Genetic variability in bloodworm (Glycera dibranchiata) populations in the Gulf of Maine

Genetic structure of eight Northwest Atlantic populations of the marine polychaeteGlycera dibranchiata Ehlers was examined with starch gel electrophoresis. Samples were collected during summer and fall 1981, and seven polymorphic and four monomorphic loci were consistently scored. Average heterozygosity (0.126) and percent polymorphic loci (59.3) were comparable to the averages reported for marine invertebrates. Minimum genetic distances between populations ranged from 0.003 to 0.093, levels typically associated with local populations of the same species in other taxa. Based on these data, inter- and intra-estuarine migration and gene flow appear to be low. Only two populations, separated by 13 km along the same river in New Brunswick, Canada, were not genetically different from each other. These findings may have relevance for management strategies in bloodworms.     

Genetic evidence for the cryptic species pair, <Emphasis Type="Italic">Lottia digitalis </Emphasis>and <Emphasis Type="Italic">Lottia austrodigitalis</Emphasis> and microhabitat partitioning in sympatry

It has been proposed that the common West Coast limpet, Lottia digitalis, is actually the northern counterpart of a cryptic species duo including, Lottia austrodigitalis. Allele frequency differences between southern and northern populations at two polymorphic enzyme loci provided the basis for this claim. Due to lack of further evidence, L. austrodigitalis is still largely unrecognized in the literature. Seven additional enzyme loci were examined from populations in proposed zones of allopatry and sympatry to determine the existence of L. austrodigitalis as a sibling species to L. digitalis. Significant allele frequency differences were found at five enzyme loci between populations in Laguna Beach, southern California, and Bodega Bay, northern California; strongly supporting the existence of separate species. Both species exhibit two microhabitat morphotypes, a gooseneck barnacle morph in the mid-intertidal zone and a rock morph in the high-intertidal zone. In sympatry, L. austrodigitalis was more abundant higher in the intertidal on rocks, whereas L. digitalis was more abundant lower in the intertidal on barnacles. This finding supports earlier claims of microhabitat partitioning in this sibling species pair. In addition to this finding, the transition zone between the species was found to have shifted substantially northward in only two decades, from Monterey Peninsula, CA to near Pigeon Point, CA, where L. digitalis previously dominated.     

Genetic variation in the neritic squid Loligo forbesi (Myopsida: Loliginidae) in the northeast Atlantic Ocean

Horizontal starch gel electrophoresis was employed to investigate levels of genetic differentiation between 13 samples of the neritic squid species Loligo forbesi Steenstrup obtained from throughout the majority of its known geographical range. Six enzyme loci identified in a preliminary study as being polymorphic were screened for variation between samples. No significant differences in allele distribution were detected between any of the samples obtained from the Faroe Bank in the north to Lisbon in the south, suggesting that squid throughout this range in the vicinity of the continental shelf are able to maintain panmixia, and effectively belong to a single population sharing a common gene pool. No clinal variation in allele distribution was detected throughout this range, a result which complements the findings of a detailed morphological companion study of the same individuals. Comparison of this homogenous European continental shelf population with squid from the Azores revealed highly significant (P<0.01) differences in allele distribution at five of the six polymorphic enzyme loci studied. A genetic identity value (I) equivalent to 0.93 over 33 loci was obtained. Analysis of F-statistics suggested migration rates between sites to be as low as one individual per five generations, a rate deemed insufficient under most models to prevent divergence by random genetic drift. The large distance and oceanic depths separating the Azores from continental Europe seem to present an effective barrier to gene flow to L. forbesi, a squid belonging to a family considered to be confined in distribution to relatively shallow, near coastal waters. The two populations of squid in the Azores and along the European continental shelf currently both ascribed to L. forbesi should therefore probably best be regarded as relative subspecies.     

