Does maternal stress influence winter survival of offspring in root voles Microtus oeconomus? A field experiment

Maternal stress can have long‐term adverse consequences on immunocompetence and disease risk of offspring, and winter survival is a crucial demographic parameter in the life‐history of an individual that can substantially affect northern rodent population dynamics. An understanding of the effects of maternal stress on winter survival of offspring may help identify mechanisms driving population fluctuations of northern small mammals. Thus, we assessed the effects of maternal stress, resulting from high population densities, on winter survival of first generation (F₁) and second generation (F₂) in root voles Microtus oeconomus. Replicate high‐ and low‐density enclosed parental populations were established, from which we obtained F₁ generation that were used to establish new enclosed, equal‐density populations. The adults of the high‐density parental populations had higher corticosterone levels, an indication of physiological stress, than did those of the low‐density parental populations. Over‐winter survival of the F₁ generation voles from the low‐density parental populations was greater than that of those from the high‐density parental populations. Over‐winter survival of F₂ generation voles did not differ between the two treatments. Our results suggest that maternal stress affected over‐winter survival of first generations but not second generations. Reduced immunocompetence, resulting from high population density stresses, transferred to offspring may be a factor in annual (winter) population declines. Because the effect is transitory, i.e. immunocompetence of F₂ voles is not affected, reduced immunocompetence resulting from high density stresses would not contribute to lengthy periods of low population densities that are characteristic of multi‐annual population fluctuations.     

The effect of maternal and paternal environments on seed characters in the herbaceous plant Campanula Americana (Campanulaceae)

Maternal environments typically influence the phenotype of their offspring. However, the effect of the paternal environment or the potential for joint effects of both parental environments on offspring characters is poorly understood. Two populations of Campanula americana, a woodland herb with a variable life history, were used to determine the influence of maternal and paternal light and nutrient environments on offspring seed characters. Families were grown in the greenhouse in three levels of light or three levels of nutrients. Crosses were conducted within each environmental gradient to produce seeds with all combinations of maternal and paternal environments. On average, increasing maternal nutrient and light levels increased seed mass and decreased percentage germination. The paternal environment affected seed mass, germination time, and percentage germination. However, the influence of the paternal environment varied across maternal environments, suggesting that paternal environmental effects should be evaluated in the context of maternal environments. Significant interactions between family and the parental environments for offspring characters suggest that parental environmental effects are genetically variable. In C. americana, the timing of germination determines life history. Therefore parental environmental effects on germination timing, and genetic variation in those parental effects, suggest that parental environments may influence life history evolution in this system.     

The influence of light on paternal plants in Campanula americana (Campanulaceae): pollen characteristics and offspring traits

Offspring trait expression is determined by the combination of parental genes and parental environments. Although maternal environmental effects have been widely characterized, few studies have focused on paternal environmental effects. To determine whether light availability influences pollen and offspring traits in the woodland herb Campanula americana, we reared clones of 12 genotypes in two light levels. In the parental generation we measured pollen number and size. Plants grown under high light produced more pollen grains per flower than those grown under low light. However, the response was genotype specific; some individuals responded little to changes in light availability while others substantially reduced pollen production. As a consequence, paternity ratios may vary between light environments if more pollen is associated with greater siring success. We crossed a subset of these plants to produce the offspring generation. The paternal and maternal light environments influenced offspring seed mass, percentage germination, and days to germination, while only maternal light levels influenced later life traits, such as leaf number and size. Maternal and paternal environmental effects had opposite influences on seed mass, percentage germination and days to germination. Finally, there was no direct relationship between light effects on pollen production and offspring trait expression.     

