Relatively weak inbreeding depression in selfing but also in outcrossing populations of North American Arabidopsis lyrata

Hermaphroditic plants can potentially self‐fertilize, but most possess adaptations that promote outcrossing. However, evolutionary transitions to higher selfing rates are frequent. Selfing comes with a transmission advantage over outcrossing, but self‐progeny may suffer from inbreeding depression, which forms the main barrier to the evolution of higher selfing rates. Here, we assessed inbreeding depression in the North American herb Arabidopsis lyrata, which is normally self‐incompatible, with a low frequency of self‐compatible plants. However, a few populations have become fixed for self‐compatibility and have high selfing rates. Under greenhouse conditions, we estimated mean inbreeding depression per seed (based on cumulative vegetative performance calculated as the product of germination, survival and aboveground biomass) to be 0.34 for six outcrossing populations, and 0.26 for five selfing populations. Exposing plants to drought and inducing defences with jasmonic acid did not magnify these estimates. For outcrossing populations, however, inbreeding depression per seed may underestimate true levels of inbreeding depression, because self‐incompatible plants showed strong reductions in seed set after (enforced) selfing. Inbreeding‐depression estimates incorporating seed set averaged 0.63 for outcrossing populations (compared to 0.30 for selfing populations). However, this is likely an overestimate because exposing plants to 5% CO₂ to circumvent self‐incompatibility to produce selfed seed might leave residual effects of self‐incompatibility that contribute to reduced seed set. Nevertheless, our estimates of inbreeding depression were clearly lower than previous estimates based on the same performance traits in outcrossing European populations of A. lyrata, which may help explain why selfing could evolve in North American A. lyrata.     

Different levels of inbreeding depression between outcrossing and selfing Serapias species

We quantified inbreeding depression for fruit production, embryo vitality and seed germination in three deceptive orchids, Serapias vomeracea, S. cordigera and S. parviflora, which do not provide any reward to their pollinators, and are predicted to experience high outcrossing. Of the three species examined only S. parviflora was autonomously selfing. Both S. vomeracea and S. cordigera showed highly significant differences in fitness between selfed and outcrossed progenies, resulting in high levels of inbreeding depression, which increased in magnitude from seed set to seed germination. Inbreeding depression may promote outcrossing in Serapias by acting as a post-pollination barrier to selfing. Cumulative inbreeding depression across three stages in S. parviflora was lower that in both outcrossing species. The large difference in germination between selfed and outcrossed seeds is an important issue in conservation biology.     

Maintenance of Inbreeding Depression in a Highly Self-Fertilizing Tree, Magnolia obovata Thunb

Inbreeding depression is a major selective force that maintains outcrossing in flowering plants. If the long life and large mature size of trees cause high inbreeding depression via mitotic mutations and half-sib competition, these characteristics may increase inbreeding depression sufficiently to maintain traits that facilitate outcrossing even with high primary selfing rates (proportion of selfed ovules). Here, I report the maintenance of inbreeding depression in a population of a tree (Magnolia obovata Thunb.) with primary selfing rates greater than 0.8 resulting from geitonogamy. The progenies exhibited inbreeding depression for germination, seedling survival, and seedling mass (δ = 0.29–0.38), but no significant difference between crossing type in seedling height. Cumulative inbreeding depression for early survival (from zygote to 2-year-old stage) estimated from these results and from prior data on embryonic survival was high (δ_e = 0.91). The fixation index at maturity based on six allozyme loci was low (F_is = 0.08), indicating that significant inbreeding depression for late survival results in a low level of inbreeding with respect to gene transmission to the next generation. From these results, I estimated that inbreeding depression for late and lifetime survival equaled 0.69 and 0.97, respectively. These results suggest that M. obovata trees maintain high inbreeding depression at both early and late life stages, resulting in a low level of inbreeding despite a high primary selfing rate. The high inbreeding depression can be explained by previous theories and is consistent with the predicted maintenance of inbreeding depression in highly self-fertilizing tree populations. The inbreeding load due to the high primary selfing rate represents a cost of this tree’s pollination system for outcrossing, which is based on automimicry and mass flowering. Co-ordinating editor: S.-M. Chang     

