Genetic divergence between Melipona quadrifasciata Lepeletier (Hymenoptera,Apidae) populations

Melipona quadrifasciata is a stingless bee widely found throughout the Brazilian territory, with two recognized subspecies, M. quadrifasciata anthidioides, that exhibits interrupted metasomal stripes, and M. quadrifasciata quadrifasciata, with continuous metasomal stripes. This study aimed to estimate the genetic variability of these subspecies. For this purpose, 127 colonies from 15 Brazilian localities were analyzed, using nine species-specific microsatellite primers. At these loci, the number of alleles ranged from three to 15 (mean: 7.2), and the observed heterozygosity (H_o) ranged from 0.03–0.21, while the expected heterozygosity (H_e) ranged from 0.23–0.47. The genetic distances among populations ranged from 0.03–0.45. The F_ST multilocus value (0.23) indicated that the populations sampled were structured, and the clustering analysis showed the formation of two subgroups and two more distant populations. The first group contained the subspecies M. quadrifasciata quadrifasciata, and the other, the subspecies M. quadrifasciata anthidioides and the two M. quadrifasciata populations with continuous metasomal stripes from northern Minas Gerais. These results confirmed that the yellow metasomal stripes alone are not a good means for correctly identifying the different subspecies of M. quadrifasciata.     

Population Genetic Structure of Astyanax scabripinnis (Teleostei, Characidae) from an Urban Stream

Despite the great anthropogenic interference on urban streams, information is still scarce about the genetic variability and structure of native fish populations inhabiting such streams. In the present study, random amplified polymorphic DNA (RAPD) markers were used to analyze genetic variability and structure of populations assigned to the Neotropical fish species Astyanax scabripinnis from an urban stream located in Londrina, Paraná State, southern Brazil. Thirty individuals of this species were collected from three sites throughout the upper Cambé stream. A total of 10 primers amplified 159 loci, of which 128 (80.5%) were polymorphic. Each of the three populations showed very similar proportions of polymorphic loci, which ranged from 63.5 to 64.8%. Unbiased genetic distances varied from 0.0612 to 0.0646. Theta_p-test values indicated moderate to high genetic differentiation among individuals from different localities. The number of migrants varied from 1.34 to 1.46, suggesting a low gene flow between populations. The genetic similarity among all individuals studied ranged from 0.424 to 0.848. The results suggest that populations of A. scabripinnis in Cambé stream are undergoing genetic differentiation.     

Parallel speciation in Astyanax cave fish (Teleostei) in Northern Mexico

We investigated differentiation processes in the Neotropical fish Astyanax that represents a model system for examining adaptation to caves, including regressive evolution. In particular, we analyzed microsatellite and mitochondrial data of seven cave and seven surface populations from Mexico to test whether the evolution of the cave fish represents a case of parallel evolution. Our data revealed that Astyanax invaded northern Mexico across the Trans-Mexican Volcanic Belt at least three times and that populations of all three invasions adapted to subterranean habitats. Significant differentiation was found between the cave and surface populations. We did not observe gene flow between the strongly eye and pigment reduced old cave populations (Sabinos, Tinaja, Pachon) and the surface fish, even when syntopically occurring like in Yerbaniz cave. Little gene flow, if any, was found between cave populations, which are variable in eye and pigmentation (Micos, Chica, Caballo Moro caves), and surface fish. This suggests that the variability is due to their more recent origin rather than to hybridization. Finally, admixture of the young Chica cave fish population with nuclear markers from older cave fish demonstrates that gene flow between populations that independently colonized caves occurs. Thus, all criteria of parallel speciation are fulfilled. Moreover, the microsatellite data provide evidence that two co-occurring groups with small sunken eyes and externally visible eyes, respectively, differentiated within the partly lightened Caballo Moro karst window cave and might represent an example for incipient sympatric speciation.     

