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Feeding pits dug by waterfowl in Zostera noltii meadows are thought to promote seedling recruitment by accumulating seeds and enhancing germination. We tested the latter hypothesis by creating a series of "treatment pits" (resembling natural feeding pits) in the center and at the edge of two meadows near the Island of Sylt (Germany). Seedling density was monitored from the autumn seed set until the following spring. Seedling density (mean, SE) in treatment pits was significantly higher (4.4, 5.3) than in manipulated (2.4, 1.9) and unmanipulated controls (1.4, 0.4), as well as significantly higher in center (2.8, 0.5) relative to edge (2.5, 1.1) locations. Results confirm a facilitating effect of waterfowl grazing on seedling recruitment in spring due to seed accumulation in feeding pits in autumn. The mechanism could provide a valuable tool for the conservation of intertidal Z. noltii meadows in the Wadden Sea. (C) 2010 Elsevier B.V. All rights reserved.

Genotypic structure and temporal dynamics of the dwarf seagrass, Zostera noltii, were studied in an intertidal meadow that has persisted since prior to 1936 near the Wadden Sea island of Sylt. Samples were collected from two 10 x 10 m plots separated by 250 m from May 2002 to June 2005 and from four 1 x 1 m plots from June 2003 to September 2004. All the samples were genotyped with nine microsatellite loci. No genotypes were shared between the plots separated by 250 m. Genetic diversity was higher in the Wadden Sea than in the other regions of its geographic range. The average clone size (genets) (SD) in the two plots was 1.38 (0.26) and 1.46 (0.4) mA(2), respectively, with a range up to 9 mA(2) and 4 years. A high genetic and genotypic diversity was maintained by annual recruitment of seedlings despite a dramatic decrease in ramet density that coincided with the severe heat stress event of 2003. Fine-scale (1 mA(2)) analysis suggested that extensive loss of seagrass cover precluded space competition among the genets, while a persistent seed bank prevented local extinction. Long-term persistence of Z. noltii meadows in the intertidal Wadden Sea was achieved by high genet turnover and frequent seedling recruitment from a seed bank, in contrast to the low diversity observed in large and long-living clones of Z. noltii and other seagrasses in subtidal habitats.

The intertidal dwarf eelgrass Zostera noltii is a dominant species in the Dutch and German Wadden Sea. Although numerous studies of its reproductive ecology have been conducted, few have examined the importance of seeds and seed banks for meadow maintenance. We investigated the contribution of a seed bank (size, genetic potential and persistence) to annual recruitment of dwarf eelgrass in the German Wadden Sea using temporal sampling of seeds from the sediment and genetic assignment tests of seedlings to populations of adult shoots from previous years. Annual sediment seed density (SD) was 487.5 m(-2) (269.4) and 367.3 m(-2) (95.5) in 2004 and 2005, respectively, and distribution of seeds in the sediment was highly aggregated. The proportion of over-wintering seeds that germinated under laboratory conditions was 16 to 25%, and field-germination revealed a 12% survival to the seedling stage. Nearly 20% of all shoots present in May 2004 were seedlings. Using 9 microsatellite loci, seedlings sampled in 2004, 2005 and 2006 were compared with adults sampled in 2002, 2003 and 2004; results revealed that 7 to 33% of seedlings could be assigned to the local adult population in Current or previous years. Although new recruitment plays an important role in the maintenance of these meadows, considerable new recruitment comes from within the meadow., itself. Seeds are viable for at least 3 yr, thereby forming a relatively short-term, but persistent, seed bank.

Understanding the flexibility of the endosymbioses between scleractinian corals and single-cell algae of the genus Symbiodinium will provide valuable insights into the future of coral reefs. Here, a real-time polymerase chain reaction (PCR) assay is presented to accurately determine the cell densities of Symbiodinium clades C and D in the scleractinian coral Acropora millepora, which can be extended to other coral-symbiont associations in the future. The assay targets single- to low-copy genes of the actin family of both the coral host and algal symbiont. Symbiont densities are expressed as the ratio of Symbiodinium cells to each host cell (S/H ratio, error within 30%), but can also be normalized to coral surface area. Greater accuracy in estimating ratios of associations involving multiple clades is achieved compared with previous real-time PCR assays based on high-copy ribosomal DNA loci (error within an order of magnitude). Healthy adult A. millepora containing 1.4 x 10(6) zooxanthellae per cm(2) (as determined by haemocytometer counts) had S/H ratios of c. 0.15, i.e. 15 symbiont cells per 100 host cells. In severely bleached colonies, this ratio decreased to less than 0.005. Because of its capacity to accurately determine both densities and ratios of multiple symbionts within one sample, the assay will open the door for novel research into the mechanisms of symbiont shuffling and switching.

