Within-population Crosses Research Articles

The conservation value of small population remnants: variability in inbreeding depression and heterosis of a perennial herb, the narrow-leaved purple coneflower (Echinacea angustifolia).

Anthropogenically fragmented populations may have reduced fitness due to loss of genetic diversity and inbreeding. The extent of such fitness losses due to fragmentation and potential gains from conservation actions are infrequently assessed together empirically. Controlled crosses within and among populations can identify whether populations are at risk of inbreeding depression and whether interpopulation crossing alleviates fitness loss. Because fitness depends on environment and life stage, studies quantifying cumulative fitness over a large portion of the lifecycle in conditions that mimic natural environments are most informative. To assess fitness consequences of habitat fragmentation, we leveraged controlled within-family, within-population, and between-population crosses to quantify inbreeding depression and heterosis in seven populations of Echinacea angustifolia within a 6400-hectare area. We then assessed cumulative offspring fitness after 14 years of growth in a natural experimental plot (N = 1136). Mean fitness of progeny from within-population crosses varied considerably, indicating genetic differentiation among source populations, even though these sites are all less than 9 km apart. The fitness consequences of within-family and between-population crosses varied in magnitude and direction. Only one of the seven populations showed inbreeding depression of high effect, while four populations showed substantial heterosis. Outbreeding depression was rare and slight. Our findings indicate that local crossings between isolated populations yield unpredictable fitness consequences ranging from slight decreases to substantial increases. Interestingly, inbreeding depression and heterosis did not relate closely to population size, suggesting that all fragmented populations could contribute to conservation goals as either pollen recipients or donors.

Development of reproductive barriers in sympatry

Freshwater zooplankter Brachionus plicatilis is able to inhabit different habitats and locally adapt to their environmental conditions. It also shows a high degree of population structuring in small geographical regions. Here we try to shed light on the evolution of reproductive isolation in populations of B. plicatilis with presumptive gene flow among locally adapted populations. We have conducted laboratory experiments on admixed pairwise populations that differ in predictability of the water regime. We have assessed the potential for within-population reproductive preferences as a deviation of genotypes from Hardy–Weinberg equilibrium in diapausing eggs, a product of sexual reproduction. We expected heterozygote deficit to increase with environmental distance. We have found signs for incipient reproductive isolation in one third of our admixed populations, however no correlation with environmental distance was found, nor with genetic or geographic predictor variables. The overall inbreeding coefficient showed a tendency for within-population crosses preferences to decrease over time.

Intraspecific variation in reproductive barriers between two closely related Arabidopsis sister species.

Reproductive isolation (RI) is a critical component of speciation and varies strongly in timing and strength among different sister taxa, depending on, for example the geography of speciation and divergence time. However, these factors may also produce variation in timing and strength among populations within species. Here we tested for variation in the expression of RI among replicate population pairs between the sister taxa Arabidopsis lyrata subsp. lyrata and A. arenicola. While the former is predominantly outcrossing, the latter is predominantly selfing. We focused on intrinsic prezygotic and postzygotic RI as both species occur largely in allopatry. We assessed RI by performing within-population crosses and interspecific between-population crosses, and by raising offspring. RI was generally high between all interspecific population pairs, but it varied in timing and strength depending on population history. Prezygotic isolation was strongest between the closest-related population pair, while early postzygotic isolation was high for all other population pairs. Furthermore, the timing and strength of RI depended strongly on cross direction. Our study provides empirical support that reproductive barriers between species are highly variable among population pairs and asymmetric within population pairs, and this variation seems to follow patterns typically described across species pairs.

