Paternal Inheritance of Mitochondrial DNA May Lead to Dioecy in Conifers.

In angiosperms cytoplasmic DNA is typically passed on maternally through ovules. Genes in the mtDNA may cause male sterility. When male-sterile (female) cytotypes produce more seeds than cosexuals, they pass on more copies of their mtDNA and will co-occur with cosexuals with a neutral cytotype. Cytoplasmic gynodioecy is a well-known phenomenon in angiosperms, both in wild and crop plants. In some conifer families (e.g. Pinaceae) mitochondria are also maternally inherited. However in some other families (e.g. Taxaceae and Cupressaceae) mtDNA is paternally inherited through the pollen. With paternal mtDNA inheritance, male cytotypes that produce more pollen than cosexuals are expected to co-occur with cosexuals. This is uncharted territory. An ESS model shows that the presence of male cytotypes selects for more female allocation in the cosexual, i.e. for sexual specialisation. An allele that switches sex from male to female can then invade. This leads to rapid loss of the neutral cytotype of the cosexual, fixation of the male cytotype and dioecy with 50% males and 50% females. The models suggest that paternal inheritance of mtDNA facilitates the evolution dioecy. Consistent with this hypothesis the Pinaceae are 100% monoecious, while dioecy is common in the Taxaceae family and in the genus Juniperus (Cupressaceae). However, no reliable data are yet available on both mode of inheritance of mtDNA and gender variation of the same species. When cosexuals benefit from reproductive assurance (high selfing rate, low inbreeding depression, low fertilisation) they maintain themselves next to males and females. This predicted pattern with three sex types present in the same population is observed in conifers in nature.

Pollination efficiency and the pollen-ovule ratio.

Pollination presents a risky journey for pollen grains. Pollen loss is sometimes thought to favour greater pollen investment to compensate for the inefficiency of transport. Sex allocation theory, to the contrary, has consistently concluded that postdispersal loss should have no selective effect on investment in either sex function. But the intuitively appealing compensation idea continues to be raised despite the lack of theoretical endorsement. We address the theoretical issue with a model that directly represents pollen loss (and ovule loss through floral demise or loss of receptivity) as rate-dependent dynamical processes. These loss rates can be varied to examine the effect of pollination efficiency on optimal sex allocation. Pollen-ovule ratios follow from the sex allocation based on the resource costs of pollen and ovule production. This model confirms conventional findings that pollen loss should have essentially no effect on sexual resource allocation in large, panmictic populations. Pollen limitation of seed set does not alter this conclusion. These results force us to rethink the empirical association of pollination efficiency with low pollen-ovule ratios. This pattern could arise if efficient pollen transport commonly results in stigmatic deposition of cohorts of related pollen. Empirical evidence of correlated paternity supports this explanation.

Self-Recognition and Allorecognition Mechanisms Exert a Significant Influence on the Sex Allocation Patterns of the Pea Aphid.

The mechanism controlling sex allocation in the pea aphid, Acyrthosiphon pisum (Harris), remains a crucial yet unresolved issue in the field of evolutionary ecology. This study aims to assess the influence of the presence of both self and non-self clones, along with juvenile hormone III (JH III) titer, on the sex allocation of aphid offspring. To this end, red and green clones were utilized as experimental subjects, and the agar method was employed. Initially, three distinct experimental treatments were established using sexuparae, and the daily offspring count and sex allocation in each treatment zone were recorded. Subsequently, an additional experimental condition involving mixed-clone treatments was introduced. This procedure entailed the transfer of a single sexupara and 20 oviparous females from either the red (1G + 20Rov) or green clone (1G + 20Gov) onto a leaf on agar medium. Simultaneously, a control setup with a new sexupara (1G) was established. Three days following sexupara production, a dose of 0, 25, or 50 ng of JH III was applied to the aphids' abdomens. Subsequently, the titers of JH III in the sexuparae across each treatment group were quantified, and the extent of sex allocation was tallied. The findings demonstrated pronounced disparities in sex allocation among the various treatments and, notably, a substantial increase in the total offspring and oviparous number in the mixed-clone treatment group. The effects of mixed-clone treatment on the sex allocation patterns of the sexupara progeny could be determined by the application of exogenous JH III, indicating that JH may mediate the effects of mixed-clone treatment on sex allocation. Consequently, it can be concluded that A. pisum sexuparae possess the capability to modulate their sex allocation in response to the nature of adjacent competitor clones, thereby demonstrating a variety of sex allocation patterns. Throughout this process, JH III plays a pivotal role.

