Data for: Synchronous seasonal plasticity in colouration, behaviour, and visual gene expression in a wild butterfly population

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.

Methods
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).