Genetic and morphological evidence for reproductive isolation between sympatric populations of Galaxias (Teleostei: Galaxiidae) in South Island, New Zealand

New Zealand's South Island houses a flock of closely related stream-resident fish taxa (Galaxias vulgaris sensu lato), including a number of species recently described on the basis of subtle morphological differences. The taxonomic status of some members of the species complex remains uncertain. This study examines the degree of reproductive isolation between recently recognized morphotypes from Southland (G. 'southern', flatheads; G. gollumoides, roundheads) which co-occur in Bushy Creek, a tributary of the Mataura R. Although these morphotypes are broadly sympatric in Southland and Stewart Island, Bushy Creek is their only documented zone of contact. Molecular (microsatellite, isozyme and mtDNA markers) and morphological analyses of 139 fish samples across a 500-m transect (seven stations) reveal a cline from predominantly G. 'southern' (N=85) to predominantly G. gollumoides (JV=54), corresponding with a gradual increase in stream gradient. Multivariate analyses of genotypic and morphological data independently reveal distinct clusters that are completely congruent with mtDNA type, suggesting an absence of mtDNA introgression. Our data support the separate species status of G. 'southern' and G. gollumoides under both biological and phylogenetic species concepts. We suggest that the speciation of these taxa occurred in allopatry through independent losses of diadromy, with sympatry resulting from secondary contact.     

Genetic differentiation between close eastern Mediterranean Dicentrarchus labrax (L.) populations

Differentiation at nine microsatellite loci revealed that a Levantine Basin sea bass Dicentrarchus labrax population probably represents a further subdivision of this species in the eastern Mediterranean.     

Gonadal morphology and difference in reproductive development of two isolated populations of Astyanax rivularis (Teleostei,Characidae)

Individuals of the same species may present different reproductive tactics depending on the environment in which they develop and mature. The present study aimed to define the gonadal development phases of males and females of Astyanax rivularis and to carry out a comparative analysis of the reproductive development of specimens captured in two isolated environments of the São Francisco River basin in Serra da Canastra, Brazil (Point 1: low vegetation and river showing calm and crystalline waters with small well formations; Point 2: current waters, and well-established areas of arboreal vegetation). Thus, the gonads of A. rivularis specimens were collected, fixed and processed with techniques for light microscopy. Five maturation phases of the females' reproductive cycle were established: immature, developing, spawning capable, regressing and regenerating. Three maturation phases of the males' reproductive cycle were observed: spawning capable, regressing, and regenerating. There are differences in the phases of gonadal development of A. rivularis between the two sampling points so that, possibly, animals upstream of the waterfall demonstrate a delay in the reproductive cycle in relation to animals downstream.     

Genetic divergence and a hybrid zone between Baltic and North Sea Mytilus populations (Mytilidae: Mollusca)

Populations of the common mussel ( Mytilus edulis ) from the North Sea area (Skakerrak-Kattegat) and those from the Baltic Sea are almost diagnostically differentiated at five out of 22 studied allozyme loci; at a further seven loci, alleles predominant or common in one area are nearly absent in the other. Genetic distance was estimated at 0.28; this is similar to the distances of these populations to the Mediterranean mussel M. galloprovincialis. The three mussel types obviously represent equal evolutionary divergence from one another, and should also be taxonomically equally separated; a semispecies rank within a more comprehensive M. edulis complex or superspecies is suggested. The age of the Baltic mussel type ( 'M. trossulus' ), as an independent evolutionary lineage, is probably far greater than that of the post-glacial Baltic Sea.
Allele frequencies change gradually and in parallel when entering from the Kattegat through the Sound into the Baltic. Only a slight Wahlund effect at the strongly diverged Gpi and Pgm loci was found in intermediate populations, indicating that extensive hybridization of the two taxa takes place in the area. However, strong interlocus genotypic associations suggest that selection against hybrids is intense in later generations; the c. 100 km wide hybrid zone is narrow relative to the dispersal distance. The genotypic structure of the Lap locus does not conform with those of the other loci studied in the hybrid zone; it cannot be viewed merely as a neutral marker of the process of hybridization.     

Determining Management Units in Southeastern Brazil: The Case of Astyanax bimaculatus (Linnaeus, 1758) (Teleostei: Ostariophysi: Characidae)

Characterization, management and protection of biodiversity are the most prominent challenges in conservation biology. Analyses on molecular similarity (Random Amplified Polymorphic DNA-Polymerase Chain Reaction, RAPD-PCR) and morphologic patterns (morphometric and meristic characters) were performed to evaluate the existence of population structuring in the Doce river basin, in a widespread small-sized characin, Astyanax bimaculatus. Three hundred (300) individuals were collected from six locations isolated by waterfalls or dams, in the Doce river basin, Minas Gerais State, Brazil. Genetic differentiation among tributaries was significant (p<0.00001) and accounted for 21% of total variance (p<0.00001). Only the Grande waterfall of the Casca river was an effective factor for differentiation of populations (p<0.00001), and meristic characters were also consistent with molecular data. We concluded that genetic and morphologic variation of this species was not necessarily associated with waterfalls and that molecular and meristic data are effective predictors of population divergence in this basin. The consequences of these findings for the protection and management of aquatic biodiversity were discussed.     

