doi: 10.1111/jeb.

12563

Receptive females mitigate costs of sexual conflict

T. HARANO*
*Department of Evolutionary Studies of Biosystems, School of Advanced Sciences, The Graduate University for Advanced Studies, Hayama, Japan
Laboratory of Ecological Science, Department of Biology, Faculty of Sciences, Kyushu University, Fukuoka, Japan

Keywords:

Abstract

artificial selection;
multiple mating;
polyandry;
seed beetle;
sexual harassment;
sexual selection.

Males typically gain fitness from multiple mating, whereas females often
lose fitness from numerous mating, potentially leading to sexual conflict
over mating. This conflict is expected to favour the evolution of female
resistance to mating. However, females may incur male harassment if they
refuse to copulate; thus, greater female resistance may increase costs
imposed by males. Here, I show that the evolution of resistance to mating
raises fitness disadvantages of interacting with males when mating is harmful in female adzuki bean beetles, Callosobruchus chinensis. Females that were
artificially selected for higher and lower remating propensity evolved to
accept and resist remating, respectively. Compared with females that
evolved to accept remating, females that evolved to resist it suffered higher
fitness costs from continuous exposure to males. The costs of a single mating
measured by the effect on longevity did not differ among selection line
females. This study indicates that receptive rather than resistant females mitigate the fitness loss resulting from sexual conflict, suggesting that even
though mating is harmful, females can evolve to accept additional mating.

Introduction
Sexual conflict, defined as conflict between evolutionary interests of individuals of the two sexes (Parker,
1979), arises when the fitness optima for males and
females cannot be achieved simultaneously (Parker,
2006). In general, optimal mating frequency is much
higher in males than in females. Males typically maximize their fitness by having as many mates as possible
because each mating offers an opportunity to sire offspring. In contrast, females seem not to require a number of mates to maximize their offspring production
(Bateman, 1948; Thornhill & Alcock, 1983). Females
may gain from multiple mating, but their fitness is usually maximized at intermediate mating frequencies,
beyond which further mating is deleterious (Arnqvist &
Nilsson, 2000). This is because mating carries a variety
of potential costs: time and energy expenditures
(Thornhill & Alcock, 1983), increased risk of predation
(Arnqvist, 1989; Kemp, 2012), risk of pathogen or paraCorrespondence: Tomohiro Harano, Department of Evolutionary Studies
of Biosystems, School of Advanced Sciences, The Graduate University
for Advanced Studies, Shonan Village, Hayama, Kanagawa 240-0193,
Japan. Tel.: +81 46 858 1617; fax: +81 46 858 1544;
e-mail: tharano518@yahoo.co.jp

320

site transmission (Hurst et al., 1995; Martinez-Padilla

et al., 2012), physical injury (Crudgington & Siva-Jothy,
2000; Blanckenhorn et al., 2002) and the toxic effects
of substances in male ejaculates (Chapman et al., 1995).
Thus, encounters between a male and a female frequently involve conflict over whether to mate, which is
a representative form of interlocus sexual conflict (Arnqvist & Rowe, 2005). This conflict is expected to select
for greater female resistance to male mating attempts,
causing females to copulate only with males that overcome this resistance. Males, in turn, should be selected
for the development of traits that seduce or coerce
females into mating (Holland & Rice, 1998; Gavrilets
et al., 2001; Kokko et al., 2003). This could lead to sexually antagonistic coevolution of female resistance and
male persistence, potentially driving reproductive isolation and speciation (Gavrilets et al., 2001; Arnqvist &
Rowe, 2005).
However, resistance to mating can also be costly to
females (Arnqvist & Rowe, 2005). Males that locate
females generally attempt to mate until females either
leave or submit to copulation (Thornhill & Alcock,
1983). Females that refuse to copulate may undergo
repeated mating attempts by males that is, sexual
harassment (Clutton-Brock & Parker, 1995). Male
harassment can cause drastic negative fitness

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Female receptivity and sexual conflict costs

