Genetic differentiation and reproductive incompatibility among Baja California populations of the copepod Tigriopus californicus

Populations of the harpacticoid copepod Tigriopus californicus were sampled from five sites from San Diego, California, to Playa Altamira, Baja California, Mexico. Allozyme analyses revealed that all the study populations are sharply differentiated genetically. At the extreme, two populations, Punta Baja and Playa Altamira, have no alleles in common at the seven allozyme loci studied. All pairwise interpopulation crosses successfully produced F₂ hybrids except those involving the Playa Altamira population. All crosses using Playa Altamira females failed to produce F₁ hybrids, while Playa Altamira males successfully produced F₁ progeny with females from all other sites. These F₁ offspring, however, were completely sterile (with San Diego and Punta Banda females) or only occasionally produced F₂ offspring (with Punta Morro and Punta Baja females). These results suggest that allopatric differentiation among Baja populations has resulted in exceptionally high levels of genetic divergence and nearly complete reproductive isolation of the Playa Altamira population, which should now be recognized at the semispecies (or perhaps sibling species) level.     

Chromosome number in ten species of the genus Tisbe (Copepoda: Harpacticoida)

Of 10 species of the genus Tisbe, 9 were shown to possess the same haploid number of 12, whereas only 1 has a haploid number of 8. The hypothesis is advanced that n=12 is the primitive condition in the genus Tisbe and that T. biminiensis with n=8, is a recent form.     

Phylogeography of the rock-pool copepod Tigriopus brevicornis (Harpacticoida) in the northern North Atlantic, and its relationship to other species of the genus

We investigated relationships among North Atlantic Tigriopus brevicornis populations and their relationships to Mediterranean T. fulvus and North American T. californicus, using crossing experiments and mitochondrial DNA sequencing. All T. brevicornis populations tested were interfertile, while interspecific crosses produced either no offspring or offspring that did not survive past the larval stage, with the exception of a few T. brevicornis × T. californicus crosses that produced mature adults. DNA sequencing of a fragment of mitochondrial cytochrome oxidase I (COI) showed that samples of T. brevicornis from Iceland, the Faroes, Ireland, Scotland and Nova Scotia formed a single shallow clade. In contrast, T. brevicornis from more southern populations in France and Portugal formed a clade with substantially greater branch lengths. Tigriopus brevicornis was monophyletic, and T. brevicornis plus Mediterranean T. fulvus were together also monophyletic. The phylogeography of T. brevicornis closely mirrored that found in T. californicus, with substantially reduced interpopulation divergence at northern latitudes. The known distribution of T. brevicornis in Iceland and the Faroes is shown and dispersal mechanisms and habitat selection briefly discussed.     

Male residence and the patterning of serum testosterone in vervet monkeys (<Emphasis Type="Italic">Cercopithecus aethiops</Emphasis>)

This study investigated hormonal and demographic processes underlying unimale and multimale mating systems in primates. Reproductive skew and challenge models of male competition provide conflicting predictions of the relationship of male residence to group composition and androgen regulation. These predictions were tested using endocrine and socioecological data from Kenyan vervet monkeys (Cercopithecus aethiops). Serum samples from 57 adult male monkeys, drawn from 19 separate groups and 4 populations, were assayed for testosterone by radioimmunoassay. Male ability to respond to conspecific challenge was assessed by their testosterone response to the capture protocol.Analyses showed that reproductive skew models were useful predictors of intergroup and interpopulation variation in male residence and T profiles. The Limited Control model of male residence was supported by positive correlations of the number of males per group with the number of females without dependent offspring, demonstrating that monopolization potential was a key determinant of male residence. Testosterone concentrations under conditions that elevated serum levels were positively correlated with infanticide risk, supporting the Concession model. Population comparisons provided evidence for increased T responsiveness where groups were predominantly unimale. Unimale populations were from sites with higher rainfall, suggesting that ecological factors contributed to population differences in male residence and T regulation.For species name, we follow the 2000 taxonomy of the IUCN/SSC Primate Specialist Groups workshop (Grubb et al. 2003)     