Why the Shaw-Mohler equation works and when it doesn't.

Fitness gain curves were introduced into the framework of the Shaw-Mohler equation, the foundation of sex allocation theory. I return to the Shaw-Mohler equation to consider how it embodies the rare-sex advantage underlying frequency-dependent selection on the sex ratio. The Shaw-Mohler formulation is based on the numbers of males and females randomly mating in a panmictic population. Gain curves are meant to describe reproductive success through male and female functions in hermaphrodites and have been inserted in place of male and female numbers in the Shaw-Mohler equation. In doing so, gain curves bypass consideration of the implicit mating process in the Shaw-Mohler argument and can lead to anomalies like unequal total male and female fitness in a population. If gain curves truly represent fitness gain, equality of total male and female fitness requires a constant sex allocation of equal resource investment into male and female functions. The blurring of input with fitness outcome has led to misinterpretation of what gain curves mean in reproductive ecology. They can describe a particular reproductive ecology, such as diminishing fitness returns on resource investment, but lack causal efficacy with respect to sex allocation.

Sublethal heat reduces overall reproductive investment and male allocation in a simultaneously hermaphroditic snail species.

The exposure to sublethally high temperature reduces reproductive performance in diverse organisms. Although this effect has been particularly emphasized for males or male reproductive functioning, it remains largely unknown whether the effect of heat on fertility is sex-specific. Here we examined the impact of sublethally high temperature on male and female functions in a simultaneously hermaphroditic snail species, Lymnaea stagnalis. Examining hermaphrodites is useful to evaluate the sex-specific impacts of heat exposure, since they possess male and female functions within a single individual, sharing genetic and environmental factors. Moreover, previously developed sex allocation theory allows us to compare the differential performance of sex functions. In this study, we exposed snails to 20°C (control), 24°C and 28°C for 14 days and assessed their egg and sperm production, sperm transfer, mating behaviour and growth. Both types of gamete production were significantly reduced by higher temperature, leading to an overall reduction of reproductive investment. By quantifying sex allocation, we furthermore revealed that the heat-stressed snails reduced the relative investment in their male function. This study illustrates that examining simultaneous hermaphrodites can provide significant insights for the impact of heat, and the proximate mechanism, on reproduction in diverse organisms.

Parental effects on offspring sex ratio in the Numbat (Myrmecobius fasciatus): does captivity influence paternal sex allocation?

Sex allocation theories predict that under different ecological conditions the production of sons and daughters will affect parental fitness differently. Skewed offspring sex ratios often occur under captive conditions where individuals are exposed to nutritional and social conditions that differ from nature. Here, we analyzed 29 years of offspring sex ratio data from a captive population of an endangered marsupial, the Numbat (Myrmecobius fasciatus). We partitioned variation in offspring sex ratio based on parental origin (captive- vs. wild-bred), parental weight, maternal age, and maternal reproductive history. Our analyses revealed no effect of parental weight or maternal origin on offspring sex ratio-however, there was a significant effect of paternal origin. Data visualization indicated that captive-bred males tended to produce male-biased litters. We discuss the result in relation to recent studies that have shown that male mammals have the capacity to be arbiters of sex allocation and highlight candidate mechanisms, but consider it with caution due to the small sample size from which the result was derived. We performed a population viability analysis (PVA) to explore the potential impact of a sex ratio skew on the sustainability of the captive Numbat population under hypothetical scenarios. Our PVA revealed that supplementation with wild individuals is critical to the persistence of the captive Numbat population and that a biased sex ratio will lead to extinction of the captive colony under certain conditions. Overall, our study demonstrates that covert sex ratio skews can persist undetected in captive populations, which have the potential to become impactful and compromise population sustainability under changed management processes.

