Effects of inbreeding and pollen donor provenance and diversity on offspring performance under environmental stress in the rare plant Cochlearia bavarica

Habitat degradation and loss can reduce size and genetic variability of natural populations, increasing individual homozygosity and average relatedness between individuals. While the resulting inbreeding depression may be reduced by natural selection under prevailing environmental conditions, it may increase again under environmental stress. To investigate the effect of environmental stress on offspring performance and the expression of inbreeding depression, we hand-pollinated maternal plants in small (< 100, n=5) and large populations (> 400 flowering plants, n=5) of the rare plant Cochlearia bavarica (Brassicaceae) and raised the offspring under experimentally manipulated water and light regimes (normal or reduced supply). In addition to considering natural variation in inbreeding levels due to population size, we manipulated pollen donor provenance and diversity. Maternal plants were pollinated with nine donors from a different population or with one or nine donors from the same population. One further inflorescence of each maternal plant was exposed to free pollination. Offspring growth and survival were monitored over 300 days. Offspring performance varied significantly among populations and maternal plants. Environmental stress interacted significantly with these factors. However, there was no general indication that offspring from small populations were more negatively affected. In seven out of 10 populations, offspring derived from between-population pollination performed better than offspring derived from within-population pollination. Also, in five out of 10 populations, average offspring size was higher after within-population pollination with nine than after pollination with one pollen donor. These results suggest low genetic diversity within C. bavarica populations, both smaller and larger ones. Interactions between environmental stress and pollination treatment indicated that using pollen donors from outside a population or increasing the number of pollen donors can reduce inbreeding depression, but that this beneficial effect is impaired under stressful conditions.     

Inbreeding depression and mating-distance dependent offspring fitness in large and small populations of Lychnis flos-cuculi (Caryophyllaceae)

The spatial structure of four Lychnis flos-cuculi populations, varying in size and degree of isolation, was studied by comparing the fitness of offspring resulting from self-pollination and pollinations by neighbouring plants, plants within the same population, and plants from other populations. Selfed offspring had the lowest fitness of the four offspring groups. No significant difference was found between the performance of offspring from pollinations by neighbouring plants and offspring pollinated by plants further apart but within the same population. A lower fitness of offspring from pollinations between neighbours would be expected if these matings, on average, yielded inbred offspring which suffered from inbreeding depression. These results imply that either a tight neighbourhood structuring is not present, or that the inbreeding depression for offspring by neighbours is too low to detect, although these are inbred. Crossings between populations produced offspring with a significantly higher fitness than offspring sired within populations. There were no significant differences in response to inbreeding among the populations, and differences in mean fitness among populations had no clear relation to the population size or degree of isolation. A reduced fitness of small populations due to inbreeding depression or a less severe response to experimental inbreeding due to purging of deleterious alleles is therefore not supported by our results.     

The inbreeding depression cost of selfing: Importance of flower size and population size in Collinsia parviflora (Veronicaceae)

Inbreeding depression should evolve with selfing rate when frequent inbreeding results in exposure of and selection against deleterious alleles. The selfing rate may be modified by plant traits such as flower size, or by population characteristics such as census size that can affect the probability of biparental inbreeding. Here we quantify inbreeding depression (δ) among different population sizes of Collinsia parviflora, a wildflower with interpopulation variation in flower size, by comparing fitness components and multiplicative fitness of experimentally produced selfed and outcrossed offspring. Selfed offspring had reduced multiplicative fitness compared to outcrossed offspring, but inbreeding depression was low in all combinations of population size and flower size (δ ≤ 0.05) except in large populations of large-flowered plants (δ = 0.45). The decrement to multiplicative fitness with inbreeding was not affected by population size nested within flower size, but differed between small- and large-flowered plants: small-flowered populations had lower overall inbreeding depression (δ = 0.04) compared to large-flowered populations (δ = 0.25). The difference in load with flower size suggests that either selection has removed deleterious recessive alleles or these alleles have become fixed in small-flowered, potentially more selfing populations, but that purging has not occurred to the same extent in presumably outcrossing large-flowered populations.     

