Steven Freedberg

CULTURAL INHERITANCE AS A MECHANISM FOR POPULATION SEX-RATIO BIAS IN REPTILES

Abstract.— Although natural populations of most species exhibit a 1:1 sex ratio, biased sex ratios are known to be associated with non‐Mendelian inheritance, as in sex‐linked meiotic drive and cytoplasmic inheritance (Charnov 1982; Hurst 1993). We show how cultural inheritance, another type of non‐Mendelian inheritance, can favor skewed primary sex ratios and propose that it may explain the female‐biased sex ratios commonly observed in reptiles with environmental sex determination (ESD). Like cytoplasmic elements, cultural traits can be inherited through one sex. This, in turn, favors skewing the primary sex allocation in favor of the transmitting sex. Female nest‐site philopatry is a sex‐specific, culturally inherited trait in many reptiles with ESD and highly female‐biased sex ratios. We propose that the association of nest‐site selection with ESD facilitates the maternal manipulation of offspring sex ratios toward females.

Nesting fidelity and molecular evidence for natal homing in the freshwater turtle, Graptemys kohnii

Numerous studies of sea turtle nesting ecology have revealed that females exhibit natal homing, whereby they imprint on the nesting area from which they hatch and subsequently return there to nest as adults. Because freshwater turtles comprise the majority of reptiles known to display environmental sex determination (ESD), the study of natal homing in this group may shed light on recent evolutionary models of sex allocation that are predicated on natal homing in reptiles with ESD. We examined natal homing in Graptemys kohnii, a freshwater turtle with ESD, using mitochondrial sequencing, microsatellite genotyping and mark and recapture of 290 nesting females. Females showed high fidelity to nesting areas, even after being transplanted several kilometres away. A Mantel test revealed significant genetic isolation by distance with respect to nesting locations (r=0.147; p<0.05), suggesting that related females nest in close proximity to one another. The patterns of fidelity and genotype distributions are consistent with homing at a scale that may affect population sex ratios.

Long-term sex reversal by oestradiol in amniotes with heteromorphic sex chromosomes

Oestradiol application during embryonic development reverses the sex of male embryos and results in normal female differentiation in reptiles lacking heteromorphic sex chromosomes, but fails to do so in birds and mammals with heteromorphic sex chromosomes. It is not clear whether the evolution of heteromorphic sex chromosomes in amniotes is accompanied by insensitivity to oestradiol, or if the association between oestradiol insensitivity and heteromorphic sex chromosomes can be attributable to phylogenetic constraints in these taxa. Turtles provide an ideal system to examine the potential relationship between oestradiol insensitivity and sex chromosome heteromorphy, since there are species with heteromorphic sex chromosomes that are closely related to species lacking heteromorphic sex chromosomes. We investigated this relationship by examining the long-term effects of oestradiol-17β application on sex determination in Staurotypus triporcatus and Staurotypus salvinii, two turtle species with male heterogamety. After raising the turtles in the lab for 3 years, we found follicular and Müllerian duct morphology in oestradiol-treated turtles that was identical to that of untreated females. The lasting sex reversal suggests that the evolutionary transition between systems lacking heteromorphic sex chromosomes and those with heteromorphic sex chromosomes is not constrained by a fundamental mechanistic difference.

Sex ratio variance and the maintenance of environmental sex determination

Although variation in population sex ratios is predicted to increase the extinction rate of clades with environmental sex determination (ESD), ESD is still seen in a wide array of natural systems. It is unclear how this common sex‐determining system has persisted despite this inherent disadvantage associated with ESD. We use simulation modelling to examine the effect of the sex ratio variance caused by ESD on population colonization and establishment. We find that an accelerating function of establishment success on initial population sex ratio favours a system that produces variance in sex ratios over one that consistently produces even sex ratios. This sex ratio variance causes ESD to be favoured over genetic sex determination, even when the mean global sex ratio under both sex‐determining systems is the same. Data from ESD populations suggest that the increase in population establishment can more than offset the increased risk of extinction associated with temporal fluctuations in the sex ratio. These findings demonstrate that selection in natural systems can favour increased variance in a trait, irrespective of the mean trait value. Our results indicate that sex ratio variation may provide an advantage to species with ESD, and may help explain the widespread existence of this sex‐determining system.