Unisexual flowers as a resolution to intralocus sexual conflict in hermaphrodites.

In dioecious populations, males and females may evolve different trait values to increase fitness through their respective sexual functions. Because hermaphrodites express both sexual functions, resolving sexual conflict is potentially more difficult for them. Here, we show that hermaphrodite plants can partially resolve sexual conflict by expressing different trait values in different male and female modules (e.g. different flowers, inflorescences, branches etc.). We analysed the flowering phenology, sex allocation and selection gradients on floral traits of flowers of the andromonoecious plant Pulsatilla alpina, which produces both bisexual and male flowers. Our results indicate that strong protogyny prevents early bisexual flowers from profiting from high siring opportunities early in the reproductive season at a time when male flowers could achieve high siring success. The production of unisexual male flowers thus resolves this sexual conflict because it allows the flowers to express their male function without waiting until after the female function has been performed. Our study illustrates the resolution of sexual conflict arising from phenological constraints via modular divergence in sex allocation. We discuss the extent to which modular variation in sex allocation in the context of other sexual systems may be similarly explained.

Long-term female bias in sex ratios across life stages of Harpy Eagle, a large raptor exhibiting reverse sexual size dimorphism.

The primary (PSR), secondary (SSR) and adult (ASR) sex ratios of sexually reproducing organisms influence their life histories. Species exhibiting reversed sexual size dimorphism (RSD) may imply a higher cost of female production or lower female survival, thus generating biases in PSR, SSR and/or ASR towards males. The Harpy Eagle is the world's largest eagle exhibiting RSD. This species is found in the Neotropical region and is currently threatened with extinction. We used molecular markers to determine the sex of 309 Harpy Eagles spanning different life stages-eaglets, subadults and adults-from 1904 to 2021 within the Amazon Rainforest and Atlantic Forest. Sex ratios for all life stages revealed a female-biased deviation across all periods and regions. Our results suggest that the population bias towards females is an evolutionary ecological pattern of this species, and SSR and ASR likely emerged from the PSR. This natural bias towards females may be compensated by an earlier sexual maturation age of males, implying a longer reproductive lifespan and a higher proportion of sexually active males. A better understanding of the Harpy Eagle's life history can contribute to understanding sex-role evolution and enable more appropriate conservation strategies for the species.

Male reproductive success is not strongly affected by phenological changes in mate availability in monoecious Sagittaria latifolia.

Many plants express their female and male sex roles at different times (dichogamy), with important consequences for mating. Dichogamy can yield mate limitation via biased floral sex ratios, particularly at the beginning and end of the flowering season when many plants simultaneously function as the same sex. This form of mate limitation should be reduced if plants adjust their allocations to female versus male sex functions in a manner that tracks seasonal variability in mating opportunities. For example, under protogyny (i.e. dichogamy with female function expressed first) plants with male-biased sex expression should have enhanced mating opportunities early in the flowering season as other plants begin to flower (in female sex phase). We quantified seasonal changes in sex allocation, patterns of mate availability and realized siring success in a population of protogynous Sagittaria latifolia. Our results were consistent with previous findings that seasonal changes in sex allocation should compensate for lost mating opportunities under the temporally variable mating environments caused by dichogamy. However, patterns of siring success in the population were inconsistent with this interpretation. We suggest that realized siring success might depend more strongly on spatial than on temporal aspects of mate availability.

