Local adaptation of an anuran amphibian to osmotically stressful environments

Abstract. Water salinity is an intense physiological stress for amphibians. However, some species, such as Bufo calamita, breed in both brackish and freshwater environments. Because selection under environmentally stressful conditions can promote local adaptation of populations, we examined the existence of geographic variation in water salinity tolerance among B. calamita populations from either fresh or brackish water ponds in Southern Spain. Comparisons were made throughout various ontogenetic stages. A combination of field transplant and common garden experiments showed that water salinity decreased survival probability of individuals in all populations, prolonged their larval period, and reduced their mass at metamorphosis. However, significant population X salinity interactions indicated that the population native to brackish water (Saline) had a higher salinity tolerance than the freshwater populations, suggesting local adaptation. Saline individuals transplanted to freshwater environments showed similar survival probabilities, length of larval period, and mass at metamorphosis than those native to freshwater. This indicates that increased tolerance to osmotic stress does not imply a loss of performance in freshwater, at least during the larval and juvenile phases. Despite the adaptive process apparently undergone by Saline, all populations still shared the same upper limit of embryonic stress tolerance (around 10 g/l), defining a window of salinity range within which selection can act. Significant differences in embryonic and larval survival in brackish water among sibships for all populations suggest the existence of a genetic basis for the osmotic tolerance.     

Adaptation to sea level rise: does local adaptation influence the demography of coastal fish populations?

This study compared the growth of two western mosquitofish Gambusia affinis populations that were previously demonstrated to have genetic adaptations that increased survival under lethal salinity exposures. The objective was to evaluate how genetic adaptations to lethal salinity stress affect population demography when exposed to sublethal salinity stress. Results indicate that chronic salinity exposure had a generally negative impact on population size, but fish originating from one of the two populations established with fish from a brackish site exhibited an increase in population size. Saltwater intrusion seems to result in reduced population size for most populations. Some populations inhabiting more saline sites, however, may develop localized adaptations, mitigating the consequences of increased salinity on population productivity.     

Stress tolerance and population stability of rock pool Daphnia in relation to local conditions and population isolation

Small fragmented populations can lose genetic variability, which reduces population viability through inbreeding and loss of adaptability. Current and previous environmental conditions can also alter the viability of populations, by creating local adaptations that determine responses to stress. Yet, most studies on stress tolerance usually consider either the effect of genetic diversity or the local environment, missing a more holistic perspective of the factors contributing to stress tolerance among natural populations. Here, we studied how salinity stress affects population growth of Daphnia longispina, Daphnia magna, and Daphnia pulex from rock pools with varying degrees of population isolation and salinity conditions. Standing variation of in situ rock pool salinity conditions explained more variation in salt tolerance than the standing variation of population isolation or genetic diversity, in both a pulse and a press disturbance experiment. This indicates that the level of stress, which these natural populations experience, influences their response to that stress, which may have important consequences for the conservation of fragmented populations. However, long-term population stability in the field decreased with population isolation, indicating that natural populations experience a variety of stresses; thus, population isolation and genetic diversity may stabilize population dynamics over larger spatiotemporal scales.     

Molecular Genetic Variation in a Clonal Plant Population of Leymus chinensis (Trin.) Tzvel.

Randomly amplified polymorphic DNA (RAPD) analysis was used to investigate the genetic variation among populations, between populations, and within populations, relationships between genetic distance and geographic distance, and the molecular variation and population size. The effects of geographic and genetic distances, as well as of genetic differentiation and population size, on genetic variations of Leymus chinensis (Trin.) Tzvel. are discussed. The present study showed that there was significant RAPD variation between the Baicheng region population and the Daqing region population, with a molecular variance of 6.35% (P ＜ 0.04), and for differentiation among area populations of the Daqing region, with a molecular variance of 8.78% (P ＜ 0.002). A 21.06% RAPD variation among all 16 populations among two regions was found (P ＜ 0.001), as well as 72.59% variation within populations (P ＜ 0.001). Molecular variation within populations was significantly different among 16 populations.     