Uptake,binding and clearance of divalent cadmium in Glycera dibranchiata (Annelida: Polychaeta)

The bloodworm Glycera dibranchiata Ehlers, 1968 accumulates cadmium through the general body surface and the intestine. Absorption through the gut accounts for cadmium which rapidly binds to coelomic proteins. Intracoelomic injection of ¹⁰⁹Cd demonstrates that cadmium binds readily to hemoglobin and other proteins. The degree of cadmium binding is pH-dependent. The apparent pK of binding sites in body wall and musculature homogenates is 5.39. Cadmium ions injected into the coelom at 7 g g^-1 tissue increase proline incorporation rates into the positively charged hemoglobin (cathode fraction) by 15-fold in 3 days. A 3-fold increase of proline incorporation was observed in the anode hemoglobin fraction over the same time period. Radioactivity in various protein fractions decreases at different rates after injection of ¹⁰⁹Cd. Comparisons between the function of mammalian metallothionein and the coelomic fluid proteins of G. dibranchiata as a detoxification mechanism are discussed.     

Genetic evidence of self-fertilization in the sea anemone Epiactis prolifera

Biochemical genetic variation provided evidence for the mode of reproduction of brooded young in the sea anemone Epiactis prolifera Verrill, 1869. Individuals of E. prolifera are female when small but hermaphroditic when large (i.e., gynodioecious); juveniles are brooded externally on the column. Brooding individuals collected from 6 intertidal sites (5 in central California and 1 in Washington State, USA) in the spring and summer of 1980 were assayed for gene-enzyme variation by starch-gel electrophoresis. Three of 12 enzyme loci were polymorphic; phosphoglucose isomerase appeared to be encoded by two, closely linked loci. Genotypic frequencies deviated markedly from expected random mating proportions. Only three heterozygotes were found; two were heterozygous at all three polymorphic loci, and the other was polymorphic at the two PGI loci. All 158 juveniles from 25 brooding individuals were assayed (2–19 juveniles per parent). Juveniles on homozygous adults were always identical to their parent. However, brooded young of heterozygous individuals were not identical to their parent. but showed 1:2:1 phenotypic segregation ratios consistent with reproduction by self-fertilization. This genetic evidence together with findings of marked heterozygote deficiencies and genetic identity of homozygous adults and their brooded young supports the conclusion that E. prolifera usually reproduces by self-fertilization, and cross-fertilization is rate.     

Genetic consequences of long larval life in the starfish Linckia laevigata (Echinodermata: Asteroidea) on the Great Barrier Reef

Gene flow between populations of the asteroid Linckia laevigata (Linnaeus) was investigated by examining over 1000 individuals collected from ten reefs throughout the Great Barrier Reef (GBR), Australia, for genetic variation at seven polymorphic enzyme loci. Despite geographic separations in excess of 1000 km, Nei's unbiased genetic distance (0 to 0.003) and standardised genetic variation between populations (F _ST) values (mean 0.0011) were small and not significant. Genetic homogeneity among L. laevigata populations is consistent with the long-distance dispersal capability of its 28 d planktonic larval phase, and is greater than that observed for other asteroid species, including another high-dispersal species, Acanthaster planci, which has a 14 d larval phase. Variation within populations was also higher than previously recorded for asteroids (mean heterozygosity=0.384; number of alleles per locus ranged from 5.1 to 6.0 in each population). Among asteroids, dispersal ability is positively correlated with gene flow and levels of variation, and negatively correlated with levels of differentiation.     

Genetic variability and differentiation of sporophytes and gametophytes in populations of Gelidium arbuscula (Gelidiaceae: Rhodophyta) determined by isozyme electrophoresis

Genetic differentiation and genetic variability of sporophytic and gametophytic populations of Gelidium arbuscula (Bory) from three localities sampled in 1989 and 1990 in the Canary Islands (Spain) were examined by isozyme electrophoresis. Twenty-three to 29 putative alleles corresponding to 22 gene loci, were compared. High deviations in Hardy-Weinberg equilibrium, and significant differences between allelic frequencies of sporophytic and gametophytic subpopulations at the same locality were found, suggesting a predominant asexual reproduction of G. arbuscula. The genetic variability (percentage of polymorphic loci, mean number of alleles per locus and average gene diversity) of haploid subpopulations was lower than that of diploid subpopulations at all three localities, being the lowest described for seaweeds. No correlation between genetic and geographical distance was found. The high genetic differentiation coefficient between all subpopulations suggests a very reduced genetic flow between subpopulations of the same and of different localities. These results suggest that the genetic structure of the populations of G. arbuscula from the Canary Islands is due to a founder-effect combined with a predominance of asexual reproduction. This is the first report comparing allelic frequencies between sporophytic and gametophytic subpopulations of seaweeds.     