Fitness consequences of targeted gene flow to counter impacts of drying climates on terrestrial-breeding frogs

Targeted gene flow (TGF) could bolster the adaptive potential of isolated populations threatened by climate change, but could also lead to outbreeding depression. Here, we explore these possibilities by creating mixed- and within-population crosses in a terrestrial-breeding frog species threatened by a drying climate. We reared embryos of the crawling frog (Pseudophryne guentheri) on wet and dry soils and quantified fitness-related traits upon hatching. TGF produced mixed outcomes in hybrids, which depended on crossing direction (origin of gametes from each sex). North-south crosses led to low embryonic survival if eggs were of a southern origin, and high malformation rates when eggs were from a northern population. Conversely, east-west crosses led to one instance of hybrid vigour, evident by increased fitness and desiccation tolerance of hybrid offspring relative to offspring produced from within-population crosses. These contrasting results highlight the need to experimentally evaluate the outcomes of TGF for focal species across generations prior to implementing management actions.

Evaluating Assisted Gene Flow in Marginal Populations of a High Mountain Species

Many species cannot either migrate or adapt at the rate of temperature increases due to climate warming. Therefore, they need active conservation strategies to avoid extinction. Facilitated adaptation actions, such assisted gene flow, aim at the increase of the evolutionary resilience of species affected by global change. In elevational gradients, marginal populations at the lower elevation edges are experiencing earlier snowmelt and higher temperatures, which force them to adapt to the new conditions by modifying their phenology. In this context, advancing the onset of flowering and seed germination times are crucial to ensure reproductive success and increase seedling survival prior to summer drought. Assisted gene flow may bring adaptive alleles and increase genetic diversity that can help throughout ontogeny. The main aim of this work is to assess the effects that different gene flow treatments could have on the desired trait changes in marginal populations. Accordingly, we established a common garden experiment in which we assayed four different gene flow treatments betweenSilene ciliataPourr. (Caryophyllaceae) populations located in similar and different elevation edges, belonging to the same and different mountains. As a control treatment, within-population crosses of low elevation edge populations were performed. The resulting seeds were sown and the germination and flowering onset dates of the resulting plants recorded, as well as the seedling survival. Gene flow between populations falling on the same mountain and same elevation and gene flow from high-elevation populations from a different mountain to low-elevation populations advanced seed germination time with respect to control crosses. No significant effects of gene flow on seedling survival were found. All the gene flow treatments delayed the onset of flowering with respect to control crosses and this effect was more pronounced in among-mountain gene flows. The results of this study highlight two important issues that should be thoroughly studied before attempting to apply assisted gene flow in practical conservation situations. Firstly, among-populations gene flow can trigger different responses in crucial traits throughout the ontogeny of plant species. Secondly, the population provenance of gene flow is determinant and plays a significant role on the effects of gene flow.

Selfish chromosomal drive shapes recent centromeric histone evolution in monkeyflowers.

Centromeres are essential mediators of chromosomal segregation, but both centromeric DNA sequences and associated kinetochore proteins are paradoxically diverse across species. The selfish centromere model explains rapid evolution by both components via an arms-race scenario: centromeric DNA variants drive by distorting chromosomal transmission in female meiosis and attendant fitness costs select on interacting proteins to restore Mendelian inheritance. Although it is clear than centromeres can drive and that drive often carries costs, female meiotic drive has not been directly linked to selection on kinetochore proteins in any natural system. Here, we test the selfish model of centromere evolution in a yellow monkeyflower (Mimulus guttatus) population polymorphic for a costly driving centromere (D). We show that the D haplotype is structurally and genetically distinct and swept to a high stable frequency within the past 1500 years. We use quantitative genetic mapping to demonstrate that context-dependence in the strength of drive (from near-100% D transmission in interspecific hybrids to near-Mendelian in within-population crosses) primarily reflects variable vulnerability of the non-driving competitor chromosomes, but also map an unlinked modifier of drive coincident with kinetochore protein Centromere-specific Histone 3 A (CenH3A). Finally, CenH3A exhibits a recent (<1000 years) selective sweep in our focal population, implicating local interactions with D in ongoing adaptive evolution of this kinetochore protein. Together, our results demonstrate an active co-evolutionary arms race between DNA and protein components of the meiotic machinery in Mimulus, with important consequences for individual fitness and molecular divergence.