Environmental correlates of population differentiation in Atlantic herring

The marine environment is characterized by few physical barriers, and pelagic fishes commonly show high migratory potential and low, albeit in some cases statistically significant, levels of genetic divergence in neutral genetic marker analyses. However, it is not clear whether low levels of differentiation reflect spatially separated populations experiencing gene flow or shallow population histories coupled with limited random genetic drift in large, demographically isolated populations undergoing independent evolutionary processes. Using information for nine microsatellite loci in a total of 1951 fish, we analyzed genetic differentiation among Atlantic herring from eleven spawning locations distributed along a longitudinal gradient from the North Sea to the Western Baltic. Overall genetic differentiation was low (theta = 0.008) but statistically significant. The area is characterized by a dramatic shift in hydrography from the highly saline and temperature stable North Sea to the brackish Baltic Sea, where temperatures show high annual variation. We used two different methods, a novel computational geometric approach and partial Mantel correlation analysis coupled with detailed environmental information from spawning locations to show that patterns of reproductive isolation covaried with salinity differences among spawning locations, independent of their geographical distance. We show that reproductive isolation can be maintained in marine fish populations exhibiting substantial mixing during larval and adult life stages. Analyses incorporating genetic, spatial, and environmental parameters indicated that isolating mechanisms are associated with the specific salinity conditions on spawning locations.     

Landscape genetic structure of Scirpus mariqueter reveals a putatively adaptive differentiation under strong gene flow in estuaries

Estuarine organisms grow in highly heterogeneous habitats, and their genetic differentiation is driven by selective and neutral processes as well as population colonization history. However, the relative importance of the processes that underlie genetic structure is still puzzling. Scirpus mariqueter is a perennial grass almost limited in the Changjiang River estuary and its adjacent Qiantang River estuary. Here, using amplified fragment length polymorphism (AFLP), a moderate‐high level of genetic differentiation among populations (range F_ST: 0.0310–0.3325) was showed despite large ongoing dispersal. FLOCK assigned all individuals to 13 clusters and revealed a complex genetic structure. Some genetic clusters were limited in peripheries compared with very mixing constitution in center populations, suggesting local adaptation was more likely to occur in peripheral populations. 21 candidate outliers under positive selection were detected, and further, the differentiation patterns correlated with geographic distance, salinity difference, and colonization history were analyzed with or without the outliers. Combined results of AMOVA and IBD based on different dataset, it was found that the effects of geographic distance and population colonization history on isolation seemed to be promoted by divergent selection. However, none‐liner IBE pattern indicates the effects of salinity were overwhelmed by spatial distance or other ecological processes in certain areas and also suggests that salinity was not the only selective factor driving population differentiation. These results together indicate that geographic distance, salinity difference, and colonization history co‐contributed in shaping the genetic structure of S. mariqueter and that their relative importance was correlated with spatial scale and environment gradient.     

Comparison of genetic diversity at microsatellite loci and quantitative traits in hatchery populations of Japanese flounder Paralichthys olivaceus

Relationships of genetic diversity at microsatellite loci and quantitative traits were examined in hatchery-produced populations of Japanese flounder using a relatively straightforward experiment. Five hatchery populations produced by wild-caught and domesticated broodstocks were used to examine the effects of different levels (one to three generations) of domestication on the genetic characteristics of hatchery populations. Allelic richness at seven microsatellite loci in all hatchery populations was lower than that in a wild population. Genetic variation measured by allelic richness and heterozygosity tended to decrease with an increase in generations of domestication. In addition, the degree of genetic differentiation from a wild population increased with an increase in generations of domestication. Significant differences in three morphometric traits (dorsal and anal fin ray counts and vertebral counts) and three physiological traits (high temperature, salinity and formalin tolerance) were observed among the hatchery populations. The degree of phenotypic difference among populations was larger in morphometric traits than in physiological traits. The divergence pattern of some quantitative traits was similar to that observed at microsatellite loci, suggesting that domestication causes the decrease of genetic variation and the increase of genetic differentiation for some quantitative traits concomitantly with those for microsatellite loci. Significant positive correlation was observed between F _ST and the degree of phenotypic difference in the three morphometric traits and formalin tolerance, indicating that genetic variation at microsatellite loci predicts the degree of phenotypic divergence in some quantitative traits.     

