DOI: 10.5061/dryad.32665
title: Data from: Postcopulatory sexual selection generates speciation phenotypes in Drosophila

description: Background: Identifying traits that reproductively isolate species, and
 the selective forces underlying their divergence, is a central goal of
 evolutionary biology and speciation research. There is growing recognition
 that postcopulatory sexual selection, which can drive rapid
 diversification of interacting ejaculate and female reproductive tract
 traits that mediate sperm competition, may be an engine of speciation.
 Conspecific sperm precedence (CSP) is a taxonomically widespread form of
 reproductive isolation, but the selective causes and divergent traits
 responsible for CSP are poorly understood. Results: To test the hypothesis
 that postcopulatory sexual selection can generate reproductive isolation,
 we expressed green (GFP) or red fluorescent protein (RFP) in sperm heads
 of recently diverged sister species, Drosophila simulans and D.
 mauritiana, to enable detailed resolution of species-specific sperm
 precedence mechanisms. Between-species divergence in sperm competition
 traits and mechanisms prompted six a priori predictions regarding
 mechanisms of CSP and degree of cross asymmetry in reproductive isolation.
 We resolved four distinct mechanisms of CSP that were highly consistent
 with predictions. These comprise interactions between multiple
 sex-specific traits, including two independent mechanisms by which females
 exert sophisticated control over sperm fate to favor the conspecific male.
 Conclusions: Our results confirm that reproductive isolation can quickly
 arise from diversifying (allopatric) postcopulatory sexual selection. This
 experimental approach to "speciation phenotypes" illustrates how
 knowledge of sperm precedence mechanisms can be used to predict the
 mechanisms and extent of reproductive isolation between populations and
 species.
description: RemateSperm numbers in space
 and time after second mating. Variables: Cross - Species identity of first
 and second male to mate with a D. simulans female, where "s"
 indicates D. simulans and "m" indicates D. mauritiana, such that
 "ss" indicates that both males were D. simulans, "ms"
 indicates that the first male was D. mauritiana and the second male was D.
 simulans, etc. MinASM – time after the start of mating at which female was
 frozen, in minutes. Remate – day after first mating (day 0) on which
 female remated. Cop1 – duration in minutes of the first copulation. Cop2 –
 duration in minutes of the second copulation. PriorProg – number of
 progeny produced by female before remating. Matings - number of successful
 matings by that female. ThoraxF - female thorax length, in arbitrary
 units. ThoraxM1 - thorax length of the first male, in arbitrary units.
 ThoraxM2 - thorax length of the second male, in arbitrary units. Bursa1 –
 number of first-male sperm in the bursa. Bursa2 – number of second-male
 sperm in the bursa. SR1 – number of first-male sperm in the SR. SR2 –
 number of second-male sperm in the SR. Sp1 – number of first-male sperm in
 the spermathecae. Sp2 – number of second-male sperm in the
 spermathecaeVirginData on sperm transfer
 and storage after single mating of D. simulans female with D. mauritiana
 male. Variables: Treat – treatment, storage or transfer. Cop – copulation
 duration in minutes. ThoraxF - female thorax length, in arbitrary units.
