Agnès Mignot

Gene flow and local adaptation in two endemic plant species

Abstract In order to detect the evolutionary potential of two endangered species, Brassica insularis (Brassicaceae) and Centaurea corymbosa (Asteraceae), within and among-population genetic variation for both quantitative traits and allozymic markers was examined. Four populations of each species were studied, representing a large proportion of extant populations. High values of θST (0.213 and 0.364 for B. insularis and C. corymbosa respectively) suggested that low amounts of gene flow occur among the study populations. In each species, the genetic distance based on allozymes (estimated by the ratio (θST/1−θST)) was positively correlated with the geographical distance, indicating isolation by distance. In contrast to previous studies in either outcrossing or selfing plant species, and especially for B. insularis, population differentiation for quantitative traits (QST) was generally found lower than differentiation for allozymes (θST), suggesting that the populations studied were experiencing similar selective forces acting upon the quantitative traits measured. Such forces would be strong enough to counteract local genetic drift. Interestingly, for both species QSTs were statistically independent of geographical distance, in contrast to the marginally significant positive isolation by distance shown by θST. Altogether, these results suggest that θSTs might not always be used as conservative estimates of QSTs, and might instead overestimate the evolutionary potential of endangered species. This would be especially expected in narrow-endemic species, whose ecological niche is often so restricted that indeed homogeneous selective forces are likely to occur, whereas small population sizes and restricted dispersal are likely to produce strong differentiation for neutral variation. In fact, knowledge of both neutral and quantitative diversity patterns allows identification of those traits undergoing natural selection, and could be useful in designing reinforcement or reintroduction programs. However, this approach might have limitations too, in the presence of outbreeding depression due to locally coevolved gene complexes.

SPATIAL AND TEMPORAL DEMOGRAPHIC VARIABILITY IN THE ENDEMIC PLANT SPECIES CENTAUREA CORYMBOSA (ASTERACEAE)

Centaurea corymbosa is an endemic plant species restricted to a 3-km2 area in southern France. This species is known from only six small populations that are highly differentiated genetically. Matrix models based on eight years of data (1994–2001) were used to assess the pattern of variation in the demographic vital rates of this species, and to investigate the causes of their variation. Asymptotic growth rates λ varied widely between years and populations (0.613–1.424). Randomization tests were developed to test for spatial and temporal variation in the asymptotic growth rates. These tests rely on individual data on both survival and fecundity. As our demographic survey only allowed us to estimate average fecundities, additional fecundity data collected from 1994 to 1996 were used to assess the distribution of individual fecundity expected within populations under demographic stochasticity or sampling error. Randomization tests showed that asymptotic growth rates were significantly different between popu...

Fine‐scale genetic structure and gene dispersal in Centaurea corymbosa (Asteraceae) I. Pattern of pollen dispersal

Pollen dispersal was characterized within a population of the narrowly endemic perennial herb, Centaurea corymbosa, using exclusion‐based and likelihood‐based paternity analyses carried out on microsatellite data. Data were used to fit a model of pollen dispersal and to estimate the rates of pollen flow and mutation/genotyping error, by developing a new method. Selfing was rare (1.6%). Pollen dispersed isotropically around each flowering plant following a leptokurtic distribution, with 50% of mating pairs separated by less than 11 m, but 22% by more than 40 m. Estimates of pollen flow lacked precision (0–25%), partially because mutations and/or genotyping errors (0.03–1%) could also explain the occurrence of offspring without a compatible candidate father. However, the pollen pool that fertilized these offspring was little differentiated from the adults of the population whereas strongly differentiated from the other populations, suggesting that pollen flow rate among populations was low. Our results suggest that pollen dispersal is too extended to allow differentiation by local adaptation within a population. However, among populations, gene flow might be low enough for such processes to occur.