A threefold genetic allee effect: population size affects cross-compatibility, inbreeding depression and drift load in the self-incompatible Ranunculus reptans

A decline in population size can lead to the loss of allelic variation, increased inbreeding, and the accumulation of genetic load through drift. We estimated the fitness consequences of these processes in offspring of controlled within-population crosses from 13 populations of the self-incompatible, clonal plant Ranunculus reptans. We used allozyme allelic richness as a proxy for long-term population size, which was positively correlated with current population size. Crosses between plants of smaller populations were less likely to be compatible. Inbreeding load, assessed as the slope of the relationship between offspring performance and parental kinship coefficients, was not related to population size, suggesting that deleterious mutations had not been purged from small populations. Offspring from smaller populations were on average more inbred, so inbreeding depression in clonal fitness was higher in small populations. We estimated variation in drift load from the mean fitness of outbred offspring and found enhanced drift load affecting female fertility within small populations. We conclude that self-incompatibility systems do not necessarily prevent small populations from suffering from inbreeding depression and drift load and may exacerbate the challenge of finding suitable mates.     

Population size,pollination and phenotypic trait selection in Phyteuma spicatum

Plants in small populations may receive fewer visits, smaller pollen loads or pollen of poorer quality and suffer from reduced reproductive success compared to plants in larger populations. Consequently, pollen limitation of plants in small populations has been suggested to result in the evolution of reduced reliance on pollinators or the enhancement of traits that attract pollinators. The main aim of this study was to experimentally quantify the strength of pollinator-mediated selection on floral display size and flowering phenology in populations of varying size, using the self-incompatible, perennial herb Phyteuma spicatum as study species. We conducted supplementary hand pollinations in six populations (ranging in size between ca. 20–3000 flowering individuals) over two consecutive years and assessed selection gradients (i.e., trait–fitness relationships) in open- and hand-pollinated plants. Our results show that some populations are pollen limited in some years, but, contrary to our expectation, the degree of pollen limitation was not significantly related to population size. We found phenotypic selection for increased inflorescence size (in most populations and in both years), but we obtained no or no strong evidence that selection was pollinator-mediated or that the strength of selection was related to population size. This may have been the result of low statistical power, an inherent problem of studies examining effects of population size that require the inclusion of populations with only few individuals. In addition, given that selection appeared to be spatially and temporally variable, abiotic or biotic factors other than pollinators may have contributed to selection on inflorescence size.     

Genetic rescue in interconnected populations of small and large size of the self-incompatible Ranunculus reptans

Small populations of our study species Ranunculus reptans have reduced fitness because of inbreeding, genetic load, and reduced mate availability; that is, they suffer from a three-fold genetic Allee effect. Here, we investigate how the effect of interpopulation outbreeding on offspring fitness depends on population size. We performed within- and between-population crosses with plants originating from 15 populations, and measured offspring performance in a common environment. Interpopulation outbreeding led to an increase in population means of clonal performance, which was defined as the number of rooted offspring rosettes produced per maternal ovule. This fitness gain mainly occurred at the life stage of seed set. It was especially pronounced for populations with a long-term history of small size inferred from their low genetic diversity, estimated from eight allozyme loci. We conclude that in a self-incompatible plant such as R. reptans, interpopulation outbreeding can lead to an immediate genetic rescue effect due to increased cross-compatibility and heterosis, and that this rescue effect is increased as population size decreases.     

Effects of population size on reproductive success of the endangered and endemic species Primula merrilliana

The reproductive ability related to the population size of the endangered and endemic species Primula merrilliana Schltr.was investigated.In 26 populations observed,only four contain more than 500 flowering individuals,whereas most of them(53.8%) consist of less than 100 flowering individuals.Though the ratio of pin and thrum plants keeps its balance at 1:1 for all populations,the frequency of pin and thrum flowers was significantly biased in most small populations.Population size strongly affected reproductive success; plants in small populations produced significantly fewer flowers and fruits per plant and fewer seeds per fruit,and therefore fewer seeds per plant.The floral density was another important factor that influenced the reproductive success of P.merrilliana,because four main reproductive success parameters (fruits per plant,seeds per fruit,seeds per plant,and the proportion of flowers setting fruit) were all positively correlated with floral density.The size of plants and the number of leaves per plant (measure of habitat quality) were not influenced by the variation of population size,suggesting that the reduced fecundity in small populations may not be a consequence of lower habitat quality,inbreeding depression and pollen limitation as a result of less attractiveness in small populations are therefore likely explanations for these patterns.     