consequences for females (Arnqvist & Rowe, 2005; Takahashi & Watanabe, 2010; Gasparini et al., 2012; Makowicz & Schlupp, 2013). A number of studies have
shown that females continuously exposed to males
have reduced fitness (Newport & Gromko, 1984; Partridge et al., 1987; M
uhlhauser & Blanckenhorn, 2002;
R
onn et al., 2006; Shuker et al., 2006). This reduction
reflects the magnitude of costs arising from sexual conflict and could result from elevated mating frequency as
well as sexual harassment (Partridge & Fowler, 1990).
In the adzuki bean beetle, Callosobruchus chinensis
(Coleoptera: Chrysomelidae: Bruchinae), mating has
been shown to reduce female longevity independently
of egg production (R
onn et al., 2006). Two factors give
rise to the costs to females of mating. First, the tip of the
male intromittent organ is covered with spines that
puncture and scar the inner reproductive tract of
females during copulation in Callosobruchus species,
including C. chinensis (Crudgington & Siva-Jothy, 2000;
R
onn et al., 2007; Hotzy & Arnqvist, 2009; Sakurai et al.,
2012). Second, seminal substances of C. chinensis males
are toxic to females (Yamane, 2013). Previous studies
have reported that remating did not enhance female
lifetime fecundity when adult females were denied food
and water (Harano, 2012) and that it reduced female
fecundity when females were given food and water
(Harano et al., 2006). Thus, a single mating seems optimal for females, and females are expected to come into
conflict over mating with males they encounter after
having already mated once. Single-mated females frequently refuse male mating attempts by kicking males
with their hind legs or by running away, whereas the
males persistently attempt to mount the females. Male
harassment has been shown to depress female lifetime
fitness in this species (Sakurai & Kasuya, 2008).
Here, I address whether the evolution of female resistance to mating reduces fitness costs to females from
sexual conflict in C. chinensis. Genetic variation in
female receptivity to remating is found in this species
(Harano & Miyatake, 2005, 2007a, 2009). It has been
suggested that this variation is due to sensitivity to
male-derived substances that induce nonreceptivity in
females (Yamane & Miyatake, 2012). Bidirectional artificial selection for female remating receptivity has produced direct responses, with females selected for higher
receptivity being less resistant to remating than those
selected for lower receptivity (Harano & Miyatake,
2009). Female remating receptivity has been shown to
be correlated with mating frequency within a given
period (Harano & Miyatake, 2007b). A previous study
showed that C. chinensis females housed with males had
lower fitness than those that were isolated from males
after mating once (R
onn et al., 2006). In this study, I
compared the extent to which fitness was reduced by
continuous exposure to males in females from different
selection lines. If the evolution of female resistance to
mating alleviates fitness costs from sexual conflict, this

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extent should be smaller in females selected for lower

receptivity to remating than in females selected for
higher receptivity to remating.
Furthermore, costs of mating itself may vary between
females with higher and lower remating receptivity. A
comparative study in Callosobruchus seed beetles found
that the costs of mating to females were reduced in species where females have evolved a thicker genital tract
wall as a counteradaptation to spiny male genitalia
(R
onn et al., 2007). In Callosobruchus maculatus, an
experimental evolution study showed that the negative
effects of genital injury inflicted during copulation on
fitness were smaller in females evolved under polygamous conditions compared with females evolved under
monogamous conditions (Gay et al., 2011). A theoretical study has suggested that females who have suffered
extensive damage from mating may refrain from further mating (Johnstone & Keller, 2000). If females that
are less vulnerable to copulation remate more readily,
per-mating costs are expected to be lower in females
selected for higher receptivity to remating than in those
selected for lower receptivity. To test this possibility, I
compared the effect of a single mating on fitness among
females of the different selection lines.

Materials and methods

Study animals
Callosobruchus chinensis is a worldwide pest of stored
legumes such as the adzuki bean, Vigna angularis, and
the cowpea, Vigna unguiculata (Tuda et al., 2006). Adult
females lay eggs on the surface of the host beans and
hatched larvae burrow into the bean, where they complete their development, pupation and eclosion. Adults
can reproduce and live about 12 weeks without drinking or feeding, but their fecundity and longevity greatly
increase when water and food are available (Harano
et al., 2006). All rearing and experiments were conducted at a temperature of 25 C with a photoperiod
cycle of 14 : 10 h light : dark.
Selection lines
I used artificial selection lines established and explained
in detail by Harano & Miyatake (2009). Briefly, two
replicate lines selected for higher female propensity to
remate (H1 and H2 lines) and two replicate lines
selected for lower female propensity to remate (L1 and
L2 lines) were established in parallel with the maintenance of two unselected control lines (C1 and C2 lines).
To assess female propensity to remate, virgin females
were mated once with males from their own line and
periodically received opportunities to remate with tester
males. Tester males were collected as virgins from a
stock culture of one strain (is C: originating from
Ishigaki City, Japan, in 1997; see Yanagi & Miyatake,