W. D. Hamilton and the golden sex ratio.

In his famous two-part paper, published in Journal of Theoretical Biology in 1964, W. D. Hamilton predicted that natural selection acting in male-haploid populations favours a ratio of males to females that is in accordance with the golden ratio. This prediction has found its way into the pages of one of the best-selling books of all time, Dan Brown's 2003 novel The da Vinci Code, and is therefore in the running for the most widely known quantitative result in the history of evolutionary biology. Unfortunately, this golden-ratio result is wrong, and was later corrected by Hamilton, who showed that natural selection actually favours an unbiased sex ratio in this setting. But it has been unclear exactly how Hamilton arrived at the golden-ratio result in the first place. Here I show that the solution to this puzzle is found in unpublished work held in the British Library's W. D. Hamilton Archive. Specifically, in addition to employing a faulty method for calculating relatedness, Hamilton had also employed a faulty method for calculating reproductive value, considering only genetic contributions to the next generation rather than to the distant future. Repeating both mistakes recovers his erroneous golden-ratio result.

Reproductive plasticity in response to the changing cluster size during the breeding period: a case study in a spider mite.

Animals living in clusters should adjust their reproductive strategies to adapt to the social environment. Theories predict that the benefits of cluster living would outweigh the costs of competition. Yet, it is largely unknown how animals optimize their reproductive fitness in response to the changing social environment during their breeding period. We used Tetranychus ludeni Zacher, a haplodiploid spider mite, to investigate how the ovipositing females modified their life-history traits in response to the change of cluster size (i.e., aggregation and dispersal) with a consistent population density (1 ♀/cm2). We demonstrate that (1) after females were shifted from a large cluster (16 ♀♀) to small ones (1 ♀, 5 and 10 ♀♀), they laid fewer and larger eggs with a higher female-biased sex ratio; (2) after females were shifted from small clusters to a large one, they laid fewer and smaller eggs, also with a higher female-biased sex ratio, and (3) increasing egg size significantly increased offspring sex ratio (% daughters), but did not increase immature survival. The results suggest that (1) females fertilize more larger eggs laid in a small population but lower the fertilization threshold and fertilize smaller eggs in a larger population, and (2) the reproductive adjustments in terms of egg number and size may contribute more to minimize the mate competition among sons but not to increase the number of inhabitants in the next generation. The current study provides evidence that spider mites can manipulate their reproductive output and adjust offspring sex ratio in response to dynamic social environments.

Adaptive Response to Gillnets Bycatch in a North Sardinia Mediterranean Shag (Gulosus aristotelis desmarestii) Population.

Mediterranean Shag (Gulosus aristotelis desmarestii) is a seabird endemic to the Mediterranean and Black Seas, recently included in the IUCN list of threatened Species. Most of the reproductive colonies are hosted in Sardinia and surrounding islets. Bycatch in fishing nets is one of the most significant threats for this population. Our work aimed to assess alterations in the sex ratio caused by bycatch and to study the adaptive response of the population to a skewed adult sex ratio. The sex ratio of Mediterranean Shags found drowned in the gillnets near the colonies and that of the nestlings of the Corcelli (northeast Sardinia) colony was determined using the sex-linked polymorphism of the gene Chromobox-Helicase-DNA-binding 1. The data of the shags found drowned in gillnets evidenced a high mortality rate (83.3%; p < 0.001) and a larger size of males (35% heavier than females, p < 0.05) compared to females, supporting the theory that heavier individuals are able to forage at great depths. With 64.8% of the nestlings being male, the sex ratio of nestlings was statistically different from parity (p < 0.05). Furthermore, it was related to the brood size. In one- and two-chick broods, 73% and 70% of nestlings, respectively, were males, while in three-chick broods, only 33% were males. Our data identify the higher rate of male shags drowned in gillnets as a factor causing an alteration of the sex ratio in the Mediterranean Shag population. According to the Sex Allocation Theory, an adaptive adjustment of sex made by adult females restores the Mendelian sex ratio in the population.

