https://doi.org/10.5061/dryad.p2ngf1w30

Authors: Colby Behrens (1), Meghan F. Maciejewski (1), Alexandra Sumarli (2), Gina Lucas (2), German Lagunas-Robles (2,3), Kieran Samuk (2), Alison M. Bell (1,4,5,6)

(1) Department of Evolution, Ecology and Behavior, School of Integrative Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USA

(2) Department of Evolution, Ecology, and Organismal Biology, University of California, Riverside, Riverside, California, USA

(3) Department of Biology, Indiana University, Bloomington, IN, USA

(4) Program in Ecology, Evolution and Conservation, University of Illinois Urbana-Champaign, Urbana, IL, USA

(5) Carl R. Woese Institute for Genomic Biology, University of Illinois Urbana-Champaign, Urbana, IL, USA

(6) Program in Neuroscience, University of Illinois Urbana-Champaign, Urbana, IL, USA

* README contents:

-Associated manuscript

-Study summary

-Software information

-Dataset contents

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Associated manuscript:

*Title: Evidence of parental care as a newly identified reproductive isolating barrier

• Authors: as above

Study summary:

This dataset contains data collected between 2021, 2022, and 2024. Three-spined stickleback (Gasterosteus aculeatus) from two Nova Scotian populations, referred to as “whites” and “commons”, were hybridized and reared in the laboratory. Behavioral measurements were then collected on whites, commons, and a full reciprocal cross of F1 hybrids. We collected data on nest architecture, courtship rates, and parenting behaviors. Rates of egg cannibalism were also collected on commons, whites, and F2 hybrids. See associated manuscript for more information

Software information:

The software R is required to run the analyses performed here. R version 4.0.2 was used in these analyses. The following R packages were loaded: blmeco, dplyr, lme4, lmerTest, tidyr, ggplot2, multcompView, EnvStats, cowplot, emmeans, car, dunn.test, betareg, fitdistrplus

Data and File Overview:

The dataset includes five data files in .csv format and a single R script (Behavior_survival_analysis.R) that is used to analyze all of the data files. Data files are described below.

File 1: 2024_Reproductive_Isolation_behavior.csv

This file includes data from parenting trials. Columns are named for the occurrence or time spent on each of the behaviors described in the manuscript.

Missing data code: NA

Specific columns are described below:

• ID: Name of the focal fish

• Fert.Date: Date (mm/dd/yyyy) that the male's clutch was fertilized as well as time (AM or PM)

• days.post.fert: The number of days after clutch fertilization that the trial occurred. Measured in whole or half days. A "0" means the trial occurred immediately after fertilization.

• date: Date (mm/dd/yyyy) of the trial

• AM_PM: Indicates if the trial was performed in the morning (AM) or afternoon (PM)

• time: time of the trial

• observer: initials of the observer

• dispersals: number of egg dispersals in the trial

• fan_bouts: number of times the male fanned the nest

• glues: number of nest glues in the trial

• nest_bouts: number of times the male attended the nest

• pokes: Number of times the male poked the nest

• retrievals: Number of times a dispersed egg was grabbed out of the water column or off the ground

• spits: Number of times the male spit sand or other debris onto the nst

• thru_nest: number of times the male swam through the nest

• fan_s: time(s) fanning the nest

• nest_s: time (s) attending the nest

• nest_min: time (min) attending the nest

• family: Clutch of the focal male

• population: Population of the focal male (common, white, or F1). If the male is an F1 hybrid, the population lists the mother's population x the father's population.

• egg_ecotype: ecotype of the mother. "C" represents common, "W" represents white.

• male_ecotype: Population of the focal male (common, white, or F1).

• mating: Mating type being analyzed. The first word and parenthetical represent the male ecotype and the final letter represents the female ecotype.

File 2: 2024_Reproductive_Isolation_nest.csv

This file includes data on nest characteristics. Raw nest characteristics include Location (where in the tank the nest was built), Flatness, Opening, Sand, and Algae_mass (part of the main algae clump or separate). NAs in these columns mean that the trait could not be measured or observed without disrupting the experiment or nest. The last four variables were combined into a single nest architecture scale (scale_omsf). To test for robustness, scales including only opening, sand, and flatness (scale_osf) were also included. Specifics on the coding and scaling are described in a previously published dataset (Behrens, Colby, Meghan F. Maciejewski, Eric Arredondo, Anne C. Dalziel, Laura K. Weir, and Alison M. Bell. 2024. “Divergence in Reproductive Behaviors Is Associated with the Evolutionary Loss of Parental Care.” American Naturalist 203 (5): 590–603. https://doi.org/10.1086/729465.)

