A striking example of the founder effect in the mollusc Littorina saxatilis

Two South African populations of Littorina saxatilis were examined by starch-gel electrophoresis at 16 enzyme loci and compared with 13 populations of North Atlantic saxatilis from both American and European coasts, and with six British populations of the closely related species Littorina arcana. The South African animals showed a severely reduced heterozygosity (– 0.052) compared with Atlantic populations of saxatilis ( = 0.181), and the mean genetic distance between the two areas was high ( = 0.203) compared with distances within the North Atlantic saxatilis populations (D = 0.034). In fact, the saxatilis from South Africa were genetically more distant from the North Atlantic samples of L. saxatilis than were the arcana from British shores. The reduced genetic heterozygosity and genetic divergence of the South African populations is attributed to founder effects following a postulated recent introduction by man.     

Microgeographic variation in allozyme and shell characters in Littorina saxatilis Olivi (Prosobranchia: Littorinidae)

Twenty-three enzymes and five shell parameters were screened in 11 subpopulations of Littorina saxatilis Olivi (= L. rudis Maton) occupying different habitats over a 1 km stretch of coastline. Shell morphology varied considerably and consistently with respect to degree of exposure, and since there is evidence that such morphology is at least partly under genetic control, it is likely that natural selection selects particular genotypes at particular locations. There was significant allozyme heterogeneity between neighbouring subpopulations, sometimes only metres apart, but little of the allozyme variability could be related directly to environmental pressures. Thus, with the exception of the Odh locus, the considerable morphological differentiation between snails from exposed and sheltered sites was not reflected in differentiation of those genes coding for electrophoretically assayed enzymes. At the Odh locus, virtually all the genetic differentiation between subpopulations was attributable to differentiation between habitat types. Two loci, Sod-1 and Aat-1, showed highly significant genetic disequilibrium, and possible reasons for this are explored. The population structure as assessed electrophoretically accords well with the stepping-stone model which permits greater differentiation of neighbouring populations than the island model, and which seems realistic in the ovoviviparously reproducing L. saxatilis, where the greater part of gene flow is likely to occur through the occasional migration of adults between contiguous populations.     

Species relationships and population structure of Littorina saxatilis Olivi and L. tenebrosa Montagu in Ireland using single-strand conformational polymorphisms (SSCPs) of cytochrome b fragments

Littorina saxatilis is a ubiquitous snail of intertidal habitats in the North Atlantic. Shell type in littorinids is extremely polymorphic and defined by habitat. Taxonomy based upon shell type has been revised in the light of anatomic and genetic information, but uncertainties remain. In this study, the population structure of L. saxatilis and L. tenebrosa was studied at 11 sites in Ireland using single-strand conformational polymorphisms of a 375-bp portion of the cytochrome b gene, and the status of L. tenebrosa, the small, fragile-shelled, brackish water type, was considered. The genetic patterns among L. saxatilis and L. tenebrosa populations were examined over varying distances and L. tenebrosa was compared with adjacent L. saxatilis populations at four sites on the west coast of Ireland and one site on the east coast. Haplotype diversity was high with 32 haplotypes present among 995 individuals. Pairwise tests suggest gene flow over small scales among and between habitat types and may reflect the stochastic legacy of postglacial recolonization over larger scales. In AMOVA tests, geography explained nearly twice as much of the variance (30%) as habitat type (18%), indicating that gene flow is more restricted by distance than by habitat type, and supporting the status of L. tenebrosa as an ecotype of L. saxatilis rather than a separate species.     

A preliminary study on genetic differentiation in Littorina saxatilis from Galway Bay,Ireland: Littorina tenebrosa Montagu – a valid species or ecotype?

