DOI: 10.5061/dryad.x95x69prp
title: Divergence in reproductive behaviors is associated with the evolutionary loss of parental care

description: The mechanisms underlying the divergence of reproductive strategies
 between closely-related species are still poorly understood. Additionally,
 it is unclear which selective factors drive the evolution of reproductive
 behavioral variation and how these traits coevolve, particularly during
 early divergence. To address these questions, we quantified behavioral
 differences in a recently diverged pair of Nova Scotian three-spined
 stickleback (Gasterosteus aculeatus) populations, which vary in parental
 care, with one population displaying paternal care and the other lacking
 this. We compared both populations, and a full reciprocal F1 hybrid cross,
 across four major reproductive stages: territoriality, nesting, courtship,
 and parenting. We identified significant divergence in a suite of
 heritable behaviors. Importantly, F1 hybrids exhibited a mix of behavioral
 patterns, some of which suggest sex-linkage. This system offers fresh
 insights into the coevolutionary dynamics of reproductive behaviors during
 early divergence and offers support for the hypothesis that coevolutionary
 feedback between sexual selection and parental care can drive rapid
 evolution of reproductive strategies.
description: # Divergence in reproductive behaviors is associated with the evolutionary
 loss of parental care *Authors: Colby Behrens (1), Meghan F. Maciejewski
 (1), Eric Arredondo (1), Anne C. Dalziel (2), Laura K. Weir (2), Alison M.
 Bell (1,3,4,5) (1) Department of Evolution, Ecology and Behavior, School
 of Integrative Biology, University of Illinois at Urbana-Champaign,
 Urbana, IL, USA (2) Department of Biology, St. Marys University, Halifax,
 Nova Scotia, Canada (3) Program in Ecology, Evolution and Conservation,
 University of Illinois Urbana?Champaign, Urbana, IL, USA\ (4) Carl R.
 Woese Institute for Genomic Biology, University of Illinois
 Urbana?Champaign, Urbana, IL, USA (5) Program in Neuroscience, University
 of Illinois Urbana Champaign, Urbana, IL, USA * README contents:
 -Associated manuscript -Study summary -Software information -Dataset
 contents --- ## Associated manuscript: *Title: Divergence in reproductive
 behaviors is associated with the evolutionary loss of parental care *
 Authors: as above ## Study summary: This dataset contains data collected
 between 2020 and 2021. 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 territorial aggression, nest architecture, courtship,
 and parenting behaviors to understand how the behaviors have diverged
 between the two natural populations. 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, plyr, dplyr, lme4, lmerTest,
 tidyr, ggplot2, multcompView, EnvStats, emmeans, cowplot, emmeans, car,
 ggpubr, corrplot, gplots, dunn.test ## Data and File Overview: The dataset
 includes five data files in .csv format and a single R script that is used
 to analyze all of the data files. Data files are from each major behavior
 timepoint (aggression, nesting, courtship, parenting) and are described
 below. An additional file contains data formatted for cross context
 comparisons. Here, data on nest architecture, the 1st courtship trial,
 post-fertilization parenting, and 4 days post-fertilization behaviors are
 organized by male ID. Individual behaviors and column names are described
 in detail in the supplementary material of the associated manuscript. A
 single R script is used to analyze all of the data files. -File 1:
 Aggression.csv - This file includes data from territorial aggression
 trials. Variables are described below: *ID: Name of the focal fish *date:
 date of the trial (mm/dd/yyyy) *observer: observer of the trial *time:
 time of the start of the trial. *intruder_family: Tank that intruder males
 were drawn from. Focal individuals never interacted with a close relative
 during a trial *intruder_section: Section of the tank that the intruder
 flask was placed *Population: Population of the focal individual *Length:
 Standard length (mm) of the focal individual at the time of the trial
 *orient_time: Seconds oriented towards the intruder flask
 *intruder_orient_bout: Number of times the focal male oriented to the
 intruder. *intruder_section_bout: number of times the focal male entered
 the section with the intruder male *intruder_section_time: Time in seconds
 spent in the intruder's section *intruder_zigags: Zigzags towards the
 intruder male *intruder_leads: Leads toward the intruder male
 *intruder_bites: Bites toward the intruder male *section1_bouts,
 section2_bouts, section3_bouts: Number of times that a focal male moved
 into each of the three named sections. *bites: total number of bites in a
 trial *charges: total number of charges in a trial *fan_bouts, glue_bouts,
 nest_bouts, leads, fan_time, and nest_time: Courtship and parental
 behaviors that could occur, and are described in the courtship and
 parenting sections, but were rare in aggression trials. Their occurrence
 is recorded here. *orient1, orient3,total_orient: Number of times the male
 oriented to a flask in section 1, section 3, and total *section1_time,
 section2_time, section3_time: Time spent in each of the three sections
 -File 2: Nesting.csv - This file includes data on nest characteristics,
 which are further descried in the manuscript. 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)
 and including only sand and flatness (scale_sf) were also included.