To mix or not to mix the sources of relocated plants? The case of the endangered Iris lortetii

Active management of endangered species is required for the persistence of many rare species. Species translocation, a common practice used to mitigate the negative effects of small population sizes, entails risks of outbreeding depression due to dilution of local adaptations, and therefore must be weighed against the costs of a hands-off conservation approach. Iris lortetii is an endangered rhizomatous plant, growing in a small number of isolated populations in northern Israel. We implemented a quasi-in-situ reintroduction program by planting 234 rhizomes from diverse origins in five new sites. All new sites were selected to be as similar as possible to those of the largest natural population. We recorded plant survival and flowering in the field after four years. Flowering plants were artificially crossed either with plants from the same population of origin (within population) or with plants from different origin (between populations). We found no differences in survival between populations of origin and only some indication of local adaptations in the form of increased flowering of the local population. Nonetheless, seed set was significantly higher (a 73% increase) in crosses between populations of origin, compared to within-population crosses, suggesting low genetic diversity within the natural populations. The ability to combine active conservation with rigid testing of theoretical hypothesis, while avoiding all risk to natural populations, highlights the value of the quasi-in-situ approach for restoration. Our results indicate that, in the case of Iris lortetii, active relocation of genotypes, seeds or pollen can enhance the survival of natural populations over time.

The genetic architecture of sexually selected traits in two natural populations of Drosophila montana.

We investigated the genetic architecture of courtship song and cuticular hydrocarbon traits in two phygenetically distinct populations of Drosophila montana. To study natural variation in these two important traits, we analysed within-population crosses among individuals sampled from the wild. Hence, the genetic variation analysed should represent that available for natural and sexual selection to act upon. In contrast to previous between-population crosses in this species, no major quantitative trait loci (QTLs) were detected, perhaps because the between-population QTLs were due to fixed differences between the populations. Partitioning the trait variation to chromosomes suggested a broadly polygenic genetic architecture of within-population variation, although some chromosomes explained more variation in one population compared with the other. Studies of natural variation provide an important contrast to crosses between species or divergent lines, but our analysis highlights recent concerns that segregating variation within populations for important quantitative ecological traits may largely consist of small effect alleles, difficult to detect with studies of moderate power.

Remnants of populations provide effective source material for reintroduction of an endangered Hawaiian plant, Schiedea kaalae (Caryophyllaceae).

Reintroductions may be essential to prevent extinction of many critically endangered species. Ideally, reintroduction efforts rely on adjacent source populations, but limited source material may necessitate crossing individuals from different and possibly distant populations. To determine the consequences of integrating multiple populations in reintroductions, we investigated levels of inbreeding depression, outbreeding depression, and heterosis for populations of Schiedea kaalae (Caryophyllaceae), an endangered species endemic to the Wai'anae and Ko'olau Mountains of O'ahu, Hawai'i. The possibility of gene flow among plants was explored through pollinator observations. Individuals from ex situ living collections of nine populations were hand-pollinated with pollen from the same plant, plants from the same population (for three populations only), or plants from different populations. Progeny were outplanted into two common gardens, one in each mountain range on O'ahu. Cumulative fitness was estimated using several independent life history stages. Inbreeding depression was minimal, and no outbreeding depression was detected. In contrast, strong heterosis was evident in progeny from between-population crosses, which had higher relative fitness than progeny from self-pollinations or within-population crosses. Observations of floral visitors provided the first evidence that biotic pollination may be important for this species. Results demonstrate the ability to conduct genetic rescue of rare species and suggest that reintroductions may be most successful using heterotic individuals from crosses between populations and at sites where pollinators are present and promote outcrossing.