Maternal provisioning is structured by species’ competitive neighborhoods

Differential maternal provisioning of offspring in response to environmental conditions has been argued as ‘the missing link’ in plant life histories. Although empirical evidence suggests that maternal provisioning responses to abiotic conditions are common, there is little understanding of how differences in maternal provisioning manifest in response to competition. Frequency manipulations are commonly employed in ecological studies to assess the strength of interspecific competition, relative to intraspecific competition, and we used frequency manipulations to test how competition in two soil moisture environments affects maternal provisioning of seed mass. Specifically, for 15 pairs comprised from 25 annual plant species that occur in California, we varied the relative frequencies of conspecific to heterospecific competitors from 90% (intraspecific competition) to 10% (interspecific competition). We found that conspecific frequency affected maternal provisioning (seed mass) in 12 of the 25 species (eight significantly (p < 0.05), four marginally significantly (p < 0.07)), and that these responses included both increased (five species) and decreased (six species) seed mass, as well as one species with opposing directions of response to conspecific frequency that depended on the soil moisture environment. Conspecific frequency also affected per capita fecundity (seed number) for 17 of the 25 species (15 significantly (p < 0.05), two marginally significantly (p < 0.09)), which generally decreased seed number as conspecific frequency increased. The direction and magnitude of frequency‐dependent seed mass depended on the identity of the competitor, even among species whose fecundity was not affected by competitor identity; the latter finding reveals competitive differences among species that would otherwise appear to be competitively equivalent. Our research demonstrates how species responses to different competitive environments manifest through maternal provisioning, and that these responses alter previous estimates of environmentally‐determined maternal provisioning and reproductive output; future study is needed to understand their combined effects on population and community dynamics.     

Parental effects in Plantago lanceolata L. III. Measuring parental temperature effects in the field

To determine the evolutionary importance of parental environmental effects in natural populations, we must begin to measure the magnitude of these effects in the field. For this reason, we conducted a combined growth chamber-field experiment to measure parental temperature effects in Plantago lanceolata. We grew in the field offspring of controlled crosses of chamber-grown parents subjected to six temperature treatments. Each treatment was characterized by a unique combination of maternal prezygotic (prior to fertilization), paternal prezygotic, and postzygotic (during fertilization and seed set) temperatures. Offspring were followed for three years to measure the effects of treatment on several life-history traits and population growth rate, our estimate of fitness. Parental treatment influenced germination, growth, and reproduction of newborns, but not survival or reproduction of offspring at least one year old. High postzygotic temperature significantly increased germination and leaf area at 17 weeks by approximately 35% and 2%, respectively. Probability of flowering and spike production in the newborn age class showed significant parental genotype x parental treatment interactions. High postzygotic temperature increased offspring fitness by approximately 50%. The strongest contributors to fitness were germination and probability of flowering and spike production of newborns. A comparison of our data with previously collected data for chambergrown offspring shows that the influence of parental environment on offspring phenotype is weaker but still biologically meaningful in the field. The results provide evidence that parental environment influences offspring fitness in natural populations of P. lanceolata and does so by affecting the life-history traits most strongly contributing to fitness. The data suggest that from the perspective of offspring fitness, natural selection favors parents that flower later in the flowering season in the North Carolina Piedmont when it is warmer. Genotypic-specific differences in response of offspring reproductive traits to parental environment suggest that parental environmental effects can influence the rate of evolutionary change in P. lanceolata.     

Adaptive responses and invasion: the role of plasticity and evolution in snail shell morphology

Invasive species often exhibit either evolved or plastic adaptations in response to spatially varying environmental conditions. We investigated whether evolved or plastic adaptation was driving variation in shell morphology among invasive populations of the New Zealand mud snail (Potamopyrgus antipodarum) in the western United States. We found that invasive populations exhibit considerable shell shape variation and inhabit a variety of flow velocity habitats. We investigated the importance of evolution and plasticity by examining variation in shell morphological traits 1) between the parental and F₁ generations for each population and 2) among populations of the first lab generation (F₁) in a common garden, full‐sib design using Canonical Variate Analyses (CVA). We compared the F₁ generation to the parental lineages and found significant differences in overall shell shape indicating a plastic response. However, when examining differences among the F₁ populations, we found that they maintained among‐population shell shape differences, indicating a genetic response. The F₁ generation exhibited a smaller shell morph more suited to the low‐flow common garden environment within a single generation. Our results suggest that phenotypic plasticity in conjunction with evolution may be driving variation in shell morphology of this widespread invasive snail.     

PLANTING DENSITY INFLUENCES THE EXPRESSION OF GENETIC VARIATION IN SEED MASS IN WILD RADISH (RAPHANUS SATIVUS L.: BRASSICACEAE)

To determine the effects of density, genotype, and their interaction on individual seed mass in Raphanus sativus L., we replicated maternal and paternal families of seed across two planting densities in an experimental garden. Seeds were produced by a nested breeding design performed in the greenhouse. Among garden-raised plants, density had a strong negative effect on the mass of seeds produced. At low density, the identity of the greenhouse-grown maternal plants had a strong effect on F₂ seed mass, while in high-density plots, there were no significant parental effects on mean seed mass. Significant parental genotype density interactions contributed to variation in F₂ seed mass. Norms of reaction for each of the 15 paternal sibships illustrate paternal family density interactions. Three sibships exhibited significant declines in mean seed mass with increasing density; 12 sibships showed no change. Maternal family density interaction effects on seed mass were also detected; among maternal sibships, mean seed mass at low density was negatively correlated with mean seed mass at high density. These results demonstrate: a) planting density has a strong effect on mean individual seed mass produced by adults; b) density influences the magnitude of maternal effects on progeny phenotype; and c) genotype density interactions influence seed mass, potentially contributing to the maintenance of maternal genetic variation in seed mass in natural populations of wild radish.     