Littorina tenebrosa is a small fragile-shelled periwinkle which lives on permanently submerged algae in coastal lagoons and non-tidal brackish pools. This periwinkle is a member of the rough periwinkle group which also comprises Littorina saxatilis, L. arcana, L. compressa and L. neglecta and is most closely related to L. saxatilis although its exact systematic status is in some doubt. Based on its unique ecology many believe L. tenebrosa to be a valid species. However, shell morphometric and allozyme analyses on Scottish and Swedish populations of L. tenebrosa and L. saxatilis have indicated that the two periwinkles are virtually identical. Preliminary results on five allozyme loci (AAT-1, GPI, PGM-1, PGM-2 and PNP) in samples of L. tenebrosa and L. saxatilis from Golam Head, Lettermullen, and other locations on the west coast of Ireland show L. tenebrosa to be genetically differentiated from L. saxatilis At Golam Head, where opportunities for gene flow occur between the two taxa, L. tenebrosa is as genetically differentiated from local L. saxatilis as it is from L. saxatilis from more geographically distant locations.     

Phylogeographic analysis reveals a deep lineage split within North Atlantic Littorina saxatilis

Phylogeographic studies provide critical insight into the evolutionary histories of model organisms; yet, to date, range-wide data are lacking for the rough periwinkle Littorina saxatilis, a classic example of marine sympatric speciation. Here, we use mitochondrial DNA (mtDNA) sequence data to demonstrate that L. saxatilis is not monophyletic for this marker, but is composed of two distinct mtDNA lineages (I and II) that are shared with sister species Littorina arcana and Littorina compressa. Bayesian coalescent dating and phylogeographic patterns indicate that both L. saxatilis lineages originated in the eastern North Atlantic, around the British Isles, at approximately 0.64 Ma. Both lineages are now distributed broadly across the eastern, central and western North Atlantic, and show strong phylogeographic structure among regions. The Iberian Peninsula is genetically distinct, suggesting prolonged isolation from northeastern North Atlantic populations. Western North Atlantic populations of L. saxatilis lineages I and II predate the last glacial maximum and have been isolated from eastern North Atlantic populations since that time. This identification of two distinct, broadly distributed mtDNA lineages further complicates observed patterns of repeated incipient ecological speciation in L. saxatilis, because the sympatric origins of distinct ecotype pairs on eastern North Atlantic shores may be confounded by admixture of divergent lineages.     

Glacial history of the North Atlantic marine snail, Littorina saxatilis, inferred from distribution of mitochondrial DNA lineages

The North Atlantic intertidal gastropod, Littorina saxatilis (Olivi, 1792), exhibits extreme morphological variation between and within geographic regions and has become a model for studies of local adaptation; yet a comprehensive analysis of the species' phylogeography is lacking. Here, we examine phylogeographic patterns of the species' populations in the North Atlantic and one remote Mediterranean population using sequence variation in a fragment of the mitochondrial cytochrome b gene (607 bp). We found that, as opposed to many other rocky intertidal species, L. saxatilis has likely had a long and continuous history in the Northwest Atlantic, including survival during the last glacial maximum (LGM), possibly in two refugia. In the Northeast Atlantic, several areas likely harboured refugial populations that recolonized different parts of this region after glacial retreat, resulting in strong population structure. However, the outlying monomorphic Venetian population is likely a recent anthropogenic introduction from northern Europe and not a remnant of an earlier wider distribution in the Mediterranean Sea. Overall, our detailed phylogeography of L. saxatilis adds an important piece to the understanding of Pleistocene history in North Atlantic marine biota as well as being the first study to describe the species' evolutionary history in its natural range. The latter contribution is noteworthy because the snail has recently become an important model species for understanding evolutionary processes of speciation; thus our work provides integral information for such endeavours.     

