Pollinator shifts between Ophrys sphegodes populations: might adaptation to different pollinators drive population divergence?

Local adaptation to different pollinators is considered one of the possible initial stages of ecological speciation as reproductive isolation is a by‐product of the divergence in pollination systems. However, pollinator‐mediated divergent selection will not necessarily result in complete reproductive isolation, because incipient speciation is often overcome by gene flow. We investigated the potential of pollinator shift in the sexually deceptive orchids Ophrys sphegodes and Ophrys exaltata and compared the levels of floral isolation vs. genetic distance among populations with contrasting predominant pollinators. We analysed floral hydrocarbons as a proxy for floral divergence between populations. Floral adoption of pollinators and their fidelity was tested using pollinator choice experiments. Interpopulation gene flow and population differentiation levels were estimated using AFLP markers. The Tyrrhenian O. sphegodes population preferentially attracted the pollinator bee Andrena bimaculata, whereas the Adriatic O. sphegodes population exclusively attracted A. nigroaenea. Significant differences in scent component proportions were identified in O. sphegodes populations that attracted different preferred pollinators. High interpopulation gene flow was detected, but populations were genetically structured at species level. The high interpopulation gene flow levels independent of preferred pollinators suggest that local adaptation to different pollinators has not (yet) generated detectable genome‐wide separation. Alternatively, despite extensive gene flow, few genes underlying floral isolation remain differentiated as a consequence of divergent selection. Different pollination ecotypes in O. sphegodes might represent a local selective response imposed by temporal variation in a geographical mosaic of pollinators as a consequence of the frequent disturbance regimes typical of Ophrys habitats.     

Population Genetic Structure of a Widespread Bat-Pollinated Columnar Cactus

Bats are the main pollinators and seed dispersers of Stenocereus thurberi, a xenogamous columnar cactus of northwestern Mexico and a good model to illustrate spatial dynamics of gene flow in long-lived species. Previous studies in this cactus showed differences among populations in the type and abundance of pollinators, and in the timing of flowering and fruiting. In this study we analyzed genetic variability and population differentiation among populations. We used three primers of ISSR to analyze within and among populations genetic variation from eight widely separated populations of S. thurberi in Sonora, Mexico. Sixty-six out of 99 of the ISSR bands (P = 66.7%) were polymorphic. Total heterozygosity for all populations sampled revealed high genetic diversity (H_sp = 0.207, H_BT = 0.224). The AMOVA showed that most of the genetic variation was within populations (80.5%). At the species level, estimates of population differentiation, θ = 0.175 and θ^B = 0.194, indicated moderate gene flow among populations. The absence of a significant correlation between genetic and geographic distances indicated little isolation by geographic distance. The large genetic variation and diversity found in S. thurberi is consistent with its open reproductive system and the high mobility of bats, a major pollinator. However, small changes in number or kind of pollinators and seed dispersal agents, in the directionality of migratory routes, and/or in the timing of flowering and fruiting among populations, can critically affect gene flow dynamics.     

Landscape genetics and spatial pattern of phenotypic variation of Eristalis tenax across Europe

A study of population connectivity of the migratory insect species, such as dronefly Eristalis tenax (Diptera, Syrphidae), has an essential importance in understanding the relative influence of the evolutionary forces and environmental features that interact in the spatial distribution of molecular and morphological diversity. However, specific study aiming to understand spatial genetic structure of dronefly populations and its migratory potential is lacking. Hence, we studied a spatial pattern of genetic and phenotypic variation of seven European populations of E. tenax incorporating landscape genetic methods using allozyme data, wing size and shape and abdominal colour pattern. Based on the observed lack of genotypic structuring, we suggested that there has been sufficient long‐distance gene flow to effectively homogenize population structuring at a broader geographical scale. Wing shape similarity among populations and an overlap of abdominal colour variation showed no clear clustering related to geography, which is in congruence with genetic data. However, genetic (F_ST values) and phenotypic (wing size) data and landscape genetics indicated subdivision between the Balkan populations (four Serbian samples) and populations from Central (Germany and Switzerland) and Northern (Finland) Europe. These findings indicated a potential connection between the Central and Northern Europe supporting the Central European origin of the flies caught in Finland. Thus, by performing spatial analysis and combining genetic–morphological approach, we shed light on the movement pattern in complex landscapes and thus provided the necessary guidelines to a broad‐scale analysis of this widespread generalist pollinator.     