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T. HARANO

2003) throughout the selection and subsequent experiments. At each generation, the 10 females with the
shortest latency to remating were selected in the H1
and H2 lines, and 10 females that did not remate at all
were selected in the L1 and L2 lines. Eggs laid by the
selected females before their remating opportunity were
used to found the next generation; thus, all offspring
were derived from females that experienced no effect of
remating, and second males sired no offspring. The C1
and C2 lines were maintained by propagating 10
females chosen randomly for every generation. Female
remating propensity diverged among the selection lines
after more than 10 generations of selection. The differences in female remating propensity may be attributable to female receptivity to remating, which is a trait
of females, and/or to male ability to inhibit female remating; however, the differences remained when
females of all lines were presented with tester males at
both the first mating and remating to standardize male
effects, indicating that female remating receptivity
responded to selection.
All lines were maintained without selection after
generation 18. In this study, I restarted selection at
generation 36 and continued until generation 57, except
for generations 4446 and 5355. Five generations from
the resumption of selection, the proportion of females
that remated diverged consistently over time between
the lines selected for higher and those selected for lower
female remating propensity (Fig. S1). After generation
58, all lines were maintained without selection and used
for the subsequent experiments. When selection was
eliminated, each line was propagated through approximately 20 pairs chosen randomly for every generation.
I established that female receptivity to remating differed between the selection lines as previously reported
(Harano & Miyatake, 2009) at generation 61. A generalized linear mixed model (GLMM) using the binomial
distribution and a logit link function, with selection
regime (higher, control or lower) as a fixed effect and
replicate line nested within the selection regime as a
random effect, showed that selection regime had a significant effect on the proportion of females that remated (Wald v2 test: v22 = 26.46, P < 0.001). Compared
with the control females, females from lines selected for
higher and lower propensity to remate had, respectively, higher and lower remating receptivity (Fig. 1).
Costs of male exposure
I established no-exposure and male-exposure treatments. In the no-exposure treatment, females were
mated once and then had no contact with males. In the
male-exposure treatment, females were mated once
and then cohabited with males, exposed to multiple
mating opportunities and to sexual harassment by
males. I used virgin females collected from each line at
generation 78. Females at 2448 h post-emergence

Fig. 1 Remating receptivity of females from selection lines. H1 and

H2 represent replicate lines selected for higher female propensity
to remate; C1 and C2 represent replicate control lines; and L1 and
L2 represent replicate lines selected for lower female propensity to
remate. All females were mated first with tester males and
received opportunities to remate with tester males to check their
remating receptivity.

were mated once with a tester male in a glass vial

(4.4 cm high, 1.7 cm in diameter). Single-mated
females were placed individually in a plastic cup
(3.8 cm high, 7.6 cm diameter) that contained adzuki
beans as the oviposition substrate, adult food (1 : 2
yeast extract : sugar) and water-soaked cotton in the
cap of a glass vial. Under these conditions, female remating has been shown to have detrimental effects on
fecundity (Harano et al., 2006). The beans were
replaced with fresh ones after 10 days. Females were
randomly allocated to the two treatments (the numbers
of females for the no-exposure and male-exposure
treatments, respectively, were 24 and 21 in H1, 23 and
25 in H2, 22 and 23 in C1, 21 and 21 in C2, 23 and 22
in L1, and 16 and 16 in L2). In the male-exposure treatment, two tester males were placed into the plastic cup,
following R
onn et al. (2006). These males were replaced
after 10 days to ensure live males. Females were kept
until 20 days after their first mating, and I recorded
their survival or death and the number of eggs they
laid. Beans were provided ad libitum, so most beans
bore up to three easily countable eggs.
Costs of mating
Most females of lines selected for lower remating propensity refused to remate when given the opportunity
(Fig. 1). It is not possible to examine the effects of

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Female receptivity and sexual conflict costs

remating on the fitness of females that do not remate

when allowed to do so. However, excluding such
females from the examination results in a nonrandom
sample of females with higher remating receptivity in
lines selected for lower remating propensity. Thus, the
effects of remating on fitness could not be adequately
compared among selection line females. However, the
effect of a single mating could be compared among
selection line females because the majority of virgin
females in all lines mated with the first male offered to
them (Harano & Miyatake, 2009). I established no-mating and single-mating treatments and evaluated the
effect of a single mating on female longevity as a measure of fitness, because female C. chinensis lay fewer
eggs without mating. Virgin C. chinensis females have
mature eggs in their ovaries at emergence and rarely
lay eggs without an oviposition substrate even if they
have mated (Umeya, 1987). By providing no oviposition substrate, it is possible to estimate the costs of mating independent of egg production (Yanagi & Miyatake,
2003; R
onn et al., 2006).
Virgin females were collected from each line at generation 65 and assigned randomly to the two treatments (the numbers of females for the no-mating and
single-mating treatments, respectively, were 10 and 11
in H1, 27 and 26 in H2, 24 and 23 in C1, 26 and 26 in
C2, 24 and 26 in L1, and 25 and 26 in L2). In the nomating treatment, females were kept as virgins, and in
the single-mating treatment, females at 2448 h postemergence were mated only once with a virgin tester
male. The females were placed individually in an
empty petri dish (1.5 cm high, 5 cm in diameter) to
exclude the effects of egg laying, feeding and male
harassment. Costs to females of a single mating have
been detected previously under these conditions (R
onn
et al., 2006). All females were checked daily to record
dates of death.