Distribution and biomass of microphytes measured by benthic chlorophyll a in a tropical lagoon (New Caledonia,South Pacific)

Starch-gel electrophoretic techniques were applied to the investigation of molecular genetic variation in populations of the rough periwinkle Littorina saxatilis. The investigation comprised two phases: a) technique development to resolve as many as possible of the allozyme loci reported in the literature as having been screened in the genus Littorina and in Melarhaphe neritoides; b) the use of these loci to assess levels of genetic variation in and patterns of genetic differentiation among populations of L. saxatilis from a relatively isolated group of populations from Galway Bay, Ireland. More than 43 allozyme loci (of which four were screened for the first time here in this species), coding for 37 enzymes, were investigated and thirteen of these loci (including two loci screened for the first time here) were found to be variable and reliably scorable. Samples from five pairs of transects were collected from Inismór, Aran Islands, from sites with known exposure levels; one transect within each pair was collected from an exposed site and the other from a nearby, but relatively sheltered site. UPGMA for eleven loci, (ARK and PGDH were excluded from cluster and FST analysis as they were unscorable in a few samples), showed that the samples cluster mostly by pair, reflecting their geographic origin and is indicative of little gene flow between populations. Levels of population differentiation were high among samples from the top of the shore, but unusually so at AAT-1 which showed nearly three times the mean FST value for the eleven loci. There was also a significant regression of frequency of AAT-1100 against level of exposure. In addition, among midshore samples, there was a consistently higher frequency of AAT-1100 in sheltered habitats. These results support the findings of others, indicating that this locus may be subject to natural selection.     

The taxonomy of the Littorina saxatilis speicies-complex, with particular reference to the systemaitc status of Littorina patula Jeffrys

Variation in shell shape and penis morphology of Littorina rudis Maton is examined using data from all parts of Britain. The shell shape variation within populations of L. rudis is shown to account for Liltorina patula Jeffrys at the only site where the latter species was recorded. In addition, the shell shape of I,. rudis varies with exposure, individuals on exposed shores having a relatively larger aperture than those on sheltered shores. Wave action and desiccation are considered the most likely factors maintaining this variation. The penis morphology of L. rudis varies within and between shores to an extent that renders the use of this character invalid for distinguishing L. patula from L. rudis. The radulae of adults of L. rudis, L. patula and Littorina nigrolineata (Gray) are similar in structure having blunt cusps, whilst adult Littorina neglecta Bean and juvenile L. rudis have pointed cusps. The possibility of a neotenous origin of L. neglecta from L. rudis is discussed. On the evidence presented here it is suggested that L. patula must be regarded as a synonym of L. rudis.     

Assortative mating between two distinct micro-allopatric populations of Littorina saxatilis (Olivi) on the northeast coast of England

Size assortative mating is a common invertebrate mating pattern and is usually accompanied by male and female sexual selection, and these three behaviours can contribute to reproductive isolation. Two distinct populations of the marine prosobranch Littorina saxatilis, H and M, occur within 15 m of each other on the same shore. Previous studies have demonstrated that these two forms have different reproductive strategies and that the rare hybrids between the two forms show evidence of reproductive dysfunction and hence are less fit than the assumed parental forms. In both populations, female shell height was shown to be a predictor of the number of embryos contained within the brood pouch. The mean shell height of the M population was significantly larger than that of the H population, and the M population matures at a larger shell size than the H population. The two populations show complete assortative mating to type in the field, and occupy different microhabitats on the same shore. Therefore, laboratory-based experiments were performed to determine if assortative mating was maintained in sympatry and also to determine the effect of population density on mate choice. The males of both populations showed sexual selection for female size, choosing to mate with females approximately 10% larger than themselves from an assortment of female sizes. The M population showed complete assortative mating to type, irrespective of the density of H and M females, whereas at low densities the H males did occasionally mate with M females. The role of assortative mating and reinforcement (due to natural selection acting against the less fit hybrids), in maintaining the partial reproductive barrier between the two populations is discussed.     

Phenotypic plasticity in two marine snails: constraints superseding life history

In organisms encountering predictable environments, fixed development is expected, whereas in organisms that cannot predict their future environment, phenotypic plasticity would be optimal to increase local adaptation. To test this prediction we experimentally compared phenotypic plasticity in two rocky-shore snail species; Littorina saxatilis releasing miniature snails on the shore, and Littorina littorea releasing drifting larvae settling on various shores, expecting L. littorea to show more phenotypic plasticity than L. saxatilis. We compared magnitude and direction of vectors of phenotypic difference in juvenile shell traits after 3 months exposure to different stimuli simulating sheltered and crab-rich shores, or wave-exposed and crab-free shores. Both species showed similar direction and magnitude of vectors of phenotypic difference with minor differences only between ecotypes of the nondispersing species, indicating that plasticity is an evolving trait in L. saxatilis. The lack of a strong plastic response in L. littorea might be explained by limits rather than costs to plasticity.     