Maternal sex and mate relatedness affect offspring quality in the gynodioecious Silene acaulis

In gynodioecious species, females sacrifice fitness by not producing pollen, and hence must have a fitness advantage over hermaphrodites. Because females are obligately outcrossed, they may derive a fitness advantage by avoiding selfing and inbreeding depression. However, both sexes are capable of biparental inbreeding, and there are currently few estimates of the independent effects of maternal sex and multiple levels of inbreeding on female advantage. To test these hypotheses, females and hermaphrodites from six Alaskan populations of Silene acaulis were crossed with pollen from self (hermaphrodites only), a sibling, a random plant within the same population, and a plant from a different population. Germination, survivorship and early growth revealed inbreeding depression for selfs and higher germination but reduced growth in sib-crosses, relative to outcrosses. Independent of mate relatedness, females germinated more seeds that grew faster than offspring from hermaphrodites. This indicates that inbreeding depression as well as maternal sex can influence breeding system evolution. The effect of maternal sex may be explained by higher performance of female genotypes and a greater abundance of female genotypes among the offspring of female mothers.     

Effects of pollen quantity and quality on reproduction and offspring vigor in the rare plant Scorzonera humilis (Asteraceae)

We studied the effects of pollinator exclusion, interparental distance, and supplementary hand pollination on reproduction and progeny vigor in Scorzonera humilis (Asteraceae), a rare plant of fragmented, nutrient-poor grasslands. Caged flowers produced no seeds and selfed flowers only very rarely, indicating that S. humilis is mainly self-incompatible. Seed production, seed mass, and seed germination following between-population crosses were consistently, but not significantly, higher than after within-population crosses. Seed set increased with local density of conspecifics, indicating that the reduced plant density in fragmented populations may reduce plant reproductive success. Seed set was pollen limited in all four populations studied. Supplementary hand-pollination strongly increased the survival of offspring, indicating that either pollinators transferred pollen from related individuals resulting in inbreeding depression in spite of the incompatibility system or that higher pollen loads increased pollen competition and the selectivity among gametes. In one of the populations, adding pollen from a different population strongly increased progeny fitness compared with both natural pollination and pollen supplementation from the same population. The results indicate that S. humilis is sensitive to inbreeding and that pollen limitation can reduce both the number and quality of offspring.     

Effects of population size and isolation on reproductive output in Aquilegia canadensis (Ranunculaceae)

It is generally expected that small, isolated populations will suffer reduced fitness due to inbreeding, yet few studies have investigated the relation between population characteristics, inbreeding and fitness. Among Ontario populations of the short‐lived, perennial plant Aquilegia canadensis, large populations (N>90 flowering plants) outcross twice as frequently as small populations (N=30–40), and inbreeding depression is extremely strong. We tested the prediction that reproductive output, a major component of population fitness, should be positively associated with population size. Data from a survey of 33 populations located on small islands in the St. Lawrence River, Canada and 23 populations on adjacent mainland areas supported this prediction. Population size correlated positively with reproductive output, measured as the number of seedlings produced per plant in 1995 (average r=+0.39 pooled P=0.019), and the number of fruits per plant in 1997 (r=+0.30, P=0.056). We also tested the prediction that fitness should decline with increasing spatial isolation between populations by measuring the distance separating all island populations. However, reproductive output did not correlate with isolation in either year. We compared island and mainland populations to test the prediction that reproductive output should be lower for populations on small islands than those occurring in more continuous mainland habitat. In contrast to our predictions, island populations exhibited, if anything, higher reproductive output than mainland populations. We also found no support for the prediction that the positive association between population size and reproductive output should be stronger for presumably isolated populations on small islands than for those on adjacent mainland areas. While the mechanisms underlying the association between population size and fitness are impossible to identify with correlations alone, our results are consistent with the hypothesis that inbreeding can significantly reduce the fitness of natural populations.     

濒危植物居群恢复的遗传学考量

本文针对濒危植物居群的遗传多样性、生殖适合度、基因流、近交和远交衰退等遗传学问题在居群恢复过程中的应用进行了探讨。濒危植物居群的回归重建,既面临遗传多样性的迅速丧失、近交衰退等遗传风险,还因回归引种地存在较多近缘种而带来远交衰退的风险,最终导致遗传适应性降低,生境适应性变窄,繁殖和竞争能力减弱。为提高濒危物种保护的质量和效率,在构建回归居群时,应分批次从同一来源居群的不同母株采集材料,确保种源的遗传纯正性和遗传组成的多样性,还应使回归居群尽可能远离近缘广布种。另外,还需要对回归种群进行持续的监测和管理才能保证回归引种的成功。     

The relationship between individual seed quality and maternal plant body size in crowded herbaceous vegetation