Consequences of Low Mate Availability in the Rare Self-Incompatible Species Brassica insularis

Self-incompatibility systems prevent self-fertilization in angiosperms. Although numerous S alleles are usually maintained by negative frequency-dependent selection, the number of S alleles can be low in small populations, which limits mate availability and reduces fecundity in endangered populations of self-incompatible plants. Despite the increasing evidence of the negative effect of self-incompatibility in small populations, the direct link between the number and the distribution of S alleles and their reproductive consequences has been rarely reported. Brassica insularis is a rare self-incompatible species with medium to very small populations. Results of a previous study showed that the smallest population has very few S alleles. We investigated whether reduced mate availability affects reproduction in this species. We compared the pollination success and the fruit set in 4 populations differing in population size and number of S alleles. Our results suggest that reproduction may be negatively affected by the low S-allele diversity in the smallest population. Nevertheless, other populations also had reduced fruit set that could not be attributed to self-incompatibility alone.

Pollen aperture heteromorphism. Variation in pollen-type proportions along altitudinal transects in Viola calcarata.

Some species produce pollen grains with different aperture numbers within a single individual (pollen aperture heteromorphism). In the pansy Viola diversifolia, aperture number is positively correlated with pollen germination speed, and negatively correlated with viability. In V. calcarata, young five-aperturate pollen grains germinate faster than four-aperturate ones. Heteromorphism could thus be favoured when pollination is unpredictable, as plants produce both very competitive and long-lived pollen grains. Depending on the efficiency of the pollinators, different proportions of pollen types will be optimal. In insect-pollinated species, such as V. calcarata, pollination efficiency generally decreases as elevation increases. We therefore expect a decrease in mean aperture number as altitude increases. This was found in four transects (out of six). Pollinator activity therefore has a potential impact on pollen morphology.Some species produce pollen grains with different aperture numbers within a single individual (pollen aperture heteromorphism). In the pansy Viola diversifolia, aperture number is positively correlated with pollen germination speed, and negatively correlated with viability. In V. calcarata, young five-aperturate pollen grains germinate faster than four-aperturate ones. Heteromorphism could thus be favoured when pollination is unpredictable, as plants produce both very competitive and long-lived pollen grains. Depending on the efficiency of the pollinators, different proportions of pollen types will be optimal. In insect-pollinated species, such as V. calcarata, pollination efficiency generally decreases as elevation increases. We therefore expect a decrease in mean aperture number as altitude increases. This was found in four transects (out of six). Pollinator activity therefore has a potential impact on pollen morphology.

Variation in dispersal traits in a narrow-endemic plant species, Centaurea corymbosa Pourret. (Asteraceae)

The existence of genetic variability for dispersal is a crucial issue for organisms facing increased habitat fragmentation and climate change. We study the genetic basis and evolutionary potential for diaspore traits related to dispersal in Centaurea corymbosa. Using diaspores collected in natural conditions in four of the six extant populations of this narrow-endemic plant species and diaspores produced in a common garden experiment, we study the variation for pappus and achene sizes, and diaspore mass. Using a sample of achenes from the common garden experiment, we find that the best predictor of terminal velocity is a linear combination of pappus length, achene width, and achene weight. We find significant differences among populations for all traits in both conditions, as well as significant differences among families within population. Although the differences among populations for some traits are not exactly the same in controlled conditions compared to natural conditions, the ranking of populations according to their mean trait values is consistent in both conditions. Our study is therefore one of the first to show a correlation between phenotypic differentiation for dispersal traits in natural conditions vs. controlled conditions. We also show evidence of genetic variation for traits commonly thought to be involved in dispersal ability, suggesting the potential for evolutionary changes following environmental change and management actions.

Marker-based investigation of inbreeding depression in the endangered species Brassica insularis

Various methods have been proposed to estimate inbreeding depression and to assess its consequences for natural populations. As an alternative to controlled crosses, the use of molecular markers has allowed direct investigation of inbreeding depression in natural populations, but usually suffers from low statistical power. Here, we investigated the effect of inbreeding depression on survival in two populations of the rare species Brassica insularis, using both controlled crosses and a marker-based approach. We compare the respective merits of the two approaches for studying inbreeding depression. We also use information from the molecular markers to dissect in detail patterns of inbreeding depression in this species. A posteriori, we find that combining the approaches was not necessary to obtain simple point estimates of inbreeding depression. However, using molecular markers may give insight into the genetic basis of inbreeding depression, such as the occurrence of epistatic interactions among deleterious alleles or purging.