Gills and size in some small Littorina

Gills were examined in Littorina saxatilis, L. arcana and L. neglecta, using animals of similar size. The form of the gills in the three groups is very similar, although there is an indication of qualitative differences between them. The relationship of number of gill lamellae to shell length shows that L. saxatilis and L. arcana share the same allometric relationship, while in L. neglecta the slope is the same but the intercept is less: at equivalent shell sizes, these animals have about three fewer lamellae.     

Rough periwinkle polymorphism on the east coast of Yorkshire: comparison of RAPD-DNA data with morphotype

A genetic analysis of morphotypes of Littorina saxatilis from two locations on the north-east coast of England (Filey and Ravenscar), using randomly amplified DNA polymorphisms (RAPD) generated with a single primer, revealed quite different patterns of variation. Thin shelled, wide-apertured (H-form) animals from Ravenscar tended to cluster separately from thick shelled (M) forms, indicating genetic differentiation of these morphs. Animals of similar morphology (H and M) from Filey (about 30 km distant) did not display such an obvious pattern, and although there was still evidence of differentiation from discriminant analysis of RAPD data, levels of correct classification were reduced at Filey. This suggests that the utility of a single RAPD primer for separation of such forms varies over a relatively small distance. L. arcana from Ravenscar, included as an outgroup, were generally well differentiated from L. saxatilis and were noted to exhibit less variation, a phenomenon that has been noted previously in some allozyme and RAPD analyses. A similar RAPD analysis undertaken on small, barnacle dwelling, brooding forms from Peak Steel, Ravenscar revealed that animals appeared to have as great a tendency to cluster together on a microgeographic scale (by collection patch) as by species ( L. neglecta or L. saxatilis b) although predominance of certain species in individual patches largely explains this. Discriminant analysis of RAPD presence/absence data did correctly place over 90% of barnacle dwelling animals to their respective species, and we consider this as evidence of separate gene pools. RAPD is taken to be a useful tool for screening genetic variation in this complex of animals on a local scale when either a pre-selected informative primer is utilised or a battery of primers is used, but its efficacy may be reduced when a single primer is employed for screening animals from different shores.     

Case studies and mathematical models of ecological speciation. 3: Ecotype formation in a Swedish snail

Formation of partially reproductively isolated ecotypes in the rough periwinkle, Littorina saxatilis , may be a case of incipient nonallopatric ecological speciation. To better understand the dynamics of ecotype formation, its timescale, driving forces and evolutionary consequences, we developed a spatially explicit, individual-based model incorporating relevant ecological, spatial and mate selection data for Swedish L. saxatilis . We explore the impact of bounded hybrid superiority, ecological scenarios and mate selection systems on ecotype formation, gene flow and the evolution of prezygotic isolation. Our model shows that ecotypes are expected to form rapidly in parapatry under conditions applicable to Swedish L. saxatilis and may proceed to speciation. However, evolution of nonrandom mating had complex behaviour. Ecotype evolution was inhibited by pre-existing mating preferences, but facilitated by the evolution of novel preferences. While in many scenarios positive assortative mating reduced gene flow between ecotypes, in others negative assortative mating arose, preferences were lost after ecotype formation, preferences were confined to one ecotype or the ancestral ecotype became extinct through sexual selection. Bounded hybrid superiority (as observed in nature) enhanced ecotype formation but increased gene flow. Our results highlight that ecotype formation and speciation are distinct processes: factors that contribute to ecotype formation can be detrimental to speciation and vice versa. The complex interactions observed between local adaptation and nonrandom mating imply that generalization from data is unreliable without quantitative theory for speciation.     