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Results
Costs of male exposure
The selection regime had no significant effect
(v22 = 0.32; P = 0.853), but the treatment (v21 = 60.22;
P < 0.001) and the interaction of the selection regime
and treatment (v22 = 7.00; P = 0.030) had significant
effects on female fecundity, indicating that cohabitation
with males affected fecundity differently according to
the females selection line (Fig. 2a). I estimated the
effect of cohabitation on fecundity separately in each
selection regime using GLMMs, with treatment as a
fixed effect and replicate line as a random effect. The
estimate ( SE) of the fixed effect was 14.2 ( 5.3)
in females with higher receptivity to remating, 24.7
( 4.0) in control females and 34.1 ( 6.6) in females
with lower receptivity to remating. This experiment
underestimates lifetime fecundity in the females that
survived for 20 days after the first mating because it
did not continue until all oviposition ceased. As with
fecundity, the selection regime had no significant effect
(v22 = 0.97; P = 0.617), but the treatment (v21 = 9.78,

(a)

(b)

Statistical analysis
All data were analysed using a GLMM, with selection
regime (higher, control or lower), treatment and their
interaction as fixed effects and replicate line nested
within the selection regime as a random effect. Interactions indicated a difference between selection regimes
in the effect of the treatment. If an interaction had no
significant effect, I tested the main effects in a reduced
model without the interaction term. When fecundity or
longevity was the response variable, I used the Gaussian distribution and an identity link function. When
the proportion of females that survived for 20 days
after the first mating was the response variable, I used
the binominal distribution and a logit link function.
The significance of the explanatory variables was
assessed by a Wald v2 test. All analyses were performed
in R 3.0.2 (R Development Core Team, 2013).

Fig. 2 The effects of continuous exposure to males on (a)

fecundity (mean  SE) and (b) survival for 20 days after the first
mating in females of selection lines. H1 and H2 represent replicate
lines selected for higher female propensity to remate; C1 and C2
represent replicate control lines; and L1 and L2 represent replicate
lines selected for lower female propensity to remate.

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T. HARANO

P = 0.002) and the interaction between the selection

regime and treatment (v22 = 15.85, P < 0.001) had significant effects on the proportion of females that survived, indicating that cohabitation with males affected
survival rate differently according to the females selection line (Fig. 2b). Applying GLMMs to each selection
regime, the estimate ( SE) of the fixed effect (treatment) on survival rate was 0.1 ( 0.5) in females with
higher receptivity to remating, 4.2 ( 1.1) in control
females and 2.0 ( 0.6) in females with lower receptivity to remating. Thus, cohabitation had a smaller
effect on survival rate in females with higher receptivity to remating than in control females and females
with lower remating receptivity. Surviving females may
have continued to lay eggs after the experimental
period ended. Therefore, differences between females
with higher and lower remating receptivity in the effect
of cohabitation on lifetime fecundity could be larger
than those observed over the limited span of this
experiment.
Costs of mating
The interaction of selection regime and treatment had
no significant effect (v22 = 2.01; P = 0.366) on female
longevity. When the interaction term was removed,
both selection regime (v22 = 30.96; P < 0.001) and treatment (v21 = 48.59; P < 0.001) had significant effects on
female longevity. A single mating consistently shortened longevity across all lines of females (Fig. 3).

Discussion
In C. chinensis, mating inflicts genital scarring (R
onn
et al., 2007) and releases toxic seminal fluids (Yamane,
2013), causing detrimental effects on female fitness
(Harano et al., 2006; R
onn et al., 2006). Thus, females

Fig. 3 The effect of a single mating on adult longevity

(mean  SE) in females of selection lines. H1 and H2 represent
replicate lines selected for higher female propensity to remate; C1
and C2 represent replicate control lines; and L1 and L2 represent
replicate lines selected for lower female propensity to remate.

