Effects of two seminal fluid transcripts on post‐mating behaviour in the simultaneously hermaphroditic flatworm Macrostomum lignano

The seminal fluid proteins (SFPs) transferred to mating partners along with sperm often play crucial roles in mediating post‐mating sexual selection. One way in which sperm donors can maximize their own reproductive success is by modifying the partner's (sperm recipient's) post‐copulatory behaviour to prevent or delay re‐mating, thereby decreasing the likelihood or intensity of sperm competition. Here, we adopted a quantitative genetic approach combining gene expression and behavioural data to identify candidates that could mediate such a response in the simultaneously hermaphroditic flatworm Macrostomum lignano. We identified two putative SFPs—Mlig‐pro46 and Mlig‐pro63—linked to both mating frequency and ‘suck’ frequency, a distinctive behaviour, in which, upon ejaculate receipt, the worm places its pharynx over its female genital opening and apparently attempts to remove the received ejaculate. We, therefore, performed a manipulative experiment using RNA interference‐induced knockdown to ask how the loss of Mlig‐pro46 and Mlig‐pro63 expression, singly and in combination, affects mating frequency, partner suck propensity and sperm competitive ability. None of the knockdown treatments impacted strongly on the mating frequency or sperm competitive ability, but knockdown of Mlig‐pro63 resulted in a significantly decreased suck propensity of mating partners. This suggests that Mlig‐pro63 may normally act as a cue in the ejaculate to trigger recipient suck behaviour and—given that other proteins in the ejaculate have the opposite effect—could be one component of an ongoing arms race between donors and recipients over the control of ejaculate fate. However, the adaptive significance of Mlig‐pro46 and Mlig‐pro63 from a donor perspective remains enigmatic.     

Control of reproduction and a survival cost to mating in female Bicyclus anynana butterflies

Abstract.  1. Given that sexual conflict is all pervasive, investigating potential costs to mating and the control of female reproduction are important issues.
2. Here, female reproductive output and longevity are investigated in relation to mating status (virgin vs. once-mated females) and host-plant availability in the tropical butterfly Bicyclus anynana .
3. Both factors significantly affected realised fecundity in a manner demonstrating that female reproduction is not entirely under control of the female herself, but is rather subject to additive contributions of the female, her male partner, and intrinsic physiological processes. As evident from the deposition of significant egg numbers by virgin, host-deprived females, B. anynana is effectively unable to completely inhibit oogenesis.
4. Mated females suffered a reduction in adult life span, which cannot be explained as a side-effect of variation in egg size, lifetime, or early fecundity.
5. Such detrimental effects of mating per se are indicative of the cooperation–conflict balance between sexes being shifted towards conflict in B. anynana .     

The structure of the bursa copulatrix in virgin and mated snails, Helisoma duryi (Mollusca): role of acid phosphatase in reproduction

Abstract. The fine structure of the bursa copulatrix of the virgin snails has been compared with that of mated snails. One of the noticeable changes after mating is an increase in the number of the Golgi and the secretory vesicles. Since some of the vesicles react positively for acid phosphatase it is suggested that this enzyme activity increases following mating. The bursa lumen of the virgin snail contains gel-like materials devoid of spermatozoa, however, following mating, the lumen is full of semen containing live spermatozoa and bacteria. The source of bacteria in the lumen is not known. Acid phosphatase activity is significantly higher in the luminal content of mated snails than in the virgin snails. The activity is higher in the lumen than in the epithelial cells, suggesting that the enzyme is secreted into the lumen where it is utilized for extracellular degradation of spermatozoa. Following mating, the spermatozoa are motile in the lumen of the bursa for ∼3–7 d, but become immobile and finally undergo extracellular digestion so that intact spermatozoa are not recognizable by day 10. The use of castrated snails in mating experiments suggest that individuals of Helisoma duryi reproduce by cross fertilization and that the bursa may act as the holding organ from where the spermatozoa are periodically transported to the carrefour over ∼7 d. At day 10 following mating, however, autosperms appear in the hermaphroditic duct awaiting the next mating.     