Specific columns are described below:

Missing data code: NA

• MaleID: Name of the focal fish

• Date: date of the measurement (mm/dd/yyyy)

• Observer: Initials of the individual recording the data

• Algae_mass: Identifies if the nest was located in the main algae mass (main) or separate (separate).

• Flatness: Score for nest flatness (ranges 1-5; scale described in paper)

• Location: The location of the nest in grid format

• Opening: Score for the nest opening (ranges 1-5)

• Sand: Score for the amount of sand on the nest (ranges 1-5)

• Algae_mass_num: Score (1 or 5) for if the nest was located in the main algae mass (described in paper)

• scale_omsf: combined opening, flatness, sand, and algae_mass score (ranges 4-20)

• scale_osf: combined opening, sand, and flatness score (ranges 3-15)

• family: family of the phenotyped individual

• population: Population of the focal fish (common, white, or F1)

File 3: 2024_Reproductive_Isolation_courtship.csv

This file includes data from courtship trials. Metadata on the focal male's ID, trial information (time, female.ID), focal male's family (family), and population (population) are included. Specific columns are described below:

Missing data code: NA

• Date:date of the trial (mm/dd/yyyy)

• Time: time of the trial

• Male.ID: Name of the focal fish

• Female.ID: Name of the female fish

• Female.Ecotype: Ecotype (white or common) of the female fish

• Observer.Initials: Initials of the individual recording the data

• Female.pre.W: weight of the female (grams) before the trial

• Female.post.W: weight of the female (grams) after the trial. If the female did not release her clutch she was not reweighed, so the cell is left as "NA".

• Fertilization.: Indicates if the courtship trial resulted in a fertilization (Y=yes, N=no).

• family: clutch name of the focal male

• population: population of the focal male

File 4: 2024_Reproductive_Isolation_survival.csv

This file includes data on the survival of clutches, their mating type, and the rearing type. Each row describes a single clutch of embryos.

Missing data code: NA

Specific columns are described below:

• Male.ID: Name of the focal fish. This fish sired the clutch of interest.

• Female.ID: Name of the mother of the clutch

• Male.Family: Family name of the father

• Mating.type: Two character representation of the father and mother pairing. The first letter represent the father and the second letter represents the mother. "C"=common, "W"=white,"Y"=F1 hybrid whose father was a common, "P" = F1 hybrid whose father was a white.

• Care_type: Indicates if the clutch received care from the father (Care) or was artificially reared (Artificial)

• Date.of.fert: Date (mm/dd/yyyy) that the male's clutch was fertilized

• Time.of.Fert: Time that the clutch was fertilized

• Date.male.removed: Date that the clutch and male were removed from their tanks. This is only relevant to "Care" clutches, so all "Artificial" clutches are left NA.

• Female.preW: weight of the female (grams) before the trial or fertilization

• Female.post.W: weight of the female (grams) after the trial or fertilization.

• Clutch_weight: weight of the clutch (grams)

• Clutch_weight_mg: weight of the clutch (mg)

• Clutch_name: ID of the clutch

• egg.count: Number of eggs in the initial clutch. If the clutch was artificially reared the eggs were counted. If the clutch received care the number could not be counted and is left NA.

• fertilized.eggs: Number of eggs fertilized 30 minutes after fertilization. If the clutch was artificially reared the eggs were counted. If the clutch received care the number could not be counted and is left NA.

• egg_count_day4_alive: Number of embryos alive after 4 days.

• egg_per_mg: The number of eggs alive after 4 days per milligrams of initial clutch weight.

• live_in_nest: Number of living embryos found in the nest after 4 days.

• live_outside: Number of living embryos found outside of the nest after 4 days.

• egg_ecotype: ecotype of the mother. C represents common, W represents W.

• male_ecotype: ecotype of the father of the clutch. Males are either common, white, F1 (CxW), or F1 (WxC).

File 5: cannibalism.csv

This file includes data on filial cannibalism. After fertilizing embryos, males were collected from their tanks and dissected and the number of cannibalized eggs were counted. More details are in the manuscript.

Missing data code: NA

• ID: Name of the focal fish

• Length: Standard length of the fish in mm

• year: year that the data was collected

• population: population of the focal fish. Individuals were either Common, White, or F2 hybrid.

• eggs_eaten: The number of eggs found in the stomach of the male

• mass of the focal fish (g)