are expected to face sexual conflict over mating when

they encounter males after mating once. I found that
cohabitating with males reduced fitness to a larger
extent in females that had evolved to resist remating
than in females that had evolved to accept remating
(Fig. 2). This indicates that when mating is harmful,
receptive females better mitigate the fitness costs associated with sexual conflict. Sexual conflict over mating is
ubiquitous (Arnqvist & Rowe, 2005). This conflict is
hypothesized to generate chase-away sexual selection,
which drives antagonistic coevolution between female
resistance and male traits used to seduce or coerce
females into mating (Holland & Rice, 1998; Gavrilets
et al., 2001). This assumes that selection favours more
resistant females, causing mating success to be biased
towards males that overcome female resistance (Holland & Rice, 1998; Gavrilets et al., 2001; Kokko et al.,
2003). My results illustrate that this assumption is not
always the case.
A potential explanation for why females selected for
higher receptivity to remating suffered less from cohabitating with males is that they were less sensitive to
mating costs. This was not supported by my finding
that the fitness costs of a single mating in terms of its
effect on adult longevity were similar among the selection line females (Fig. 3). However, sensitivity to costs
of the first mating may not reflect sensitivity to further
mating if per-mating costs change over successive matings (Kuijper et al., 2006) and the pattern of change differs among selection line females.
Females cohabiting with males and allowed to mate
multiple times should experience sexual harassment if
they are reluctant to copulate when courted by males.
Thus, the effect of cohabitation on female fitness could
be due to additional mating and/or to male harassment.
In C. chinensis, male harassment depresses female fitness
(Sakurai & Kasuya, 2008). The costs to females may
result from energy expenditure while fending off
harassing males or from lost opportunities to oviposit.
Male C. chinensis that court females generally mount
from the rear, and reluctant females kick males with
their hind legs or run away. If females flee, males frequently pursue them. While being mounted or running
away, females are unable to oviposit. Whether females
with lower remating receptivity are harassed more than
females with higher receptivity remains to be elucidated, although costs of sexual harassment that outweigh the costs of copulation are the most plausible
explanation for the disadvantages for females of
increased resistance to remating.
In C. chinensis, adult longevity has previously been
shown to be longer in females that remate readily than
in those that refuse to remate (Harano et al., 2006).
This indicates a positive association between high remating receptivity and longevity at the phenotypic
level. In contrast, females selected for higher remating
receptivity died younger than females selected for lower

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Female receptivity and sexual conflict costs

remating receptivity (Fig. 3). This suggests that the phenotypic association is inconsistent with a genetic correlation between female remating receptivity and
longevity.
Females are theoretically expected to ensure adequate fertilization in a single mating (Bateman, 1948;
Thornhill & Alcock, 1983), although multiple mating in
females is found in most animal species (Birkhead &
Mller, 1998; Taylor et al., 2014). This discrepancy has
generated a number of hypotheses to account for the
evolution of female multiple mating (Yasui, 1998; Arnqvist & Nilsson, 2000; Simmons, 2001; Pitnick & Hosken, 2010). In C. chinensis, some hypotheses have been
empirically tested, but the reason for female multiple
mating remains an open question (Harano et al., 2006;
Harano & Miyatake, 2007b; Harano, 2012; Harano &
Katsuki, 2012). The results of this study are consistent
with the convenience polyandry hypothesis, which
states that the fitness benefits to females of multiple
mating come from their avoiding the costs of sexual
harassment (Thornhill & Alcock, 1983). On the other
hand, resistant females may acquire genetic benefits
from increased offspring fitness by screening males for
persistence or vigour (Cordero & Eberhard, 2003; Kokko et al., 2003). However, no significant effects of
female resistance on offspring fitness have been found
in C. chinensis (Maklakov & Arnqvist, 2009).
Experimental evolution approaches have been used
to investigate the evolutionary consequences of sexual
conflict. When sexual conflict was removed by enforcing monogamy, females evolved to be more susceptible
to costs from cohabitating with males in the fruit fly
Drosophila melanogaster (Holland & Rice, 1999) and in
the mite Rhizoglyphus robini (Tilszer et al., 2006). The
intensity of sexual conflict has also been manipulated
by altering the adult sex ratio, under the assumption
that sexual conflict should increase or decrease under
male- or female-biased sex ratios, respectively. Females
that were allowed to evolve under female-biased sex
ratio conditions suffered larger costs from male exposure than did females that were allowed to evolve
under male-biased sex ratios in D. melanogaster (Wigby
& Chapman, 2004; Nandy et al., 2014) and the flour
beetle Tribolium castaneum (Michalczyk et al., 2011).
These studies provide evidence that the ability of
females to reduce male-induced costs has evolved due
to sexual conflict. Genetic variation among females in
the degree to which female fecundity or longevity is
reduced by male exposure was detected in D. melanogaster (Friberg, 2005; Linder & Rice, 2005; Lew et al.,
2006). A lower remating rate was shown to be correlated with decreased costs of male exposure in female
D. melanogaster (Linder & Rice, 2005; Lew et al., 2006).
This contrasts with the results for C. chinensis from the
present study. This inconsistency is likely attributable to
the magnitude of costs to females of sexual harassment
relative to copulation. The balance between costs of