Patterns of sperm precedence are not affected by female mating history in Drosophila melanogaster

In promiscuously mating species, there is strong selection on males to maximize their share of paternity through both defensive and offensive means. This has been most extensively examined using the Drosophila melanogaster model system. In these studies, sperm competition has been examined by mating a virgin female to two consecutive males and then determining the fertilization success of both the first male (defending, P1) and the second male (offending, P2). Recent evidence suggests that male defense may be influenced by female mating history (i.e., virgin versus nonvirgin). Here, by mating females to males with three different genotypes, we show that female mating history does not affect male defensive or offensive abilities in sperm competition. We also show that, although female lifetime fecundity was not correlated with the number of times that she mated, it was reduced by increased exposure to males. These data indicate that measures of P1 and P2 previously reported in D. melanogaster may be robust to the specific mating history of the females used in these studies.     

The effects of prey lipid on female mating and reproduction of a wolf spider

As predators, the macronutrients spiders extract from their prey play important roles in their mating and reproduction. Previous studies of macronutrients on spider mating and reproduction focus on protein, the potential impact of prey lipid content on spider mating and reproduction remains largely unexplored. Here, we tested the influence of prey varying in lipid content on female mating, sexual cannibalism, reproduction, and offspring fitness in the wolf spider Pardosa pseudoannulata. We acquired 2 groups of fruit fly Drosophila melanogaster that differed significantly in lipid but not protein content by supplementing cultural media with a high or low dose of sucrose on which the fruit flies were reared (HL: high lipid and LL: low lipid). Subadult (i.e., 1 molt before adult) female spiders that fed HL flies matured with significantly higher lipid content than those fed LL flies. We found that the mated females fed with HL flies significantly shortened pre-oviposition time and resulted in a significantly higher fecundity. However, there was no significant difference in female spiders varying in lipid content on other behaviors and traits, including the latency to courtship, courtship duration, mating, copulation duration, sexual cannibalism, offspring body size, and survival. Hence, our results suggest that the lipid content of prey may be a limiting factor for female reproduction, but not for other behavioral traits in the wolf spiders P. pseudoannulata.     

Cross-generational fitness benefits of mating and male seminal fluid

In many species, the physical act of mating and exposure to accessory gland proteins (Acps) in male seminal fluid reduces female survival and offspring production. It is not clear what males gain from harming their sexual partners or why females mate frequently despite being harmed. Using sterile strains of Drosophila melanogaster that differ in their production of Acps, we found that both the physical act of mating and exposure to male seminal fluid in mothers increase the fitness of daughters. We show that the changes in daughter fitness are mediated by parental effects, not by sexual selection involving good genes or owing to variation in maternal egg production. These results support the idea that male harm of females might partly evolve through cross-generational fitness benefits.     

The effects of spatial habitat structure on the evolution of density-dependent growth and reproduction in freshwater snails

We examined the growth and reproductive rates of freshwater snails, Physa acuta, in two habitat types. In the Asabata habitat, snails lived in isolated water pools, which occasionally joined to form a single large pool; in the Kakegawa habitat, they lived in a slow-running water way. Genetic structure assessments using three microsatellite loci supports the idea that a stable panmictic population occupies the Kakegawa habitat. The Asabata habitat, however, is occupied with an alternate mixing population as revealed by microsatellite data. The Asabata population might alternate between localized mating within isolated pools (as revealed by high F _IS and F _IT values) when the water levels are low and panmixia (as revealed by the low F _ST values and AMOVA analysis) when the habitat is flooded. Laboratory experiments, using snails collected from the two habitats, showed that juvenile snails grew faster, laid more eggs, and laid them earlier in the Asabata habitat than in the Kakegawa habitat. Growth rates were lower at high density than at low density in the Kakegawa habitat; the inverse was true in the Asabata habitat. Density-dependent response of individual snail reproduction was higher in the Kakagawa habitat than in the Asabata habitats. The results support the hypothesis that spatial structure affects the evolution of density-dependent growth rates and of timing for reproduction.     

Higher rate of tissue regeneration in polyploid asexual versus diploid sexual freshwater snails

Characterizing phenotypic differences between sexual and asexual organisms is a critical step towards understanding why sexual reproduction is so common. Because asexuals are often polyploid, understanding how ploidy influences phenotype is directly relevant to the study of sex and will provide key insights into the evolution of ploidy-level variation. The well-established association between genome size and cell cycle duration, evidence for a link between genome size and tissue regeneration rate and the growing body of research showing that ploidy influences growth rate and gene expression led us to hypothesize that healing and tissue regeneration might be affected by ploidy-level variation. We evaluated this hypothesis by measuring the rate of regeneration of antenna tissue of Potamopyrgus antipodarum, a New Zealand snail characterized by frequent coexistence between diploid sexuals and polyploid asexuals. Antennae of triploid and presumptive tetraploid asexuals regenerated more rapidly than the antennae of diploid sexuals, but regeneration rate did not differ between triploids and tetraploids. These results suggest either that ploidy elevation has nonlinear positive effects on tissue regeneration and/or that factors associated directly with reproductive mode affect regeneration rate more than ploidy level. The results of this study also indicate that the lower ploidy of sexual P. antipodarum is unlikely to confer advantages associated with more rapid regeneration.     