Gene flow among giant clam (Tridacna gigas) populations in Pacific does not parallel ocean circulation

Tridacna gigas (L.) populations were sampled from 13 locations throughout the west Pacific in 1990–1991 and screened for allozyme variation at seven polymorphic loci. Significant genetic differences were found between the few large stocks of T. gigas remaining in the world. Cluster analysis identified two major regional groups of populations: a Central Pacific group consisting of Kiribati and the Marshall Islands, and a West Pacific group comprising all other populations. Within the latter group, the Great Barrier Reef populations were significantly differentiated from those in the Solomon Islands. The main routes of gene flow did not run parallel with known major surface currents as might be expected, but crossed them. Gene exchange was limited east-west between the Central and West Pacific groups and the GBR and the Solomon Islands. The lack of correspondence between the major surface currents and the routes of gene flow suggests that the genetic structure of T. gigas reflects historical patterns of migration that no longer occur, rather than presentday dispersal. These findings raise fundamental questions concerning the origin and maintenance of genetic diversity in Pacific faunas, and have important implications for the mariculture and conservation of T. gigas.     

Tisbe (Copepoda: Harpacticoida) species from the Lagoon of Venice. I. Seasonal fluctuations and ecology

Distribution and abundance of Tisbe species were studied throughout a period of 3 years at 3 stations in the Lagoon of Venice (Italy). At 2 other stations samples were taken occasionally, as also at the Lido station (open sea). Twelve species may be considered as common inhabitants of the lagoon, and 3 species appear to be occasional transients, compared to the 9 species found at the Lido. A comparison of the physical data shows that the stations were similar in temperature and salinity, but differed primarily in pH values, sediment characteristics, algal substratum and faunal benthic community. The fact that remarkable differences in species composition and distribution of Tisbe could be observed between the various parts of the lagoon and the lagoon and the open sea, indicates that temperature and salinity are not so important for species diversity as are biotic factors. A particularly strong difference existed between samples taken at the bottom and from pilings, which constitute a very special biotope for the benthic communities in the lagoon and seem to be a preferred habitat for T. lagunaris and T. cucumariae. The data suggest that there is a certain interaction between T. holothuriae and the other species. Whenever T. holothuriae was abundant, the other species were rare, but T. clodiensis, T. dobzhanskii (Stations 1 and 2) and Tisbe sp. (Station 3) attained high relative abundance whenever T. holothuriae decreased considerably in number. T. holothuriae represents certainly the hardiest species of the lagoon, displaying a great ability to cope with stress conditions, even those present in the interior lagoon where strong fluctuations in salinity, temperature, pH and oxygen prevail.     

Reproductive isolation and reproductive output in two sympatric mussel species (Mytilus edulis, M. trossulus) and their hybrids from Newfoundland

The mussels Mytilus edulis L. and M. trossulus Gould are found sympatrically in most areas of Newfoundland, with a low frequency of hybrids. To assess the potential for reproductive isolation, we sampled mussels from three sites in an eastern Newfoundland Bay from May–October 1996 to determine if there were differences in the reproductive cycles of the two species and their natural hybrids. In mussels with sheil lengths of 38–42 mm, males and females with mature gametes were dominant in June for M. edulis and hybrids, while M. trossulus showed a lower frequency of individuals with mature gametes. M. trossulus and hybrids spawned over a prolonged period (from late spring to early autumn) compared with most M. edulis individuals that spawned over a period of 2–3 weeks in July. This asynchrony in spawning activity between the two species may partially explain the low frequency of hybrids found in previous studies of these mussel populations. Female and male hybrids between M. edulis and M. trossulus showed normal gonad development, ripening and spawning, providing an opportunity for the introgression of genes between the two species. M. trossulus had a higher reproductive output than M. edulis of similar shell length, while hybrids showed intermediate values of reproductive output. M. trossulus females produced smaller eggs than either M. edulis or hybrids. Differences in reproductive traits may partially explain the maintenance of the mussel hybrid zone in Newfoundland. Published online: 13 August 2002     