Outbreeding Depression in Atlantic Salmon Revealed by Hypoxic Stress During Embryonic Development

The fitness consequences of outbreeding in wild populations are extremely variable. Heterosis and outbreeding depression can both be observed but the effect of environmental stressors on the occurrence of these phenomena is still poorly understood. We tested the influence of oxygen stress during embryonic development on consequences of outbreeding in wild populations of Atlantic salmon (Salmo salar). We used a common garden experiment and performed crosses within and between salmon populations to study performances of embryos reared under normal and hypoxic conditions. We detected both heterosis and outbreeding depression depending on traits but irrespective of divergence between parental populations. Nevertheless, outbreeding depression was observed almost exclusively under hypoxic conditions and prevailed over heterosis regarding survival during the whole embryonic development. Notably, the post-hatching survival of all between population crosses was approximately 15 % lower than the survival of within-population crosses under hypoxic conditions. Different hypoxia reaction norms for post-hatching survival, length and time to hatch were also noticed among within and between populations crosses further indicating outbreeding depression. These results demonstrate that consequences of outbreeding can dramatically vary depending on environmental conditions with outbreeding depression being possibly stronger under stressful conditions.

Rapid range expansion is not restricted by inbreeding in a sexually cannibalistic spider.

Few studies investigated whether rapid range expansion is associated with an individual's short-term fitness costs due to an increased risk of inbred mating at the front of expansion. In mating systems with low male mating rates both sexes share potential inbreeding costs and general mechanisms to avoid or reduce these costs are expected. The spider Argiope bruennichi expanded its range recently and we asked whether rapid settlement of new sites exposes individuals to a risk of inbreeding. We sampled four geographically separated subpopulations, genotyped individuals, arranged matings and monitored hatching success. Hatching success was lowest in egg-sacs derived from sibling pairs and highest in egg-sacs derived from among-population crosses, while within-population crosses were intermediate. This indicates that inbreeding might affect hatching success in the wild. Unlike expected, differential hatching success of within- and among-population crosses did not correlate with genetic distance of mating pairs. In contrast, we found high genetic diversity based on 16 microsatellite markers and a fragment of the mitochondrial COI gene in all populations. Our results suggest that even a very recent settlement secures the presence of genetically different mating partners. This leads to costs of inbreeding since the population is not inbred.

The Battle of the Sexes over Seed Size: Support for Both Kinship Genomic Imprinting and Interlocus Contest Evolution

Outcrossing creates a venue for parental conflict. When one sex provides parental care to offspring fertilized by several partners, the nonproviding sex is under selection to maximally exploit the caring sex. The caring sex may counteradapt, and a coevolutionary arms race ensues. Genetic models of this conflict include the kinship theory of genomic imprinting (parent-of-origin-specific expression of maternal-care effectors) and interlocus conflict evolution (interaction between male selfish signals and female abatement). Predictions were tested by measuring the sizes of seeds produced by within-population crosses (diallel design) and between-population crosses in outcrossing and selfing populations of Arabidopsis lyrata. Within-population diallel crosses revealed substantial maternal variance in seed size in most populations. The comparison of between- and within-population crosses showed that seeds were larger when pollen came from another outcrossing population than when pollen came from a selfing or the same population, supporting interlocus contest evolution between male selfish genes and female recognition genes. Evidence for kinship genomic imprinting came from complementary trait means of seed size in reciprocal between-population crosses independent of whether populations were predominantly selfing or outcrossing. Hence, both kinship genomic imprinting and interlocus contest are supported in outcrossing Arabidopsis, whereas only kinship genomic imprinting is important in selfing populations.

Parental Influences on Pathogen Resistance in Brown Trout Embryos and Effects of Outcrossing within a River Network

Phenotypic plasticity can increase tolerance to heterogeneous environments but the elevations and slopes of reaction norms are often population specific. Disruption of locally adapted reaction norms through outcrossing can lower individual viability. Here, we sampled five genetically distinct populations of brown trout (Salmo trutta) from within a river network, crossed them in a full-factorial design, and challenged the embryos with the opportunistic pathogen Pseudomonas fluorescens. By virtue of our design, we were able to disentangle effects of genetic crossing distance from sire and dam effects on early life-history traits. While pathogen infection did not increase mortality, it was associated with delayed hatching of smaller larvae with reduced yolk sac reserves. We found no evidence of a relationship between genetic distance (W, FST) and the expression of early-life history traits. Moreover, hybrids did not differ in phenotypic means or reaction norms in comparison to offspring from within-population crosses. Heritable variation in early life-history traits was found to remain stable across the control and pathogen environments. Our findings show that outcrossing within a rather narrow geographical scale can have neutral effects on F1 hybrid viability at the embryonic stage, i.e. at a stage when environmental and genetic effects on phenotypes are usually large.