Genetic variability and structure of the water vole Arvicola amphibius across four metapopulations in northern Norway

Water vole Arvicola amphibius populations have recently experienced severe decline in several European countries as a consequence of both reduction in suitable habitat and the establishment of the alien predator American mink Neovison vison. We used DNA microsatellite markers to describe the genetic structure of 14 island populations of water vole off the coast of northern Norway. We looked at intra‐ and inter‐population levels of genetic variation and examined the effect of distance among pairs of populations on genetic differentiation (isolation by distance). We found a high level of genetic differentiation (measured by F_ST) among populations overall as well as between all pairs of populations. The genetic differentiation between populations was positively correlated with geographic distance between them. A clustering analysis grouped individuals into 7 distinct clusters and showed the presence of 3 immigrants among them. Our results suggest a small geographic scale for evolutionary and population dynamic processes in our water vole populations.     

Molecular Genetic Variation in a Clonal Plant Population of Leymus chinensis （Trin.） Tzvel.

Randomly amplified polymorphic DNA （RAPD） analysis was used to investigate the genetic variation among populations, between populations, and within populations, relationships between genetic distance and geographic distance, and the molecular variation and population size. The effects of geographic and genetic distances, as well as of genetic differentiation and population size, on genetic variations of Leymus chinensis （Trin.） Tzvel. are discussed. The present study showed that there was significant RAPD variation between the Baicheng region population and the Daqing region population, with a molecular variance of 6.35% （P 〈 0.04）, and for differentiation among area populations of the Daqing region, with a molecular variance of 8.78% （P 〈 0.002）. A 21.06% RAPD variation among all 16 populations among two regions was found （P 〈 0.001）, as well as 72.59% variation within populations （P 〈 0.001）. Molecular variation within populations was significantly different among 16 populations.     

Effects of salinity on the life history and fitness of Daphnia magna: variability within and between populations

The life history traits of Daphnia magna were studied in laboratory experiments under freshwater and brackish (5 salinity) conditions. The variability of responses within and between populations was examined by comparing 11 clones from a brackish lake and 10 clones from a freshwater pond. Experimental clones were hatched from ephippia collected from the sediment and thus represent random samples of the clone banks of each population.Most clones with a high salinity tolerance were from the population of the brackish habitat, but some were also found in the freshwater population. Thus, freshwater populations appear to have the potential to invade brackish habitats. A proportion of clones from the brackish population had very low fitness (measured as e^r) under freshwater conditions. This unexpected result means that freshwater adaptation can be lost by the freshwater cladoceran Daphnia magna. The effects of unfavourable conditions on growth and reproduction varied among clones and were not correlated. This clonal variation in growth and reproduction indicates that the environmental sensitivities of these traits are independent. The pattern of fitness reaction norms showed no trade-off between fitness under brackish and under freshwater conditions for either population. Thus, euryhaline generalists should be favoured in habitats with salinity fluctuations between freshwater and brackish conditions.     

Genetic Evaluation of the Efficacy of In Situ and Ex Situ Conservation of <Emphasis Type="Italic">Parashorea chinensis</Emphasis> (Dipterocarpaceae) in Southwestern China