 ThoraxM - thorax length of the second male, in arbitrary units. Bursa –
 number of sperm in the bursa. SR – number of sperm in the seminal
 receptacle. Sp – number of sperm in the spermathecaeEjection1Timing and number of sperm ejected after a single mating of a D. simulans female with a D. mauritiana male. Variables: CopDur - copulation duration in minutes. EjTime - minutes after start of copulation when female ejected sperm mass. Sperm – number of sperm in the ejected sperm massEjection2Timing and numbers of first- and second-male sperm ejected after remating, and numbers of first- and second-male sperm remaining in the female reproductive tract after ejection. Variables: Cross – Species identity of first and second male to mate with a D. simulans female, where "s" indicates D. simulans and "m" indicates D. mauritiana, such that "ss" indicates that both males were D. simulans, "ms" indicates that the first male was D. mauritiana and the second male was D. simulans, etc. Remate – day after first mating (day 0) on which female remated. Eject - 1 if female ejected, 0 if she did not eject. EjStart - Minutes after start of mating when female began ejecting a sperm mass (sometimes it got stuck). EjEnd - Minutes after start of mating when female finished ejecting a sperm mass. PriorProg - presence of prior progeny, where 0 = none, 1 = a few, 2 = a lot. Eject1 – number of first-male sperm in ejected sperm mass. Eject2 - number of second-male sperm in ejected sperm mass. ThoraxF - female thorax length, in arbitrary units. ThoraxM - thorax length of the second male, in arbitrary units. Bursa1 – number of first-male sperm in the bursa. Bursa2 – number of second-male sperm in the bursa. SR1 – number of first-male sperm in the SR. SR2 – number of second-male sperm in the SR. Sp1 – number of first-male sperm in the spermathecae. Sp2 – number of second-male sperm in the spermathecaeVelocitySperm velocity and sperm numbers in the SR. Variables: Treat - sxm indicates single mating between D. simulans female and D. mauritiana male, NR indicates "no remate" treatment, other treatments are rematings defined by the species identity of first and second male to mate with a D. simulans female, where "s" indicates D. simulans and "m" indicates D. mauritiana, such that "ss" indicates that both males were D. simulans, "ms" indicates that the first male was D. mauritiana and the second male was D. simulans, etc. Region – region of the female reproductive tract, bursa or SR. Male – species and color tag of male whose sperm velocity is recorded. Order - first or second male to mate. Remate - for remating treatments, day after first mating (day 0) on which female remated. Velmps – instantaneous linear velocity in microns per second, averaged for each sperm and for all sperm recorded in a movie. N – number of sperm tracked per movie. SR.G – number of GFP sperm in the SR. SR.R – number of RFP sperm in the SR.mxsNumbers of sperm transferred and stored for D. mauritiana females mating with D. simulans males. Variables: Cross - mxs = D. mauritiana female mating once with a D. simulans male; mxsm = D. mauritiana female mating first with a D. simulans male followed by a D. mauritiana male; mxms = D. mauritiana female mating first with a D. mauritiana male followed by a D. simulans male. Cop - copulation duration in minutes, for second copulation for mxsm and mxms crosses. Bursa1 – number of first-male sperm in the bursa. Bursa2 – number of second-male sperm in the bursa. SR1 – number of first-male sperm in the SR. SR2 – number of second-male sperm in the SR. Sp1 – number of first-male sperm in the spermathecae. Sp2 – number of second-male sperm in the spermathecaeP2Paternity and sperm count data. Variables: Cross - Species identity of first and second male to mate with a D. simulans female, where "s" indicates D. simulans and "m" indicates D. mauritiana, such that "ss" indicates that both males were D. simulans, "ms" indicates that the first male was D. mauritiana and the second male was D. simulans, etc. Treat – time after mating for which paternity and sperm numbers in the female reproductive tract were quantified. ThoraxF - female thorax length, in arbitrary units. ThoraxM1 - thorax length of the first male, in arbitrary units. ThoraxM2 - thorax length of the second male, in arbitrary units. Cop1 – duration in minutes of the first copulation. Cop2 – duration in minutes of the second copulation. Remate – day after first mating (day 0) on which female remated. PriorProg – number of progeny produced by female before remating. P2Prog1 - number of progeny produced after remating sired by the first male. P2Prog2 - number of progeny produced after remating sired by the second male. P2 – proportion of progeny produced after remating sired by the second male. Bursa1 – number of first-male sperm in the bursa. Bursa2 – number of second-male sperm in the bursa. SR1 – number of first-male sperm in the SR. SR2 – number of second-male sperm in the SR. Sp1 – number of first-male sperm in the spermathecae. Sp2 – number of second-male sperm in the spermathecae.