Genetic diversity in Dactylorhiza majalis subsp. majalis populations (Orchidaceae) of northern Poland

We analysed 16 populations of Dactylorhiza majalis subsp. majalis from northern Poland, simultaneously utilizing both morphological and molecular data. Genetic differentiation was examined using five microsatellite loci, and morphological variation was assessed for 23 characters. At the species level, our results showed a moderate level of genetic diversity (A = 6.00; A_e = 1.86; H_o = 0.387; F_IS = 0.139) which varied between the studied populations (A = 2.60–4.20; A_e = 1.68–2.39; H_o = 0.270–0.523; F_IS = ?0.064–0.355). A significant excess of homozygotes was detected in five population, while excess of heterozygotes was observed in four populations, but the latter values were statistically insignificant. Moderate, but clear between population genetic differentiation was found (F_ST = 0.101; p < 0.001). Considering pairwise‐F_ST and number of migrants among populations, we recognized three population groups (I, II, III), where the first could be further divided into two subgroups (Ia, Ib). These three groups differed with respect to gene flow values (N_m = 0.39–1.12). The highest number of migrants per generation was noticed among populations of subgroup Ia (8.58), indicative of a central panmictic population with free gene flow surrounded by peripatric local populations (Ib) with more limited gene flow. Geographic isolation, habitat fragmentation and limited seed dispersal are inferred to have caused limitations to gene flow among the three indicated population groups. There was a significant correlation between the morphological and genetic distance matrices. A weak but significant pattern of isolation by distance was also observed (r = 0.351; p < 0.05).     

Genetic diversity of natural populations of Taxus mairei

Taxus mairei (Lemée & Lév.) S.Y. Hu ex T. S. Liu is a vulnerable tree species, and it is also a precious timber species in China. We used 13 microsatellites to assess the genetic diversity and differentiation of 665 T. mairei samples from 18 natural populations. A total of 291 alleles were detected. The average number of alleles (N_a?=?22.39), expected heterozygosity (H_e?=?0.74), polymorphic information content (PIC?=?0.86) and Shannon diversity index (I?=?1.66) of the loci indicated a high level of genetic diversity in natural T. mairei populations. Moreover, gene flow was more active among populations (N_m?=?1.62) than within populations. Among the 18 populations, the Xinfeng population in Jiangxi Province has the highest genetic diversity. Although each of the studied populations should be protected from further deforestation and agricultural expansion, the Xinfeng population deserves the highest conservation priority. The results based on analysis of molecular variance showed that genetic variation occurred mainly within populations (84.90%; P?<?0.001), which indicated that the degree of genetic differentiation of the natural populations of T. mairei was low. Based on UPGMA, the 18 populations were categorized into two groups. A Mantel test showed that there was no significant correlation between standard genetic distance and geographical distance or altitude differences among the populations. The genetic clustering results also indicated that there are varying degrees of gene penetration among natural populations of T. mairei. The information presented here forms the basis for the development of genetic guidelines for appropriate conservation programs.

     

Landscape pattern and genetic structure of a yellow-necked mouse <Emphasis Type="Italic">Apodemus flavicollis</Emphasis> population in north-eastern Poland

Reduced connectivity among local populations inhabiting a spatially heterogeneous landscape may restrict gene flow and thus contribute to diminished genetic variation within a population. The aim of this study was to determine the role of geographic distance and habitat barriers in developing genetic structure of a yellow-necked mouse Apodemus flavicollis (Melchior, 1834) population, taking into consideration the spatial organization of the landscape. A field study was carried out in two plots located in NE Poland that differed considerably in terms of the scale of habitat fragmentation: (1) a continuous forest complex, and (2) a mosaic of smaller forest habitats. The plots were separated by a water barrier comprised of a chain of lakes. DNA samples from a total of 654 individuals were examined by microsatellite analysis (5 loci). The results showed that the yellow-necked mouse population was characterized by a poorly pronounced genetic structure throughout the study area, although the statistical significance of F _ST for most location pairs indicated that gene flow in the area was not free. The division of the mouse population into three genetically distinct groups clearly demonstrated the significant role of water bodies as a natural barrier effectively hindering free movement of animals and thus gene flow. Analysis of the genetic structure of the mouse population throughout the study area and also within the distinguished groups indicated that the entire study population may be considered as a single metapopulation. Our results suggest that geographic distance alone is not the predominant factor affecting the genetic structure of population, but in the mosaic landscape the relative isolation of individual forest fragments, and barriers hindering movements of individuals and limiting gene flow among local populations played a much more important role.     

Measures of gene flow in the Columbian ground squirrel

From analyses of published data and a review of the literature, I studied indirect and direct measures of gene flow among populations of Columbian ground squirrels, Spermophilus columbianus. New analyses were used to examine an allozyme data set (seven polymorphic loci) that had been collected by Zammuto and Millar (1985a) from six populations of ground squirrels that were spread over 183 km. G-tests indicated significant variation in allele frequencies among populations, but F-statistics revealed relatively little population differentiation (average F _ST=0.026). F _ST values were used to estimate rates of gene flow indirectly and indicated fairly high rates of gene flow (average N _e m=13.5). Recorded dispersal distances of individual ground squirrels were fairly short (most<4 km, maximum recorded distance was 8.5 km), and the minimum distance between populations used to create the allozyme data set was about 25 km. Thus, direct dispersal among the populations in the allozyme data set was highly unlikely. Small genetically effective populations may have experienced high rates of migration over short distances (about 43% of adults in local populations were immigrants), however, resulting in homogeneous allele frequencies over the geographic range. This explanation provides an alternative to invoking gene flow in the recent past to explain discrepancies between dispersal distances in the field and homogenization of allele frequencies over large ranges, Mammalian species that have virtually complete dispersal of subadult males from the natal area might be expected to exhibit relatively high rates of gene flow, regardless of actual dispersal distances. Genetically effective populations may be much smaller than more extensive ecological populations and experience higher rates of gene flow.     