OUTCROSSING RATE AND INBREEDING DEPRESSION IN TWO ANNUAL MONOECIOUS HERBS,BEGONIA HIRSUTA AND B. SEMIOVATA

Most models of mating-system evolution predict inbreeding depression to be low in inbred populations due to the purging of deleterious recessive alleles. This paper presents estimates of outcrossing rates and inbreeding depression for two highly selfing, monoecious annuals Begonia hirsuta and B. semiovata. Outcrossing rates were estimated using isozyme polymorphisms, and the magnitude of inbreeding depression was quantified by growing progeny in the greenhouse produced through controlled selfing and outcrossing. The estimated single-locus outcrossing rate was 0.03 ± 0.01 (SE) for B. hirsuta and 0.05 ± 0.02 for B. semiovata. In both species, the seed production of selfed flowers was on average 12% lower than that of outcrossed flowers (B. hirsuta P = 0.07, B. semiovata P < 0.05, mixed model ANOVAs). There was no significant effect of crosstype on germination rate or survival, but selfed offspring had a lower dry mass than outcrossed offspring 18 weeks after planting in both species (on average 18% lower in B. hirsuta and 31% lower in B. semiovata). Plants that were the products of selfing began flowering later than plants produced through outcrossing in B. semiovata, but not in B. hirsuta. The effects of crosstype on seed production (B. semiovata), days to first flower and offspring dry mass (both species) varied among maternal parents, as indicated by significant crosstype x maternal parent interactions for these characters. Both species showed significant inbreeding depression for total fitness (estimated as the product of seed production, germination rate, survival and dry mass at 18 weeks). In B. hirsuta, the average total inbreeding depression was 22% (range -57%-98%; N = 23 maternal parents), and in B. semiovata, it was 42% (-11%-84%; N = 21). This study demonstrates that highly selfing populations can harbor substantial inbreeding depression. Our findings are consistent with the hypothesis that a high mutation rate to mildly deleterious alleles contributes to the maintenance of inbreeding depression in selfing populations.     

Severe outbreeding and inbreeding depression maintain mating system differentiation in Epipactis (Orchidaceae)

In hermaphroditic plants, theory for mating system evolution predicts that populations will evolve to either complete autonomous selfing (AS) or complete outcrossing, depending on the balance between automatic selection favouring self‐fertilization and costs resulting from inbreeding depression (ID). Theory also predicts that selection for selfing can occur rapidly and is driven by purging of genetic load and the loss of ID. Therefore, selfing species are predicted to have low levels of ID or even to suffer from outbreeding depression (OD), whereas predominantly outcrossing species are expected to have high levels of ID. To test these predictions, we related the capacity of AS to the magnitude of early‐acting inbreeding or OD in both allogamous and autogamous species of the orchid genus Epipactis. For each species, the level of AS was assessed under controlled greenhouse conditions, whereas hand‐pollinations were performed to quantify early costs of inbreeding or OD acting at the level of fruit and seed production. In the autogamous species, the capacity of AS was high (> 0.72), whereas in the allogamous species AS was virtually absent (< 0.10). Consistent with our hypothesis, allogamous Epipactis species had significantly higher total ID (average: 0.46) than autogamous species, which showed severe costs of OD (average: ?0.45). Overall, our findings indicate that strong early‐acting ID represents an important mechanism that contributes to allogamy in Epipactis, whereas OD may maintain selfing in species that have evolved to complete selfing.     

Inbreeding depression in Vriesea gigantea,a perennial bromeliad from southern Brazil

Inbreeding depression is a reduction of fitness in the progeny of closely related individuals and its effects are assigned to selfing or biparental inbreeding. Vriesea gigantea is a self‐compatible bromeliad species distributed in the Brazilian Atlantic rainforest and habitat destruction and fragmentation and collection have decreased the natural populations. We aim to describe the occurrence of inbreeding depression (δ) in three natural populations of V. gigantea and to correlate this phenomenon with previous studies of fertility, genetic diversity, population genetic structure, gene flow, mating system and seed dispersal in this species. Fifty‐four adult plants were sampled and 108 flowers were used for pollination treatments (selfing, outcrossing and control). For adult plants, we analysed plant and inflorescence height, flower numbers and seed set. In the progenies, evaluated parameters included seed germination and seedling survival rate. The results indicated low to moderate levels of inbreeding depression in V. gigantea (δ = 0.02 to 0.39), in agreement with molecular data from a previous study. Vriesea gigantea populations tolerate some degree of inbreeding, which is consistent with previous results on fertility, mating system, genetic diversity and gene flow. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 169 , 312–319.     