325

mating and avoiding mating may be species dependent

but may also vary according to ecological circumstances
within a species.
Sexual conflict over mating should generate selection
for an increased male capacity to persuade or coerce
females to mate that is, male offence (Rice, 1998).
This could result in persistent mating attempts by males
and sexual harassment towards females (Clutton-Brock
& Parker, 1995). If male harassment escalates and its
cost to females exceeds that of mating itself, then
females should gain a net fitness benefit from accepting
additional mating, putting females interests in conflict
with those of their previous mates, as female remating
reduces the paternity of previous mates (Simmons,
2001; Arnqvist & Rowe, 2005). This conflict should
generate selection for an increased male capacity to
deter females from remating that is, male defence
(Rice, 1998). Thus, my results suggest that conflict
between females and males willing to copulate with
them can trigger a separate conflict between females
and their previous mates. Consequently, a three-way
conflict between male offense, male defence and female
traits potentially drives the antagonistic coevolution of
males and females (Rice, 1998; Gavrilets & Hayashi,
2006).
As a consequence of sexual conflict, females living in
the presence of males would often be pressed to compromise between the costs of mating and sexual harassment. My results present evidence that it is better for
female C. chinensis entangled in sexual conflict to incur
the costs of mating. This demonstrates that evolution
does not always strengthen female resistance to mating,
even though mating is harmful to females.

Acknowledgments
I thank anonymous referees for valuable comments on
the manuscript. This study was partly supported by a
Grant-in-Aid for JSPS Fellows (203976) from the Japan
Society for the Promotion of Science.

References
Arnqvist, G. 1989. Multiple mating in a water strider: mutual
benefits or intersexual conflict? Anim. Behav. 38: 749756.
Arnqvist, G. & Nilsson, T. 2000. The evolution of polyandry:
multiple mating and female fitness in insects. Anim. Behav.
60: 145164.
Arnqvist, G. & Rowe, L. 2005. Sexual Conflict. Princeton University Press, Princeton, NJ.
Bateman, A.J. 1948. Intra-sexual selection in Drosophila. Heredity 2: 349368.
Birkhead, T.R. & Mller, A.P. 1998. Sperm Competition and Sexual Selection. Academic Press, San Diego, CA.
Blanckenhorn, W.U., Hosken, D.J., Martin, O.Y., Reim, C.,
Teuschl, Y. & Ward, P.I. 2002. The costs of mating in the
dung fly Sepsis cynipsea: I. Costs of copulation. Behav. Ecol.
13: 353358.

2014 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY. J. EVOL. BIOL. 28 (2015) 320327
JOURNAL OF EVOLUTIONARY BIOLOGY 2014 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

326

T. HARANO

Chapman, T., Liddle, L.F., Kalb, J.M., Wolfner, M.F. & Partridge, L. 1995. Cost of mating in Drosophila melanogaster
females is mediated by male accessory-gland products. Nature 373: 241244.
Clutton-Brock, T.H. & Parker, G.A. 1995. Sexual coercion in
animal societies. Anim. Behav. 49: 13451365.
Cordero, C. & Eberhard, W.G. 2003. Female choice of sexually
antagonistic male adaptations: a critical review of some current research. J. Evol. Biol. 16: 16.
Crudgington, H.S. & Siva-Jothy, M.T. 2000. Genital damage,
kicking and early deaththe battle of the sexes takes a sinister turn in the bean weevil. Nature 407: 855856.
Friberg, U. 2005. Genetic variation in male and female reproductive characters associated with sexual conflict in Drosophila melanogaster. Behav. Genet. 35: 455462.
Gasparini, C., Devigili, A. & Pilastro, A. 2012. Cross-generational effects of sexual harassment on female fitness in the
guppy. Evolution 66: 532543.
Gavrilets, S. & Hayashi, T.I. 2006. The dynamics of two and
three-way sexual conflicts over mating. Philos. Trans. R. Soc.
Lond. B Biol. Sci. 361: 345354.
Gavrilets, S., Arnqvist, G. & Friberg, U. 2001. The evolution of
female mate choice by sexual conflict. Proc. R. Soc. Lond. B
268: 531539.
Gay, L., Hosken, D.J., Eady, P., Vasudev, R. & Tregenza, T.
2011. The evolution of harmeffect of sexual conflicts and
population size. Evolution 65: 725737.
Harano, T. 2012. Water availability affects female remating in
the seed beetle, Callosobruchus chinensis. Ethology 118: 925
931.
Harano, T. & Katsuki, M. 2012. Female seed beetles, Callosobruchus chinensis, remate more readily after mating with relatives. Anim. Behav. 83: 10071010.
Harano, T. & Miyatake, T. 2005. Heritable variation in polyandry in Callosobruchus chinensis. Anim. Behav. 70: 299304.
Harano, T. & Miyatake, T. 2007a. Interpopulation variation in
female remating is attributable to female and male effects in
Callosobruchus chinensis. J. Ethol. 25: 4955.
Harano, T. & Miyatake, T. 2007b. No genetic correlation
between the sexes in mating frequency in the bean beetle,
Callosobruchus chinensis. Heredity 99: 295300.
Harano, T. & Miyatake, T. 2009. Bidirectional selection for
female propensity to remate in the bean beetle, Callosobruchus chinensis. Popul. Ecol. 51: 8998.
Harano, T., Yasui, Y. & Miyatake, T. 2006. Direct effects of
polyandry on female fitness in Callosobruchus chinensis. Anim.
Behav. 71: 539548.
Holland, B. & Rice, W.R. 1998. Chase-away sexual selection:
antagonistic seduction versus resistance. Evolution 52: 17.
Holland, B. & Rice, W.R. 1999. Experimental removal of sexual selection reverses intersexual antagonistic coevolution
and removes a reproductive load. Proc. Natl. Acad. Sci. USA
96: 50835088.
Hotzy, C. & Arnqvist, G. 2009. Sperm competition favors
harmful males in seed beetles. Curr. Biol. 19: 404407.
Hurst, G.D.D., Sharpe, R.G., Broomfield, A.H., Walker, L.E.,
Majerus, T.M.O., Zakharov, I.A. et al. 1995. Sexually-transmitted disease in a promiscuous insect, Adalia bipunctata.
Ecol. Entomol. 20: 230236.
Johnstone, R.A. & Keller, L. 2000. How males can gain by
harming their mates: sexual conflict, seminal toxins, and the
cost of mating. Am. Nat. 156: 368377.