Non-optical releasers for aggressive behavior in blind and blinded Astyanax (Teleostei, Characidae)

Prior to this study, it was believed that epigean and hypogean Astyanax differ markedly in their display of agonistic behavior. Research suggested that surface-dwelling individuals were extremely aggressive whereas their blind, cave-dwelling counterparts tended to show little or no aggressive behavior. Aggression in Astyanax was thought to be triggered by visual stimuli because surface fish in a dark environment or surface fish blinded late in life did not show aggression. Here, we demonstrate that surface fish blinded early on in their embryonic development are highly aggressive as adults. We also report the first case of a population of blind cave-dwelling Astyanax that is highly aggressive. We conclude that reduced aggression is not the only evolutionary pathway for troglobitic Astyanax and that there is some degree of developmental plasticity in the releaser of aggression and in the selection of its triggering stimuli.     

Characterization of microsatellite loci for the Amazonian rummy‐nose tetra,Hemigrammus bleheri (Teleostei,Characidae)

The rummy‐nose tetra, Hemigrammus bleheri, is a small fish found in forest streams of the Rio Negro floodplain, central Amazonia. This species is popular in aquaria and used as an ornamental fishery resource by the riverine people from middle Rio Negro. Here we describe eight microsatellite DNA loci for rummy‐nose tetras. Number of alleles and heterozygosity per locus in a sample of 30 fish ranged from five to 22 and from 0.52 to 0.96, respectively. These highly variable genetic markers provide important tools for investigating population history and identifying conservation units in rummy‐nose tetras.     

Genetic admixture of burbot (Teleostei: Lota lota) in Lake Constance from two European glacial refugia

The burbot, Lota lota, is the only freshwater species of the codfish family and has a Holarctic distribution. Pleistocene glaciations caused significant geographical differentiation in the past, but its life history characterized by winter spawning migrations over large distances is likely to homogenize populations by contemporary gene flow. We investigated the population genetic structure of 541 burbots from Lake Constance and adjacent Rhine and Danube tributaries in Europe using the entire mitochondrial DNA (mtDNA) control region and 11 microsatellites. Microsatellites revealed considerable population divergence (F(ST) = 0.26) and evidenced recent bottlenecks in two Central European rivers. In accordance to previous evidence two main phylogeographic lineages (Atlantic and Danubian) were found co-occurring at similar frequencies in Lake Constance, where they currently undergo random mating as indicated by microsatellites. The Danubian lineage contributed only a small proportion to the lake's mtDNA diversity, and probably expanded within the lake shortly after its formation approximately 10,000-15,000 BP. The larger Atlantic haplotype diversity suggested a population expansion older than the lake itself. Levels of admixture at microsatellite loci were less obvious due to their high variability, and coalescence methods were used to estimate past admixture proportions. Our results reinforce a model of a two-step colonization of Europe by burbot from an ancestral Danubian refuge, and confirm the persistence of a secondary Atlantic refuge, as proposed to exist for other freshwater fish. We conclude that the present-day burbot population in Lake Constance bears the genetic signature of both contemporary gene flow and historical separation events.     

Genetic changes associated to declining populations of Formica ants in fragmented forest landscape

We monitored populations of two wood ant species, Formica aquilonia and Formica lugubris, through annual mapping of the colonies in a fragmenting forest landscape from 1966 to 1998. The genetic population structure was studied at the end of the study period by using 12 microsatellite loci. Fragmentation of forest led to a decline and spatial redistribution of populations. Changes in the spatial distribution were particularly pronounced in the highly polygynous (many queens in a single nest) species F. aquilonia, whose local populations declined or became extinct, or relocated themselves and colonized new patches. The genetic relationships of the remaining subpopulations indicated the historical developments, revealing the boundaries of the historical populations (high values of genetic differentiation, F(ST)), recolonization histories (genetic affinities revealed by Bayesian analyses) and population decline (reduced variation). Big genetic differences could be detected over short distances, so differentiation also depended on social factors. Our results showed that a genetic study can be reliably used to dissect the recent historical changes underlying the present population structure, and that species with different social structures can respond differently to habitat changes. Combining our demographic and genetic results suggests that habitat fragmentation forms a clear threat on a local scale with large negative effects on ant population viability.     