We characterized 37 single nucleotide polymorphism (SNP) makers for eelgrass Zostera marina. SNP markers were developed using existing EST (expressed sequence tag)-libraries to locate polymorphic loci and develop primers from the functional expressed genes that are deposited in The ZOSTERA database (V1.2.1). SNP loci were genotyped using a single-base-extension approach which facilitated high-throughput genotyping with minimal optimization time. These markers show a wide range of variability among 25 eelgrass populations and will be useful for population genetic studies including evaluation of population structure, historical demography, and phylogeography. Potential applications include haplotype inference of physically linked SNPs and identification of genes under selection for temperature and desiccation stress.

There is worldwide concern about the aquarium strain of the green alga Caulerpa taxifolia (Vahl) C. Agardh that was introduced to the Mediterranean Sea in 1984. Since that time, it has flourished and now covers thousands of hectares of near-shore waters. More recently, aquarium strains of C. taxifolia invaded southern California and Australian waters. Our goal was to evaluate potential invasion of C. taxifolia to Florida’s coastal waters. We looked for evidence of C. taxifolia—aquarium strain, as well as the present distribution of all species of Caulerpa, in Florida’s near-shore waters. We surveyed 24 areas in six zones along the Floridian coastline, and evaluated the association of potential indicators for the presence of Caulerpa. Latitude, presence of seagrass beds, human population density, and proximity to marinas were the four variables simultaneously considered. Caulerpa taxifolia—aquarium strain was not found at any of our survey locations. However, 14 species of Caulerpa were found at 31 of the 132 sites visited. Percent correct for our model was 61.5% for presence and 98.1% for absence. There was a positive correlation between Caulerpa spp. and seagrass beds and proximity to marinas. There was a negative correlation with latitude and human population density. The parameters in the logistic regression model assessing the association of Caulerpa occurrence with the measured variables were then used to predict current and future probabilities of Caulerpa spp. presence throughout the state. This prediction model will allow resource managers to focus their efforts in future surveys.

Rays and skates are an unavoidable part of the by-catch in demersal fisheries. Over the past 40 years, the thornback ray (Raja clavata) has decreased in numbers and even disappeared in some areas, leading to concerns about genetic risk. For this reason, the effective population size (N-e), the migration rate (m) and temporal changes in the genetic diversity were estimated for the population of thornback rays in the Irish Sea and Bristol Channel. Using genotyped, archived and contemporary samples (1965 and 2003-2004), N-e was estimated at 283 individuals (95% CI = 145-857), m at 0.1 (95% CI 0.03-0.25) and the N-e/N ratio between 9 x 10(-5) and 6 x 10(-4). Although these results must be treated with caution, due to the small sample sizes, this is the first attempt to estimate N-e in an elasmobranch species. The low N-e/N ratio suggests that relatively few individuals contribute to the next generation. The combined effect of sex bias, inbreeding, fluctuations in population size and, perhaps most important, the variance in reproductive success may explain the low N-e/N ratio. In addition, the relatively high gene flow between Irish Sea population and other source populations is likely to have had an impact on our estimate, which may be more relevant at the metapopulation scale. No significant loss of genetic diversity was found over the 40-year timeframe and long-term maintenance of the genetic diversity could be due to gene flow.

Reef corals form associations with an array of genetically and physiologically distinct endosymbionts from the genus Symbiodinium. Some corals harbor different clades of symbionts simultaneously, and over time the relative abundances of these clades may change through a process called symbiont shuffling. It is hypothesized that this process provides a mechanism for corals to respond to environmental threats such as global warming. However, only a minority of coral species have been found to harbor more than one symbiont clade simultaneously and the current view is that the potential for symbiont shuffling is limited. Using a newly developed real-time PCR assay, this paper demonstrates that previous studies have underestimated the presence of background symbionts because of the low sensitivity of the techniques used. The assay used here targets the multi-copy rDNA ITS1 region and is able to detect Symbiodinium clades C and D with > 100-fold higher sensitivity compared to conventional techniques. Technical considerations relating to intragenomic variation, estimating copy number and non-symbiotic contamination are discussed. Eighty-two colonies from four common scleractinian species (Acropora millepora, Acropora tenuis, Stylophora pistillata and Turbinaria reniformis) and 11 locations on the Great Barrier Reef were tested for background Symbiodinium clades. Although these colonies had been previously identified as harboring only a single clade based on SSCP analyses, background clades were detected in 78% of the samples, indicating that the potential for symbiont shuffling may be much larger than currently thought.