Hidden responses to environmental variation: maternal effects reveal species niche dimensions

Species responses to fluctuating environments structure population and community dynamics in variable ecosystems. Although offspring number is commonly used to measure these responses, maternal effects on offspring quality may be an important but largely unrecognised determinant of long‐term population growth. We selected 29 species across a Mediterranean annual plant phylogeny, and grew populations of each species in wet and dry conditions to determine responses in seed number and maternal effects (seed size, seed dormancy, and seedling growth). Maternal effects were evident in over 40% of species, but only 24% responded through seed number. Despite a strong trade‐off between seed size and seed number among species, there was no consistent trade‐off within species; we observed correlations that ranged from positive to negative. Overall, species in this plant guild show a complex range of responses to environmental variation that may be underestimated when only seed number responses are considered.     

Maternal effects and maternal selection arising from variation in allocation of free amino acid to eggs

Maternal provisioning can have profound effects on offspring phenotypes, or maternal effects, especially early in life. One ubiquitous form of provisioning is in the makeup of egg. However, only a few studies examine the role of specific egg constituents in maternal effects, especially as they relate to maternal selection (a standardized selection gradient reflecting the covariance between maternal traits and offspring fitness). Here, we report on the evolutionary consequences of differences in maternal acquisition and allocation of amino acids to eggs. We manipulated acquisition by varying maternal diet (milkweed or sunflower) in the large milkweed bug, Oncopeltus fasciatus. Variation in allocation was detected by examining two source populations with different evolutionary histories and life‐history response to sunflower as food. We measured amino acids composition in eggs in this 2 × 2 design and found significant effects of source population and maternal diet on egg and nymph mass and of source population, maternal diet, and their interaction on amino acid composition of eggs. We measured significant linear and quadratic maternal selection on offspring mass associated with variation in amino acid allocation. Visualizing the performance surface along the major axes of nonlinear selection and plotting the mean amino acid profile of eggs from each treatment onto the surface revealed a saddle‐shaped fitness surface. While maternal selection appears to have influenced how females allocate amino acids, this maternal effect did not evolve equally in the two populations. Furthermore, none of the population means coincided with peak performance. Thus, we found that the composition of free amino acids in eggs was due to variation in both acquisition and allocation, which had significant fitness effects and created selection. However, although there can be an evolutionary response to novel food resources, females may be constrained from reaching phenotypic optima with regard to allocation of free amino acids.     

Parental environmental effects on life history traits in Arabidopsis thaliana (Brassicaceae)

Environmentally induced maternal effects on offspring phenotype are well known in plants. When genotypes or maternal lineages are replicated and raised in different environmental conditions, the phenotype of their offspring often depends on the environment in which the parents developed. However, the degree to which such maternal effects are maintained over subsequent generations has not been documented in many taxa. Here we report the results of a study designed to assess the effects of parental environment on vegetative and reproductive traits, using glasshouse-raised maternal lines sampled from natural populations of Arabidopsis thaliana . Replicates of five highly selfed lines from each of four wild populations were cultivated in two abiotic environments in the glasshouse, and the quality and performance of seeds derived from these two environments were examined over two generations. We found that offspring phenotype was strongly influenced by parental environment, but because the parental environments differed with respect to the time of seed harvest, it was not possible to distinguish clearly between parental environmental effects and the possible (but unlikely) effects of seed age on offspring phenotype. We observed a rapid decline in the expression of ancestral environmental effects, and no main environmental effects on progeny phenotype persisted in the second generation. The mechanism of transmission of environmental effects did not appear to be associated with the quantity or quality of reserves in the seeds, suggesting that environmental effects may be transmitted across subsequent generations via some mechanism that generates environment-specific gene expression.     