A conversation with Mariana Wolfner,newly inducted member of the National Academy of Sciences

Molecular Reproduction and Development is delighted to announce that editorial board member Mariana F. Wolfner has been elected to the National Academy of Sciences. Here, Dr Wolfner is interviewed by two of her former postdocs. She discusses her path to studying reproduction and her career as a researcher and mentor.     

Arabinogalactan proteins in plant sexual reproduction

Summary Arabinogalactan proteins (AGPs) are a class of plant extracellular-matrix proteins believed to participate in a broad range of processes involving the plant cell surface. They are extremely abundant in female reproductive tissues and in pollen tubes, the haploid male structures that traverse the diploid female reproductive tissues to deliver sperms to the egg cells. The prevalence of AGPs in reproductive tissues has led to speculations that they play significant functional roles ranging from serving as nutrient resources to cell-cell recognition in plant reproduction. Recent research from several laboratories demonstrated functional participation by AGPs in reproductive processes and began to examine the mechanisms underlying these functional roles. An overview of these recent studies will be discussed with a historical perspective as well as with a view towards future studies in establishing the significance of AGPs that, as a class, they have prominent roles in plant sexual reproduction in multiple and diverse ways.     

Foodborne uptake and sublethal effects of copper and zinc to freshwater snails

Snail habitat selection was strongly influenced in thisresearch by measured copper (Cu) contribution from a powerplant discharge. Laboratory feeding studies showed that thesnail Leptoxis praerosa significantly bioconcentrated Cuwhen fed aufwuchs containing 564 (±269) µg Cu g^-1but did not have significant cellulolytic enzyme activityimpairment. No foodborne zinc (Zn) bioconcentration was foundfrom aufwuchs containing up to 20 000 (±18 400) µg Zn g^-1. These results suggest that water column Cuconcentrations leading to impairment via feeding mechanismscan be an order of magnitude higher than concentrations causingimpairments via waterborne exposures. Therefore, foodborneuptake might be expected to contribute to toxicity rather thandominate toxicity in most conditions where waterborneconcentrations of metals occur. Changes in food qualityappeared to have a greater overall effect on snail populationsthan foodborne uptake of metals alone. Snail populations took2 years to recover following intensive metal treatment on theplant discharge and subsequent reduction of measured metalconcentrations in both water and aufwuchs.     

桦纤孔菌有性生殖相关特征及交配型位点分析

对桦纤孔菌菌株MDJCBS88的显微形态、菌丝及担孢子核相进行了观察。采用棉籽壳培养基对担孢子萌发形成的菌株进行栽培试验,筛选出不形成子实体或子实体发育不完整的菌株,将这些菌株在平板上进行了亲和试验,分析桦纤孔菌的有性生殖方式;并基于基因组序列进行交配型基因克隆验证,分析桦纤孔菌的交配型位点结构。显微观察发现,桦纤孔菌菌丝没有锁状联合结构,菌丝细胞无核到多核;子实层担孢子可含0-4个不等的细胞核,不同时期弹射的担孢子含有的细胞核数量不同。桦纤孔菌担孢子萌发率极低,能萌发的担孢子多为早期弹射的担孢子;培养基也影响担孢子的萌发率,与PDA培养基和CYM培养基相比,桦木屑培养基最适合桦纤孔菌担孢子萌发,萌发率为4.55%。从担孢子萌发的96个菌株中获得了2个不结实菌株和9个结实不产孢菌株,占11.5%,这些菌株间亲和试验出现不同的表现特征,包括形成产孢子实体,产生菌丝纽结,相互融合和相互拮抗等现象,认为桦纤孔菌的有性生殖以次级同宗结合为主,并受交配型基因控制。交配型位点克隆测序后分析发现,桦纤孔菌交配型A位点共14 034 bp,含有一个MIP基因和两组HD1和HD2基因;交配型B位点包含3个疑似信息素受体基因和1个信息素前体编码基因。     

Sexual reproduction in Daphnia pulex (Crustacea: Cladocera): observations on male mating behaviour and avoidance of inbreeding