The last glacial maximum (20 000-18 000 years ago) dramatically affected extant distributions of virtually all northern European biota. Locations of refugia and postglacial recolonization pathways were examined in Fucus serratus (Heterokontophyta; Fucaceae) using a highly variable intergenic spacer developed from the complete mitochondrial genome of Fucus vesiculosus. Over 1500 samples from the entire range of F. serratus were analysed using fluorescent single strand conformation polymorphism. A total of 28 mtDNA haplotypes was identified and sequenced. Three refugia were recognized based on high haplotype diversities and the presence of endemic haplotypes: southwest Ireland, the northern Brittany-Hurd Deep area of the English Channel, and the northwest Iberian Peninsula. The Irish refugium was the source for a recolonization sweep involving a single haplotype via northern Scotland and throughout Scandinavia, whereas recolonization from the Brittany-Hurd Deep refugium was more limited, probably because of unsuitable soft-bottom habitat in the Bay of Biscay and along the Belgian and Dutch coasts. The Iberian populations reflect a remnant refugium at the present-day southern boundary of the species range. A generalized skyline plot suggested exponential population expansion beginning in the mid-Pleistocene with maximal growth during the Eems interglacial 128 000 - 67 000 years ago, implying that the last glacial maximum mainly shaped population distributions rather than demography.

The phylogeography of thornback rays (Raja clavata) was assessed from European waters, using five nuclear microsatellite loci and mitochondrial cytochome b sequences. Strong regional differentiation was found between the Mediterranean basin, the Azores and the European continental shelf. Allelic and haplotype diversities were high in Portuguese populations, consistent with the existence of a refugium along the Iberian Peninsula. Unexpectedly, high diversity was also found in the English Channel/North Sea area. The lowest genetic diversity was found in the Black Sea. Populations sampled from the Mediterranean, Adriatic and Black Seas were characterized by a single mitochondrial haplotype. This haplotype was also the most ancestral and widespread outside of the Mediterranean basin except for the Azores. Populations from the Azores were dominated by a second ancestral haplotype which was shared with British populations. Results from multidimensional scaling, amova and nested clade analysis indicate that British waters are a secondary contact zone recolonized from at least two refugia--one around the Iberian Peninsula and one possibly in the Azores. Links to a potential refugium known as the Hurd Deep, between Cornwall and Brittany, are discussed. Finally, a historical demographic analysis indicates that thornback ray populations started to expand between 580,000 and 362,000 years ago, which suggests that the Last Glacial Maximum (20,000 years ago) had mainly affected the distribution of populations rather than population size.

Species of Fucus are among the dominant seaweeds along Northern Hemisphere shores, but taxonomic designations often are confounded by significant intraspecific morphological variability. We analyzed intra- and inter-specific phylogenetic relationships within the genus (275 individuals representing 16 taxa) using two regions of the mitochondrion: a variable intergenic spacer and a conserved portion of the 23S subunit. Bayesian ML and MP analyses verified a shallow phylogeny with two major lineages (previously reported) and resolved some intra-lineage relationships. Significant species-level paraphyly/polyphyly was observed within lineages 1A and 2. Despite higher species richness in the North Atlantic, a North Pacific origin of the genus is supported by a gradient of decreasing haplotype and nucleotide diversities in F distichus from the North Pacific to the East Atlantic. (c) 2006 Elsevier Inc. All rights reserved.

