DOI: 10.5061/dryad.7m0cfxpqt
title: Novelty and emergent patterns in sperm: morphological diversity and evolution of spermatozoa and sperm conjugation in ground beetles (Coleoptera: Carabidae)

description: The beetle family Carabidae, with about 40,000 species, exhibits enough
 diversity in sperm structure and behavior to be an excellent model system
 for studying patterns and processes of sperm evolution. We explore their
 potential, documenting sperm form in 177 species of ground beetles using
 light microscopy and collecting data on 1 qualitative and 7 quantitative
 sperm phenotypic traits. Our sampling captures 61% of the tribal-level
 diversity of ground beetles. These data highlight the notable
 morphological diversity of sperm in ground beetles and suggest that sperm
 in the group have dynamic evolutionary histories with much morphological
 innovation and convergence. Sperm vary among species in total length
 (48–3,400mm), head length (0.5–270mm), and head width (0.2–6.3mm). Most
 ground beetles make sperm with heads that are indistinct from the flagella
 at the gross morphological level. However, some or all
 Omophron,Trachypachus, and Dyschiriini make broad-headed sperm that show
 morphological differences between species. Most ground beetles package
 their sperm into groups of sperm, termed conjugates, and ground beetles
 show variation in conjugate form and in the number and arrangement of
 sperm in a conjugate. Most ground beetles make sperm conjugates by
 embedding their sperm in a hyaline rod or spermatostyle. The spermatostyle
 is remarkably variable among species and varies in length from
 17–41,000mm. Several unrelated groups of ground beetles make only
 singleton sperm, including Nebriinae, Cicindelinae, many Trechinae, and
 the tribe Paussini. In order to study patterns in sperm evolution, we
 combine these data with a low-resolution phylogeny of ground beetles.
 Results from modern comparative analyses suggest the following: sperm
 differ from conjugates in some aspect of their underlying evolutionary
 process, sperm have influenced conjugate evolution and vice versa, and
 conjugation with a spermatostyle likely evolved early within the history
 of Carabidae and it has been lost independently at least three times.
description: These data include sperm data recorded for 177 species of the
 beetle family Carabidae, trees dervied from a low-resolution phylogeny of
 the group, R scripts and data, and online supporting
 information. The sperm data was collected from live
 beetles using light microscopy and image analysis. The trees were derived
 from the literature and software programs designed for evolutionary
 analyses (Mesquite and R). The data were analyzed using
 a comparative phylogenetic framework using various R packages.
description: <strong>Supplemental figure</strong>
 Figure S1 SuppInfo.pdf (a sperm conjugate
 morphospace for 55 species of ground beetles showing trait loadings for
 PC1 and PC2)  
 <strong>Media</strong> 
 16 movies of carabid sperm of 8 different species viewed in
 saline on slides  
 <strong>Methods S1 SuppInfo.docx</strong>
 Supplemental methods describing additional details of
 our methods for sperm data collection  
 <strong>R data</strong> 
 Various data files used for comparative analyses in R
 1000trees.phy (1000 fully dichotomous random
 resolutions of our low-resolution phylogeny of Carabidae in phylip
 format) colors.txt (a text file with discrete
 conjugation type data for carabid beetles for use in plotting)
 ConjugateDataCarabidR.txt (a text file containing continouous
 trait data for our studied conjugate traits across carabid
 beetles) discretetraitsRdata.txt (a text file
 containing discrete character data for carabid beetles)
 JustSpermDataCarabidR.txt (a text file containing continuous
 trait data for our studied sperm traits across carabid beetles)
 Species.Pairs.Data.txt (a text file containing the quotients of
 species traits values for 7 continouous sperm traits between 20 species
 pairs of ground beetles) SpermDataCarabid.nex (a
 nexus file for Mesquite containing our low-resolution phylogeny and 7
 continous sperm trait data for 177 species of ground beetles)
 SpermDataCarabid.phy (a low-resolution phylogeny of Carabidae in
 phylip format with branch lengths arbitrarily ultrametricized in
 Mesquite) SpermDataCarabidR.txt (a text file
 containing continuous trait data for our 7 continuous sperm traits across
 carabid beetles)  
 <strong>R scripts</strong> 
 CompareUnivariateModels.2.2.R (R script that fits
 several models of evolution for sperm head length and the number of sperm
 in a conjugate using phytools function make.simmap to reconstruct
 ancestral regimes and OUwie for model fitting and data
 simulation) construct.morphospace.1.0.R (R script for
 reconstructing sperm phylomorphospaces for 101 species of ground beetles
 and a morphospace of all studied sperm data for 55 species of ground
 beetles) pANOVA.PGLS.spermtraits.1.0.R (R script for
 performing pANOVAs and PGLS on ground beetle sperm data)
 Species.Pairs.Comparison.1.0.R (R script that performs a one-way
 ANOVA and a post-hoc Tukey HSD test of sperm and conjugate data for 20
 species pairs of ground beeltes) StochasticMaps.2.0.R
 (R script that performs ancestral character estimation using stochastic
 character mapping of conjugate type data for ground beetles using various
 sets of trees with different branch length schemes) 
   <strong>Spreadsheet S1
 SuppInfo.xslx</strong> Excel
 spreadsheet with sperm trait data recorded for 397 specimens of ground
 beetles.  
 <strong>Supplemental References Cited
 SuppInfo.docx</strong> Microsoft word
 document containing our supplemental references cited
   <strong>Table S1
 SuppInfo.docx</strong> A supplemental
 table containing data summarizing the state of knowledge of ground beetle
 sperm phenotype.  
 <strong>Table S2 SuppInfo.docx</strong>
 A supplemental table containing taxon and specimen
 sampling for sperm data.  
 <strong>Table S3 SuppInfo.docx</strong>
 A supplemental table containing data locality data of
 studied specimens.  
 <strong>Table S4 SuppInfo.docx</strong>
 A supplemental table containing sperm data summarized
 across specimens for 177 species of ground beetles.
   <strong>Table S5
 SuppInfo.docx</strong> A supplemental
 table containing results of a one-way ANOVA of data for 20 species pairs
 of ground beetles recorded for all 7 studied continuous traits.
   <strong>Taxon-by-taxon results S1
 SuppInfo.docx</strong> Supplemental
 results containing detailed taxon-by-taxon observations of sperm taken
 from this study and the literature.