No genetic diversity at molecular markers and strong phenotypic plasticity in populations of Ranunculus nodiflorus, an endangered plant species in France

BACKGROUND AND AIMS: Although conservation biology has long focused on population dynamics and genetics, phenotypic plasticity is likely to play a significant role in population viability. Here, an investigation is made into the relative contribution of genetic diversity and phenotypic plasticity to the phenotypic variation in natural populations of Ranunculus nodiflorus, a rare annual plant inhabiting temporary puddles in the Fontainebleau forest (Paris region, France) and exhibiting metapopulation dynamics. METHODS: The genetic diversity and phenotypic plasticity of quantitative traits (morphological and fitness components) were measured in five populations, using a combination of field measurements, common garden experiments and genotyping at microsatellite loci. KEY RESULTS: It is shown that populations exhibit almost undetectable genetic diversity at molecular markers, and that the variation in quantitative traits observed among populations is due to a high level of phenotypic plasticity. Despite the lack of genetic diversity, the natural population of R. nodiflorus exhibits large population sizes and does not appear threatened by extinction; this may be attributable to large phenotypic plasticity, enabling the production of numerous seeds under a wide range of environmental conditions. CONCLUSIONS: Efficient conservation of the populations can only be based on habitat management, to favour the maintenance of microenvironmental variation and the resulting strong phenotypic plasticity. In contrast, classical actions aiming to improve genetic diversity are useless in the present case.     

The endangered Tunbridge buttercup (Ranunculus prasinus): Ecology,conservation status and introduction to the Township Lagoon Nature Reserve,Tasmania

Abstract Ranunculus prasinus is a mat-forming species of the marginal herbfields and tussock grasslands of brackish wetlands. It is only known from four wetlands, all on private land, in the driest part of Tasmania. The species has been successfully introduced from divisions from three of the populations into a secure reserve with similar vegetation to that of the natural populations. This is considered to be desirable and morally sound in that it increases the chances of survival of native biological diversity as a whole. However, ex situ measures and a gaining of security for natural populations would also be prudent.     

Preferred habitat and effective population size drive landscape genetic patterns in an endangered species

Landscape genetics provides a framework for pinpointing environmental features that determine the important exchange of migrants among populations. These studies usually test the significance of environmental variables on gene flow, yet ignore one fundamental driver of genetic variation in small populations, effective population size, N_e. We combined both approaches in evaluating genetic connectivity of a threatened ungulate, woodland caribou. We used least-cost paths to calculate matrices of resistance distance for landscape variables (preferred habitat, anthropogenic features and predation risk) and population-pairwise harmonic means of N_e, and correlated them with genetic distances, F_ST and D_c. Results showed that spatial configuration of preferred habitat and N_e were the two best predictors of genetic relationships. Additionally, controlling for the effect of N_e increased the strength of correlations of environmental variables with genetic distance, highlighting the significant underlying effect of N_e in modulating genetic drift and perceived spatial connectivity. We therefore have provided empirical support to emphasize preventing increased habitat loss and promoting population growth to ensure metapopulation viability.     

Survival of Ranunculus repens L. (creeping buttercup) in an amphibious habitat

The turlough form of Ranunculus repens is subjected to several months' complete inundation with hard groundwater. Experimental flooding to the level of the soil surface had no effect on turlough or ruderal populations relative to drained controls. Experimental submergence resulted in direct tissue death of the ruderal population but did not affect the turlough population relative to drained controls. There was no detectable difference in the proportion of aerenchyma in drained, flooded and submerged roots of plants from either population. The proportion of aerenchyma increased with root age in the ruderal population. Up to twice the proportion of aerenchyma occurred in the lower third of the root in the turlough population relative to the middle and upper thirds. Submergence in artificially hardened tap water increased the amount of tissue death in the ruderal population, whereas it appeared to enhance the growth of plants from the turlough population relative to that of plants submerged in tap water. Only the ruderal population demonstrated a depth accommodation response in submerged conditions. Root concentrations of ethanol-soluble carbohydrates were up to three times higher in a field- collected turlough population during winter and autumn months than those in a ruderal population. Low levels of ethanol-insoluble carbohydrates were present in the turlough population but were absent from the ruderal population. Starch concentrations fluctuated greatly in the turlough population and were generally higher than those in the ruderal population. These results, together with those from previous investigations, suggest that the turlough population survives prolonged submergence by maintaining low levels of submerged photosynthesis, which may circulate oxygen within the plant tissues, and by utilizing storage carbohydrates for maintenance respiration.     