Phylogenetic evidence of mitochondrial DNA introgression among pocket gophers in New Mexico (family Geomyidae)

Mitochondrial DNA (mtDNA) variation in the cytochrome b gene was determined for two divergent taxa of pocket gophers, Thomomys bottae actuosus and T. b. ruidosae . These two taxa hybridize in a narrow contact zone, but introgression of nuclear markers such as allozymes or chromosomes does not extend much beyond the hybrid zone (Patton et al. 1979). We found that despite their distinctness, the two subspecies shared very similar mtDNA haplotypes. By a comparison of phylogenetic histories derived from nuclear markers (allozymes) and from mtDNA haplotypes sampled in different populations of T. bottae from New Mexico, we show that apparent similarity is due to an introgression of T. b. ruidosae mtDNA into T. b. actuosus nuclear background. Evidence of introgression is not limited to the present-day contact zone between these two taxa, but extends at least 75 km away from it. The actuosus haplotype coexists along with the ruidosae mtDNA in the Gallinas Mts., which are inhabited by otherwise pure T. b. actuosus , while further north only typical actuosus haplotypes were detected. Of several potential mechanisms which could lead to such a geographical pattern of variation, we argue that a combination of range shifts due to climatic fluctuations, and genetic drift are most likely. Horizontal gene transfers due to hybridization are historical events which seem rather common among pocket gophers. Although they can be identified with careful phylogenetic study using independent data sets, the potential for misinterpreting a gene tree as an organismal tree is great in this and other groups of animals.     

Gill‐derived glands in species of Astyanax (Teleostei: Characidae)

The presence of a gill‐derived gland is herein reported for the first time in males of species of Astyanax and related genera; they are described through histological cuts and SEM. The gill‐derived glands described for the Characidae, when fully developed, present a similar structure in different species. The main external feature of gill‐derived glands is the fusion of anteriormost gill filaments on the ventral branch of first gill arch. This fusion is caused by squamous stratified epithelial tissue that covers adjacent filaments, forming a series of chambers. In the region where the gill‐derived gland develops, the secondary lamellae of the gill filaments are much reduced or completely atrophied being characterized by the presence of glandular cells forming nests.     

Population genetic patterns revealed by microsatellite data challenge the mitochondrial DNA based taxonomy of Astyanax in Mexico (Characidae, Teleostei)

Astyanax has become an important model system for evolutionary studies of cave animals. We investigated correlations of population genetic patterns revealed by microsatellite data and phylogeographic patterns shown by mitochondrial DNA sequences in Mexican cave and surface fish of the genus Astyanax (Characidae, Teleostei) to improve the understanding of the colonization history of this neotropical fish in Central and North America and to assess a recent taxonomic classification. The distribution of nuclear genotypes is not congruent with that of the mitochondrial clades. Admixture analyses suggest there has been nuclear gene flow between populations defined by different mitochondrial clades. The microsatellite data indicate that there was mitochondrial capture of a cave population from adjacent populations. Furthermore, gene flow also occurred between populations belonging to different nuclear genotypic clusters. This indicates that neither the nuclear genotypic clusters nor the mitochondrial clades represent independent evolutionary units, although the mitochondrial divergences are high and in a range usually characteristic for different fish species. This conclusion is supported by the presence of morphologically intermediate forms. Our analyses show that the Trans-Mexican Volcanic Belt limited gene flow, but has been crossed by Astyanax several times. In Yucatán, where obvious geographic barriers are missing, the incongruence between the distribution of nuclear and mitochondrial markers reflects random colonization events caused by inundations or marine transgressions resulting in random phylogeographic breaks. Thus, conclusions about the phylogeographic history and even more about the delimitation of species should not be based on single genetic markers.     