Kemp, D.J. 2012. Costly copulation in the wild: mating

increases the risk of parasitoid-mediated death in swarming
locusts. Behav. Ecol. 23: 191194.
Kokko, H., Brooks, R., Jennions, M.D. & Morley, J. 2003. The
evolution of mate choices and mating biases. Proc. R. Soc. B
270: 653664.
Kuijper, B., Stewart, A.D. & Rice, W.R. 2006. The cost of mating rises nonlinearly with copulation frequency in a laboratory population of Drosophila melanogaster. J. Evol. Biol. 19:
17951802.
Lew, T.A., Morrow, E.H. & Rice, W.R. 2006. Standing genetic
variance for female resistance to harm from males and its
relationship to intralocus sexual conflict. Evolution 60: 97
105.
Linder, J.E. & Rice, W.R. 2005. Natural selection and genetic
variation for female resistance to harm from males. J. Evol.
Biol. 18: 568575.
Maklakov, A.A. & Arnqvist, G. 2009. Testing for direct and
indirect effects of mate choice by manipulating female
choosiness. Curr. Biol. 19: 19031906.
Makowicz, A.M. & Schlupp, I. 2013. The direct costs of living
in a sexually harassing environment. Anim. Behav. 85: 569
577.
Martinez-Padilla, J., Vergara, P., Mougeot, F. & Redpath, S.M.
2012. Parasitized males increase infection risk for partners.
Am. Nat. 179: 811820.
Michalczyk, L., Millard, A.L., Martin, O.Y., Lumley,
A.J., Emerson, B.C. & Gage, M.J.G. 2011. Experimental
evolution exposes female and male responses to sexual
selection and conflict in Tribolium castaneum. Evolution 65:
713724.
M
uhlhauser, C. & Blanckenhorn, W.U. 2002. The costs of
avoiding matings in the dung fly Sepsis cynipsea. Behav. Ecol.
13: 359365.
Nandy, B., Gupta, V., Udaykumar, N., Samant, M.A., Sen, S. &
Prasad, N.G. 2014. Experimental evolution of female traits
under different levels of intersexual conflict in Drosophila
melanogaster. Evolution 68: 412425.
Newport, M.E. & Gromko, M.H. 1984. The effect of experimental design on female receptivity to remating and its
impact on reproductive success in Drosophila melanogaster.
Evolution 38: 12611272.
Parker, G.A. 1979. Sexual selection and sexual conflict. In: Sexual Selection and Reproductive Competition in Insects (M.S. Blum
& N.A. Blum, eds), pp. 123166. Academic Press, New York,
NY.
Parker, G.A. 2006. Sexual conflict over mating and fertilization: an overview. Philos. Trans. R. Soc. Lond. B Biol. Sci. 361:
235259.
Partridge, L. & Fowler, K. 1990. Non-mating costs of exposure
to males in female Drosophila melanogaster. J. Insect Physiol.
36: 419425.
Partridge, L., Green, A. & Fowler, K. 1987. Effects of egg-production and of exposure to males on female survival in Drosophila melanogaster. J. Insect Physiol. 33: 745749.
Pitnick, S. & Hosken, D.J. 2010. Postcopulatory sexual selection. In: Evolutionary Behavioral Ecology (D.F. Westneat &
C.W. Fox, eds), pp. 379399. Oxford University Press,
Oxford.
R Development Core Team. 2013. R: A Language and Environment for Statistical Computing. R Foundation for Statistical
Computing, Vienna, Austria.