Close‐kin mating,but not inbred parents,reduces hatching rates and offspring quality in a threatened tortoise

Inbreeding depression, the reduction in fitness due to mating of related individuals, is of particular conservation concern in species with small, isolated populations. Although inbreeding depression is widespread in natural populations, long‐lived species may be buffered from its effects during population declines due to long generation times and thus are less likely to have evolved mechanisms of inbreeding avoidance than species with shorter generation times. However, empirical evidence of the consequences of inbreeding in threatened, long‐lived species is limited. In this study, we leverage a well‐studied population of gopher tortoises, Gopherus polyphemus, to examine the role of inbreeding depression and the potential for behavioural inbreeding avoidance in a natural population of a long‐lived species. We tested the hypothesis that increased parental inbreeding leads to reduced hatching rates and offspring quality. Additionally, we tested for evidence of inbreeding avoidance. We found that high parental relatedness results in offspring with lower quality and that high parental relatedness is correlated with reduced hatching success. However, we found that hatching success and offspring quality increase with maternal inbreeding, likely due to highly inbred females mating with more distantly related males. We did not find evidence for inbreeding avoidance in males and outbred females, suggesting sex‐specific evolutionary trade‐offs may have driven the evolution of mating behaviour. Our results demonstrate inbreeding depression in a long‐lived species and that the evolution of inbreeding avoidance is shaped by multiple selective forces.     

Forest Fragmentation and Seed Germination of Native Species from the Chaco Serrano Forest

Habitat fragmentation is a widespread phenomenon that alters pollination and plant reproductive processes. These effects have demographic and genetic implications that determine offspring fitness and the long‐term viability of plant populations in fragmented systems. We evaluated fragmentation effects on early plant offspring fitness traits, individual seed mass, and percentage of seed germination in five native plant species (Acacia caven, Celtis ehrenbergiana, Croton lachnostachyus, Rivina humilis, Schinus fasciculatus) from the Chaco Serrano forest, a subtropical highly fragmented ecosystem. We found evidence of strong negative fragmentation effects on germination in the shrub C. lachnostachyus and the perennial herb R. humilis, after 30 d of controlled tests. No fragmentation effects were found in the studied traits on the remaining three tree species. We found significant maternal effects in offspring fitness traits in all five species. We discuss the relative magnitude of maternal vs. fragmentation effects taking into account both plant species' lifespan and the time elapsed in fragmentation conditions. We emphasize the need to increase the study of early and late plant offspring fitness produced in fragmented habitats coupled with analyses of genetic parameters and the pollination process in order to evaluate the conservation value of remnant forest fragments. Abstract in Spanish is available at http://www.blackwell‐synergy.com/loi/btp .     

Fitness Traits and Dispersal Ability in the Herb Tragopogon pratensis (Asteraceae): Decoupling the Role of Inbreeding Depression and Maternal Effects

Abstract: Inbreeding depression can decrease several fitness traits and maternal effects can strongly influence the amount of inbreeding depression. Understanding the effects of inbreeding depression on plant fitness is especially important in the context of habitat fragmentation, where plant populations become smaller and more isolated, exhibiting increasing levels of inbreeding depression. We examined the joint influence of inbreeding depression and maternal effects on life cycle traits and dispersal ability in the herb Tragopogon pratensis that grows in fragmented populations in Europe. We conducted experimental crosses to obtain selfed and outcrossed progeny in two contrasted environments. In particular, we produced a first generation of seeds and plants that were self-pollinated again to produce a second generation of seeds. Individual seeds were weighed and their pappuses measured to estimate the dispersal potential. Pollination treatment only had a significant effect on seed mass and dispersal ability. Coefficients of inbreeding depression did not differ between selfed and outcrossed plants. Seed mass had a significant effect on germination date. Environment had a significant effect on mass of the second generation of seeds and the interaction between pollination treatment and family was significant for six traits, indicating the existence of strong maternal effects in T. pratensis. Results suggest population differentiation. Overall, T. pratensis populations exhibited a good performance under selfing, in terms of life cycle traits and dispersal ability, which would allow the species to cope with problems associated with fragmentation.     