Genetic variability in five populations of Partamona helleri (Hymenoptera, Apidae) from Minas Gerais State, Brazil

Partamona is a Neotropical genus of stingless bees that comprises 33 species distributed from Mexico to southern Brazil. These bees are well-adapted to anthropic environments and build their nests in several substrates. In this study, 66 colonies of Partamona helleri from five localities in the Brazilian state of Minas Gerais (São Miguel do Anta, Teixeiras, Porto Firme, Viçosa and Rio Vermelho) were analyzed using nine microsatellite loci in order to assess their genetic variability. Low levels of observed (H_o = 0.099-0.137) and expected (H _e = 0.128-0.145) heterozygosity were encountered and revealed discrete genetic differentiation among the populations (F _ST = 0.025). AMOVA further showed that most of the total genetic variation (94.24%) in P. helleri was explained by the variability within local populations.     

Genetic structure of Mesoamerican populations of Big-leaf mahogany (Swietenia macrophylla) inferred from microsatellite analysis

While microsatellites have been used to examine genetic structure in local populations of Neotropical trees, genetic studies based on such high-resolution markers have not been carried out for Mesoamerica as a whole. Here we assess the genetic structure of the Mesoamerican mahogany Swietenia macrophylla King (big-leaf mahogany), a Neotropical tree species recently listed as endangered in CITES which is commercially extinct through much of its native range. We used seven variable microsatellite loci to assess genetic diversity and population structure in eight naturally established mahogany populations from six Mesoamerican countries. Measures of genetic differentiation (FST and RST) indicated significant differences between most populations. Unrooted dendrograms based on genetic distances between populations provide evidence of strong phylogeographic structure in Mesoamerican mahogany. The two populations on the Pacific coasts of Costa Rica and Panama were genetically distant from all the others, and from one another. The remaining populations formed two clusters, one comprised of the northern populations of Mexico, Belize and Guatemala and the other containing the southern Atlantic populations of Nicaragua and Costa Rica. Significant correlation was found between geographical distance and all pairwise measures of genetic divergence, suggesting the importance of regional biogeography and isolation by distance in Mesoamerican mahogany. The results of this study demonstrate greater phylogeographic structure than has been found across Amazon basin S. macrophylla. Our findings suggest a relatively complex Mesoamerican biogeographic history and lead to the prediction that other Central American trees will show similar patterns of regional differentiation.     

Genetics and hybridization in surface and cave Astyanax (Teleostei): a comparison of regressive and constructive traits

Change in ecological conditions, as seen in surface and cave populations of Astyanax (Teleostei), has caused the divergent evolution of a large number of traits like eyes, coloration, taste, lateral line, and different kinds of behaviour like schooling, sleep or feeding posture. Because of the interfertility of surface and cave forms these fish are an exceptional object to study the morphological and genetic basis of the evolution of such complex regressive and constructive traits. Classical crossing analyses and genomic studies are contributing to growing understanding. Both kinds of traits mostly rely on multiple genetic bases and the phenotypic manifestation in the various crosses is similar. The gene effect underlying the phenotypic manifestation may exhibit an exponential increase at differing amounts in the various traits and crosses. Missing or presence of such genetic interaction helps determine whether the variability of eyes or pigmentation exhibited by Astyanax cave fish populations like Micos, is due to a more recent origin or to secondary hybridization with the surface fish. Neither crossing analysis nor QTL mapping revealed that eye reduction is pleiotropically antagonistically related to the increase of taste buds or lateral line sense. Independent inheritance of traits suggests that Astyanax cave fish are subjected to mosaic evolution.     

Genetic variability and divergence between populations and species of Nesticus cave spiders

Genetic variability and divergence at 21 enzyme loci were studied in and between Italian populations of the cave spiders Nesticus eremita (13 populations), N. menozzii and N. sbordonii (one population each). The three species differ with respect to the degree of specialization to cave life, dispersion ability, isolation of populations, abundancy, extent of the distribution area, and range from the troglophilic and widespread N. eremita to the troglobitic N. sbordonii, endemic to a single cave in the Central Appennines.Heterozygosity ranges from 0.05 to 0.15 in N. eremita populations and appears to be largely controlled by the occurrence and the extent of gene flow among populations. The relatively low polymorphism levels of N. menozzii (H=0.081) and N. sbordonii (H=0.106) are also associated with reduced gene flow and small population sizes.Genetic distances between N. eremita populations vary considerably and are strictly related to the geographical distances involved, again indicating a major role of gene flow in determining the patterns of genetic differentiation between populations. This view is strongly supported by the results of a principal component analysis applied to the gene-frequency data. Estimates of genetic divergence between species suggest that the major cladogenetic events leading to complete separation of these three Nesticus species occurred in the Middle-late Pliocene.