Reproductive biology and spawning habitat supplementation of the relict darter, Etheostoma chienense, a federally endangered species

We investigated the reproductive biology of Etheostoma chienense in the Bayou du Chien drainage of western Kentucky. Etheostoma chienense is similar in its ecology and reproductive biology to other members of the E. squamiceps complex. However, E. chienense is opportunistic in its choice of egg deposition substrates, as nests were found on a variety of natural and anthropogenic items. Due to an apparent lack of suitable spawning substrates, we added half-cylindrical ceramic tiles to several stretches of stream to increase potential nest productivity. Egg-clutches attached to artificial substrates were twice the size of egg clutches attached to naturally occurring materials. Laboratory experiments were conducted to determine nest rock size and mate choice preferences. Both the size of the male and of the nest rock appeared to be important parameters to spawning females. The life history information gathered here has significant management implications. Artificial spawning substrates should be placed in appropriate microhabitats (i.e., shallow, low-flow reaches in headwaters). Nest cavity vertical height should be about 3.0 cm and tiles should be spaced at least 0.5 m apart. We feel the use of surrogates (e.g., E. oophlyax) to investigate other types of spawning cover, and restoration of riparian buffer zones among other actions, would particularly benefit recovery efforts for this endangered species.     

Micropropagation of Turbinicarpus laui Glass et Foster, an endemic and endangered species

Summary In a short period of time, a large number of adventitious shoots were regenerated from longitudinal sections of in vitro-germinated seedlings of the endangered Mexican cactus, Turbinicarpus laui. The induction medium consisted of Murashige and Skoog salts, supplemented with 6-benzylaminopurine (BA) and α-naphthaleneacetic acid (NAA) in a wide range of combinations. The most effective concentrations of growth regulators for shoots initiation were 8.8–13.32 μM BA with 0–2.68 μM NAA. After 3–4 mo., individualized shoots were rooted on half-strength Murashige and Skoog medium and then transferred to soil to acclimatize under greenhouse conditions. In vitro strategies play a key role in the conservation and propagation of this commercially important endangered species of cactus.     

Effective population sizes and migration rates in fragmented populations of an endangered insect (Coenagrion mercuriale: Odonata)

1. Effective population sizes (N(e)) and migration rates (m) are critical evolutionary parameters that impact on population survival and determine the relative influence of selection and genetic drift. While the parameter m is well-studied in animal populations, N(e) remains challenging to measure and consequently is only rarely estimated, particularly in insect taxa. 2. We used demographic and genetic methods to estimate N(e) and m in a fragmented population of the endangered damselfly Coenagrion mercuriale to better understand the contrast between genetic and field estimates of these parameters and also to identify the spatial scale over which populations may become locally adapted. 3. We found a contrast between demographic- and genetic-based estimates of these parameters, with the former apparently providing overestimates of N(e), owing to substantial underestimation of the variance in reproductive success, and the latter overestimating m, because spatial genetic structure is weak. 4. The overall N(e) of sites within the population network at Beaulieu Heath, the largest C. mercuriale site in the UK, was estimated to vary between approximately 60 and 2700. 5. While N(e) was not correlated with either the total numbers of adults (N) or the area of habitat, this parameter was always less than N, because of substantial variance in reproductive success. The ratio N(e)/N varied between 0.006 and 0.42 and was generally larger in smaller populations, possibly representing some 'genetic compensation'. 6. From a simple genetic model and these data on N(e) and m, it seems that populations of C. mercuriale have the potential to respond to localized spatial variation in selection and this would need to be considered for future genetic management of this endangered species.     

Patterns, sources and ecological implications of clonal diversity in apomictic Ranunculus carpaticola (Ranunculus auricomus complex, Ranunculaceae)

Sources and implications of genetic diversity in agamic complexes are still under debate. Population studies (amplified fragment length polymorphisms, microsatellites) and karyological methods (Feulgen DNA image densitometry and flow cytometry) were employed for characterization of genetic diversity and ploidy levels of 10 populations of Ranunculus carpaticola in central Slovakia. Whereas two diploid populations showed high levels of genetic diversity, as expected for sexual reproduction, eight populations are hexaploid and harbour lower degrees of genotypic variation, but maintain high levels of heterozygosity at many loci, as is typical for apomicts. Polyploid populations consist either of a single AFLP genotype or of one dominant and a few deviating genotypes. genotype/genodive and character incompatibility analyses suggest that genotypic variation within apomictic populations is caused by mutations, but in one population probably also by recombination. This local facultative sexuality may have a great impact on regional genotypic diversity. Two microsatellite loci discriminated genotypes separated by the accumulation of few mutations ('clone mates') within each AFLP clone. Genetic diversity is partitioned mainly among apomictic populations and is not geographically structured, which may be due to facultative sexuality and/or multiple colonizations of sites by different clones. Habitat differentiation and a tendency to inhabit artificial meadows is more pronounced in apomictic than in sexual populations. We hypothesize that maintenance of genetic diversity and superior colonizing abilities of apomicts in temporally and spatially heterogeneous environments are important for their distributional success.     