Interaction of climate, demography and genetics: a ten-year study of Brassica insularis, a narrow endemic Mediterranean species

Long-term demographic surveys, needed to obtain accurate information on population dynamics and efficiently manage rare species, are still very scarce. Matrix population models are useful tools to identify key demographic transitions and thus help setting up conservation actions. Furthermore, the combination of ecological, demographic and genetic data is likely to improve the identification of the threats acting upon populations and help conservation decisions. In this paper we illustrate the power of this approach on Brassica insularis, a Mediterranean endemic plant species, rare and endangered in Corsica (France). In four populations of this species, a long-term demographic survey (2000–2009), genetic analyses (in 2000 and 2009) and survey of ecological variables (climatic variables, competition and herbivory) were performed. By using both deterministic and stochastic matrix model analyses, we assessed the viability of each population and tested for both spatial and temporal variations in demographic vital rates. Populations exhibited differing demographic behaviours and environmental stochasticity occurred in populations. Significant correlations between climatic variables and vital rates were detected. Stochastic simulations suggested that three out of the four populations studied might present a high risk of extinction on the short-term and should actively be managed, or at least surveyed. It could be, however, that two of these populations are experiencing density-dependent regulation, rather than being declining. Microsatellite diversity was slightly reduced in a single population and similar in the three others, consistently with expectations based on population census size and geographic area, as well as with diversity at the S-locus observed in 2000. The combination of all data led to specific recommendations for managing each population. We discuss the implications for conservation of such a general approach.

Lower selfing rates in metallicolous populations than in non-metallicolous populations of the pseudometallophyte Noccaea caerulescens (Brassicaceae) in Southern France

Background and Aims The pseudometallophyte Noccaea caerulescens is an excellent model to study evolutionary processes, as it grows both on normal and on heavy-metal-rich, toxic soils. The evolution and demography of populations are critically impacted by mating system and, yet, information about the N. caerulescens mating system is limited. Methods Mean selfing rates were assessed using microsatellite loci and a robust estimation method (RMES) in five metallicolous and five non-metallicolous populations of N. caerulescens in Southern France, and this measure was replicated for two successive reproductive seasons. As a part of the study, the patterns of gene flow among populations were analysed. The mating system was then characterized at a fine spatial scale in three populations using the MLTR method on progeny arrays. Key Results The results confirm that N. caerulescens has a mixed mating system, with selfing rates ranging from 0·2 to 0·5. Selfing rates did not vary much among populations within ecotypes, but were lower in the metallicolous than in the non-metallicolous ecotype, in both seasons. Effective population size was also lower in non-metallicolous populations. Biparental inbreeding was null to moderate. Differentiation among populations was generally high, but neither ecotype nor isolation by distance explained it. Conclusions The consequences of higher selfing rates on adaptation are expected to be weak to moderate in non-metallicolous populations and they are expected to suffer less from inbreeding depression, compared to metallicolous populations.

Why evolution matters for species conservation: perspectives from three case studies of plant metapopulations

We advocate the advantage of an evolutionary approach to conservation biology that considers evolutionary history at various levels of biological organization. We review work on three separate plant taxa, spanning from one to multiple decades, illustrating extremes in metapopulation functioning. We show how the rare endemics Centaurea corymbosa (Clape Massif, France) and Brassica insularis in Corsica (France) may be caught in an evolutionary trap: disruption of metapopulation functioning due to lack of colonization of new sites may have counterselected traits such as dispersal ability or self‐compatibility, making these species particularly vulnerable to any disturbance. The third case study concerns the evolution of life history strategies in the highly diverse genus Leucadendron of the South African fynbos. There, fire disturbance and the recolonization phase after fires are so integral to the functioning of populations that recruitment of new individuals is conditioned by fire. We show how past adaptation to different fire regimes and climatic constraints make species with different life history syndromes more or less vulnerable to global changes. These different case studies suggest that management strategies should promote evolutionary potential and evolutionary processes to better protect extant biodiversity and biodiversification.