DOI: 10.5061/dryad.p5hqbzkxz
title: Data from: Human-driven evolution of insect color

description: Adaptation is thought critical for the survival of species under global
 change, but our understanding of human-induced evolution in the wild
 remains limited. Here we show that widespread deforestation has
 underpinned repeated color shifts in wild insects. Loss of forest cover
 has led to drastic color evolution across lineages that mimic the warning
 coloration of a toxic forest stonefly. Predation experiments indicate that
 the relative fitness of color phenotypes varies markedly between forested
 and deforested habitats. Genomic and coloration analyses of 1200 wild
 specimens reveal repeated selection at the ebony locus controlling color
 polymorphism across lineages. These findings represent a compelling case
 of rapid, replicated anthropogenic evolution linking both the agents and
 targets of natural selection, revealing the potential for wild populations
 to adapt in the wake of human-driven change.
description: <strong>Colour measurements</strong>
 Digital photography was used to obtain color measurements as
 reflectance in the visible spectrum adequately captures variance in
 melanism (<em>60</em>), which underpins the mimetic
 resemblance between <em>Zelandoperla </em>and
 <em>Austroperla</em>. Dry-frozen stoneflies (866
 <em>Zelandoperla</em>, 68 <em>Austroperla</em>)
 were photographed on an 18% neutral grey reflectance standard at a
 standardized distance and angle. Uniform lighting was attained using a
 fiber optic illuminator split through two guides diffused through a
 polystyrene foam cylinder 60mm in diameter. Photos were taken in RAW
 format using a Nikon D5200 with a Nikon AF-S DX 40mm micro lens under
 fixed ISO sensitivity (320), aperture (f/5) and shutter speed (1/8) using
 automatic focus. Raw image files were subsequently normalized to the
 neutral grey reflectance standard and linearized using the Multispectral
 Image Calibration and Analysis (MICA) toolbox  and DCRAW plugins in
 ImageJ, generating a multispectral image file. Color
 measurements were extracted from the calibrated multispectral images using
 the RGB Measure function in ImageJ across three regions of interest
 (ROIs): (1) head, (2) pronotum, and (3) femora. The ROIs are
 representative of overall body melanism underpinning mimetic resemblance.
 The mean absolute normalized reflectance (0 – 100%) of the red, green, and
 blue (RGB) channels were sampled for each ROI using the circular selection
 tool in ImageJ, with a diameter of 30 pixels for the head and femora, and
 40 for the pronotum. RGB values for each ROI were subsequently converted
 to achromatic greyscale values or “brightness”, ranging from 0% (black) to
 100% (white) which was quantified as the arithmetic mean of the three
 channels. ROI brightness variables were standardized (centered on zero
 with a unit variance) and we performed a principal component analysis
 (PCA) using R version 4.0.5 (<em>63</em>) to obtain a
 univariate measure of overall body melanism. Overall, 73.94% of the total
 variance in brightness was attributed to the first principal component,
 with approximately equal loadings for each ROI (z<sub>1</sub>
 = 0.5813head + 0.5968pronotum + 0.5531femora).<em> </em>We
 also included the Leith <em>Zelandoperla</em> population from
 Foster et al. 2023. RAW image files (338
 <em>Zelandoperla</em>, 22 <em>Austroperla</em>)
 were processed using the methodology outlined above (PCA:
 z<sub>1</sub> = 0.5899head + 0.5747pronotum + 0.5672femora;
 82.44%).  <strong>Predation
 experiments</strong> We constructed
 410 <em>Zelandoperla</em> color-models from odorless,
 non-toxic polymer modelling clay (Du-kit). The bodies of melanic models
 were constructed entirely from black clay, whereas non-melanic models were
 constructed from a mixture of two parts dark-brown to one part yellow
 clay. Each model (body length ~25 mm) comprised distinct ‘head’,
 ‘thoracic’ and ‘abdominal’ segments shaped and assembled by hand. Model
 legs were shaped by hand from black or brown solid core hookup cable (0.75
 mm; Duratech, Electus Distribution; WH3032), and paired antennae/cerci
 from the plastic bristles of a Raven Hearth Brush. The distinctive white
 (wing-base) and yellow (tibiae) ‘warning’ color-features of
 <em>Zelandoperla </em> were added to models with a fine paint
 brush, using white and yellow acrylic paint, respectively. In summary,
 black and brown models were constructed from identical materials , and
 identical proportions, differing only in the <em>color</em> of
 modelling clay used for the body and cable used for the legs.
 Using not-toxic glue, models were attached to cleaned, flat river
 cobble stones (typical dimensions ~80 mm x ~50 mm) of either schist or
 greywacke, both of which are common in rivers of southeastern New Zealand.