2014 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY. J. EVOL. BIOL. 28 (2015) 320327
JOURNAL OF EVOLUTIONARY BIOLOGY 2014 EUROPEAN SOCIETY FOR EVOLUTIONARY BIOLOGY

Female receptivity and sexual conflict costs

Rice, W.R. 1998. Intergenomic conflict, interlocus antagonistic

coevolution, and the evolution of reproductive isolation. In:
Endless Forms: Species and Speciation (D.J. Howard & S.H. Berlocher, eds), pp. 261270. Oxford University Press, New
York, NY.
R
onn, J., Katvala, M. & Arnqvist, G. 2006. The costs of mating
and egg production in Callosobruchus seed beetles. Anim.
Behav. 72: 335342.
R
onn, J., Katvala, M. & Arnqvist, G. 2007. Coevolution
between harmful male genitalia and female resistance in
seed beetles. Proc. Natl. Acad. Sci. USA 104: 1092110925.
Sakurai, G. & Kasuya, E. 2008. The costs of harassment in the
adzuki bean beetle. Anim. Behav. 75: 13671373.
Sakurai, G., Himuro, C. & Kasuya, E. 2012. Intra-specific variation in the morphology and the benefit of large genital sclerites of males in the adzuki bean beetle (Callosobruchus
chinensis). J. Evol. Biol. 25: 12911297.
Shuker, D.M., Ballantyne, G.A. & Wedell, N. 2006. Variation
in the cost to females of the sexual conflict over mating in
the seed bug Lygaeus equestris (Hemiptera: Lygaeidae). Anim.
Behav. 72: 313321.
Simmons, L.W. 2001. Sperm Competition and Its Evolutionary
Consequences in the Insects. Princeton University Press, Princeton, NJ.
Takahashi, Y. & Watanabe, M. 2010. Female reproductive success is affected by selective male harassment in the damselfly
Ischnura senegalensis. Anim. Behav. 79: 211216.
Taylor, M.L., Price, T.A. & Wedell, N. 2014. Polyandry in nature: a global analysis. Trends Ecol. Evol. 29: 376383.
Thornhill, R. & Alcock, J. 1983. The Evolution of Insect Mating
Systems. Harvard University Press, Cambridge.
Tilszer, M., Antoszczyk, K., Salek, N., Zajac, E. & Radwan, J.
2006. Evolution under relaxed sexual conflict in the bulb
mite Rhizoglyphus robini. Evolution 60: 18681873.
Tuda, M., R
onn, J., Buranapanichipan, S., Wasano, N. & Arnqvist, G. 2006. Evolutionary diversification of the bean beetle

327

genus Callosobruchus (Coleoptera: Bruchidae): traits associated with stored-product pest status. Mol. Ecol. 15: 3541
3551.
Umeya, K. 1987. Biology of Bruchids. Tukizi-Shokan, Tokyo (in
Japanese).
Wigby, S. & Chapman, T. 2004. Female resistance to male
harm evolves in response to manipulation of sexual conflict.
Evolution 58: 10281037.
Yamane, T. 2013. Intra-specific variation in the effect of male
seminal substances on female oviposition and longevity in
Callosobruchus chinensis. Evol. Biol. 40: 133140.
Yamane, T. & Miyatake, T. 2012. Evolutionary correlation
between male substances and female remating frequency in
a seed beetle. Behav. Ecol. 23: 715722.
Yanagi, S. & Miyatake, T. 2003. Costs of mating and egg production in female Callosobruchus chinensis. J. Insect Physiol. 49:
823827.
Yasui, Y. 1998. The genetic benefits of female multiple mating reconsidered. Trends Ecol. Evol. 13: 246250.

Supporting information
Additional Supporting Information may be found in the
online version of this article:
Figure S1 Changes in the proportion of females that
remated in lines selected for higher (H1 and H2) and
lower (L1 and L2) female remating propensity.
Data deposited at Dryad: doi:10.5061/dryad.46t7q
Received 6 August 2014; revised 14 November 2014; accepted 24
November 2014

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