Effects of a recent founding event and intrinsic population dynamics on genetic diversity in an ungulate population

Maintenance of genetic diversity has recently become a management goal for a number of species, due to its importance for present and future population viability. Genetic drift, primarily through differential reproductive success and inbreeding, can accelerate the loss of genetic diversity in recently recovered populations. We attempt to quantify the consequences of these factors on the genetic diversity contained in a small, recently founded wood bison (Bison bison athabascae) population by examining the genetic variation in this conservation herd, the calves born therein, and its large source population. The Hook Lake Wood Bison Recovery Project was initiated to found a disease-free herd of wood bison containing a representative amount of the genetic diversity present in the Wood Buffalo National Park metapopulation. Levels of diversity in the Hook Lake Wood Bison Recovery Project founders are higher than in previous salvage attempts. To examine the effects of differential reproductive success on this population, we monitored parentage of the calves born in the Hook Lake Wood Bison Recovery Project for 3 years since the founders reached sexual maturity. Two of the male founders sired over 90% of the offspring born in this population, which has led to a reduction in diversity in their calves. Monitoring of reproductive success, and incorporation of selective breeding strategies will be required to reduce the rate at which genetic diversity is lost from this small, isolated population. These steps should occur in other recovery projects, particularly when a small number of individuals are capable of dominating reproduction.     

濒危植物大果木莲种群格局及濒危原因分析

采用径级结构代替年龄结构以及方差均值比率法对木兰科(Magnoliaceae)木莲属(Manglietia Bl.)濒危植物大果木莲(Manglietia grandis Hu et Cheng)种群的年龄结构和种群格局进行了研究,并编制了大果木莲种群的特定时间生命表和存活曲线;结合生殖生物学特征以及遗传多样性研究结果,分析了导致大果木莲濒危的主要原因.根据株高和胸径可分别将大果木莲种群的年龄结构分为5级、高度结构分为6级;在大果木莲的5个年龄结构分级中,成年个体较多,幼年个体较少;其高度结构完整,个体高度主要在20 m以下.种群的方差均值比率为0.838 3,其空间分布格局属于随机分布.根据特定时间生命表可将大果木莲种群的发育分为3个阶段:幼树阶段(年龄级为Ⅱ～Ⅲ级)、成树阶段(年龄级为Ⅲ～Ⅳ级)、老树阶段(年龄级为Ⅳ～Ⅴ级),其中成树阶段个体死亡率最低.大果木莲种群存活曲线接近Deevey Ⅰ型,属于衰退型种群.种群自我更新能力差、种子生产力低下、有性生殖困难、生境片断化导致的基因流受限以及人为干扰是大果木莲濒危的主要原因.针对大果木莲濒危现状和致危原因,提出了相应的保护对策和建议.     

Patterns of male reproductive success in a highly promiscuous whale species: the endangered North Atlantic right whale

Parentage analyses of baleen whales are rare, and although mating systems have been hypothesized for some species, little data on realized male reproductive success are available and the patterns of male reproductive success have remained elusive for most species. Here we combine over 20 years of photo-identification data with high-resolution genetic data for the majority of individual North Atlantic right whales to assess paternity in this endangered species. There was significant skew in male reproductive success compared to what would be expected if mating was random (P < 0.001). The difference was due to an excess of males assigned zero paternities, a deficiency of males assigned one paternity, and an excess of males assigned as fathers for multiple calves. The variance in male reproductive success was high relative to other aquatically mating marine mammals, but was low relative to mammals where the mating system is based on resource- and/or mate-defence polygyny. These results are consistent with previous data suggesting that the right whale mating system represents one of the most intense examples of sperm competition in mammals, but that sperm competition on its own does not allow for the same degree of polygyny as systems where males can control access to resources and/or mates. The age distribution of assigned fathers was significantly biased towards older males (P < 0.05), with males not obtaining their first paternity until approximately 15 years of age, which is almost twice the average age of first fertilization in females (8 years), suggesting that mate competition is preventing younger males from reproducing. The uneven distribution of paternities results in a lower effective population size in this species that already has one of the lowest reported levels of genetic diversity, which may further inhibit reproductive success through mate incompatibility of genetically similar individuals.     