Gametic incompatibility and genetic divergence of Pacific and Kumamoto oysters,Crassostrea gigas and C. sikamea

The biological and taxonomic separation of the Pacific oyster Crassostrea gigas (Thunberg, 1793) from the Kumamoto oyster C. sikamea (Amemiya, 1928) is affirmed by three concordant lines of evidence: (1) fixed differences in 2% of a mtDNA sequence coding for large subunit rRNA; (2) a genetic distance of 0.440 based on 19 allozyme loci, including 5 diagnostic loci (Aat-1, Idh-1, Idh-2, Mpi-1, Mdh-2); (3) one-way gametic incompatibility resulting in partial reproductive isolation in interspecific crosses. C. gigas sperm x C. sikamea egg fertilizations form viable hybrid offspring, but C. sikamea sperm do not fertilize C. gigas eggs. Divergence between these two species is mediated by differing peaks in the periods for gamete release and by one-way sperm/egg incompatibility. Two attempts to recover C. sikamea from its place of origin in southern Japan have yielded only individuals with the mitochondrial haplotype that characterizes C. gigas. We thus identify a crucial need for careful screening, management, and conservation of the cultivated populations of C. sikamea on the US west coast.     

Sex pheromones and their potential role in the evolution of reproductive isolation in small ermine moths (Yponomeutidae)

Summary Sex pheromone communication in the nine European species of small ermine moths (Yponomeuta) is reviewed in regard to the potential role of pheromones in the speciation process. Six of the nine species studied (viz.,Y. evonymellus, Y. cagnagellus, Y. padellus, Y. irrorellus, Y. plumbellus, andY. vigintipunctatus) use a mixture of (E)-11-and (Z)-11-tetradecenyl acetate in different ratios as primary pheromone components, with combinations of tetradecyl acetate, (Z)-9-tetradecenyl acetate, (Z)-11-hexadecenyl acetate and the corresponding alcohols of the acetates as additional pheromone components. Analysis of (Z)- to (E)-11-tetradecenyl acetate ratios produced by individual females of these species demonstrated significant variation among females of all species. However, the ranges of ratios produced byY. cagnagellus, Y. irrorellus, andY. plumbellus, sharing the same host-plant species, spindle tree, did not overlap. Niche separation of all six species mentioned required consideration of at least one additional pheromone component or of temporal aspects. The remaining three species,i.e. Y. malinellus, Y. mahalebellus andY. rorellus, have pheromones that differ qualitatively.Biosynthetic routes to the pheromone components identified are proposed on the basis of fatty acid pheromone precursors found in the pheromone glands. A phylogenetic tree for the genus is constructed based on allozyme frequency data and changes in pheromone composition are superimposed on this tree. We suggest that the ancestral ermine moth pheromone is a mixture of (Z)-11- and (E)-11-tetradecenyl acetate and the corresponding alcohols, and a scenario of how present-day patterns evolved is outlined. The pheromone differences among the three species using spindle tree as their host-plant might have evolved throughreproductive character displacement upon secondary contact between populations that had already diverged genetically in allopatry. Pheromone differences within the so-calledpadellus-complex (includingY. cagnagellus, Y. mahalebellus, Y. malinellus, Y. padellus, andY. rorellus) in which species might have originated sympatrically, may have evolved byreinforcing selection as these species still hybridise and produce viable offspring when confined in cages. The role of pheromones in reproductive isolation amongYponomeuta species is emphasised by (1) the function of pheromone components of some of the species as behavioural antagonists to other species, (2) the cross-attraction under experimental conditions between allochronic species with similar pheromones, and (3) the formation of hybrids in the laboratory between species that are isolated in nature by pheromone differences.     