Polymorphism of mannose phosphate isomerase in North Sea and Baltic Sea populations of the amphipods Gammarus zaddachi and G. salinus

Genetic variation at the mannosephosphate isomerase (MPI) locus was examined in the euryhaline sibling species Gammarus zaddachi Sexton and G. salinus Spooner. Both crustacean amphipods share identical enzyme mobilities, following electrophoresis on vertical starch gels. The MPI locus turned out to be highly polymorphic; it is encoded by 6 alleles in G. zaddachi and 7 alleles in G. salinus. Geographic variation of allelic diversity was studied in samples from 9 G. zaddachi and 10 G. salinus populations, primarily obtained from Baltic Sea and North Sea sites. Patterns of inter- and intraspecific heterogeneity are described. Differences in allelic composition exist between Baltic and North Sea samples of G. salinus. In G. zaddachi, levels of polymorphism are higher in North Sea populations than in those from Baltic Sea areas. The significance of these findings is discussed in the light of previous biochemical genetic investigations on the population structure of the two amphipods considered.     

Separation of the two Atlantic species of Pomatoceros,P. lamarckii and P. triqueter (Annelida: Serpulidae) by means of biochemical genetics

In order to confirm the status of two proposed species of the serpulid Pomatoceros, the electrophoretic mobilities of 13 enzymes encoded at 19 loci in P. lamarckii (Quatrefages) and P. triqueter (L.) were compared using horizontal starch gel electrophoresis. The serpulids were sampled from Menai Bridge (North Wales), Plymouth (England) and the Isle of Man in the winter of 1981/1982. Six loci were monomorphic or nearly so and showed no useful or significant difference between species. At a further 7 loci, some of the allozymes common to both species were present, though usually at significantly different frquencies, while others were species-specific. The remaining 6 loci presented species-specific allozymes. In P. lamarckii, 61% of the loci were polymorphic compared with 50% in P. triqueter. The mean observed heterozygosity was 0.169 for P. lamarckii and 0.208 for P. triqueter. Calculation of genetic distance and identity gave values of 0.821 and 0.440, respectively. This study, therefore, confirms the existence of two separate species of Pomatoceros as has been previously proposed on the basis of opercular structure and supported by ecological separation where these species occur sympatrically.     

Genetic differentiation between populations of Talitrus saltator and Talorchestia deshayesii (Crustacea: Amphipoda) from coastal areas of the north-western European continent

Electrophoretic studies of gene-enzyme variation in the littoral talitrids Talitrus saltator (Montagu) and Talorchestia deshayesii (Audouin) were undertaken to estimate the amount of divergence among geographically separated populations. Samples of both species were taken from sandy beaches over a transect of approximately 3 500 km along the coast of the European continent including Baltic, North Sea and Atlantic locations. A total of 22 T. saltator and 15 T. deshayesii populations were analysed for genetic variation at various enzyme loci. Both amphipods revealed relatively low levels of polymorphism and heterozygosity. Among the loci studied, phosphoglucose isomerase (Pgi) and phosphoglucomutase (Pgm) were highly polymorphic. Patterns of micro- and macrogeographic variation in terms of distributions of allele frequencies at these particular loci are compared. Interpopulation allozymic variation was shown to be lower in T. deshayesii than in T. saltator. As demonstrated by T. saltator populations sampled in coastal sites ranging from Denmark to western France, clinal variation in frequencies of two alleles became evident at the PGI locus, exhibiting a steady increase in the level of polymorphism with decreasing latitudes. It is argued that limited gene flow and, to some extent, random genetic drift may account for the gene pool structure of the talitrid species investigated.     