We used Single Strand Conformation Polymorphism (SSCP) of mtDNA control region to assess the population structure of the flatfish Pleuronectes platessa (plaice), to compare these data with a previous study based on microsatellite loci, and to test for possible sex-biased dispersal. From 461 individuals, 163 haplotypes were identified across 11 locations. Diversity was higher with mtDNA (h = 0.776 to 0.981; pi = 0.0178 to 0.0298) as compared to microsatellite loci using the same samples (H-e = 0.721 to 0.77). Genetic diversity was lower in samples from Iceland and Faroe, as compared to the continental shelf samples. Although both classes of markers revealed a relatively strong differentiation between shelf and off-shelf populations (theta = 0.1015 and theta = 0.0351, respectively), only the mtDNA data were able to detect differentiation within the continental shelf, i.e., a North Sea-Irish Sea group which was weakly distinguishable from Norway (theta = 0.0046), the Baltic (theta = 0.0136) and the Bay of Biscay (theta = 0.0162). No evidence was obtained for isolation by distance, nor for sex-biased dispersal. This study demonstrates the importance of using more than one class of markers, especially for species such as plaice, with large populations, high dispersal and recent colonisation histories. (C) 2004 Elsevier B.V. All rights reserved.

The seaweed Fucus serratus is hypothesized to have evolved in the North Atlantic and present populations are thought to reflect recolonization from a southern refugium since the last glacial maximum 18 000-20 000 years bp. We examined genetic structure across several spatial scales by analysing seven microsatellite loci in populations collected from 21 localities throughout the species' range. Spatial auto-correlation analysis of seven microsatellite loci revealed no evidence for spatial clustering of alleles on a scale of 100 m despite limited gamete dispersal in F. serratus of approximate to 2 m from parental individuals. Pairwise theta analysis suggested that the minimal panmictic unit for F. serratus was between 0.5 and 2 km. Isolation by distance was significant along some contiguous coastlines. Population differentiation was strong within the Skagerrak-Kattegat-Baltic Seas (SKB) (global theta = 0.17) despite a short history of approximate to 7500 years. A neighbour-joining tree based on Reynold's distances computed from the microsatellite data revealed a central assemblage of populations on the Brittany Peninsula surrounded by four well-supported clusters consisting of the SKB, the North Sea (Ireland, Helgoland), and two populations from the northern Spanish coast. Samples from Iceland and Nova Scotia were most closely aligned with northwest Sweden and Brittany, respectively. When sample sizes were standardized (N = 41), allelic diversity was twofold higher for Brittany populations than for populations to the north and threefold higher than southern populations. The Brittany region may be a refugium or a recolonized area, whereas the Spanish populations most likely reflect present-day edge populations that have undergone repeated bottlenecks as a consequence of thermally induced cycles of recolonization and extinction.

The internal transcribed spacer (ITS) region was sequenced in symbiotic dinoflagellates (zooxanthellae) from five morphospecies in the genus Madracis. The phylogeny of the symbionts is congruent with a companion phylogeny of the coral host. Comparison with known clade B symbiont ITS types reveals that M. mirabilis contains the B13 symbiont and that the other morphospecies contain the B7 symbiont. Madracis formosa also contains a previously undescribed type. The B7 and B13 symbionts appear to be highly specific to morphospecies in the genus Madracis. The host specificity between the B13 symbionts and its coral host may be the result of co-evolution of the coral-symbiont association and suggests that the brooding species, M. mirabilis, is reproductively isolated. Microhabitat differentiation associated with light utilization independent of depth is discussed.

For many corals, the existence of morphologically distinct yet sympatric populations/species implies reproductive isolation. Conversely, the presence of many intermediate and overlapping morphologies combined with synchronous, mass spawning suggests incomplete reproductive isolation. In Madracis (Scleractinia: Astrocoeniina: Pocilloporidae), high levels of morphological plasticity among the five most commonly recognized species (M. mirabilis, M. senaria, M. decactis, M. formosa and M. pharensis) on Caribbean reefs led us to question species boundaries. Phylogenetic relationships were investigated at the intra-individual, inter-individual and inter-specific levels using the ITSI-5.8S-ITS2 region (ca. 613 bp) of the ribosomal DNA cistron. Inter-specific divergence was ca. 6%, while intra-individual and intra-specific divergences ranged from 0% to 4.9% and 3.3% to 3.5%, respectively. M. senaria and M. mirabilis formed monophyletic groups. M. formosa, M. decactis and M. pharensis formed a paraphyletic complex. High levels of intra-individual and intra-specific ITS polymorphism in the decactis-formosa-pharensis cluster may be the result of very recent speciation within the clade (i.e. maintenance of ancestral polymorphism and incomplete lineage sorting), or the result of repeated introgressive hybridization among the three taxa. Polymorphism parsimony of 89 sites, including nine that showed additivity, revealed a phylogenetic topology more consistent with inter-taxal hybridization. Results are discussed in terms of weak reproductive barriers, and phylogenetic fission and fusion under Veron's model of reticulate speciation in corals. Ecological studies involving Madracis should consider AT decactis, M. formosa and M. pharensis as a complex.