Inbreeding depression is high in a self‐incompatible perennial herb population but absent in a self‐compatible population showing mixed mating

High inbreeding depression is thought to be one of the major factors preventing evolutionary transitions in hermaphroditic plants from self‐incompatibility (SI) and outcrossing toward self‐compatibility (SC) and selfing. However, when selfing does evolve, inbreeding depression can be quickly purged, allowing the evolution of complete self‐fertilization. In contrast, populations that show intermediate selfing rates (a mixed‐mating system) typically show levels of inbreeding depression similar to those in outcrossing species, suggesting that selection against inbreeding might be responsible for preventing the transition toward complete self‐fertilization. By implication, crosses among populations should reveal patterns of heterosis for mixed‐mating populations that are similar to those expected for outcrossing populations. Using hand‐pollination crosses, we compared levels of inbreeding depression and heterosis between populations of Linaria cavanillesii (Plantaginaceae), a perennial herb showing contrasting mating systems. The SI population showed high inbreeding depression, whereas the SC population displaying mixed mating showed no inbreeding depression. In contrast, we found that heterosis based on between‐population crosses was similar for SI and SC populations. Our results are consistent with the rapid purging of inbreeding depression in the derived SC population, despite the persistence of mixed mating. However, the maintenance of outcrossing after a transition to SC is inconsistent with the prediction that populations that have purged their inbreeding depression should evolve toward complete selfing, suggesting that the transition to SC in L. cavanillesii has been recent. SC in L. cavanillesii thus exemplifies a situation in which the mating system is likely not at an equilibrium with inbreeding depression.     

EVOLUTION OF THE MAGNITUDE AND TIMING OF INBREEDING DEPRESSION IN PLANTS

Estimates of inbreeding depression obtained from the literature were used to evaluate the association between inbreeding depression and the degree of self-fertilization in natural plant populations. Theoretical models predict that the magnitude of inbreeding depression will decrease with inbreeding as deleterious recessive alleles are expressed and purged through selection. If selection acts differentially among life history stages and deleterious effects are uncorrelated among stages, then the timing of inbreeding depression may also evolve with inbreeding. Estimates of cumulative inbreeding depression and stage-specific inbreeding depression (four stages: seed production of parent, germination, juvenile survival, and growth/reproduction) were compiled for 79 populations (using means of replicates, N = 62) comprising 54 species from 23 families of vascular plants. Where available, data on the mating system also were collected and used as a measure of inbreeding history. A significant negative correlation was found between cumulative inbreeding depression and the primary selfing rate for the combined sample of angiosperms (N = 35) and gymnosperms (N = 9); the correlation was significant for angiosperms but not gymnosperms examined separately. The average inbreeding depression in predominantly selfing species (δ = 0.23) was significantly less (43%) than that in predominantly outcrossing species (δ = 0.53). These results support the theoretical prediction that selfing reduces the magnitude of inbreeding depression. Most self-fertilizing species expressed the majority of their inbreeding depression late in the life cycle, at the stage of growth/reproduction (14 of 18 species), whereas outcrossing species expressed much of their inbreeding depression either early, at seed production (17 of 40 species), or late (19 species). For species with four life stages examined, selfing and outcrossing species differed in the magnitude of inbreeding depression at the stage of seed production (selfing δ = 0.05, N = 11; outcrossing δ = 0.32, N = 31), germination (selfing δ = 0.02, outcrossing δ = 0.12), and survival to reproduction (selfing δ = 0.04, outcrossing δ = 0.15), but not at growth and reproduction (selfing δ = 0.21, outcrossing δ = 0.27); inbreeding depression in selfers relative to outcrossers increased from early to late life stages. These results support the hypothesis that most early acting inbreeding depression is due to recessive lethals and can be purged through inbreeding, whereas much of the late-acting inbreeding depression is due to weakly deleterious mutations and is very difficult to purge, even under extreme inbreeding.     