Aims In most natural plant populations, there is a strong right-skewed distribution of body sizes for reproductive plants—i.e. the vast majority are relatively small, suppressed weaklings that manage not just to survive effects of crowding/competition and other hazards but also to produce offspring. Recent research has shown that because of their relatively large numbers, these relatively small resident plants collectively contribute most of the seed offspring production available for the population in the next generation. However, the success of these offspring will depend in part on their quality, e.g. reflected by seed size and resource content. Accordingly, in the present study, we used material from natural populations of herbaceous species to test the null hypothesis that there is no significant relationship between body size variation in resident plants—resulting from between-site variation in the intensity of crowding/competition—and variation in the mass or N content of their individual seeds.Methods Using populations of 56 herbaceous species common in eastern Ontario, total above-ground dry plant mass, mean mass per seed and mean nitrogen (N) content per seed were recorded for a sample of the largest resident plants and also for the smallest reproductive plants growing in local neighbourhoods with the most severe crowding/competition from near neighbours.Important findings Mass per seed was numerically smaller from the smallest resident plants for most study species, but with few exceptions, this was not significantly different (P> 0.05) from mass per seed from the largest resident plants. The results therefore showed no general effect of maternal plant body size on individual seed mass, or N content. This suggests that the reproductive output of the smaller half of the resident plant size distribution within these populations is likely to contribute not just most of the seed production available for the next generation but also seed offspring that are just as likely—on a per individual basis—to achieve seedling/juvenile recruitment success as the seed offspring produced by the largest resident plants. This conflicts with the traditional 'size-advantage' hypothesis for predicting plant fitness under severe competition, and instead supports the recent 'reproductive-economy-advantage' hypothesis, where competitive fitness is promoted by capacity to produce offspring that—despite severe body size suppression imposed by neighbour effects—in turn have capacity to produce grand-offspring.     

Effect of cross proximity on progeny fitness in a rare and a common species of Eupatorium (Asteraceae)

Offspring from matings between near neighbors may exhibit lower fitness relative to offspring from more distant matings due to spatial structuring of populations resulting from limited dispersal of pollen and seed. This response, which can be interpreted as inbreeding depression, is studied in the rare species, Eupatorium resinosum, and a closely related congener, E. perfoliatum, through the use of hand pollinations representing three distance classes (near-within a population, far-within a population, and between populations) and an assay of the offspring in an experimental plot. Early traits such as seed mass and first-year stem length were not significantly affected by the cross type, although they were affected by maternal parentage. Size and reproduction in the second field season increased with increased pollen donor distance from the maternal plant. Cross type was significant for many traits in the second field season, indicating inbreeding depression in crosses of neighbors (for E. resinosum) and hybrid vigor between populations (both species). This suggests that the rare species, E. resinosum, had a more spatially structured population than E. perfoliatum. The implications of these results for conservation of rare species indicate that protection of habitats sufficient for large populations is necessary to maintain genetic diversity since each population likely consists of many smaller subpopulations.     

Pollen limitation and inbreeding depression in an 'old rare' bumblebee-pollinated grassland herb

Habitat fragmentation and reduction of population size have been found to negatively affect plant reproduction in 'new rare' species that were formerly common. This has been attributed primarily to effects of increased inbreeding but also to pollen limitation. In contrast, little is known about the reproduction of 'old rare' species that are naturally restricted to small and isolated habitats and thus may have developed strategies to cope with long-term isolation and small population size. Here we study the effects of pollen source and quantity on reproduction of the 'old rare' bumblebee-pollinated herb, Astragalus exscapus. In two populations of this species, we tested for pollen autodeposition, inbreeding depression and outbreeding depression. Caged plants were left unpollinated or were pollinated with pollen from the same plant, from the same population or from a distant population (50 km). Additionally, we tested for pollen limitation by pollen supplementation in four populations of different size and density. In the absence of pollinators, plants did not produce seed whereas self-pollinated plants did. This indicates a self-compatible breeding system dependent on insect pollination. Both self-pollination and, in one of the two populations, cross-pollination with pollen from plants from the distant population resulted in a lower number of seeds per flower than cross-pollination with pollen from plants from the resident population, indicating inbreeding and outbreeding depression. Pollen addition enhanced fruit set and number of seeds per flower in three of the four populations, indicating pollen limitation. The degree of pollen limitation was lowest in the smallest but densest population. Our results suggest that, similar to 'new rare' plant species, also 'old rare' species may be at risk of inbreeding depression and pollen limitation.     