Genetic variation between geographic populations of the amphipods Gammarus zaddachi and G. salinus

Spatial and temporal patterns of gene-enzyme variation were estimated in the sibling species Gammarus zaddachi Sexton and G. salinus Spooner by starch gel electrophoresis. Twenty-one G. zaddachi and 18 G. salinus populations from coastal and estuarine areas in the Baltic Sea, North Sea and other localities of north-western Europe were surveyed. Both amphipods display similar electrophoresis patterns of the enzyme systems studied. Considerable interspecific and interpopulational differences were detected in allele frequencies at three highly polymorphic loci, phosphoglucose isomerase (PGI), glutamate oxalacetate transaminase (GOT) and arginine phosphokinase (APK). G. zaddachi exhibits a pronounced genetic heterogeneity in most areas of the sampled range. Populations from northern French and western English coasts differ significantly from the other samples in allele frequencies at the PGI or APK locus, respectively. Baltic populations are widely uniform in their genetic composition but can be distinguished from samples taken at North Sea sites in allele frequencies at the APK locus. The latter reveal a clinal variation, ranging from the Danish to the French coast. In contrast to G. zaddachi, a low degree of genetic differentiation was observed among the G. salinus populations examined. This indicates that migration and interregional mixing may be more important in maintaining the genetic structure than in G. zaddachi which, compared to G. salinus, prefers habitats of lower salinity levels. Evidently, less extensive dispersal capabilities owing to the confinement of G. zaddachi to brackish waters of dilute salt concentrations may account for a diminished gene flow and considerable genetic separation of local populations. This assumption is supported by the genetic homogeneity documented in Baltic G. zaddachi populations. In view of the low and constant salinities in wide areas of this brackish-water sea such barriers do not exist. Survey studies performed with selected populations over a 3-yr period demonstrated a general pattern of temporal constancy in the allozyme variation observed.     

Genetic structure of the common sole Solea vulgaris at different geographic scales

The genetic structure of the flatfish Solea vulgaris was investigated on several spatial scales and at the temporal level through analysis of electrophoretic variation at 8 to 12 polymorphic enzyme loci. No differentiation was apparent at the temporal scale. Some differentiation was detected at and above the regional scale. Isolation by distance was evidenced by the significant correlation between genetic and geographic distances, and by the consistency of the results of multiple-locus correspondence analysis with geographic sampling patterns. The analysis suggested that the geographic unit of population structure (i.e. a geographical area corresponding to a panmictic or nearly panmictic population) lies within a radius of the order of 100 km. The isolation-by-distance pattern in S. vulgaris contrasted with the known genetic structures of other flatfish species of the northeastern Atlantic and Mediterranean in a way that may be related to the range of their respective temperature tolerances for eggs and larvae.     

Genetic differentiation of bicolor damselfish (Eupomacentrus partitus) populations in the Florida Keys

Electrophoretic variation in proteins encoded by 23 loci revealed substantial genetic differentiation among populations of bicolor damselfish (Eupomacentrus partitus) collected from four coral reefs in the Florida Keys, USA, during 1986–1988. Genetic differentiation was concentrated between a sample collected from Little Grecian Rocks Reef (LGR) and the remaining samples, including fish from a reef only 600 m distant (Grecian Rocks Reef). Genetic distinction of the LGR sample derived from significantly heterogenous allelic frequencies at six of eight polymorphic loci. Aco-1 (aconitase); Ada (adenosine deaminase); Gpi-2 (glucosephosphate isomerase); Ldh-2 and Ldh-3 (lactate dehydrogenase); and Me-1 (malic enzyme); nevertheless, differentiation at cytosolic aconitase (Aco-1) far exceeded that observed for other loci (fixation index, F _ST=0.482), and differences in Aco-1 allele frequencies were largely responsible for large genetic distances (0.20) between LGR and the other reefs. Paradoxically, estimates of numbers of migrants exchanged between reefs per generation (mN _e=17.47) indicated the potential for extensive gene flow. The extent of genetic differentiation among these populations is evaluated relative to models of population genetic structure based on equilibrium between gene flow and natural selection or genetic drift.     