Effect of inbreeding depression on outcrossing rates among populations of a tropical pine

Inbreeding depression is common among plants and may distort mating system estimates. Mating system studies traditionally ignore this effect, nonetheless an assessment of inbreeding depression that may have occurred before progeny evaluation could be necessary. In the neotropical Pinus chiapensis inbreeding depression was evaluated using regression analysis relating progeny F-values with seed germinability, the mating system was analysed in three populations with contrasting size, using isozymes, obtained a corrected outcrossing rate. Selfing decreased seed viability by 19%, relative to an outcrossed plant. Multilocus outcrossing rates, t(m), varied widely among populations. In the two smallest populations t(m) congruent with 1. Therefore, inbreeding depression did not affect the estimates, but overestimated t(m) by 10% in the third population, which has a true mixed mating system (selfing was the major source of inbreeding), and an unusually low t(m) for pines (t(m) = 0.54, uncorrected, t(m) = 0.49, corrected). Inbreeding depression may be an uneven source of bias for outcrossing estimates even at the infraspecific level. Accuracy [corrected] but not precision [corrected] may be gained by including inbreeding depression in outcrossing estimates. Therefore, caution should be taken when comparing t(m) among species or even populations within the same species.     

Significant differences in outcrossing rate,self‐incompatibility,and inbreeding depression between two widely hybridizing species of Geum

Geum urbanum and Geum rivale are two widely hybridizing perennial herbs. Estimation of the breeding systems of these taxa using nuclear microsatellite markers scored in mother–progeny arrays demonstrated that, in pure populations, G. urbanum is predominantly selfing (outcrossing rate, t = 0.058 to 0.177), whereas G. rivale is predominantly outcrossing (t = 0.686–0.775). Theory suggests that hybridization between inbreeding and outcrossing species can potentially generate novel inbreeding lineages. However, the establishment of such lineages may be restricted either by self‐incompatibility loci or deleterious recessive alleles derived from the outcrossing parent. To assess the likelihood that hybridization between G. urbanum and G. rivale will generate novel inbreeding lineages, self‐incompatibility and inbreeding depression were investigated in the two taxa. Seed set in the absence of pollinators, and after controlled self‐ and cross‐pollination, was measured to study self‐incompatibility. Inbreeding depression was measured by estimating the relative fitness of offspring from controlled self‐and cross‐pollinations. Geum urbanum was fully self‐compatible [self‐compatibility index (SCI) = 1] and bagged flowers showed full seed set. By contrast, only 3% of bagged flowers set seed in G. rivale and controlled self‐pollinations showed a 60–80% reduction in seed set compared to controlled outcross pollinations (SCI = 0.28). There was no evidence for inbreeding depression in G. urbanum, although significant, albeit low levels of inbreeding depression were detected in one of two G. rivale populations (δ = 0.33). The implication of these results is that if genetic material from G. rivale was incorporated into a hybrid with a selfing morphology, the establishment of this selfing lineage could be compromised by self‐incompatibility and inbreeding depression. The wider implications of these results for evolution in hybrid swarms between G. urbanum and G. rivale are discussed. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101 , 977–990.     

Breeding systems of hermaphroditic and gynodioecious populations of the colonizing species Trifolium hirtum All. in California

Summary A multilocus procedure was used to estimate outcrossing rates in ten roadside populations of Trifolium hirtum in California. Three groups of populations were studied: cultivars, hermaphroditic, and gynodioecious (sexually dimorphic) populations. The multilocus outcrossing rate (t_m) varied from 0.05 to 0.43 among populations. Population level t_m estimates were significantly correlated with the observed heterozygosity in gynodioecious populations but not in hermaphroditic populations. The outcrossing rate of hermaphrodites and females was estimated in three gynodioecious populations; the estimates of t_m varied from 0.09 to 0.23 for hermaphrodites and from 0.73 to 0.80 for females. The distribution of outcrossing rates in gynodioecious populations is bimodal. Our results indicate that for the levels of selfing observed among hermaphrodites, inbreeding depression is likely to be a major factor in the maintenance of females in gynodioecious populations.     