Direct and cross-generational effects of reproduction on fitness and behavioral variability in male-biased environments.

Population structure determines individuals' interactions and trade-offs with evolutionary consequences. Male-biased populations increase intrasexual competition and intersexual harassment, reducing female resource acquisition, and thus, resources availability for the following generation. We analyzed direct and cross-generational effects of male harassment in two generations of damselflies (Odonata). We exposed adult females to treatments with different sex-ratio and density (balanced and male-biased) to modify the male harassment level. We analyzed female fecundity, fertility, and number of faecal deposits as an indirect measure of resources acquisition. We studied female flight performance after repeated exposures to males. We analyzed survivorship, development, exploration, thigmotaxis, and feeding latency of larvae produced by the experimental females. In both generations, we analyzed four metrics of behavior: mean value, interindividual differences in plasticity, intra-individual unpredictability, and repeatability. Mating duration increased in male-biased treatment, whereas female resources acquisition and fertility decreased. Females that mated longer showed higher fecundity when they were exposed to balanced treatment, but not if they were exposed to male-biased treatment. Females from the male-biased treatment showed interindividual differences in plasticity and no repeatability in flight performance. Offspring showed balanced sex-ratio and similar survivorship, development, and feeding latency independently of the parental treatment; however, females exposed to male-biased treatment produced offspring with higher differences in exploration plasticity and daughters less explorative and with higher unpredictable thigmotaxis. We propose prolonged copulation as courtship at balanced sex-ratio but a cost to females under male-biased sex-ratio. Cross-generational effects in behavioral variability may be a mechanism to cope with predicted future environments.

Effects of Glyphosate on Female Reproductive Output in the Marine Polychaete Worm Ophryotrocha diadema.

Glyphosate is a broad-spectrum herbicide widely employed in agriculture. Exposure to this genotoxic and endocrine-disrupting compound has adverse effects on terrestrial and aquatic organisms and on humans as well. Here, we explored the effects of glyphosate on female reproductive output and somatic growth rate in the marine polychaete worm, Ophryotrocha diadema. Adult focal individuals were exposed to different concentrations of pure glyphosate (0.0, 0.125 0.250, 0.500, 1.000 µg/mL) administered once a week for 3 weeks. Toxic effects and mortalities were observed at the three higher concentrations, whereas only a decrease in growth rate was noted after exposure to 0.125 µg/mL, which did not affect female allocation. An area of focus in future studies should be the effects of contaminants, their metabolites, and ecologically relevant human-driven stressors in the context of global warming.

Variance in offspring sex ratio and maternal allocation in a highly invasive mammal.

Skewed sex ratios at birth are widely reported in wild populations, however, the extent to which parents are able to modulate the sex ratio of offspring to maximize their own fitness remains unclear. This is particularly true for highly polytocous species as maximizing fitness may include trade-offs between sex ratio and the size and number of offspring in litters. In such cases, it may be adaptive for mothers to adjust both the number of offspring per litter and offspring sex to maximize individual fitness. Investigating maternal sex allocation in wild pigs (Sus scrofa) under stochastic environmental conditions, we predicted that under favorable conditions, high-quality mothers (larger and older) would produce male-biased litters and invest more in producing larger litters with more males. We also predicted sex ratio would vary relative to litter size, with a male-bias among smaller litters. We found evidence that increasing wild boar ancestry, maternal age and condition, and resource availability may weakly contribute to male-biased sex ratio, however, unknown factors not measured in this study are assumed to be more influential. High-quality mothers allocated more resources to litter production, but this relationship was driven by adjustment of litter size, not sex ratio. There was no relationship between sex ratio and litter size. Collectively, our results emphasized that adjustment of litter size appeared to be the primary reproductive characteristic manipulated in wild pigs to increase fitness rather than adjustment of offspring sex ratio.

Multiple aspects of the maternal reproductive investment in a polytocous species: What do mothers really control?

