mean of the three reflectance measurements **Sheet 3: Dorsal Wing Reflectance** This contains dorsal wing reflectance for *J. coenia* from data with associated metadata. Details about columns: ID: Wing ID Year: Collection year Week: Collection week of the year WeekPlus: Week after collection week Date: Month/Day/Year, or for animals where only Week & Year is known, Month'Year or YearCollectionWeek Sex: Sex of butterfly Notes: Annotated if wings were worn/damage (Blanks indicate a wing was undamaged) Columns H-K: Area under the % reflection curve for a dorsal area of the wing, including a blue eyespot, a white eyespot, a white stripe, and a brown basal area. **Sheet 4: Point Counts** This contains behavioral data from point counts of *J. coenia* with associated metadata. Details about columns: ID: Individual animal observance ID Week: Week of year observed in point counts Year: Year observed in point counts Date: Month/Day/Year (NA indicates only Week & Year were known) Site: Observation location Point #: Point count location ID at site (5 per site) Behavior: Observed behavior during point count **Sheet 5: Focal Watches** Contains observations of focal watches of individual *Junonia coenia* with associated metadata and seasonal conditions Details about columns: Year: Year observed Week: Week of year observed WeekMinus: Week prior to observation Week.avgF: Average weekly temperature of observation week (in Fahrenheit) Week.avgC: Average weekly temperature of observation week (converted to Celsius) Week.avgP: Average weekly temperature of of week prior to observation week (in Fahrenheit) Week.avgPC: Average weekly temperature of of week prior to observation week (converted to Celsius) Obs_Date: Date of focal watch observation (Month/Day/Year) Pweek_Start: Start date of the week prior to observation week (Month/Day/Year) PWeek_Minutes: Number of minutes of daylight for each day in week prior to observation week, starting with "Pweek_Start" Avg_Minutes_DL: Average number of minutes of daylight/day for week prior to observation week. Calculated from PWeek_Minutes Total_Seconds: Length of watch in seconds Total_minutes: Length of watch in minutes (Total_Seconds x 60) Behav.Count:Number of different kinds of behaviors observed during a focal watch Columns P-BK: Column starts with observed behavior x (e.g. "rest" or "chase") x_sum: Total duration of behavior x observed during a focal watch x_count: Number of instances of uninterupted bouts of behavior x x_secs/min: x_sum / Total_minutes x_proportion: x_sum/ Total_seconds **Sheet 6: qPCR Results** Contains Ct values from real-time quantitative PCR. Samples were cDNA reverse transcipted from mRNA of *Junonia coenia* eye tissues. Butterflies were wild and caught through-out the year. Ct values were used to later find 2-ΔΔCt. Details about columns: Plate: 96-well plate ID Cell: Well ID ID: Butterfly ID Month: Month butterfly was collected Sex: Sex of butterfly Redo Status: Signifies if this sample had to be rerun (due to known handling problems or indications of contamination (i.e. secondary amplification curve) (Blanks indicate samples from a butterfly were not rerun) Columns G-H: Ct values of genes including Blue sensitive opsin gene (Blue), Long wavelength sensitive opsin (LW), Ultraviolet sensitive opsin (UV), *Period*, and *Elongation factor 1 alpha 2* (*Ef1a2*). *Ef1a2* was used as the reference gene for subsequent 2-ΔΔCt calculations (2).(Blanks indicate samples from a butterfly were rerun and appear in a different row.) **Sheet 7: Primer Efficiencies** RT-qPCR primer efficiences for *J. coenia* opsin, *period*, and *Ef1a-2* genes. Details about columns: Gene: Gene name CT: Ct values Log: Log of Sample Quantity ## Code/software Datasheets can be saved as *.csv files and run in R 4.2.2. or JMP 17.0.0. ## Access information **Other publicly accessible locations of the data:** Average weekly average temperature were found online at wunderground.com that were recorded at the Northwest Arkansas Regional Airport. We used NOAA’s Solar Calculations spreadsheet ([https://gml.noaa.gov/grad/solcalc/calcdetails.html](https://gml.noaa.gov/grad/solcalc/calcdetails.html)) to find the average number of minutes per day per week. ## References 1\)Smith, K. C. (1991). The effects of temperature and daylength on the Rosa polyphenism in the buckeye butterfly, *Precis coenia* (Lepidoptera Nymphalidae). *Journal of Research on the Lepidoptera*, 30(3–4), 225–236. 2\)Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real- time quantitative pcr and the 2−ΔΔCT method. *Methods*, 25(4), 402–408.