The evolution of strong reproductive isolation between sympatric intertidal snails

The evolution of strong reproductive isolation (RI) is fundamental to the origins and maintenance of biological diversity, especially in situations where geographical distributions of taxa broadly overlap. But what is the history behind strong barriers currently acting in sympatry? Using whole-genome sequencing and single nucleotide polymorphism genotyping, we inferred (i) the evolutionary relationships, (ii) the strength of RI, and (iii) the demographic history of divergence between two broadly sympatric taxa of intertidal snail. Despite being cryptic, based on external morphology, Littorina arcana and Littorina saxatilis differ in their mode of female reproduction (egg-laying versus brooding), which may generate a strong post-zygotic barrier. We show that egg-laying and brooding snails are closely related, but genetically distinct. Genotyping of 3092 snails from three locations failed to recover any recent hybrid or backcrossed individuals, confirming that RI is strong. There was, however, evidence for a very low level of asymmetrical introgression, suggesting that isolation remains incomplete. The presence of strong, asymmetrical RI was further supported by demographic analysis of these populations. Although the taxa are currently broadly sympatric, demographic modelling suggests that they initially diverged during a short period of geographical separation involving very low gene flow. Our study suggests that some geographical separation may kick-start the evolution of strong RI, facilitating subsequent coexistence of taxa in sympatry. The strength of RI needed to achieve sympatry and the subsequent effect of sympatry on RI remain open questions. This article is part of the theme issue ‘Towards the completion of speciation: the evolution of reproductive isolation beyond the first barriers'.

Karyotype Characterization of Nine Periwinkle Species (Gastropoda, Littorinidae)

Periwinkles of the family Littorinidae (Children, 1834) are common members of seashore littoral communities worldwide. Although the family is composed of more than 200 species belonging to 18 genera, chromosome numbers have been described in only eleven of them. A molecular cytogenetic analysis of nine periwinkle species, the rough periwinkles Littorina arcana, L. saxatilis, and L. compressa, the flat periwinkles L. obtusata and L. fabalis, the common periwinkle L. littorea, the mangrove periwinkle Littoraria angulifera, the beaded periwinkle Cenchritis muricatus, and the small periwinkle Melarhaphe neritoides was performed. All species showed diploid chromosome numbers of 2n = 34, and karyotypes were mostly composed of metacentric and submetacentric chromosome pairs. None of the periwinkle species showed chromosomal differences between male and female specimens. The chromosomal mapping of major and minor rDNA and H3 histone gene clusters by fluorescent in situ hybridization demonstrated that the patterns of distribution of these DNA sequences were conserved among closely related species and differed among less related ones. All signals occupied separated loci on different chromosome pairs without any evidence of co-localization in any of the species.

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.

Seasonal patterns in the reproductive activity of barnacle-dwelling littorinids

The reproductive activity of a population of barnacle-dwelling littorinids at Peak Steel, on the north-east coast of England, was investigated. Variation in the number of mature males and females, expressed as a percentage of the whole population, showed a marked difference among the barnacle-dwelling littorinids. Littorina neglecta displayed a distinct seasonal component to its reproductive activity, with a peak in early summer. Littorina arcana B also displayed a distinct seasonal component, with a lull in the summer months. However, L. saxatilis B maintained its reproductive activity all year, but with an increase in output in the summer. The reproductive activity of both L. arcana B and L. saxatilis B follow similar patterns of reproductive activity as their typical non barnacle-dwelling forms, suggesting that these are true barnacle-dwelling ecotypes. The controversy surrounding the specific status of L. neglecta is also discussed.

Synchronous Penis Shedding in the Rough Periwinkle, Littorina Arcana

Penis size and morphology of an oviparous species of rough winkle Littorina arcana Hannaford Ellis (Gastropoda: Prosobranchia) was studied at Robin Hoods Bay on the English east coast. Monthly collections were made over a two year period from August 1987 to August 1989 and again from May to December 1993. Number of penial glands was recorded and a six point scale was used to quantify penis maturity. Results indicate that male L. arcana shed their penes in summer and regenerate a new one during the following 12–20 weeks. Male L. arcana in captivity shed penes in July but not in November. The significance of penis shedding and the differences in reproductive seasonality between L. arcana and its ovoviviparous sibling species L. saxatilis is discussed.

Patterns of variation in Aat-1 allele frequencies in rough periwinkles (Littorina) suggest similar selection regimes rather than conspecificity

It has been argued by Johannesson & Johannesson (1990b) that the pattern of common genetic variation in Littorina saxatilis (Olivi) and Littorina neglecta Bean indicates gene flow in sympatric populations of these ‘species’. They suggest that L. neglecta is not a good species. We report that a gene diversity analysis of polymorphism in Aat-1 suggests that L. saxatilis and Littorina arcana Hannaford Ellis also show common features in allele frequency distribution. This casts doubt on the argument of Johannesson & Johannesson (1990b) from enzyme polymorphism data on Aat-1 that L. neglecta may simply be an ecotype of L. saxatilis. It seems likely that the similarities are due to operation of the same selection factors on all these species.