The accidental introduction of Caulerpa taxifolia into the Mediterranean is no longer under dispute. What has eluded researchers until now is definitive evidence for the original, biogeographical source population. Here we present two independent lines of evidence that support an Australian origin for the Mediterranean populations of C. taxifolia. First, we reanalysed algal rDNA-internal transcribed spacer (rDNA-ITS) sequences, combining previously published sequences from different studies with 22 new sequences. The ITS sequence comparison showed that the Australian sample is the sister group of the Mediterranean-aquarium clade. Second, cloned bacterial 16S rDNA gene sequences were analysed from the associated microflora of C. taxifolia collected from Australia, Tahiti, the Philippines and the Mediterranean. Five bacterial lineages were identified, of which three were dominant. alpha Proteobacteria were the most abundant and were found in all samples. In contrast, members of the beta Proteobacterial line and Cytophaga-Flexibacter-Bacteroides line (CFB) were mainly associated with Mediterranean and Australian samples. Frequency distributions of the five bacterial lineages were significantly different among biogeographical locations. Phylogenetic analyses of the 54 bacterial sequences derived from the four C. taxifolia individuals resulted in a well-resolved tree with high bootstrap support. The topologies of the beta Proteobacteria and CFB mirror the geographical sources of their algal hosts. Bacterial-algal associations provide an identification tool that may have wide application for the detection of marine invasions.

In plant populations alleles often deviate from a random distribution and reveal positive autocorrelation at short distances. In species with both clonal and sexual reproduction, such clustering may be because ramets of the same genet were sampled at nearby locations. Alternatively, clustering may be the result of limited gene flow through pollen or seeds (isolation-by-distance). Were, we modify a conventional spatial autocorrelation analysis using the join-count statistic in order to differentiate between these two causes of genetic structure. We examined the distribution of seven microsatellite loci representing 37 alleles in a 20 x 80 m plot of a perennial population of eelgrass Zostera marina L. In analysing join-counts between all like genotypes we found significant genetic autocorrelation among ramets at distances between 1 and 7 m (P < 0.001). We then excluded joins between clonemates which were identified from the expected likelihood of their seven-locus genotypes. Without joins within genets, no autocorrelation was evident, indicating that most of the significant genetic clustering was caused by clonal spread. At distances up to 27 m, alleles were distributed at random, indicating a panmictic population at this spatial scale. These results illustrate the need for an a priori estimation of genet-ramet structure in clonally reproducing plants in order to avoid erroneous inferences about putative gene flow at various spatial scales

We examined the spatial distribution of genotypes in a perennial population of eelgrass, Zostera marina L., at two spatial scales. We mapped and sampled 80 ramets in a subtidal area of 20 x 80 m, and an additional 15 ramets in two 1-m(2) sub-quadrats. Ramets were genotyped for seven polymorphic microsatellite loci. Among a total number of 54 genotypes detected, 12 occurred more than once. The ramets of ten of these genotypes occurred in clusters and represented genets based on their expected multi-locus genotype frequencies. The size distribution of genets was uneven with estimated ramet numbers ranging from 2 to 5000. Whereas some areas displayed a high genet diversity, which may indicate past disturbances, large genets (greater than or equal to 10 m(2)) predominated in other locations of the sampled plot. Spatial heterogeneity in clone distribution was also obvious at the smaller sampling scale (15 ramets sam.. pled within m(2)) were the clonal diversity (genets identified/ramets sampled) was 0.24 in one quadrat, and 0.077 in the ether. Ramets belonging to the largest clone were maximally 17 In apart, which corresponds to a genet age of 67 yr based on annual rhizome growth rates. We conclude that the spatial arrangement of clones in seagrasses allows inferences about the past demography and the disturbance regime at a given site which may prove useful for coastal zone management of ecologically valuable seagrass meadows.