Population size and identity influence the reaction norm of the rare,endemic plant Cochlearia bavarica across a gradient of environmental stress

Habitat degradation and loss can result in population decline and genetic erosion, limiting the ability of organisms to cope with environmental change, whether this is through evolutionary genetic response (requiring genetic variation) or through phenotypic plasticity (i.e., the ability of a given genotype to express a variable phenotype across environments). Here we address the question whether plants from small populations are less plastic or more susceptible to environmental stress than plants from large populations. We collected seed families from small (<100) versus large natural populations (>1,000 flowering plants) of the rare, endemic plant Cochlearia bavarica (Brassicaceae). We exposed the seedlings to a range of environments, created by manipulating water supply and light intensity in a 2 x 2 factorial design in the greenhouse. We monitored plant growth and survival for 300 days. Significant effects of offspring environment on offspring characters demonstrated that there is phenotypic plasticity in the responses to environmental stress in this species. Significant effects of population size group, but mainly of population identity within the population size groups, and of maternal plant identity within populations indicated variation due to genetic (plus potentially maternal) variation for offspring traits. The environment x maternal plant identity interaction was rarely significant, providing little evidence for genetically- (plus potentially maternally-) based variation in plasticity within populations. However, significant environment x population-size-group and environment x population-identity interactions suggested that populations differed in the amount of plasticity, the mean amount being smaller in small populations than in large populations. Whereas on day 210 the differences between small and large populations were largest in the environment in which plants grew biggest (i.e., under benign conditions), on day 270 the difference was largest in stressful environments. These results show that population size and population identity can affect growth and survival differently across environmental stress gradients. Moreover, these effects can themselves be modified by time-dependent variation in the interaction between plants and their environment.     

Exploring the relationship between tychoparthenogenesis and inbreeding depression in the Desert Locust,Schistocerca gregaria

Tychoparthenogenesis, a form of asexual reproduction in which a small proportion of unfertilized eggs can hatch spontaneously, could be an intermediate evolutionary link in the transition from sexual to parthenogenetic reproduction. The lower fitness of tychoparthenogenetic offspring could be due to either developmental constraints or to inbreeding depression in more homozygous individuals. We tested the hypothesis that in populations where inbreeding depression has been purged, tychoparthenogenesis may be less costly. To assess this hypothesis, we compared the impact of inbreeding and parthenogenetic treatments on eight life‐history traits (five measuring inbreeding depression and three measuring inbreeding avoidance) in four laboratory populations of the desert locust, Schistocerca gregaria, with contrasted demographic histories. Overall, we found no clear relationship between the population history (illustrated by the levels of genetic diversity or inbreeding) and inbreeding depression, or between inbreeding depression and parthenogenetic capacity. First, there was a general lack of inbreeding depression in every population, except in two populations for two traits. This pattern could not be explained by the purging of inbreeding load in the studied populations. Second, we observed large differences between populations in their capacity to reproduce through tychoparthenogenesis. Only the oldest laboratory population successfully produced parthenogenetic offspring. However, the level of inbreeding depression did not explain the differences in parthenogenetic success between all studied populations. Differences in development constraints may arise driven by random and selective processes between populations.     

Effects of population size on performance and inbreeding depression in <Emphasis Type="Italic">Lupinus perennis</Emphasis>

We investigated the relationships among population size, offspring performance, and inbreeding depression (δ) in Lupinus perennis by examining the effect of population size category (large vs. small) on seed production and offspring performance for three pollination treatments (open pollination, hand crossing and hand selfing). In each of our four pairs of populations, one member of the pair was substantially larger than the other. We then grew seeds from this factorial design (2 sizes × 4 pairs × 3 pollination treatments) in the greenhouse to investigate whether population size affects offspring performance in a common environment, and how small size affects purging of the inbreeding load. Multiplicative performance across four early life-stage components (seed production, seedling emergence, seedling survival and seedling growth) of smaller populations was not significantly lower, although biomass of seedlings declined in smaller populations. Self-pollination reduced seed production, seedling emergence and seedling growth, reflecting substantial inbreeding depression (δ = 0.404 ± 0.043). Population size categories did not consistently differ in levels of inbreeding depression, suggesting that purging of genetic load in smaller populations has been limited, and that all populations still harbor inbreeding load. We also found a significant decrease in log performance with increases in the population inbreeding coefficient. These results indicate that even in seemingly large populations, lupines are susceptible to considerable fitness declines through both inbreeding load within populations, and drift load via genetic erosion and fixation of deleterious alleles between populations.     