Male mating success, reproductive success and multiple paternity in a natural population of sand lizards: behavioural and molecular genetics data

Sand lizard Lacerta agilis females characteristically mate with several males which, in staged mating experiments, results in multiple paternity of the offspring. In order to investigate multiple paternity in a natural population and interpret male reproductive behaviours in terms of sired young, we sampled the blood of females, potential fathers and hatchlings, and determined paternity using multilocus DNA fingerprinting as well as the variation at a single locus detected by the probe (TC) _n . The paternity analyses were preceded by a laboratory experiment in which we established that the parental alleles identified by the single locus probe were inherited in a Mendelian way. Our molecular data demonstrated that 12 out of 13 males (92%) that sired offspring were correctly identified from the 56 sexually mature males in the population. Also smaller males were accepted as sexual partners by the females, but sired fewer young in competition with larger males and were less able to maintain prolonged post-copulatory mate guarding. This may result in that some sexually successful males are only observed inside a female's home range, but never in pair-association with the female.     

Breeding system, flower visitors and seedling survival of two endangered species of Helianthemum (Cistaceae)

BACKGROUND AND AIMS: Helianthemum marifolium and H. caput-felis are two endangered plant species of the western Mediterranean. Several aspects of the reproduction of both species were examined to determine whether their rarity could be related to factors causing reproductive limitation. METHODS: The flowering and fruiting phenology of both species in two non-sympatric island populations (Mallorca, Balearic Islands, western Mediterranean) were compared. Hand-pollination experiments were conducted to determine their fruit and seed production under different pollen sources. The composition of the pollinator assemblage and the effect of temporal variation and sun exposure on reproductive output and seedling survival were also investigated. KEY RESULTS: Flowering periods were longer for H. marifolium than for H. caput-felis in the populations studied. Helianthemum marifolium is mostly an outbreeder, i.e. fruit and seed set was three-fold higher when pollen came from other plants. In H. caput-felis, neither fruit nor seed set was affected by pollination treatments. Flower visitors were more diverse for H. caput-felis than for H. marifolium. In both species, most floral visits were made by hymenopterans. The total number of pollinator visits varied significantly between years, decreasing more than two-fold from 2001 to 2002, in both species. In agreement with its outbreeder character, variation in reproductive output between years was also observed in H. marifolium. It showed a 50 % decrease in fruit set in 2002, a pollinator-poor year. Finally, seedling survival increased three- to six-fold from 2001 to 2002. A correlation between seedling size and survival had also been detected. CONCLUSIONS: Reproductive limitations were detected for neither species (i.e. fruit and seed set, pollination service and seedling survival on natural populations). Hence, the increasing rarity of these two species is probably a direct result of the destruction of their habitat.     

Pronounced reproductive skew in a natural population of green swordtails, Xiphophorus helleri

For many species in nature, a sire's progeny may be distributed among a few or many dams. This poses logistical challenges--typically much greater across males than across females--for assessing means and variances in mating success (number of mates) and reproductive success (number of progeny). Here we overcome these difficulties by exhaustively analyzing a population of green swordtail fish (Xiphophorus helleri) for genetic paternity (and maternity) using a suite of highly polymorphic microsatellite loci. Genetic analyses of 1476 progeny from 69 pregnant females and 158 candidate sires revealed pronounced skews in male reproductive success both within and among broods. These skews were statistically significant, greater than in females, and correlated in males but not in females with mating success. We also compare the standardized variances in swordtail reproductive success to the few such available estimates for other taxa, notably several mammal species with varied mating systems and degrees of sexual dimorphism. The comparison showed that the opportunity for selection on male X. helleri is among the highest yet reported in fishes, and it is intermediate compared to estimates available for mammals. This study is one of a few exhaustive genetic assessments of joint-sex parentage in a natural fish population, and results are relevant to the operation of sexual selection in this sexually dimorphic, high-fecundity species.     