The majority of research in genetic diversity yields recommendations rather than actual conservation achievements. We assessed the efficacy of actual in situ and ex situ efforts to conserve Parashorea chinensis (Dipterocarpaceae) against the background of the geographic pattern of genetic variation of this species. Samples from seven natural populations, including three in a nature reserve, and one ex situ conservation population were studied. Across the natural populations, 47.8% of RAPD loci were polymorphic; only 20.8% on average varied at the population level. Mean population genetic diversity was 0.787 within natural populations and 1.410 for the whole species. Significant genetic differentiation among regions and isolation by distance were present on larger scales (among regions). AMOVA revealed that the majority of the among-population variation occurred among regions rather than among populations within regions. Regression analysis, Mantel test, principal coordinates analysis, and cluster analysis consistently demonstrated increasing genetic isolation with increasing geographic distance. Genetic differentiation within the region was quite low compared to that among regions. Multilocus spatial autocorrelation analysis of these three populations revealed random distribution of genetic variation in two populations, but genetic clustering was detected in the third population. The ex situ conserved population contained a medium level of genetic variation compared with the seven natural populations; it contained 77.1% of the total genetic variation of this species and 91% of the moderate to high frequency RAPD fragments (f > 0.05). Exclusive bands were detected in natural populations, but none were found in the ex situ conserved population. The populations protected in the nature reserve contained most of the genetic variation of the whole species, with 81.4% of the total genetic variation and 95.7% of the fragments with moderate to high frequency (f > 0.05) of this species conserved. The results show that the ex situ conserved population does not contain enough genetic variation to meet the need of release in the future, and that more extensive ex situ sampling in natural populations TY, NP, HK, and MG is needed. The in situ conserved population contains representative genetic variation to maintain long-term survival and evolutionary processes of P. chinensis.     

Response to long- and short-term salinity in populations of the C4 nonhalophyte Andropogon glomeratus Walter B.S.P.

Summary This research was undertaken to investigate differences in salt tolerance under conditions in which salinity is increased gradually and maintained for long periods or increased rapidly and maintained for shorter periods. The responses of populations of a C₄ nonhalophytic grass, Andropogon glomeratus, to long- and short-term salinity were measured under controlled environment conditions. Additionally, plants from a salt marsh population and an inland population were transplanted into a salt marsh and their survival compared. The relative growth reductions in the salt marsh and the inland populations under long-term salinity were similar. Survival of seedlings of 4 populations inundated with full-strength seawater over a relatively short period indicated differential capacities to tolerate soil salinities imposed in a manner similar to tidal inundation in a salt marsh. The greater survival of plants from the marsh population transplanted into the salt marsh further indicated genetic differentiation between the populations. These results indicate that genetic differentiation to salt tolerance in A. glomeratus is better reflected by survival after shortterm salinity events, rather than growth inhibition due to long-term salinity imposed gradually.     

基于细胞色素氧化酶亚基Ⅰ基因的中缅树鼩地理种群遗传分化

中缅树鼩广泛分布于东南亚地区,在我国主要分布于西南地区及海南岛。本研究以中国10个地理种群共112只中缅树鼩为研究对象, PCR扩增得到1398 bp的细胞色素氧化酶亚基Ⅰ（COⅠ）基因序列,并对其进行分析。 结果显示：112个中缅树鼩COⅠ基因共定义了64个单倍型,其单倍型多样性（Hd）平均值为0.9730,核苷酸多样性（Pi ）平均值为0.04494;AMOVA方差分析显示种群间的变异占总变异的93.09%,说明中缅树鼩地理种群变异主要发生在种群间;整体遗传分化固定指数（F_ST）为0.93091,说明各地理种群中缅树鼩已出现明显的遗传分化;结合中性检验与碱基错配分布图结果表明中缅树鼩在历史进程中未经历过种群扩张现象; 基于单倍型构建的系统进化树与NETWORK网络图显示10个地理种群的中缅树鼩聚为4支：海南种群一支,大新种群一支,片马种群一支,其他种群一支。结果表明：中国不同地理种群的中缅树鼩具有较高的遗传多样性,各地理种群间已经出现了较为明显的遗传分化,地理阻隔作用可能是其分化的主要原因。     