Patterns of self compatibility, inbreeding depression, outcrossing, and sex allocation in a marine bryozoan suggest the predominating influence of sperm competition

Sex allocation by simultaneous hermaphrodites is theoretically influenced by selfing rate, which is in turn influenced by the benefits of enhanced genomic transmission and reproductive assurance relative to the cost of inbreeding depression. The experimental investigation of these influences in seed plants has a rich pedigree, yet although such an approach is equally relevant to colonial invertebrates, which globally dominate subtidal communities on firm substrata, such studies have been scarce. We reared self‐compatible genets of the marine bryozoan Celleporella hyalina s.l. in the presence and absence of allosperm, and used molecular genetic markers for paternity analysis of progeny to test theoretical predictions that: (1) genets from focal populations with high selfing rates show less inbreeding depression than from focal populations with low selfing rates; (2) genets whose selfed progeny show inbreeding depression prefer outcross sperm (allosperm); and (3) genets bias sex allocation toward female function when reared in reproductive isolation. Offspring survivorship and paternity analysis were used to estimate levels of inbreeding depression and preference for outcrossing or selfing. Sex allocation was assessed by counting male and female zooids. As predicted, inbreeding depression was severe in selfed progeny of genets derived from the populations with low self‐compatibility rates, but, with one exception, was not detected in selfed progeny of genets derived from the populations with higher self‐compatibility rates. Also, as predicted, genets whose selfed progeny showed inbreeding depression preferred outcrossing, and a genet whose selfed progeny did not show inbreeding depression preferred selfing. Contrary to prediction, sex allocation in the majority of genets was not influenced by reproductive isolation. Lack of economy of male function may reflect the over‐riding influence of allosperm‐competition in typically dense breeding populations offering good opportunity for outcrossing. We suggest that hermaphroditism may be a plesiomorphic character of the crown group Bryozoa, prevented by phylogenetic constraint from being replaced by gonochorism and therefore not necessarily adaptive in all extant clades. © 2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 98 , 519–531.     

STRONG INBREEDING DEPRESSION IN TWO SCANDINAVIAN POPULATIONS OF THE SELF‐INCOMPATIBLE PERENNIAL HERB ARABIDOPSIS LYRATA

Inbreeding depression is a key factor influencing mating system evolution in plants, but current understanding of its relationship with selfing rate is limited by a sampling bias with few estimates for self‐incompatible species. We quantified inbreeding depression (δ) over two growing seasons in two populations of the self‐incompatible perennial herb Arabidopsis lyrata ssp. petraea in Scandinavia. Inbreeding depression was strong and of similar magnitude in both populations. Inbreeding depression for overall fitness across two seasons (the product of number of seeds, offspring viability, and offspring biomass) was 81% and 78% in the two populations. Chlorophyll deficiency accounted for 81% of seedling mortality in the selfing treatment, and was not observed among offspring resulting from outcrossing. The strong reduction in both early viability and late quantitative traits suggests that inbreeding depression is due to deleterious alleles of both large and small effect, and that both populations experience strong selection against the loss of self‐incompatibility. A review of available estimates suggested that inbreeding depression tends to be stronger in self‐incompatible than in self‐compatible highly outcrossing species, implying that undersampling of self‐incompatible taxa may bias estimates of the relationship between mating system and inbreeding depression.     

FLORAL CORRELATES AND FITNESS CONSEQUENCES OF MATING-SYSTEM VARIATION IN TURNERA ULMIFOLIA

Outcrossing rates varied from 0% to 69% among Jamaican populations of Turnera ulmifolia. A correlation between increasing herkogamy and outcrossing rate occurred among populations. Predictions from sex-allocation theory were tested by estimating allocation to reproductive functions. Significant differences in allocation patterns occurred among populations, but they were not correlated with outcrossing rates. The fitness consequences of inbreeding were assessed in high- and low-density greenhouse experiments for nine populations with variable outcrossing rates. No evidence for inbreeding depression occurred in early portions of the life history, but multiplicative fitness functions provide evidence for inbreeding depression. We tested the prediction that selfing populations have lower levels of inbreeding depression than outcrossing populations but found no significant correlation.     

