Data for: Synchronous seasonal plasticity in colouration, behaviour, and visual gene expression in a wild butterfly population
Data files
Oct 24, 2025 version files 112.43 KB
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README.md
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WildJunonia_Data_Dryad.xlsx
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Abstract
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.
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) 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.
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).