 The black and brown color models for each site were attached to a random
 assortment of stone types, but with each model pair on matching stones
 (fig. S5). Glued specimens were left to dry in the laboratory for 48 hours
 to ensure that any glue odors were absent during subsequent field
 experiments. Unique codes were added to the underside of each stone using
 a permanent marker pen. Once attached to stones, models were photographed
 and transported to and from the field unstacked in the base layer of
 storage boxes. Both <em>Zelandoperla
 </em>and <em>Austroperla </em>stoneflies have lengthy
 adult emergence seasons extending at least from early spring to late
 autumn. To ensure that predation experiments were ecologically relevant,
 we conducted them at four of our <em>Zelandoperla</em>
 sampling sites, during mid-Autumn 2024 (mid April to early May), at the
 tail end of the stonefly emergence season. We selected two forested
 (Leith: elevation 100-200 m; Whisky:elevation 200-220 m) and two
 deforested (Pigroot: elevation 500-520 m; Black Jacks: elevation 70-100 m)
 streams. At each site, approximately 50 pairs
 of <em>Zelandoperla</em> color models mounted on cobble stones
 were placed on riparian rocks/boulders at intervals of ~5-10 m, in total
 spanning several hundreds of metres of riparian habitat. For each model
 pair, black and brown models were placed 30-50 cm apart on a large
 stone/boulder. Models were left at their field locations for 72 hours
 prior to their recollection and returned to the laboratory for inspection
 and photography. Longer-running field trials were considered unsuitable
 given the potential risks of model damage associated with precipitation
 and fluctuating river levels linked to New Zealand’s oceanic
 climate. Models were inspected for damage and
 photographed, with distinctive damage attributed to candidate predator
 groups (birds, rodents, or insects) based on the approach of Zvereva and
 Kozlov 2021 and Guerra et al 2024. We recorded attack data for each model,
 and tested for associations between attack rates and mimicry/coloration
 across all predator categories, sites, and habitat types, using Fisher’s
 exact test. <strong>Statistical analysis and
 code</strong> Phylogenetic logistic regression
 (phylogenetic generalized linear mixed model) was fit using the Bayesian
 software JAGS and executed from R via the R interface
 <em>rjags</em>, where parameters were estimated using Markov
 Chain Monte Carlo (MCMC) sampling with the Gibbs sampler. The number of
 individuals classified as mimics from population  was modelled as a
 binomial distribution with total number of individuals  and frequency of
 mimics.
description: # Data from: Human-driven evolution of insect color
 [https://doi.org/10.5061/dryad.p5hqbzkxz](https://doi.org/10.5061/dryad.p5hqbzkxz) ## Description of the data and file structure ### Files and variables #### File: Mimicry\_algorithm\_robustphylocov.docx **Description:** Code used to create a phylogenetic correlation matrix from Zelandoperla fenestrata GBS data #### File: Mimicry\_Robustphylomatrix.xlsx **Description:** Phylogenetic matrix based on GBS data indicated the genetic relationships among populations ##### Variables * Each cell indicates the genetic distance between two Zelandoperla fenestrata stonefly populations. Site codes: Bj = Black Jacks, Chloris = Chloris Pass, Fg = Frasers Gully, Hf = Hoods Creek, Jim - Jimmys Creek, Kel = Kelceys Bush, Lee2a = Lee Stream, Matai = Matai Creek, Mb = MacAndrew Bay, MePass = Meyers Pass, Mim = Leith River, Mn = Maungatua, Owaka = Owaka Creek, Pass = Pass Creek, Pigroot = Pigroot Creek, Sm = Six Mile Creek, Stoney = Stoney Creek, Whiskey = Whiskey Creek #### File: Mimicry\_Bayesian\_Inference.docx **Description:** Code used for phylogenetic logistic regression  #### File: Bird\_Predation.xlsx **Description:** Raw data for bird predation experiments. Each tab represents a different location (Leith, Whiskey, Pig Root and Black Jacks) ##### Variables * Pickup Date = date when models were collected. Location = site location of models. Codes = model codes (black model/brown model) . Black = damage to the black model, and location of damage (none = no damage, bird = bird damage, insect = insect damage, rodent = rodent damage, . = models not found/recollected). #### File: Mimicry\_RGB.xlsx **Description:** Raw data for phenotypic analyses of color using digital photography ##### Variables * Stonefly species (Z = Zelandoperla fenestrata, A = Austroperla cyrene). Site = sample site. ID = sample ID at specific site. Head R = Red colouration on head of specimen. Head G = Green colouration on head of specimen. Head B = Blue colouration on head of specimen. Pronotum R = Red colouration on pronotum of specimen. Pronotum G = Green colouration on pronotum of specimen. Pronotum B = Blue colouration on pronotum of specimen. Leg R = Red colouration on leg of specimen. Leg G = Green colouration on leg of specimen. Leg B = Blue colouration on leg of specimen. ## Code/software Datasheets **MimicryRGB.xlsx**, **Bird Predation.xlsx,** and **MimicryRobustPhylogeny.xlsx** can be opened in Microsoft excel. The code in **Mimicry_algorithm_robustphylocov.doc** and **Mimicry_Bayesian_Inference.doc** can be analyzed in R.