基于ITS序列的灵芝遗传多样性与群体遗传分化研究

本研究基于ITS序列对中国8个不同地理群体的168份灵芝样本的分子变异、遗传多样性、群体遗传分化程度进行了分析。结果表明：168个样品中共检测到39个单倍型（H1-H39）,表现出较低的遗传多样性水平（H_d=0.867±0.018,P_i=0.00304±0.00024）,各群体的遗传多样性水平相差不大;TCS单倍型网络图和基于ML、NJ法构建的单倍型系统发育树显示,各地理群体的遗传距离与地理距离之间没有形成对应关系,二者没有明显的相关性;AMOVA分析结果显示不同地理群体间的遗传变异占4.33%,群体内的变异占95.67%,遗传分化主要来自于群体内部。总体和各地理群体的中性检验结果显示,群体扩张遵循中性进化,群体大小保持相对稳定。整体的遗传分化指数F_st为0.04331,基因流N_m为2.99,表明各地理群体间存在频繁的基因交流,群体遗传分化较小。推测原因可能为种群扩张历史较短,分布范围较窄,造成遗传多样性偏低,各群体间遗传分化不大。     

Genotype-by-environment interaction for salinity tolerance in the freshwater-invading copepod Eurytemora affinis

This study examined the extent of phenotypic plasticity for salinity tolerance and genetic variation in plasticity in the invasive copepod Eurytemora affinis. Euryemora affinis is a species complex inhabiting brackish to hypersaline environments but has invaded freshwater lakes and reservoirs within the past century. Reaction norm experiments were performed on a relatively euryhaline population collected from a brackish lake with fluctuating salinity. Life history traits (hatching rate, survival, and development time) were measured for 20 full-sib clutches that were split and reared at four salinities (fresh, 5, 10, and 27 practical salinity units [PSU]). On average, higher salinities (10 and 27 PSU) were more favorable for larval growth, yielding greater survival and faster development rate. Clutches differed significantly in their response to salinity, with a significant genotype-by-environment interaction for development time. In addition, genetic (clutch) effects were evident in response to low salinity, given that survival in fresh (lake) water was significantly positively correlated with survival at 5 PSU for individual clutches. Clutches raised in fresh water could not survive beyond metamorphosis, suggesting that acclimation to fresh water could not occur in a single generation. Results suggest the importance of natural selection during freshwater invasion events, given the inability of plasticity to generate a freshwater phenotype, and the presence of genetic variation for plasticity upon which natural selection could act.     

Rapid Divergent Evolution of Male Genitalia Among Populations of Drosophila buzzatii

Increasing evidence from multiple animal systems suggests that genital evolution and diversification are driven by rapid and strong evolutionary forces. Particularly, the morphology of male genital structures is considered to be among the fastest evolving traits in animal groups with internal fertilization. In this study, we investigated patterns of male genital variation within and between natural populations of the cactophilic fly Drosophila buzzatii in its original geographic distribution range in the Neotropics. We detected significant morphological differences among populations and distinguished five differentiated groups. Moreover, among population differentiation in genital morphology was associated with the degree of geographic isolation among populations and clearly contrasted with the general homogeneity detected for the putatively neutral mitochondrial gene COI. Integrating our present data with previous molecular population genetic surveys, our results suggest that male genital morphology has rapidly diverged after the recent demographic expansion that D. buzzatii has undergone in the arid zones of South America. Because the “lock and key” hypothesis failed to explain the present pattern, we explored alternative explanations for the observed pattern of genital diversification including drift-facilitated sexual selection.     

Dwindling genetic diversity in European ground squirrels?

The European ground squirrel (Spermophilus citellus) is endangered and in decline. Populations are increasingly fragmented, and only a coordinated conservation effort at the European level may guarantee its long-term survival. To obtain a general population genetic picture on a larger geographic scale, we screened 117 individuals from seven local populations in Hungary, Romania, and Austria for allelic variation at eleven microsatellite loci. We found a high (23.4%) proportion of private alleles, and a moderate to somewhat elevated level (15.27%) of partitioning of genetic diversity among populations, compared to that found in many other terrestrial mammals. Genetic variability was significantly higher than in earlier studied Czech populations that are considered genetically depleted, but significantly lower than in undisturbed populations of S. suslicus and S. brunneus, that are similar to the European ground squirrel in their ecological requirements, reproductive biology, and social organization. Genetic diversity was also lower than in most presumably “undisturbed” populations of other Sciurid species. This, together with the observed level and pattern of genetic differentiation among populations, such as no significant increase of genetic differentiation with geographic distance and similar variance of genetic differentiation between populations independent of geographic distance, indicated the prevalence of relatively strong drift effects for all populations. A Bayesian STRUCTURE analysis and a factorial correspondence analysis concordantly revealed a fairly complex genetic composition of local populations, but no major geographic trend in the pattern of the genetic composition. Overall, the results suggest disintegration of local colonies that might earlier have been more connected genetically. The STRUCTURE analysis also suggested anthropogenic translocations among single Hungarian populations. Our data on genetic diversity and its distribution do not object to such conservation measures. Translocation of individuals particularly from nearby populations may increase the chances of survival of small and isolated populations and counteract inbreeding at low densities.     