Genetic diversity,sexual condition,and microhabitat preference determine mating patterns in Sphagnum (Sphagnaceae) peat‐mosses

In bryophytes, the possibility of intragametophytic selfing creates complex mating patterns that are not possible in seed plants, although relatively little is known about patterns of inbreeding in natural populations. In the peat‐moss genus Sphagnum, taxa are generally bisexual (gametophytes produce both sperm and egg) or unisexual (gametes produced by separate male and female plants). We sampled populations of 14 species, aiming to assess inbreeding variation and inbreeding depression in sporophytes, and to evaluate correlations between sexual expression, mating systems, and microhabitat preferences. We sampled maternal gametophytes and their attached sporophytes at 12–19 microsatellite loci. Bisexual species exhibited higher levels of inbreeding than unisexual species but did generally engage in some outcrossing. Inbreeding depression did not appear to be common in either unisexual or bisexual species. Genetic diversity was higher in populations of unisexual species compared to populations of bisexual species. We found a significant association between species microhabitat preference and population genetic diversity: species preferring hummocks (high above water table) had populations with lower diversity than species inhabiting hollows (at the water table). We also found a significant interaction between sexual condition, microhabitat preference, and inbreeding coefficients, suggesting a vital role for species ecology in determining mating patterns in Sphagnum populations. © 2015 The Linnean Society of London, Biological Journal of the Linnean Society, 2015, 115 , 96–113.     

MUTATIONAL MELTDOWN IN SELFING ARABIDOPSIS LYRATA

The majority of plant species and many animals are hermaphrodites, with individuals expressing both female and male function. Although hermaphrodites can potentially reproduce by self‐fertilization, they have a high prevalence of outcrossing. The genetic advantages of outcrossing are described by two hypotheses: avoidance of inbreeding depression because selfing leads to immediate expression of recessive deleterious mutations, and release from drift load because self‐fertilization leads to long‐term accumulation of deleterious mutations due to genetic drift and, eventually, to extinction. I tested both hypotheses by experimentally crossing Arabidopsis lyrata plants (self‐pollinated, cross‐pollinated within the population, or cross‐pollinated between populations) and measuring offspring performance over 3 years. There were 18 source populations, each of which was either predominantly outcrossing, mixed mating, or predominantly selfing. Contrary to predictions, outcrossing populations had low inbreeding depression, which equaled that of selfing populations, challenging the central role of inbreeding depression in mating system shifts. However, plants from selfing populations showed the greatest increase in fitness when crossed with plants from other populations, reflecting higher drift load. The results support the hypothesis that extinction by mutational meltdown is why selfing hermaphroditic taxa are rare, despite their frequent appearance over evolutionary time.     

The effect of inbreeding on defence against multiple enemies in Datura stramonium

The ability of plants to respond to natural enemies might depend on the availability of genetic variation for the optimal phenotypic expression of defence. Selfing can affect the distribution of genetic variability of plant fitness, resistance and tolerance to herbivores and pathogens. The hypothesis of inbreeding depression influencing plant defence predicts that inbreeding would reduce resistance and tolerance to damage by natural enemies relative to outcrossing. In a field experiment entailing experimentally produced inbred and outcrossed progenies, we assessed the effects of one generation of selfing on Datura stramonium resistance and tolerance to three types of natural enemies, herbivores, weevils and a virus. We also examined the effect of damage on relative growth rate (RGR), flower, fruit, and seed production in inbred and outcrossed plants. Inbreeding significantly reduced plant defence to natural enemies with an increase of 4% in herbivore damage and 8% in viral infection. These results indicate inbreeding depression in total resistance. Herbivory increased 10% inbreeding depression in seed number, but viral damage caused inbred and outcrossed plants to have similar seed production. Inbreeding and outcrossing effects on fitness components were highly variable among families, implying that different types or numbers of recessive deleterious alleles segregate following inbreeding in D. stramonium. Although inbreeding did not equally alter all the interactions, our findings indicate that inbreeding reduced plant defence to herbivores and pathogens in D. stramonium.     