Parental effects on seed mass: seed coat but not embryo/endosperm effects

Many biologists studying environmentally induced parental effects have indirectly suggested that the parental environment alters seed mass by altering the amount of endosperm or embryo tissue in the seed. We tested this hypothesis by measuring the effects of parental temperature on total seed mass, seed coat mass, and embryo/endosperm mass in offspring of Plantago lanceolata. Parental temperature significantly affected total seed and coat mass but not endosperm/embryo mass. Thus, larger seeds do not contain more resources in the embryo or endosperm than do small seeds. Rather they have more coat mass, which probably strongly influences germination. These results suggest caution when making assumptions about the pathways by which environmentally induced parental effects are transmitted in plant species. We also observed that controlled crosses differed significantly in their response to parental temperature, which provides evidence for genetic variation in environmentally induced parental effects, i.e., intergenerational phenotypic plasticity, in natural populations of P. lanceolata.     

Are species differences in maternal effects arising from maternal care adaptive?

Parental care benefits offspring through maternal effects influencing their development, growth and survival. However, although parental care in general is likely the result of adaptive evolution, it does not follow that specific differences in the maternal effects that arise from care are also adaptive. Here, we used an interspecific cross‐fostering design in the burying beetle species Nicrophorus orbicollis and N. vespilloides, both of which have elaborate parental care involving direct feeding of regurgitated food to offspring, to test whether maternal effects are optimized within a species and therefore adaptive. Using a full‐factorial design, we first demonstrated that N. orbicollis care for offspring longer regardless of recipient species. We then examined offspring development and mass in offspring reared by hetero‐ or conspecific parents. As expected, there were species‐specific direct effects independent of the maternal effects, as N. orbicollis larvae were larger and took longer to develop than N. vespilloides regardless of caregiver. We also found significant differences in maternal effects: N. vespilloides maternal care caused more rapid development of offspring of either species. Contrary to expectations if maternal effects were species‐specific, there were no significant interactions between caretaker and recipient species for either development time or mass, suggesting that these maternal effects are general rather than optimized within species. We suggest that rather than coadaptation between parents and offspring performance, the species differences in maternal effects may be correlated with direct effects, and that their evolution is driven by selection on those direct effects.     

Effects of increased nitrogen deposition and precipitation on seed and seedling production of Potentilla tanacetifolia in a temperate steppe ecosystem

Background

The responses of plant seeds and seedlings to changing atmospheric nitrogen (N) deposition and precipitation regimes determine plant population dynamics and community composition under global change.

Methodology/Principal Findings

In a temperate steppe in northern China, seeds of P. tanacetifolia were collected from a field-based experiment with N addition and increased precipitation to measure changes in their traits (production, mass, germination). Seedlings germinated from those seeds were grown in a greenhouse to examine the effects of improved N and water availability in maternal and offspring environments on seedling growth. Maternal N-addition stimulated seed production, but it suppressed seed mass, germination rate and seedling biomass of P. tanacetifolia. Maternal N-addition also enhanced responses of seedlings to N and water addition in the offspring environment. Maternal increased-precipitation stimulated seed production, but it had no effect on seed mass and germination rate. Maternal increased-precipitation enhanced seedling growth when grown under similar conditions, whereas seedling responses to offspring N- and water-addition were suppressed by maternal increased-precipitation. Both offspring N-addition and increased-precipitation stimulated growth of seedlings germinated from seeds collected from the maternal control environment without either N or water addition. Our observations indicate that both maternal and offspring environments can influence seedling growth of P. tanacetifolia with consequent impacts on the future population dynamics of this species in the study area.

Conclusion/Significance

The findings highlight the importance of the maternal effects on seed and seedling production as well as responses of offspring to changing environmental drivers in mechanistic understanding and projecting of plant population dynamics under global change.     

Maternal and direct effects of elevated CO2 on seed provisioning, germination and seedling growth in Bromus erectus

Elevated CO₂ can affect plant fitness not only through its effects on seed production but also by altering the quality of seeds and therefore germination and seedling performance. We collected seeds from mother plants of Bromus erectus grown in field plots at ambient and elevated CO₂ (m-CO₂, maternal CO₂) and germinated them in the greenhouse in a reciprocal design under ambient and elevated CO₂ (o-CO₂, offspring CO₂). This design allowed us to examine both the direct effects of elevated CO₂ on germination and seedling growth and the indirect (maternal) effects via altered seed quality. Elevated m-CO₂ significantly increased seed mass and increased the C:N ratio of seeds from field-grown plants. Percentage and rate of germination were not affected by the m-CO₂ or o-CO₂ treatments. Similarly, elevated m-CO₂ had no significant effect on seedling size as estimated by the total leaf length. When differences in seed mass were adjusted by using seed mass as a covariate in ANOVA, a negative effect of m-CO₂ on seedling size appeared which increased with increasing seed mass (significant covariate×m-CO₂ interaction). This may indicate that the advantage of increased seed mass at elevated m-CO₂ was offset by the reduced concentration of nitrogen (and possibly other nutrients) in these seeds. In contrast to m-CO₂, elevated o-CO₂ greatly increased seedling size, and this stimulatory effect of elevated o-CO₂ was found to increase with increasing seed mass (significant covariate×o-CO₂ interaction). Taken together, these results suggest that in B. erectus transgenerational effects of elevated CO₂ are relatively small. However, other factors (genetic and environmental) that contribute to variation in seed provisioning can critically influence the responsiveness of seedlings to elevated CO₂. Received: 10 May 1999 / Accepted: 6 January 2000     