Genetic studies in marine fishes

Nine polymorphic loci were found among 42 presumptive protein-coding gene loci surveyed among 474 red drum (Sciaenops ocellatus) sampled in 1987 from 13 nearshore and 1 offshore localities from the Atlantic coast of the southeastern USA and the northern Gulf of Mexico. The mean number of alleles over the polymorphic loci was 3.8, and the average heterozygosity over all loci examined was estimated as 0.047. These data indicate that red drum have normal levels of genetic variability. Wright'sF1 _ST values (the standardized variance of allele frequencies between samples) over all polymorphic loci ranged from 0.009 to 0.027 (meanF1 _ST =0.019), and estimates of the effective number of migrants (N _e m) per generation using Wright's island model ranged from 9.0 to 27.5. High levels of gene flow among the red drum samples were also indicated by Slatkin's qualitative analysis using conditional average allele frequencies. Nei's estimates of genetic distance between pairs of samples ranged from 0.000 to 0.009, indicating a high degree of nuclear gene similarity among all samples. Highly significant heterogeneity in allele frequencies at the locus for adenosine deaminase was detected between red drum sampled from the Atlantic and those sampled from the Gulf and among red drum sampled from the Gulf.     

Genetic differentiation of populations of the planktonic copepod Labidocera aestiva

Planktonic populations of the calanoid copepod Labidocera aestiva show significant biochemical genetic heterogeneity along the Atlantic coast of the USA. In summer, 1981, copepods were collected by surface tows at Beaufort Inlet, North Carolina; Fort Pierce Inlet, Florida; and Vineyard Sound, Massachusetts. Genetic variation within each population and genetic differentiation among the three populations were studied by micro-acrylamide gel electrophoresis of six loci encoding four enzymes. All six enzyme loci were polymorphic when all populations were considered together, but the North Carolina population was monomorphic at two loci. High genetic variability was indicated by the following: (1) the number of alleles per locus averaged over all loci was 2.57±0.26 SD; (2) the average proportion of loci for which the frequency of the most common allele was not greater than 0.95 was 0.78±0.10; (3) the frequency of heterozygous individuals was 0.25±0.07. Genetic differentiation among population samples in the three regions was demonstrated in several ways: allele frequencies at one aminopeptidase-I locus, Lap-1, differed significantly among samples of the three populations, and there were unique alleles of high frequency at this locus in two population samples. Values of the statistic of genetic distance (D) averaged 0.20±0.08 for pairwise comparisons between all samples. Compared to expected heterozygosity if individuals across the entire range sampled mated at random, there were highly significant heterozygote deficiencies at five of the six loci. Genetic differentiation of populations of L. aestiva may result from (1) differential selection on populations in the three regions, or (2) restricted gene flow between the populations. Gene flow may be limited by geographic separation or differences in life history, such as seasonal presence in the plankton and diapause egg production.Contribution No. 5810 of Woods Hole Oceanographic Institution     

High genetic variability and patchiness in a common Great Barrier Reef zoanthid (Palythoa caesia)

Allozyme electrophoretic analysis of seven polymorphic enzyme loci suggested that 1261 samples of Palythoa collected along 1765 km of the Great Barrier Reef during 1992–1993 were members of a single taxon, identified as Palythoa caesia Dana, 1846, with high external morphological variability and possibly encompassing several previously described species. Populations were slightly genetically differentiated (standardized genetic variance, F _ST=0.010, p<0.05), but there was no evidence of isolation by distance, or of the particular genetic distinction of geographic sets of reefs such as the Swains, as has been observed in other invertebrates. Differentiation was thought to be the result of random selection acting on patches of larvae, not the consequences of long-term reproductive isolation of populations.     

Genetic structure and allozyme variation of sea bass (Dicentrarchus labrax and D. punctatus) in the Mediterranean Sea