Gender-specific inbreeding depression in a gynodioecious plant, Geranium maculatum (Geraniaceae)

In gynodioecious species, females coexist with hermaphrodites in natural populations even though hermaphrodites attract more pollinators, are capable of reproducing through pollen, and can self-fertilize. This study tests the hypothesis that inbreeding depression helps to maintain females in natural populations. It also examines whether gender lineages that differ in selfing rates might experience different levels of inbreeding depression. Female and hermaphroditic lineages of the gynodioecious species Geranium maculatum were used in self, sib-cross and outcross experiments to examine inbreeding depression levels and to determine whether these levels differ between hermaphroditic and female lineages. Six fitness correlates were measured in the greenhouse and compared among pollination types and between genders. Severe inbreeding depression was found for both individual fitness traits and cumulative fitness in early life history stages. Inbreeding depression levels were slightly higher in hermaphroditic than in female lineages, but this difference was not statistically significant. Because females are unable to self-pollinate and are less likely to experience inbreeding than hermaphrodites under natural conditions, these results suggest that severe inbreeding depression could confer a selective advantage for females that could help to maintain females in natural populations.     

Mating system instability in Schiedea menziesii (Caryophyllaceae)

We investigated inbreeding depression and selfing in hermaphroditic Schiedea menziesii to assess the stability of the breeding system. A combination of high selfing rates and strong inbreeding depression suggests that the mating system is unstable. The population-level selfing rate measured in three years ranged considerably from 0.425 (SE = 0.138) to 0.704 (0.048); family measures of selfing rate varied from zero to one in all three years. Inbreeding coefficients did not differ from zero, suggesting that inbred plants do not survive to reproduction in the field. Average inbreeding depression measured in two greenhouse experiments was 0.608-0.870, with values for individual plants ranging from -0.170 to 0.940. The magnitude of inbreeding depression expressed at different life-history stages depended on experimental conditions. When plants were grown during the winter, inbreeding depression was expressed at early and late life-history stages. When plants were grown during the summer, inbreeding depression was detected for germination but not for later life-history stages. Inbreeding depression for vegetative and inflorescence biomass was also measured using field-collected seeds where cross status was assigned using genotypes determined electrophoretically. We did not detect a relation between inbreeding depression and the selfing rate at the level of the individual plant. We saw no evidence for intrafloral selfing, suggesting that the evolution of increased selfing through autogamy is unlikely, despite high selfing rates. A more likely outcome of breeding system instability is the evolution of gynodioecy, which occurs in species of Schiedea closely related to S. menziesii. Females have been detected in progeny of S. menziesii that have been raised in the greenhouse. In the absence of biotic pollen vectors, the failure of these females to establish in the natural population may result from the absence of adaptations for wind pollination.     

Prior selfing and the selfing syndrome in animals: an experimental approach in the freshwater snail Biomphalaria pfeifferi.

Inbreeding species of hermaphroditic animals practising copulation have been characterized by few copulations, no waiting time (the time that an isolated individual waits for a partner before initiating reproduction compared with paired individuals) and limited inbreeding (self-fertilization) depression. This syndrome, which has never been fully studied before in any species, is analysed here in the highly selfing freshwater snail Biomphalaria pfeifferi. We conducted an experiment under laboratory conditions over two generations (G1 and G2) using snails sampled from two populations (100 individuals per population). G1 individuals were either isolated or paired once a week (potentially allowing for crosses), and monitored during 29 weeks for growth, fecundity and survival. Very few copulations were observed in paired snails, and there was a positive correlation in copulatory activity (e.g. number of copulations) between the male and female sexual roles. The waiting time was either null or negative, meaning that isolated individuals initiated reproduction before paired ones. G2 offspring did not differ in hatching rate and survival (to 28 days) between treatments, but offspring from paired individuals grew faster than those from isolated individuals. On the whole, the self-fertilization depression was extremely low in both populations. Another important result is that paired G1 individuals began laying (selfed) eggs several weeks prior to initiating copulation: this is the first characterization of prior selfing (selfing initiated prior to any outcrossing) in a hermaphroditic animal. A significant population effect was observed on most traits studied. Our results are discussed with regard to the maintenance of low outcrossing rates in highly inbreeding species.