LOCAL ADAPTATION MAINTAINS CLINAL VARIATION IN MELANIN‐BASED COLORATION OF EUROPEAN BARN OWLS (TYTO ALBA)

Ecological parameters vary in space, and the resulting heterogeneity of selective forces can drive adaptive population divergence. Clinal variation represents a classical model to study the interplay of gene flow and selection in the dynamics of this local adaptation process. Although geographic variation in phenotypic traits in discrete populations could be remainders of past adaptation, maintenance of adaptive clinal variation requires recurrent selection. Clinal variation in genetically determined traits is generally attributed to adaptation of different genotypes to local conditions along an environmental gradient, although it can as well arise from neutral processes. Here, we investigated whether selection accounts for the strong clinal variation observed in a highly heritable pheomelanin‐based color trait in the European barn owl by comparing spatial differentiation of color and of neutral genes among populations. Barn owl's coloration varies continuously from white in southwestern Europe to reddish‐brown in northeastern Europe. A very low differentiation at neutral genetic markers suggests that substantial gene flow occurs among populations. The persistence of pronounced color differentiation despite this strong gene flow is consistent with the hypothesis that selection is the primary force maintaining color variation among European populations. Therefore, the color cline is most likely the result of local adaptation.     

Variation in gene expression along a salinity gradient in wild populations of the euryhaline black-chinned tilapia Sarotherodon melanotheron

This study evaluated variation in expression of 11 genes within and among six wild populations of the black-chinned tilapia Sarotherodon melanotheron distributed along a salinity gradient from 0 to 100. Previous laboratory studies had shown that expression of these genes was sensitive to water salinity; the current study confirmed that a number of them also varied in expression in wild populations along the salinity gradient. Principal component analysis (PCA) first distinguished two, not mutually exclusive, sets of genes: trade-off genes that were highly expressed at one or other extreme of the salinity gradient and stress genes that were up-regulated at the two salinity extremes (i.e. a U-shaped expression pattern). The PCA clearly partitioned the populations into three groups based on their gene expression patterns and their position along the salinity gradient: a freshwater (GL; 0) population, four brackish and seawater (GB, HB, SM, SF; ranging from 20 to 50) populations and a hypersaline (SK, 100) population. Individual variation in gene expression was significantly greater within the populations at the extreme compared to intermediate salinities. These results reveal phenotypically plastic regulation of gene expression in S. melanotheron, and greater osmoregulatory and plasticity costs at extreme salinities, where fitness-related traits are known to be altered.     

Adaptation or exaptation? An experimental test of hypotheses on the origin of salinity tolerance in Bufo calamita

The natterjack toad (Bufo calamita) shows variation in embryonic and larval salinity tolerance across populations in southern Spain. However, its aquatic/terrestrial biphasic life cycle, together with remarkable differences in salinity tolerance between Spanish and UK freshwater populations suggest an alternative hypothesis to local adaptation. Drought resistance during the terrestrial phase and salinity tolerance during the aquatic phase are both related to osmotic stress tolerance, and if there were an association between them, one could have evolved as an exaptation from the other. To test such an association, we reared B. calamita juveniles from three populations known to differ genetically in their salinity tolerance, under either dry or humid conditions. Drought decreased growth rate, enhanced burying behaviour, and decreased foraging activity and efficiency. No significant population x treatment interaction was found for any variable, i.e. populations were equally affected by drought. These results do not support the hypothesis of a genetic association between salinity and drought tolerance.