This paper reports data on 28 allozyme loci in wild and artificially reared sea bass (Dicentrarchus labrax) samples, originating from either coastal lagoon or marine sites in the Mediterranean Sea. F _ST analysis (θ estimator) indicated strong genetic structuring among populations; around 34% of the overall genetic variation is due to interpopulation variation. Pairwise θ estimates showed that, on average, the degree of genetic structuring was much higher between marine populations than between samples from lagoons. Six polymorphic loci showed differences in allele frequencies between marine and lagoon samples. Multivariate analyses of individual allozymic profiles and of allele frequencies suggested that different arrays of genotypes prevail in lagoons compared to marine samples, particularly at those loci that, on the basis of previous acclimation experiments, had been implicated in adaptation to freshwater. On the other hand, variation at “neutral” allozyme loci reflects to a greater extent the geographic location of populations. Allozyme differentiation was also studied in a D. labrax population from the Portuguese coast. Average genetic distance between this population and the Mediterranean populations was quite high (Nei's D = 0.236) and calls into question the taxonomic status of the Portuguese population. Finally, genetic relationships between D. labrax and D. punctatus were evaluated. Average Nei's D was 0.648, revealing high genetic differentiation between the two species, even for two sympatric populations of these species in Egypt; thus gene flow was not indicated between species. Received: 24 October 1996 / Accepted: 27 November 1996     

Systematic status of two oyster populations of the genus Tiostrea from New Zealand and Chile

A systematic examination of Tiostrea chilensis endemic to the Pacific coast of South America and of T. lutaria endemic to new Zealand was conducted using protein-gel electrophoresis. Also, ecological and life-history traits were compared between these two documented species. Our study revealed that although three different ecomorphs could be distinguished from various environmental and ecological conditions, the life-history traits between the two species were remarkably similar. Estimates were made of levels of genetic variation and similary based on 29 structural loci among individuals from each documented species. The percentage of loci polymorphic was estimated at 17.2 and 17.6, while the percentage of loci heterozygous was 2.1 and 6.7 for T. chilensis and T. lutaria, respectively. The genetic similarity between the two species was computed as 0.991, where 1.0 indicates no genetic difference among structural loci. The overwhelming evidence of this study indicates that T. chilensis and T. lutaria are different geographical populations of one species and by priority this species should be T. chilensis.     

Allozyme variation in populations of the dog-whelkNucella lapillus (Prosobranchia: Muricacea) from the South West peninsula of England

Four populations of the predatory gastropodNucella lapillus were sampled at sites around the South West Peninsula of England in 1986, and analysed for allozyme variation at 18 enzyme loci. Two of these loci, Gpd-1 andHk-1, exhibited sex-specific phenotypes. An absolute locus association was observed between two other loci,Mdh-1 andEst-3. This association was only found at one site (Prawle), and it is suggested that the presence of chromosomal polymorphisms could explain this finding. As a measure of overall similarity, Nei's genetic identity statistic,I, was calculated; the mean for all populations was 0.989, with values ranging from 0.981 to 0.997. Although similar on this gross level, considerable interpopulation variation was evident. Observed mean heterozygosity (per locus) ranged from 0.043 to 0.104 (mean 0.074). Populations differed also in the loci at which significant heterozygote deficits were seen (of the seven deficits recorded only those at thePep-1 locus were consistent across sites) and in the presence of rare alleles undetected elsewhere. The variation observed showed no correlation to shell morphology or geographical distance and confirmed the conclusion that species of the genusNucella show considerable disjunct variation.     

Population genetics of the family ostreidae. I. Intraspecific studies of Crassostrea gigas and Saccostrea commercialis

The levels of genetic variation and estimates of genetic similarity and distance were determined for populations of Crassostrea gigas (Thunberg) and Saccostrea commercialis (Iredale and Roughley). The proportion of polymorphic loci in Japanese populations of C. gigas ranged from 0.58 to 0.63, while the individual heterozygosities ranged from 0.20 to 0.22. The proportion of polymorphic loci in Australian populations of S. commercialis ranged from 0.43 to 0.46, the individual heterozygosities from 0.17 to 0.19. The genetic similarity between different geographical populations of both species was approximately 99%. The genetic distance and similarity data derived from this investigation has resulted in some provisional reclassifications. The Portuguese oyster C. angulata (Lamarck) appears to be more appropriately regarded as a recent colonized isolate of C. gigas. The New Zealand oyster S. glomerata (Gould) is considered a subspecies of the eastern Australian rock oyster S. commercialis. The Japanese Kumamoto population of C. gigas warrants reclassification as the non-sibling species C. sikamea.