Nuclear DNA restriction site polymorphisms and the phylogeny and population structure of an intertidal snail species complex (Littorina)

Primers for amplification of four novel, unlinked nuclear DNA loci, the first reported for the rough periwinkles of the genus Littorina, are described. Patterns of restriction site polymorphism for these loci are detailed within the rough periwinkles. RFLPs are not found to be diagnostic for any of the currently accepted species within this group, nor for any of the contentious subspecies, or forms, whose taxonomic status is uncertain. However, there are important differences in allele frequencies between these taxa and certain of these mirror differences detected in a previous study of the mitochondrial DNA. These allele frequency data are used to construct a phylogeny in which groupings of the three recognised species are obvious when either Nei's genetic distances or Reynold's distances are clustered. Contentious forms (L. neglecta, L. saxatilis 'b' and L. tenebrosa) do not cluster as distinct taxa, although populations of L. neglecta have important allele frequency differences from L. saxatilis. These four loci have confirmed the consensus view of Littorina phylogeny and provided important information on population structure-however four loci is insufficient for reaching definitive conclusions. Since analysis of nuclear DNA polymorphisms such as these is invaluable for analysis of phylogeny, population structure and phylogeography, identification of additional loci is considered imperative.     

Morphologic and genetic variability in the marine planktonic ciliate Laboea strobila Lohmann, 1908 (Ciliophora, Oligotrichia), with notes on its ontogenesis

Laboea strobila Lohmann, 1908 is a conspicuous oligotrich ciliate in the marine plankton. In order to compare different populations, the morphology of specimens from the Mediterranean Sea, North Sea, and Irish Sea was investigated using live observation, protargol impregnation, and scanning electron microscopy. Furthermore, the PCR-amplified products of the SSrRNA gene from a monoclonal culture of L. strobila from the Mediterranean Sea were sequenced and aligned with sequences of other oligotrichs, including a population of L. strobila from the Atlantic coast of the USA. Finally, the data from the ecological literature were summarized and the cultivation methods were described. The SSrRNA gene sequences of the two distantly located L. strobila populations from the North Atlantic are identical. Likewise, the morphometrics of most populations so far investigated after protargol impregnation (i.e. from the North Atlantic) do not show obvious differences. In all computed phylogenetic trees, L. strobila groups with Strombidium species, forming a monophyletic taxon corresponding to the subclass Oligotrichia. These results are corroborated by the ontogenetic comparison. Since no type species was fixed for Laboea Lohmann, 1908, L. strobila was designated in the present paper.     

South African populations of Littorina rudis

Littorina rudis was collected from stands of Spartina capensis in saltmarsh habitats within Langebaan and knysna lagoon, Cape Province. It was absent from rocky sores and from saltmarshes east of Knysna. The shell, radular and penial morphology are described. L. rudis from both localities bore tessellated colour patterns similar to those of British saltmarsh ecotypes. The Knysna population contained individuals with a unique amber background colour. The shells from Knysna and Langebaan were very thin in accordance with the sheltered habitats. The number of embryos carried in the brood puch was similar in both countries. Sexual maturity was attained at about 4 mm shell length; the male and female modal sizes were 4.9 mm and 5.1 mm respectively at Langebaan. No. published records were found for L. rudis in South West Africa. Some possible origins of the South African L. rudis populations are discussed.     