Quantitative genetics of seed size variation inPhlox

Summary Because seed size is often associated with survival and reproduction in plant populations, genetic variation for seed size may be reduced or eliminated by natural selection. To test this hypothesis we assessed genetic sources of variation in seed size in a population ofPhlox drummondii to determine whether genetic differences among seeds influence the size they attain. A diallel cross among 12 plants from a population at Bastrop, Texas, USA allowed us to partition variance in the mass of seeds among several genetic and parental effects. We found no evidence of additive genetic variance or dominance genetic variance for seed mass in the contribution of plants to their offspring. Extranuclear maternal effects accounted for 56% of the variance in seed mass. A small interaction was observed between seed genotype and maternal plant. Results of this study support theory that predicts little genetic variation for traits associated with fitness.     

Investigating the genetic load of an emblematic invasive species: the case of the invasive harlequin ladybird Harmonia axyridis

Introduction events can lead to admixture between genetically differentiated populations and bottlenecks in population size. These processes can alter the adaptive potential of invasive species by shaping genetic variation, but more importantly, they can also directly affect mean population fitness either increasing it or decreasing it. Which outcome is observed depends on the structure of the genetic load of the species. The ladybird Harmonia axyridis is a good example of invasive species where introduced populations have gone through admixture and bottleneck events. We used laboratory experiments to manipulate the relatedness among H. axyridis parental individuals to assess the possibility for heterosis or outbreeding depression in F₁ generation offspring for two traits related to fitness (lifetime performance and generation time). We found that inter‐populations crosses had no major impact on the lifetime performance of the offspring produced by individuals from either native or invasive populations. Significant outbreeding depression was observed only for crosses between native populations for generation time. The absence of observed heterosis is indicative of a low occurrence of fixed deleterious mutations within both the native and invasive populations of H. axyridis. The observed deterioration of fitness in native inter‐population crosses most likely results from genetic incompatibilities between native genomic backgrounds. We discuss the implications of these results for the structure of genetic load in H. axyridis in the light of the available information regarding the introduction history of this species.     

Genetic Diversity and the Reintroduction of Meadow Species

Abstract: Restoration of formerly nutrient‐poor and species‐rich grasslands generally leads to an increase in species diversity. However, species without a persistent seed bank and with poor dispersal ability often do not re‐establish spontaneously. Here, reintroduction is an option. If existing populations are comparable in their genetic composition, any population will do. This is not the case if populations have local adaptations. Unfortunately, whether populations are adapted locally is not easily determined, in contrast to assessing differentiation using neutral genetic markers. We used AFLP to study genetic diversity of Cirsium dissectum and Succisa pratensis within and among several Junco‐Molinion plant communities in the Netherlands (up to 200 km apart) that were potential source populations, and followed the reintroduction using seeds from these populations. Also, vegetative growth phase characteristics of three populations of C. dissectum were analyzed under controlled conditions. Most of the genetic variation in these cross‐fertilizing species was found within populations. Small but significant genetic differences in band frequencies were found among populations (F_st 0.100 ‐ 0.135). The first generation of reintroduced plants contained less polymorphic bands than the source populations. The genetic differences caused by reintroduction using a limited number of seeds (founder effects) were significant in all except one case (F_st 0.012 ‐ 0.101 between source and corresponding reintroduced population), but the magnitude was smaller than the source population differentiation. In assignment tests, reintroduced populations resembled their source population more than any other population, but all populations contained sizeable proportions of plants that were assigned to most similar plants from other populations, indicating that the populations are only marginally distinct. Calculations show that reintroduction from more than one source population introduces significantly more polymorphic bands into the new population, capitalizing on the existence of band frequency differences among populations.