One of the factors facilitating the expansion and proliferation of wild boar Sus scrofa is the plasticity of its reproductive biology. Nevertheless, the real influence of maternal and environmental factors on number and sex of the offspring is still controversial. While the litter size was shown to be related with the maternal condition, the strength of this relation remains to be understood, together with the possible role played by environmental conditions. Analogously, it is unclear whether wild boar females can adjust their offspring sex. We investigated multiple aspects of wild boar maternal investment by means of a 10-year dataset of female reproductive traits and a set of biologically meaningful environmental variables. The maternal condition slightly affected the litter size but not the offspring sex, and environment did not affect the litter size or the offspring sex. Moreover, mothers did not cope with the higher costs entailed by producing sons by placing them in the most advantageous intrauterine position, nor by allocating less resources on daughters. Our set of results showed that the female reproductive investment is quite rigid in comparison with other aspects of wild boar reproductive biology. Wild boar females seem to adopt a typical r-strategy, producing constantly large litters and allocating resources on both sexes regardless of internal and external conditions. Such strategy may be adaptive to cope with environmental unpredictability and an intense human harvest, contributing to explain the extreme success of wild boar within human-dominated landscapes.

The rarer-sex effect.

The study of sex allocation-that is, the investment of resources into male versus female reproductive effort-yields among the best quantitative evidence for Darwinian adaptation, and has long enjoyed a tight and productive interplay of theoretical and empirical research. The fitness consequences of an individual's sex allocation decisions depend crucially upon the sex allocation behaviour of others and, accordingly, sex allocation is readily conceptualized in terms of an evolutionary game. Here, I investigate the historical development of understanding of a fundamental driver of the evolution of sex allocation-the rarer-sex effect-from its inception in the writing of Charles Darwin in 1871 through to its explicit framing in terms of consanguinity and reproductive value by William D. Hamilton in 1972. I show that step-wise development of theory proceeded through refinements in the conceptualization of the strategy set, the payoff function and the unbeatable strategy. This article is part of the theme issue 'Half a century of evolutionary games: a synthesis of theory, application and future directions'.

The presence of a guard vicariously drives split sex ratios in a facultatively social bee.

Split sex ratios provide broad insights into how reproductive strategies evolve, and historically have special relevance to the evolution of eusociality. Yet almost no attention has been directed to situations where split sex ratios may potentially decrease the payoffs for worker-like behaviour, increasing selective thresholds for eusociality. We examined sex ratios in a facultatively social colletid bee, Amphylaeus morosus. Sex ratios in this bee vary strongly with the presence of a nest guard and in a pattern that does not conform to assumptions of previous models in which split sex ratios facilitate altruism. While the production of daughters was constant across social and solitary nests, mothers produced more brood when a non-reproductive guard was present, but these extra brood were all male. This leads to split sex ratios, vicariously driven by guards that are unable to manipulate sex ratios in their favour. Importantly, if guarding becomes more common in a population this would lead to an excess of males and lower the genetic value of these extra males to guards, effectively putting a brake on selection for worker-like behaviour.

Technical comment on "sex ratios when helpers stay at the nest".

I contributed a paper to volume 60 of the journal. The paper reported on my study of sex-ratio evolution when one sex (females) is helpful but the other sex (males) suffers less from kin competition. I had based my study on a kin-selection model, and so I was dismayed to discover an error in the relatedness calculations therein. Specifically, relatedness coefficients that should have been calculated using a sampling-without-replacement scheme were instead calculated using sampling with replacement. Here, I correct my error and show how it impacts my original findings. I argue that my main conclusions are unchanged. Furthermore, only two new findings contrast with those I presented earlier. First, changing those model details unrelated to the marginal fitness benefits of help does not, in turn, impact substantially the conflict that occurs between mates over the brood sex ratio (I had previously reported some noteworthy impact was possible). Second, help can reduce sex-ratio conflict between mates more effectively when breeders occur in smaller groups (previously, I had said this occurred in larger groups).