Estimating the population size of an endangered shorebird, the Madagascar plover, using a habitat suitability model

The Madagascar plover Charadrius thoracicus is a shorebird endemic to western Madagascar, currently classified as globally vulnerable. It is restricted to specialized wetland habitats that are increasingly threatened by humans. To inform future conservation measures for this poorly known species, we develop a predictive habitat suitability map and use this map to estimate the size of the Madagascar plover population. We integrate spatially referenced presence-only observations of Madagascar plovers with Landsat data, elevation data and measures of distance to settlements and the coast to produce a habitat suitability model using ecological niche factor analysis. Validation of this model using a receiver operating characteristic plot suggests that it is at least 84% accurate in predicting suitable sites. We then use our estimate of total area of suitable habitat above a critical suitability threshold and data on Madagascar plover density in suitable sites to estimate the total population size to derive a total population estimate of 3100±396 standard error individuals. Finally, we explore the conservation applications of our model.     

Extensive contemporary pollen-mediated gene flow in two herb species,Ranunculus bulbosus and Trifolium montanum,along an altitudinal gradient in a meadow landscape

Background and Aims

Genetic connectivity between plant populations allows for exchange and dispersal of adaptive genes, which can facilitate plant population persistence particularly in rapidly changing environments.

Methods

Patterns of historic gene flow, flowering phenology and contemporary pollen flow were investigated in two common herbs, Ranunculus bulbosus and Trifolium montanum, along an altitudinal gradient of 1200–1800 m a.s.l. over a distance of 1 km among five alpine meadows in Switzerland.

Key Results

Historic gene flow was extensive, as revealed by F_st values of 0·01 and 0·007 in R. bulbosus and T. montanum, respectively, by similar levels of allelic richness among meadows and by the grouping of all individuals into one genetic cluster. Our data suggest contemporary pollen flow is not limited across altitudes in either species but is more pronounced in T. montanum, as indicated by the differential decay of among-sibships correlated paternity with increasing spatial distance. Flowering phenology among meadows was not a barrier to pollen flow in T. montanum, as the large overlap between meadow pairs was consistent with the extensive pollen flow. The smaller flowering overlap among R. bulbosus meadows might explain the slightly more limited pollen flow detected.

Conclusions

High levels of pollen flow among altitudes in both R. bulbosus and T. montanum should facilitate exchange of genes which may enhance adaptive responses to rapid climate change.     

Local genetic population structure in an endangered plant species, Silene tatarica (Caryophyllaceae)

Genetic substructuring in plant populations may evolve as a consequence of sampling events that occur when the population is founded or regenerated, or if gene dispersal by pollen and seeds is restricted within a population. Silene tatarica is an endangered, perennial plant species growing along periodically disturbed riverbanks in northern Finland. We investigated the mechanism behind the microspatial genetic structure of S. tatarica in four subpopulations using amplified fragment length polymorphism markers. Spatial autocorrelation revealed clear spatial genetic structure in each subpopulation, even though the pattern diminished in older subpopulations. Parentage analysis in an isolated island subpopulation indicated a very low level of selfing and avoidance of breeding between close relatives. The mean estimated pollen dispersal distance (24.10 m; SD = 10.5) was significantly longer and the mean seed dispersal distance (9.07 m; SD = 9.23) was considerably shorter than the mean distance between the individuals (19.20 m; SD = 13.80). The estimated indirect and direct estimates of neighbourhood sizes in this subpopulation were very similar, 32.1 and 37.6, respectively. Our results suggested that the local spatial genetic structure in S. tatarica was attributed merely to the isolation-by-distance process rather than founder effect, and despite free pollen movement across population, restricted seed dispersal maintains local genetic structure in this species.     

Effects of individual and population parameters on reproductive success in three sexually deceptive orchid species

Reproductive success (RS) in orchids in general, and in non-rewarding species specifically, is extremely low. RS is pollinator and pollination limited in food deceptive orchids, but this has rarely been studied in sexually deceptive orchid species. Here, we tested the effects of several individual (plant height, inflorescence size, nearest neighbour distance and flower position) and population (patch geometry, population density and size) parameters on RS in three sexually deceptive Ophrys (Orchidaceae) species. Inter-specific differences were observed in RS of flowers situated in the upper versus the lower part of the inflorescence, likely due to species-specific pollinator behaviour. For all three species examined, RS increased with increasing plant height, inflorescence size and nearest neighbour distance. RS generally increased with decreasing population density and increasing patch elongation. Given these results, we postulate that pollinator availability, rather than pollinator learning, is the most limiting factor in successful reproduction for sexually deceptive orchids. Our results also suggest that olfactory 'display' ( i.e. versus optical display), in terms of inflorescence size (and co-varying plant height), plays a key role in individual RS of sexually deceptive orchids. In this regard, several hypotheses are suggested and discussed.