Two sibling species of Tisbe (Copepoda,Harpacticoida): Tisbe reluctans and T. persimilis n. sp. Research on their morphology and population dynamics

Two new forms of the copepod genus Tisbe, one of which was found in the Lagoon of Venice near Chioggia (Italy), and the other in the port of Malta, have been studied. Although showing a striking resemblance in morphology, all attempts to cross representatives of the 2 populations have failed. They may, therefore, be considered as sibling species. Even on closer study, no qualitative structural differences between them were found. However, careful analysis of some biometrical traits revealed significant differences. A certain differentiation of another kind emerges from the comparison of the life-cycle and population dynamics. Under standard laboratory conditions the following main parameters have been considered: (1) survival; (2) mean and minimum generation time, T; (3) net reproduction rate, R ₀; (4) intrinsic rate of natural increase, r _m. The significance of the findings is discussed.     

Inbreeding and Outbreeding Depression in Natural Populations of Chamaecrista fasciculata (Fabaceae)

Abstract: The deleterious consequences of inbreeding have been well documented. There are, however, few empirical studies that have examined the consequences of restoring heterozygosity and hence the fitness of inbred populations by conducting interpopulation crosses and measuring the performance of later-generation hybrids under field conditions. We conducted interpopulation crosses of 100 m to 2000 km, which spans the range of Chamaecrista fasciculata ( Fabaceae) in eastern North America. We then contrasted the performance of the F1 and later-segregating F3 hybrids with the parental generation. We found almost universal F1 superiority over the parents. The F3 hybrids suffered a loss of fitness compared to the F1 hybrids. The drop off in fitness of the F3 reflects both the loss of heterozygosity and the disruption of coadapted gene complexes. The F3 performance, however, was still often equal to that of the parents, suggesting that heterosis can outweigh the loss of coadaptation except for the longest-distance crosses. In a subset of environments, the F3 performance of long-distance (≥1000 km) interpopulation crosses was less than that of both parents and indicated true outbreeding depression. For C. fasciculata , it appears that crossing populations of up to intermediate distances of hundreds of kilometers has a short-term beneficial effect on progeny performance through F1, and that longer-term effects are not necessarily disruptive of fitness, at least relative to parental performance. The degree of F1 heterosis and F3 outbreeding depression varied between site and year, however, indicating an important role for the environment in the expression of these effects.     

Genetic variation among life-history stages of mussels in a<Emphasis Type="Italic"> Mytilus edulis–M</Emphasis>.<Emphasis Type="Italic"> trossulus</Emphasis> hybrid zone

The mussel species Mytilus edulis L. and M. trossulus Gould coexist and hybridize throughout a large area that includes the north coast of Maine and Atlantic Canada. Previous studies provided genetic evidence for limited hybridization between the two species for mussels >15 mm. The present study used two genetic markers (ITS, Glu-5) to examine the genetic composition of early life-history stages by sampling veliger and pediveliger larvae, juveniles (<2.0 to 15.0 mm shell length) and adults (>15 mm shell length) in Trinity Bay, Newfoundland, during three consecutive years (1995–1997) to determine if differential mortality limits the relative abundance of hybrids. The relative frequency of the two species and the different hybrid genotypes was similar among the larvae, juveniles and small adult mussels. The double hybrid genotype (F₁-like) was the rarest genotype observed. There was no evidence for differential mortality during the early life-history stages, and factors limiting production of hybrids appear to operate before the late larval stage. The observed frequency of hybrids is probably due to a combination of pre- and postzygotic reproductive isolating mechanisms operating early in the mussels life history. M. trossulus dominated the early life-history stages, possibly due to a higher population density and a greater reproductive output than M. edulis. Differential mortality may explain the observed decrease in frequency of M. trossulus and increase in frequency of M. edulis with increasing shell length. A similar frequency of hybrid mussels from larvae to the size class of 55 mm shell length may indicate a rate of mortality intermediate between the two parental species. The M. edulis–M. trossulus hybrid zone appears to be maintained by reproductive isolating mechanisms limiting the production of hybrids and life-history differences that allow the two species to coexist.Communicated by J.P. Grassle, New Brunswick     