Incubation Environment Affects Immune System Development in a Turtle with Environmental Sex Determination

Abstract The developmental environment can have lasting effects on posthatching phenotype in oviparous animals. Innate immune response is one important component of fitness in vertebrates because it provides a generalized defense against infection. In addition, because male vertebrates are at a higher risk of infection than females, males may benefit more from increased innate immunity than females. We determined the effects of incubation temperature on the innate immune response of hatchling map turtles (Graptemys) by incubating eggs at a range of male and female producing-temperatures and assessing plasma complement activity in the resulting hatchlings. We found a significant effect of incubation environment on circulating complement in hatchling Graptemys ouachitensis, with male-producing temperatures yielding the highest innate immune response. Most important, these results demonstrate that immune response is affected by developmental environment in a species with environmental sex determination, potentially resulting in sex differences in the ability to fend off pathogens.

Male combat favours female-biased sex ratios under environmental sex determination

Fisher predicted that sex ratios should be even when the cost of producing each sex is equal (Fisher 1930). Unbiased sex ratios are predicted to evolve because frequency-dependent selection favours the rarer sex. In contrast to this Fisherian expectation, many reptilian species with environmental sex determination (ESD) apparently manifest female-biased sex ratios (Table 1; reviewed in Bull & Charnov 1988, 1989). Several models predict that sex differences in the quality of the developmental environment can cause deviations from an even sex ratio under ESD (Bull 1981; Frank & Swingland 1988; Bull & Charnov 1989; Charnov & Bull 1989a, b). Specifically, the models predict that ‘the population primary sex ratio . may deviate substantially from 1/2 at equilibrium . (and) should favor the sex that is overproduced in the patches of poorer quality’ (Bull & Charnov 1988, page 107). However, the models do not predict any directional trend to the skew in sex ratio. Thus, the predominance of female-biased sex ratios is not predicted by sex ratio theory in species with ESD. Trivers & Willard (1973) proposed that when maternal condition varies, mothers that can control the gender of their offspring benefit from producing sons when in good condition. They argued that, in species with strong sexual selection, a male’s reproductive success is more dependent on its juvenile condition than is a female’s reproductive success. When maternal condition is good, a female could increase ultimate reproductive success of her lineage by investing in sons. Their model assumes that small differences in the condition of the young can persist until adulthood or be augmented throughout adulthood. Charnov (1982) notes that while the main prediction is valid, there may exist limitations to a mother’s ability to control the sex of her offspring. Although differential mortality or

Estradiol-17β Induces Lasting Sex Reversal at Male-Producing Temperatures in Kinosternid Turtles

Abstract In turtles with temperature-dependent sex determination (TSD), estradiol applied to eggs incubated at male-producing temperatures produces female hatchlings. In other vertebrate taxa, estradiol administration during embryogenesis often induces only temporary or incomplete feminization, and sensitivity to estradiol can vary among taxonomic families. In the present study, we treated eggs of Kinosternon scorpiodes and Sternotherus odoratus incubated at male-producing temperatures with estradiol-17β and examined reproductive morphology of offspring. Furthermore, to determine whether sex reversal resulting from estradiol application in turtles with TSD is lasting, we maintained juvenile K. scorpiodes in the lab for up to three years prior to dissection. We found normal ovarian morphology and Müllerian duct development in a significantly greater proportion of the hatchling S. odoratus and juvenile K. scorpiodes that had been treated with estradiol during embryogenesis than in the untreated group. Results further suggest that estradiol-induced sex reversal seen in TSD turtles is both permanent and ubiquitous and may shed light on the evolution of sex-determining mechanisms in vertebrates.