Drift load in populations of small size and low density

According to theory, drift load in randomly mating populations is determined by past population size, because enhanced genetic drift in small populations causes accumulation and fixation of recessive deleterious mutations of small effect. In contrast, segregating load due to mutations of low frequency should decline in smaller populations, at least when mutations are highly recessive and strongly deleterious. Strong local selection generally reduces both types of load. We tested these predictions in 13 isolated, outcrossing populations of Arabidopsis lyrata that varied in population size and plant density. Long-term size was estimated by expected heterozygosity at 20 microsatellite loci. Segregating load was assessed by comparing performance of offspring from selfings versus within-population crosses. Drift load was the heterosis effect created by interpopulation outbreeding. Results showed that segregating load was unrelated to long-term size. However, drift load was significantly higher in populations of small effective size and low density. Drift load was mostly expressed late in development, but started as early as germination and accumulated thereafter. The study largely confirms predictions of theory and illustrates that mutation accumulation can be a threat to natural populations.

Neo-Allopatry and Rapid Reproductive Isolation

Over the past 3 centuries, many species have been dispersed beyond their natural geographic limits by humans, but to our knowledge, reproductive isolation has not been demonstrated for such neo-allopatric species. We grew seeds from three species of Centaurea (Centaurea solstitialis, Centaurea calcitrapa, and Centaurea sulphurea) that are native to Spain and have been introduced into California, and we tested to what extent seed production was affected by pollen source. Compared with within-population crosses, seed production decreased by 52% and 44%, respectively, when C. solstitialis and C. sulphurea from California were pollinated with conspecific pollen from native populations in Spain. This implies rapid evolution of reproductive isolation between populations in their native and nonnative ranges. Whether reproductive isolation has evolved following the introduction of other species is unknown, but additional cases are likely, considering the large number of neo-allopatric species.

Inbreeding and outbreeding depression in Stylidium hispidum: implications for mixing seed sources for ecological restoration

The benefits of composite rather than local seed provenances for ecological restoration have recently been argued, largely on the basis of maximizing evolutionary potential. However, these arguments have downplayed the potentially negative consequences of outbreeding depression once mixed provenances interbreed. In this study, we compared intraspecific F1 hybrid performance and molecular marker differentiation among four populations of Stylidium hispidum, a species endemic to Southwestern Australia. Multivariate ordination of 134 AFLP markers analyzed genetic structure and detected two clusters of paired sites that diverged significantly for marker variation along a latitudinal boundary. To test for outbreeding depression and to determine the consequences of molecular population divergence for hybrid fitness, we conducted controlled pollinations and studied germination and survival for three cross categories (within-population crosses, short- and long-distance F1 hybrids) for paired sites distributed within and between the two genetically differentiated regions. We found evidence of outbreeding depression in long-distance hybrids (111–124 km), and inbreeding depression among progeny of within-population crosses, relative to short-distance (3–10 km) hybrids, suggesting an intermediate optimal outcrossing distance in this species. These results are discussed in light of the evolutionary consequences of mixing seed sources for biodiversity restoration.

Back from the brink: potential for genetic rescue in a critically endangered tree