Biosystematic studies of the red algal genus Eucheuma. I. Electrophoretic variation among Florida populations

Electrophoretic techniques were used to assess the degree of genetic differentiation among 4 species of the seaweed Eucheuma; E. isiforme, E. nudum, E. gelidium and E. acanthocladum. Nine Floridian populations were surveyed for 5 enzymes (14 loci), and compared using both phenotypic identity (I _p )and Nei's genetic identity (I) statistics. Results suggest there is a very high degree of genetic similarity between E. isiforme and E. nudum (=0.904), and between E. gelidium and E. acanthocladum (=0.857), but a very low degree of similarity between these two pairs of species (=0.372). These findings are in agreement with an earlier report which recommended the removal of E. gelidium and E. acanthocladum from the genus based on differences in reproductive anatomy. Greater genetic similarity was observed among the E. isiforme populations than among the E. nudum populations, the latter being ecologically more diverse and geographically or hydrographically more isolated. The proportion of polymorphic loci per population ranged from 0.25 to 0.36. Heterozygote deficiencies were observed at two acid phosphatase loci in offshore populations of E. nudum which showed a propensity for vegetative reproduction.     

Gonadal development in the coral reef damselfish<Emphasis Type="Italic"> Dascyllus flavicaudus</Emphasis> from Moorea,French Polynesia

The present study investigated the sexual pattern of the yellow-tailed dascyllus, Dascyllus flavicaudus (Randall and Allen), through histological examination of the process of gonadal differentiation and maturation and through male-removal experiments in the laboratory. The study was conducted in Moorea, French Polynesia (17°32S; 149°50W), from 31 December 1994 to 11 March 1995. For gonadal histology, two populations were sampled, one consisting of large aggregations (n=54) and the other consisting of small- to medium-sized groups around isolated corals (n=55). An additional small sample (n=21) was also collected from a population that consisted of small groups around isolated corals. After an initially undifferentiated state, gonads of D. flavicaudus developed an ovarian lumen, followed by oocytes in the primary growth stage. From this ovarian state or from more developed ovaries with cortical-alveolus stage oocytes, some gonads developed into testes through degeneration of oocytes and development of spermatogenic tissue. In all three populations, spermatogenic tissue developed only in gonads that contained pre-vitellogenic oocytes (termed as mixed stage 1–2 gonads). The two main populations did not differ in their expression of sex change despite differences in their social organization. In both populations, size of individuals with mixed stage 1–2 gonads overlapped mainly with the size range of immature females, which suggests that functional female-to-male sex change was rare. The hypothesis that D. flavicaudus is primarily gonochoristic is further supported by removal experiments in laboratory groups, in which removal of a dominant male(s) failed to induce sex change in any of the high-ranking females. Yet, in all three field populations, some fish with mixed stage 1–2 gonads were found within the size ranges of mature females, which indicates possible occurrences of functional sex change. These conflicting results indicate that it may be premature to draw any definitive conclusions about the sexual pattern of D. flavicaudus.Communicated by T. Ikeda, Hakodate     

Evolutionary genetics of marine Mysis spp. (Crustacea: Mysidacea)