Sexual selection and assortative mating by size and their roles in the maintenance of a polymorphism in Swedish Littorina saxatilis populations

Two independent components of mating behaviour, sexual selection and assortative mating, were studied in two allopatric morphs, one sheltered boulder shore form (S-morph) and one exposed cliff shore form (E-morph), of Littorina saxatilis from the west coast of Sweden. Sexual selection was studied by comparing the sizes of copulating and non-copulating snails in the field. Size assortative mating was studied by collecting copulating pairs in the field, while assortative mating between morphs was investigated by bringing the pure morphs together in intermediary habitats and then noting the matings. The S-morph mated randomly in relation to size in two of the studied populations and exhibited a trend towards size assortative mating in a third, while the E-morph showed size assortative mating in both studied populations. The microdistribution of sizes of snails on the shores could not explain all the size assortative mating found, and instead it is argued that a size-based mate rejection behaviour also contributes to the assortative mating in at least some of these populations. There was sexual selection on size in both males and females in the S-morph, with large individuals being favoured as mates. In contrast, copulating snails of the E-morph were smaller than non-copulating ones. The significantly different sexual selection intensities between the two morphs may help to explain the size differences between them. There was random mating between the E- and the S-morphs of L. saxatilis, which suggests no incipient reproductive isolation between morphs on Swedish rocky shores. This is in agreement with earlier studies of Swedish populations, but is in contrast to the situation found in other geographical areas.     

Genetic and morphological divergence between populations of the flatfish Platichthys flesus (L.) (Pleuronectidae)

Populations of the flounder, Platichthys flesus , were screened for electrophoretically detectable protein variation at up to 37 loci. Atlantic and North Sea populations (subspecies flesus ) were genetically very similar to one another ( I >0.99) but different from Adriatic (subspecies italicus ) and Black Sea (subspecies luscus ) populations. The values for genetic identity between subspecies were around 0.9. Diagnostic loci enabled specimens of flesus, italicus and luscus to be differentiated from one another. Samples of flesus showed two to three times the heterozygosity levels of italicus or luscus , consistent with the greater population size of flesus . Morphological comparisons enabled populations to be categorized to subspecies. Platichthys flesus italicus and luscus are valid subspecies: the terms are not synonyms. The genetic distance data provide an estimate of divergence time of the three subspecies at about 2 My ago, an estimate that seems reasonable in the light of present knowledge concerning the recent geological histories of the Mediterranean and Black Sea basins.     

Microgeographic variation in shell characters of Littorina saxatilis Olivi—a question mainly of size?

The question of whether there are shape differences between populations of Littorina saxatilis living in different environments is examined by multivariate analyses of 13 morphological characters. Principal component analysis reveals that morphologic differences between populations from habitats with contrasting degrees of wave exposure are mainly due to a general size factor, including shell thickness. Utilizing the group structure among the snails, canonical variate analysis discloses that the main character excluding size that influences subpopulation differentiation is pointedness.     

Allozyme and shell variation in two marine snails (Littorina, Prosobranchia) with different dispersal abilities

As adults Littorina littorea and L. saxatilis have overlapping distributions, both on the shore and on a geographical scale, this implies that they are affected by similar selective forces to a large extent. The two species have, however, different reproductive strategies. Littorina littorea has a planktonic larval stage lasting 4–6 weeks, whereas L. saxatilis has direct development. This influences dispersal rates of the two species, and gene flow and effective population size are both assumed to be much smaller in L. saxatilis than in L. littorea. Intraspeciflc variation in morphological and allozyme characters differs between the two species. Littorina littorea is homogeneous for both types of characters, both within and between populations, while populations of L. saxatilis may show pronounced differences in shell characters with micro-environment changes, and allozyme variation is markedly higher in this species too. The polymorphism in shell characters of L. saxatilis and the monomorphism of L. littorea, may be expected from theory as a consequence of the different dispersal strategies. Also the lack of divergence between geographically separate populations of L. littorea, but the high degree of between population differentiation in L. saxatilis, could be attributed to the high and low dispersal rates, respectively. However, the larger allozyme variation within subpopulations of L. saxatilis compared to that of L. littorea, is surprising as the former species has a smaller effective population size than the latter. Furthermore, there is no correlation between dispersal ability and overall allozyme heterozygosity, when 10 additional species of Littorina are included in the comparison. It is suggested that different evolutionary backgrounds of different species will possibly explain this, as neither selectionist nor neutralist models alone account for the observation. That is, an earlier passage through a population bottleneck may, for example, influence the genetic variation for a long period of time.