1.  Mating behaviour in Daphnia appears to rely on random contact between males and sexual females rather than diffusible pheromones. Males may be able to discriminate sexually receptive females from females in other developmental stages and increase their mating efficiency. Males may also use chemical signals to avoid mating with females from the same clone and avoid the severe inbreeding depression that has been documented for intraclonal mating. The present study used experiments to test for the avoidance of intraclonal mating and assess male mating efficiency in D. pulex .
2.  Three clones were examined for the avoidance of intraclonal mating by providing males with an opportunity to mate with females of the same or two different clones. The proportion of intraclonal matings did not differ from the proportion of interclonal matings, suggesting that D. pulex males do not use kin discrimination to avoid mating with females from the same clone.
3.  The proportion of mated females decreased with increasing numbers and density of sexual females when exposed to a single male. This observation suggests that a male spends more time pursuing and copulating with sexually receptive females than non-receptive females and there is insufficient time to mate with all sexual females. The decrease in proportion of females mated could also be the result of sperm depletion in the male. Sperm depletion is unlikely to occur in nature because sexually receptive females are much rarer than in the experimental conditions.     

甘草有性生殖特性研究

本文利用扫描电子显微镜对乌拉尔甘草、光果甘草和胀果甘草花粉的超微结构进行观察;利用荧光显微镜确定乌拉尔甘草的授粉方式、花粉生活力和柱头活性以及授粉后受精时间。结果表明花粉超微结构为甘草的鉴别提供了一定的形态学依据;甘草的授粉方式属于闭花受精;花粉生活力在每天12:00时最强,去雄后超过四天的柱头已经不具备接受花粉的能力;对授粉后不同时间的雌蕊进行研究得出授粉后6h花粉管进入胚珠进行受精。本文主要对甘草的有性生殖特性进行了研究,为未来制定正确的选、育种策略提供理论依据。     

甘草有性生殖特性研究

本文利用扫描电子显微镜对乌拉尔甘草、光果甘草和胀果甘草花粉的超微结构进行观察;利用荧光显微镜确定乌拉尔甘草的授粉方式、花粉生活力和柱头活性以及授粉后受精时间。结果表明花粉超微结构为甘草的鉴别提供了一定的形态学依据;甘草的授粉方式属于闭花受精;花粉生活力在每天12:00时最强,去雄后超过四天的柱头已经不具备接受花粉的能力;对授粉后不同时间的雌蕊进行研究得出授粉后6h花粉管进入胚珠进行受精。本文主要对甘草的有性生殖特性进行了研究,为未来制定正确的选、育种策略提供理论依据。     

Natural female mating rate maximizes hatchling size in a marine invertebrate

1. Males and females often differ in their optimal mating rates, resulting potentially in conflicts over remating. In species with separate sexes, females typically have a lower optimal mating rate than males, and can regulate contacts with males accordingly. The realized mating rate may therefore be closer to the female's optimum. In simultaneous hermaphrodites, however, it has been suggested that the intraindividual optimization between 'male' and 'female' interests generates more 'male'-driven mating rates. 2. In order to assess the consequences of variation in mating rate on 'female' reproductive output, we exposed the simultaneously hermaphroditic sea slug Chelidonura sandrana to four mating rate regimes and recorded the effects on a variety of fitness components. 3. In focal 'females', we found (i) a slight but significant linear decrease in fecundity with mating rate, whereas (ii) maternal investment in egg capsule volume peaked at an intermediate mating rate. 4. Combining the observed fecundity cost with the apparent benefits of larger offspring size suggests that total female fitness is maximized at an intermediate mating rate. With the latter being close to the natural mating rate of C. sandrana in the field, our findings challenge the assumption of 'male'-driven mating systems in simultaneous hermaphrodites. 5. Our study provides experimental evidence for various mathematical models in which female fitness is maximized at intermediate mating rates.     

The energetic cost of mating in a promiscuous cephalopod

Costs that individuals incur through mating can play an important role in understanding the evolution of life histories and senescence, particularly in promiscuous species. Copulation costs, ranging from energy expenditure to reduced longevity, are widely studied in insects but have received substantially less attention in other taxa. One cost of mating, the energetic cost, is poorly studied across all taxa despite its potential importance for the many species where copulation is physically demanding and/or frequent. Here, we investigated the energetic cost of mating in both male and female dumpling squid (Euprymna tasmanica). In this species, copulation can last up to 3 h and requires that the male physically restrains the female. We report that the act of copulation halves the swimming endurance of both sexes, and that they take up to 30 min to recover. Such a reduction in post-copulatory performance may have important implications for predator avoidance, foraging ability and energy allocation. Therefore, quantifying this cost is essential to understand the evolution of reproductive strategies and behaviours such as female receptivity and male and female mating frequency.     