Inter-and intraspecific allozyme differentiation in the mysid crustacean genus Mysis in the North Atlantic region was studied in order to evaluate earlier concepts of evolutionary and systematic relationships and to assess patterns of subdivision within widespread taxa. The results support a relatively ancient divergence of the marine and non-marine species of the genus, and are generally in line with the current subgeneric tridivision into Mysis s.str., Michteimysis and Auricomysis. However, the North American littoral species M. gaspensis should be returned to subgenus Mysis s.str. from its present position in Michteimysis with M. mixta. The closest observed affinities within Mysis s.str. were between M. gaspensis and the freshwater M. relicta group, and between M. oculata and M. litoralis. Intraspecific differentiation among North European coastal populations of M. oculata and M. litoralis was moderately strong (F _ST0.1), suggesting population bottlenecks and limited dispersal in the post-glacial time. On the other hand, ransoceanic differences were not essentially greater, indicating the systematic homogeneity and long-term dispersal capacity in the marine species. This contrasts with the strong genetic and systematic fragmentation earlier found within the circumboreal M. relicta species group.     

Mate choice and fitness in a hybrid frog: Rana esculenta females prefer Rana lessonae males over their own

Summary The evolution and maintenance of female choice based on purely genetic differences is still a controversial issue, not only for theoretical reasons, but also because of the practical difficulty of demonstrating the fitness consequences of preferences and heritability of and genetic variability in the chosen traits. We argue that hybrid systems (broadly defined) offer suitable models for studying mate choice according to genetic differences. We present such a study for European water-frogs of the hybridogenetic Rana lessonae/Rana esculenta complex (L/E complex). R. esculenta, originally a hybrid between R. lessonae and R. ridibunda, eliminates the L genome premeiotically and only produces eggs and sperm containing only the R. ridibunda (R) genome. Consequently, the hybrid will only persist when it lives and mates with R. lessonae in mixed populations where it can regain the lost L genome. In such mixed populations, there is strong selection against E x E matings. because these will produce no viable offspring. We tested whether females of the hybrid R. esculenta do indeed avoid their own R. exculenta males and choose males of the parental species R. lessonae instead. Eleven E females were offered a simultaenous choice between one L and one E male. Females exhibited a significant preference for L males that was determined by the type of male, rather than by its size or activity. This choice is in the direction predicted from genetics. The question of why L males agree to mate with E females, but L females only rarely mate with E males, is answered by a sexual asymmetry in the cost/benefit ratios of mating with the wrong type and the right size. Our results are consistent with the mating pattern found in natural populations, but further studies are needed to show that female choice really causes this pattern.Correspondence to: H.-U. Reyer     

Analysis of individual year-classes of a marine fish reveals little evidence of first-generation hybrids between cryptic species in sympatric regions

As settled juveniles and adults, blue rockfish (Sebastes mystinus) are nonmigratory inhabitants of kelp and rocky reef habitats along the California coast, USA, and prior to settlement, they possess a pelagic larval and juvenile stage lasting 3–5 months. A previous study of adults revealed two cryptic species within S. mystinus and evidence of reproductive isolation in a region where both cryptic adults co-occur. Given this pattern of reproductive isolation, we investigated the degree of hybridization or introgression in individual year-classes shortly after juvenile settlement in two different years (2001 and 2002). Using microsatellite markers, we found little indication of hybridization in new juvenile year-classes despite an adult population that comprised both cryptic species. However, we found an average of two percent of hybrid or introgressed individuals in regions with a low frequency of one of the two species. Therefore, while the lack of hybrids or introgression supports the hypothesis of reproductive isolation between the cryptic species within S. mystinus, the age-structured analysis also revealed a spatial pattern of low-frequency differences in the number of introgressed individuals. These results suggest that reproductive barriers may breakdown when one of the two species predominates the regional adult gene pool.     