 Specific columns are described below: *MaleID: Name of the focal fish
 *Date: date of the measurement (mm/dd/yyyy) *Observer: Initials of the
 individual recording the data *Flatness: Score for nest flatness
 (described in paper) *Location: The location of the nest in grid format
 *Population: Population of the focal fish *Opening: Score for the nest
 opening (described in paper) *Sand: Score for the amount of sand on the
 nest (described in paper) *Algae_mass: 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 *scale_osf: combined
 opening, sand, and flatness score *scale_sf: combined sand and flatness
 score *Male_length: Standard length (mm) of the focal male. Cells with NA
 do not have available data. *Male_weight: Weight in grams of the focal
 male. Cells with NA do not have available data. -File 3: Courtship.csv -
 This file includes data from courtship trials. Metadata on the focal
 male's ID (ID), trial information (time, femaleID,
 Male_Female_Date_trial) the female interest level for the trial
 (finterest), focal male's family (family), population (population),
 and the order of the trial (trial_order) are included. Remaining columns
 represent the occurrence or time spent on each of the behaviors described
 in the manuscript. Specific columns are described below: Missing data
 code: NA *ID: Name of the focal fish *date:date of the trial (mm/dd/yyyy)
 *time: time of the trial *femaleID: Name of the female fish *finterest:
 the female interest level for the trial, with progressively more interest
 from "No Interest" to "Low Interest" to "High
 Interest" *family: clutch name of the focal male *population:
 population of the focal male *ID_interest: merged male ID and
 "finterest" column *trial_order: order of the courtship trials
 for a particular male for each specific female interest (finterest) level
 *trial_order_total: The overall order of the courtship trials for a
 particular male *length_of_trial: length of the trial in minutes
 *observer: initials of the observer *Male_length: standard length of the
 male (mm) *Male_weight: weight of the male (g) *Male_Female_trial: The ID
 of the male followed by "_", followed by the ID of the female
 *Female.pre.W: Weight of the female (g) before the trial started
 *angle_ups: Number of times the female angled her belly up to show
 interest in the male *bites: Number of times the focal male bit the female
 *chase: Number of times the focal male chased the female *dorsal_pricks:
 Number of times the focal male performed a dorsal prick aginst the female
 *enter_nest_bouts: Number of times the female entered the nest *fan_bouts:
 number of times the male fanned his nest *glues: number of times the male
 glued the nest *inspects: number of times the female inspected the
 male's nest *leads: number of times the male led the female to the
 nest *nest_bouts: Number of times the male attended his nest *follows:
 number of times the female followed the male *pokes: number of times the
 male poked the nest *quivers: number of times the male quivered against
 the female *shows: number of times the male showed the female his nest
 *thru_nest: number of times the male swam through his nest *zigzags:
 number of zigzags the male performed towards the female *quiver_time: time
 (s) the male quivered against the female during the trial *fan_s: time (s)
 the male spent fanning the nest *nest_min: time (min) the male spent
 nesting during the trial -File 4: Parenting.csv - This file incudes data
 from parenting trials. As in other files, the 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 *AM.PM: Time of the day (AM or PM) that
 the clutch was fertilized *days.post.fert: The number of days after clutch
 ferilization that the trial occurred. Measured in whole or half days
 *date: Date (mm/dd/yyyy) of the trial *time: time of the trial *observer:
 initials of the observer *dispersal: 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_time:
 time(ms) fanning the nest *nest_time: time (ms) attending the nest
 *family: Clutch of the focal male *Population: Population of the focal
 male *fan_s: number of seconds spent fanning the nest *nest_s: number of
 seconds spend attending the nest *nest_min: number of minutes spent
 attending the nest *ID_fertdate: merged column of the male's ID and
 the fertilization date of the clutch *Male_length: Standard length (mm) of
 the focal male *Male_weight: weight (g) of the focal male -File 5:
 Cross-context_data.csv - This file includes data from Nesting, Courtship,
 and Parenting trials. It has been merged across contexts so each male has
 data on nest architecture, courtship (only from the first trial with an
 interested female), parenting behavior immediately after fertilization,
 and parenting 4 days after fertilization, if available. It has been saved
 separately from other files for ease of analysis. All columns have a
 prefix (nest, courtship, parent0, parent4) to describe the context of the
 behavior. "parent0" is from parenting trials immediately after
 fertilization while "parent4" is from parenting trials 4 days
 after fertilization. All column names after the prefix have been described
 for the above files. Missing data code: NA