Nuclear introgression without mitochondrial introgression in two turtle species exhibiting sex-specific trophic differentiation.

Abstract Despite the presence of reproductive barriers between species, interspecific gene introgression has been documented in a range of natural systems. Comparing patterns of genetic introgression in biparental versus matrilineal markers can potentially reveal sex‐specific barriers to interspecific gene flow. Hybridization has been documented in the freshwater turtles Graptemys geographica and G. pseudogeographica, whose ranges are largely sympatric. Morphological differentiation between the species is restricted to females, with female G. geographica possessing large heads and jaws compared to the narrow heads of G. pseudogeographica females. If hybrid females are morphologically intermediate, they may be less successful at exploiting parental feeding niches, thereby limiting the introgression of maternally inherited, but not biparental, molecular markers. We paired sequence data with stable isotope analysis and examined sex‐specific genetic introgression and trophic differentiation in sympatric populations of G. geographica and G. pseudogeographica. We observed introgression from G. pseudogeographica into G. geographica at three nuclear loci, but not at the mitochondrial locus. Analysis of ∂15N and ∂13C was consistent with species differences in trophic positioning in females, but not males. These results suggest that ecological divergence in females may reduce the opportunity for gene flow in this system.

Weakened purifying selection leads to elevated mutation load under environmental sex determination

In many gonochoristic taxa, sex is influenced by developmental environment, a system that can lead to temporal fluctuations in offspring sex ratio. Demographic models suggest that only short‐lived species with environmental sex determination (ESD) are negatively impacted by sex‐ratio fluctuations, yet these models fail to account for the potential mutation load associated with reductions in genetically effective population sizes. In this study, we developed a series of individual‐based simulation models that explore the fixation rates of mildly deleterious alleles under different sex‐determining systems and examine the impacts of variation in lifespan and offspring sex ratio. Populations with ESD exhibited increases in fixation rates in both short‐ and long‐lived populations, but substantial increases were limited to populations characterized by a combination of high sex‐ratio variation and short lifespan. Fixation rates were negatively associated with effective population size, indicating that purifying selection operates less efficiently under ESD relative to genotypic sex determination. Reductions in effective population size could be attributed to both intragenerational forces (unequal sex ratio) and intergenerational forces (variable census population sizes). Levels of temporal sex‐ratio variation calculated from wild populations of ESD species were capable of yielding large increases in fixation rates, although this relationship was strongly mediated by lifespan. Our results may help to explain the limited phylogenetic distribution of ESD in short‐lived taxa.

Intragenomic conflict produces sex ratio dynamics that favor maternal sex ratio distorters.

Abstract Maternal sex ratio distorters (MSDs) are selfish elements that enhance their transmission by biasing their hosts sex allocation in favor of females. While previous models have predicted that the female‐biased populations resulting from sex ratio distortion can benefit from enhanced productivity, these models neglect Fisherian selection for nuclear suppressors, an unrealistic assumption in most systems. We used individual‐based computer simulation modeling to explore the intragenomic conflict between sex ratio distorters and their suppressors and explored the impacts of these dynamics on population‐level competition between species characterized by MSDs and those lacking them. The conflict between distorters and suppressors was capable of producing large cyclical fluctuations in the population sex ratio and reproductive rate. Despite fitness costs associated with the distorters and suppressors, MSD populations often exhibited enhanced productivity and outcompeted non‐MSD populations in single and multiple‐population competition simulations. Notably, the conflict itself is beneficial to the success of populations, as sex ratio oscillations limit the competitive deficits associated with prolonged periods of male rarity. Although intragenomic conflict has been historically viewed as deleterious to populations, our results suggest that distorter–suppressor conflict can provide population‐level advantages, potentially helping to explain the persistence of sex ratio distorters in a range of taxa.