Reproductive isolation with little genetic divergence between Urticina (=Tealia) felina and U. eques (Anthozoa: Actiniaria)

There has long been doubt as to whether there are one or two British species of the sea anemone genus Urticina. In the present study, populations of both putative species, which occur sympatrically in the Isle of Man, have been compared by electrophoresis of isozymes and nematocyst analysis. The analysis of fourteen isozyme loci exhibited a high genetic identity between the two taxa (I=0.907), but four loci showed significant differences in gene frequencies; thus indicating reproductive isolation. The populations displayed highly significant differences in the sizes of the various nematocyst types. The individual analysis of data for each type of nematocyst revealed that the basitrics from the actinopharynx and from the tentacles contribute most to the observed difference. Consequently, U. felina (L.) and U. eques (Gosse) are assumed to be valid species, as suggested by Manuel (1981). The mean heterozygosities for each species (0.410 and 0.436, respectively) are the highest reported for cnidarians.     

Mating systems in the sea anemone genus Epiactis

Four morphologically similar species in the sea anemone genus Epiactis exhibit overlapping distributions on the Pacific coast of North America; E. prolifera, E. lisbethae, E. ritteri and E. fernaldi. All brood their offspring up to the juvenile stage, but each has a different combination of internal versus external brooding and hermaphroditism versus gonochory (separate sexes). Specimens were collected from sites ranging from British Columbia to southern California between December 1988 and July 1992. Mating systems were inferred from genetic comparisons of mothers and offspring histological analyses of sex expression and observations on brooding and spawning behavior. Allozyme and multilocus DNA fingerprint analyses of the gynodioecious hermaphrodite E. prolifera showed that offspring were all identical to their mothers, a result consistent with either asexual reproduction, self-fertilization or extreme biparental inbreeding. In the gonochore E. lisbethae, mothers and offspring were also electrophoretically identical, but variation in DNA fingerprints indicated cross-fertilization. Similar DNA fingerprint differences between mother and offspring in the gonochore E. ritteri implied that cross-fertilization also occurs in this species. No mother-offspring comparisons were performed on E. fernaldi, as this species was not observed brooding offspring during this study. Although incomplete, the results of this study increase our knowledge of the very unusual combination of reproductive modes in the genus Epiactis, and argue for further investigations of the evolution and genetic consequences of mating systems in these species.     

Variation in fertilisation abilities between hemiclonal hybrid and sexual parental males of sympatric water frogs (<Emphasis Type="Italic">Rana lessonae</Emphasis>,<Emphasis Type="Italic"> R. esculenta</Emphasis>,<Emphasis Type="Italic"> R. ridibunda</Emphasis>)

In many species, males and females mate with multiple partners, which gives rise to sperm competition and multiple paternity. The experiments on water frogs presented here demonstrate that such sperm competition can affect the structure and dynamics of mixed-species communities. The hybrid frog Rana esculenta (LR) mates with one of its parental species, usually R. lessonae (LL), although in some areas R. ridibunda (RR), to regain the premeiotically eliminated parental genome ("hybridogenesis"). Mixed LL/LR-populations are stable although hybrid numbers should continuously increase at the expense of parental animals, because of differences in female fecundity and other factors. This would finally lead to the extinction of the sexual host, followed by that of the sexual parasite, unless the reproductive superiority of R. esculenta is reduced by other factors, such as lower hybrid male fertility. Eggs from LL- and LR-females were fertilised in vitro by single- and multi-male sperm suspensions of LL-, LR- and RR-males. In all experiments, the proportion of offspring sired by R. esculenta sperm was significantly lower than that sired by R. lessonae or R. ridibunda sperm. Gonad mass, sperm morphology, sperm swimming velocity, and sperm survival did not explain these differences in fertilisation success; nor did gamete recognition and compatibility. Sperm density was the only trait that paralleled fertilisation success, but it offers no explanation either, because densities were equalised for the in-vitro fertilisations. In natural LL/LR populations, the significantly smaller amount, poorer competitive ability and lower long-term survival of R. esculenta compared to R. lessonae sperm will reduce the initial reproductive superiority of hybrids and contribute to the stabilisation of mixed water-frog populations. Differences in fertilisation ability are also likely to be relevant for the structure and dynamics of several other systems with encounters between eggs and sperm from different genotypes, ecotypes, ploidy levels and/or species.