Rare plant species are vulnerable to genetic erosion and inbreeding associated with small population size and isolation due to increasing habitat fragmentation. The degree to which these problems undermine population viability remains debated. We explore genetic and reproductive processes in the critically endangered long-lived tropical tree Medusagyne oppositifolia, an endemic to the Seychelles with a naturally patchy distribution. This species is failing to recruit in three of its four populations. We evaluate whether recruitment failure is linked to genetic problems associated with fragmentation, and if genetic rescue can mitigate such problems. Medusagyne oppositifolia comprises 90 extant trees in four populations, with only the largest (78 trees) having successful recruitment. Using 10 microsatellite loci, we demonstrated that genetic diversity is high (H(E) : 0.48-0.63; H(O) : 0.56-0.78) in three populations, with only the smallest population having relatively low diversity (H(E) : 0.26 and H(O) : 0.30). All populations have unique alleles, high genetic differentiation, and significant within population structure. Pollen and seed dispersal distances were mostly less than 100 m. Individuals in small populations were more related than individuals in the large population, thus inbreeding might explain recruitment failure in small populations. Indeed, inter-population pollination crosses from the large donor population to a small recipient population resulted in higher reproductive success relative to within-population crosses. Our study highlights the importance of maintaining gene flow between populations even in species that have naturally patchy distributions. We demonstrate the potential for genetic and ecological rescue to support conservation of plant species with limited gene flow.

Between-Population Outbreeding Affects Plant Defence

Between-population crosses may replenish genetic variation of populations, but may also result in outbreeding depression. Apart from direct effects on plant fitness, these outbreeding effects can also alter plant-herbivore interactions by influencing plant tolerance and resistance to herbivory. We investigated effects of experimental within- and between-population outbreeding on herbivore resistance, tolerance and plant fitness using plants from 13 to 19 Lychnis flos-cuculi populations. We found no evidence for outbreeding depression in resistance reflected by the amount of leaf area consumed. However, herbivore performance was greater when fed on plants from between-population compared to within-population crosses. This can reflect outbreeding depression in resistance and/or outbreeding effects on plant quality for the herbivores. The effects of type of cross on the relationship between herbivore damage and plant fitness varied among populations. This demonstrates how between-population outbreeding effects on tolerance range from outbreeding depression to outbreeding benefits among plant populations. Finally, herbivore damage strengthened the observed outbreeding effects on plant fitness in several populations. These results raise novel considerations on the impact of outbreeding on the joint evolution of resistance and tolerance, and on the evolution of multiple defence strategies.

Gene flow from foreign provenances into local plant populations: Fitness consequences and implications for biodiversity restoration

Long-distance transplantation of seed material as done in restoration programs has raised concerns about the risks associated with the introduction of maladapted genotypes that may hybridize with neighboring native conspecifics and decrease local population fitness (outbreeding depression). We studied the consequences of gene flow from foreign provenances into local populations in the common grassland species Plantago lanceolata (Plantaginaceae). Three generations of intraspecific hybrids (F(1), F(2), and backcross to the local plants) were produced by controlled crossings between local plants and plants from geographically or environmentally distant populations. Their performance was compared to that of within-population crosses in a field experiment. Early growth in some interpopulation hybrids was significantly reduced, and this decrease in performance was higher in progeny of crosses with the local population from a different habitat than with geographically distant populations. At the end of the growing season, most fitness-related traits of the interpopulation hybrids were close to the average of their parents. Crosses with low-performing foreign parents therefore resulted in reduced fitness of the hybrids compared to the local plants and dilution of local adaptation. We conclude that the introduction of maladapted populations from distant or ecologically distinct environments might, at least temporarily, decrease the fitness of neighboring local plants.

Consequences of near and far between‐population crosses for offspring fitness in a rare herb

Crosses between plants from different populations may result in heterosis or outbreeding depression. However, despite its importance for conservation, little is known about the spatial scale over which these effects may arise. To investigate the consequences of between-population crosses at two distinct spatial scales, we conducted reciprocal crosses between four populations from two regions in the rare perennial herb Aster amellus. We assessed seed set and offspring fitness in a common garden experiment. Overall, between-population crosses within regions (10 km) resulted in 8% lower seed set than within-population crosses, while between-region crosses (70 km) resulted in 17% higher seed set than within-population crosses. Moreover, offspring from between-population crosses produced 18% more flower heads than offspring from within-population crosses. We conclude that hybridisation between A. amellus plants from different populations did not lead to immediate outbreeding depression and, thus, could represent a valid conservation option to increase genetic diversity. Moreover, our results suggest that the distance between populations affects the outputs of between-population crosses and therefore needs to be taken into account when promoting gene flow between populations.