Patterns of Genetic Variation in Swertia przewalskii, an Endangered Endemic Species of the Qinghai-Tibet Plateau

Swertia przewalskii Pissjauk. (Gentianaceae) is a critically endangered and endemic plant of the Qinghai-Tibet Plateau in China. RAPD and ISSR analyses were carried out on a total of 63 individuals to assess the extent of genetic variation in the remaining three populations. Percentage of polymorphic bands was 94% (156 bands) for RAPD and 96% (222 bands) for ISSR. A pairwise distance measure calculated from the RAPD and ISSR data was used as input for analysis of molecular variance (AMOVA). AMOVA indicated that a high proportion of the total genetic variation (52% for RAPD and 56% for ISSR) was found among populations; pairwise Φ _ST comparisons showed that the three populations examined were significantly different (p < 0.001). Significant genetic differentiation was found based on different measures (AMOVA and Hickory θ_B) in S. przewalskii (0.52 on RAPD and 0.56 on ISSR; 0.46 on RAPD and 0.45 on ISSR). The differentiation of the populations corresponded to low average gene flow (0.28 based on RAPD and 0.31 based on ISSR), whereas genetic distance-based clustering and coalescent-based assignment analyses revealed significant genetic isolation among populations. Our results indicate that genetic diversity is independent of population size. We conclude that although sexual reproduction and gene flow between populations of S. przewalskii are very limited, they have preserved high levels of genetic diversity. The main factors responsible for the high level of difference among populations are the isolation and recent fragmentation under human disturbance.     

Diversity and structure of fragmented populations of a threatened endemic cyprinodontid (Aphanius sophiae) inferred from genetics and otolith morphology: Implications for conservation and management

The assessment of population structure and genetic diversity is crucial for the management and conservation of threatened species. Natural and artificial barriers to dispersal (i.e., gene flow) increase populations’ differentiation and isolation by reducing genetic exchange and diversity. Freshwater ecosystems are highly fragmented because of human activities. Threatened species with small population sizes are more sensitive to habitat fragmentation effects. Here, we investigate the genetic population structure and gene flow among seven populations of Aphanius sophiae in the Kor Basin by using sequences of the complete Cyt b gene and otolith morphometry. The Cyt b gene showed low level of genetic variation, only 4.12% of the identified sites were variable, and 2.42% were parsimony informative. Overall, haplotype diversity was low to moderate and nucleotide diversity was low to extremely low. Fish populations exhibited high levels of genetic differentiation, suggesting limited gene flow among them. These differences were obtained not only among geographically distant populations, but also among neighboring localities. Genetic population structure was supported by the AMOVA analysis and by the haplotype network (only one of 21 haplotypes were shared by two localities). Otolith morphometric analysis was in agreement with genetic results, the two most distant and isolated populations were clearly separated, and genetically close populations showed less differences in morphometry. A significant pattern of isolation by distance was also detected among A. sophiae populations, with genetic distance more correlated with hydrological distance than with geographic distance. Results suggested that limited gene flow due to habitat fragmentation is an important factor contributing to genetic structuring and to the loss of genetic variation of A. sophiae populations. Aphanius sophiae population structure seems to be the result of habitat fragmentation and water pollution, but other factors such as introduced species should be considered. Given the high degree of genetic structuring, the definition of conservation groups is of particular importance for A. sophiae, which should be considered endangered according to the IUCN criteria. Conservation plans must recognize the genetic independence of populations and manage separately preventing the loss of locally adapted genotypes.     

Genetic structure of Korean populations of bumblebees Bombus ignitus (Hymenoptera: Apidae) as revealed by microsatellite markers

Bumblebee Bombus ignitus, which is indigenous to Korea, Japan, and China, has been recognized as a valuable pollinator for both crops and wild plants. Bombus ignitus has now become commercially important as a pollinator because of its use in the agricultural industry, particularly for greenhouse pollination. For long‐term management and effective conservation of B. ignitus, an understanding of the genetic structure of its naturally occurring populations is practically important. In this study, the genetic structure among the five populations of B. ignitus in South Korea was assessed using nine microsatellite loci. These markers showed high levels of genetic variability, with the number of alleles ranging from 6 to 22 (mean = 13.4) and the frequency of the most common allele ranging from 0.11 to 0.66. Only the Sabuk (SB) population showed a genetic signature of a recent bottleneck, which was further supported by the lowest level of allelic richness (AR) (mean AR = 3.944). Genetic differentiation was highly significant among all pairs of populations (P < 0.001) across the nine microsatellite markers, suggesting a lack of gene flow among those populations. Interestingly, F_ST (and R_ST and D_est) values were always greater for the Taebaek population than for the four remaining populations. The phylogenetic analysis showed evidence supporting our hypothesis that the Taebaek population is genetically more divergent than the other populations. Overall, our results suggest that the Korean populations of B. ignitus might have undergone geographic isolation and have become highly separated spatially from one another, thereby having a limited range of migration among their local habitats.