Parental genetic distance and patterns in nonrandom mating and seed yield in predominately selfing Arabidopsis thaliana

In this study, we ask two questions: (1) Is reproductive success independent of parental genetic distance in predominately selfing plants? (2) In the absence of early inbreeding depression, is there substantial maternal and/or paternal variation in reproductive success in natural populations? Seed yield in single pollinations and proportion of seeds sired in mixed pollinations were studied in genetically defined accessions of the predominately selfing plant Arabidopsis thaliana by conducting two diallel crosses. The first diallel was a standard, single pollination design that we used to examine variance in seed yield. The second diallel was a mixed pollination design that utilized a standard pollen competitor to examine variance in proportion of seeds sired. We found no correlation between reproductive success and parental genetic distance, and self-pollen does not systematically differ in reproductive success compared to outcross pollen, suggesting that Arabidopsis populations do not experience embryo lethality due to early-acting inbreeding or outbreeding depression. We used these data to partition the contributions to total phenotypic variation from six sources, including maternal contributions, paternal contributions and parental interactions. For seed yield in single pollinations, maternal effects accounted for the most significant source of variance (16.6 %). For proportion of seeds sired in mixed pollinations, the most significant source of variance was paternal effects (17.9 %). Thus, we show that population-level genetic similarities, including selfing, do not correlate with reproductive success, yet there is still significant paternal variance under competition. This suggests two things. First, since these differences are unlikely due to early-acting inbreeding depression or differential pollen viability, this implicates natural variation in pollen germination and tube growth dynamics. Second, this strongly supports a model of fixation of pollen performance genes in populations, offering a focus for future genetic studies in differential reproductive success.     

Offspring fitness in relation to population size and genetic variation in the rare perennial plant species Gentiana pneumonanthe (Gentianaceae)

Seeds were sampled from 19 populations of the rare Gentiana pneumonanthe, ranging in size from 5 to more than 50,000 flowering plants. An analysis was made of variation in a number of life-history characters in relation to population size and offspring heterozygosity (based on seven polymorphic isozyme loci). Life-his-tory characters included seed weight, germination rate, proportion of seeds germinating, seedling mortality, seedling weight, adult weight, flower production per plant and proportion of plants flowering per family. Principal component analysis (PCA) reduced the dataset to three main fitness components. The first component was highly correlated with adult weight and flowering performance, the second with germination performance and the third component with seed and seedling weight and seedling mortality. The latter two components were considered as being maternally influenced, since these comprised life-history traits that were significantly correlated with seed weight. Multiple regression analysis showed that variation in the first fitness component was mainly associated with heterozygosity and not with population size, while the third fitness component was only correlated with population size and not with heterozygosity. The latter relationship appeared to be non-linear, which suggests a stronger loss of fitness in the smallest populations. The second (germination) component was neither correlated with population size nor with genetic variation. There was only a weak association between population size, heterozygosity and the population coefficients of variation for each life history character. Most correlation coefficients were negative, however, which suggests that there is more variation among progeny from smaller populations. We conclude that progeny from small populations of Gentiana pneumonanthe show reduced fitness and may be phenotypically more variable. One of the possible causes of the loss of fitness is a combination of unfavourable environmental circumstances for maternal plants in small populations and increased inbreeding. The higher phenotypic variation in small populations may also be a result of inbreeding, which can lead to deviation of individuals from the average phenotype through a loss of developmental stability.     

Allee effects within small populations of Aconitum napellus ssp. lusitanicum, a protected subspecies in northern France

Plants growing at low density can suffer from Allee effects as a result of pollen limitation. Previous studies of Allee effects have focused on the effects of variation among populations in size or density on reproduction. Here, the effects of plant distribution within populations on fitness components are explored in a rare plant, Aconitum napellus ssp. lusitanicum, and ecological and genetic mechanisms underlying these effects are identified. To detect pollen limitation, seed production was compared under natural versus hand-supplemented pollinations on inflorescences of different sizes in natural patches differing both in flower density and in isolation from other patches. Germination rate and juvenile survival of seeds produced in low- and high-density patches were also compared. Pollen-supplemented flowers always produced more seeds than open-pollinated flowers, especially among small plants and plants growing at low density. Offspring produced in low-density patches exhibited lower fitness that those produced in high-density patches. This could have been caused by post-fertilization mechanisms, including inbreeding depression or differential maternal resource allocation. These results show that Allee effects on fitness components (ecological and genetic Allee effects) occur within A. napellus populations at different spatial scales. The spatial distribution of plants seems to be a crucial factor affecting reproductive output and fitness.