萼花臂尾轮虫有性生殖、种群增长和休眠卵产量间的关系

利用种群指数增长和Logistic增长模型,通过计算机模拟研究了萼花臂尾轮虫有性生殖发生的频率,后代中的混交雌体发率对种群增长和休眠卵产量的影响。在所模拟的参数范围内,随有性生殖发生频率由100％减小到20％,获得了大休眠卵产量所需的混交雌体百分率由9％增大到69％,随密度制约作用的增大（环境容纳量K值由1000减小到100）,该混交雌体百分率由18％增大到69％,休眠卵产量由1072．10降低至133．67,种群的内禀增长率与获得最大休眠卵产量所需的混交雌体百分率间呈曲线相关。当有性生殖发生得越频繁,种群增长所受的密度制约作用较小时,后代中10％－30％的个体为混交雌体时的种群中休眠卵产量较大。     

Similar gender dimorphism in the costs of reproduction across the geographic range of Fraxinus ornus

BACKGROUND AND AIMS: The reproductive costs for individuals with the female function have been hypothesized to be greater than for those with the male function because the allocation unit per female flower is very high due to the necessity to nurture the embryos until seed dispersal occurs, while the male reproductive allocation per flower is lower because it finishes once pollen is shed. Consequently, males may invest more resources in growth than females. This prediction was tested across a wide geographical range in a tree with a dimorphic breeding system (Fraxinus ornus) consisting of males and hermaphrodites functioning as females. The contrasting ecological conditions found across the geographical range allowed the evaluation of the hypothesis that the reproductive costs of sexual dimorphism varies with environmental stressors. METHODS: By using random-effects meta-analysis, the differences in the reproductive and vegetative investment of male and hermaphrodite trees of F. ornus were analysed in 10 populations from the northern (Slovakia), south-eastern (Greece) and south-western (Spain) limits of its European distribution. The variation in gender-dimorphism with environmental stress was analysed by running a meta-regression between these effect sizes and the two environmental stress indicators: one related to temperature (the frost-free period) and another related to water availability (moisture deficit). KEY RESULTS: Most of the effect sizes showed that males produced more flowers and grew more quickly than hermaphrodites. Gender differences in reproduction and growth were not minimized or maximized under adverse climatic conditions such as short frost-free periods or severe aridity. CONCLUSIONS: The lower costs of reproduction for F. ornus males allow them to grow more quickly than hermaphrodites, although such differences in sex-specific reproductive costs are not magnified under stressful conditions.     

High level of sperm competition may increase transfer of accessory gland products carried by the love dart of land snails

Postcopulatory adaptations that increase reproductive success compared to rivals, like the transfer of accessory gland products that promote paternity, are common when sperm competition occurs among males. In land snails, the dart shooting behavior and its adaptive significance, in promoting individual fitness through enhanced paternity of the successful dart shooter, have been considered such an adaptation. The fitness result gained is mediated by the transfer of mucus components on the love dart capable of altering the physiology of the receiver's reproductive tract. In this context, dart shooting and mucus transfer could be considered as processes targeted by sexual selection. While the effect of dart mucus is beneficial for the dart user, so far it has remained unknown whether its transport is greater when snails experience a higher level of sperm competition. Here, we report results of a study on inter‐ and intraspecific variations of dart and mucus gland morphometry, considered to be traits reflecting the ability of snails to adjust the production and transfer of mucus under varying sperm competition scenarios. We investigated four populations with different densities from four dart‐bearing species, Arianta arbustorum, Cepaea nemoralis, Cornu aspersum, and Helix lucorum. The results indicate that different adaptations of these traits occur among the studied species that all seem to achieve the same goal of transferring more mucus when sperm competition is higher. For example, the presence of longer and more branched mucous glands or an increase in dart surface most likely reflect increased mucus production and enhanced ability of mucus transport, respectively. Interestingly, the species for which the use of the dart is reported to be facultative, A. arbustorum, did not show any variation among the examined traits. To conclude, sexual selection in the form of sperm competition intensity seems to be an important selective force for these simultaneously hermaphroditic dart‐bearing snails, driving differences in sexual traits.