Caulerpa (Chlorophyta, Ulvophyceae) is a common marine tropical-subtropical genus of about 70 species, inhabiting the eulittoral zone on rocks and corals as well as mangroves. The genus is particularly diverse and abundant along southern Australian coasts, where it is thought to have originated. Here, we compare sequences from the nuclear ribosomal cistron among five species of Caulerpa, including nine populations of Caulerpa filiformis from two biogeographic regions : five from Australia and four from South Africa. Species relationships were well resolved by internal transcribed spacer (ITS) sequences and supported by high bootstrap values as follows: (C. geminata (C. simpliciuscula (C. trifaria (C. scalpelliformis (C. filiformis – Australia, C. filiformis – South Africa ))))). Nucleotide divergence within C. filiformis was low with four and five nucleotide differences present in ITS1 and ITS2 respectively. Unexpectedly, the 5.8S rRNA gene showed eight nucleotide differences between the Australian and So...

Amino acid sequences of alpha- and beta-chains of human hemoglobin and of hemoglobins of coelacanth and 24 teleost fish species, including 11 antarctic and two temperate Notothenioidei, were analyzed using maximum parsimony. Trees were derived for the alpha- and beta-chains separately and for tandemly arranged sequences, using the human and coelacanth sequences as outgroups in all analyses. The topologies of the trees of the alpha- and beta-chains are highly congruent and indicate a specific pattern of gene duplications and gene expression of teleost hemoglobins which has not yet been investigated into more detail. The Notothenioid fish generally contain a single major hemoglobin and often a second minor component. The alpha- and beta-chains of the major components form a monophyletic group in all investigated trees, with the nonantarctic Pseudaphritis as their sister taxon. The minor chains also are a monophyletic group and form an unresolved cluster with the major chains and the hemoglobins of tuna and red gurnard. The Notothenioid families Nototheniidae and Bathydraconidae appear to be paraphyletic.

Postelsia palmaeformis Ruprecht is an annual species, occuring from southern California to Vancouver Island Canada, in upper intertidal sites exposed to extreme wave shock. Because of its limited spore dispersal, discrete and inbred populations are likely on the local scale, yet dispersal of drifting and fertile thalli raises the possibility of outbred populations on a regional scale. M13 minisatellite DNA fingerprinting and random amplified polymorphic DNA (RAPD) markers were used in a complementary fashion to investigate genetic variability among 24 individuals on scales of clusters (= coalesced holdfasts).

Phylogenetic relationships in the Desmarestiales (Phaeophyceae) were inferred among the monotypic Arthrocladia (Arthrocladiaceae) and 27 isolates from Desmarestiaceae, representing 17 taxa of Desmarestia and the monotypic Antarctic genera Himantothallus and Phaeurus. Phaeurus and Arthrocladia were used as outgroups. Parsimony analyses of nuclear ribosomal DNA internal transcribed spacer (ITS1 and ITS2) sequences, in which gaps were both included and excluded yielded well-resolved trees with a consistent general branching pattern. A parallel analysis of nine morphological and life-history characters and three ecological characters yielded a similar tree but provided little resolution in the terminal clades. The position of the monotypic Arthrocladia villosa within the Desmarestiales is consistent with monophyly for the order, but its position as the most primitive desmarestialean is not resolvable from the molecular data set. The basal position of Phaeurus, the Antarctic Desmarestia species, and Himantothallus is consistent with the hypothesis of a Southern Hemisphere origin for the family Desmarestiaceae. The more recent Northern Hemisphere ''aculeata'' clade evolved from an Antarctic ancestor. A ''D. aculeata-like'' species was ancestral to a lineage characterized by annual sporophytes with high sulfuric acid content, which radiated into many species, widely distributed in both hemispheres. Mapping of morphological and ecological characters onto the molecular tree confirm the informativeness of sulfuric acid-containing vacuoles and unilocular sporangial types. There is good congruence between phylogenetic tree topology and temperature imprints in relation to biogeographic distribution, supporting the theory that temperature tolerance is a conservative trait.

The thornback ray, Raja clavata , is an elasmobranch (cartilaginous fish). Since the 1950s, its stock has severely declined. In order to investigate the genetic population structure of the species, we developed microsatellite loci. The five loci reported here have eight to 48 alleles per locus and display an observed heterozygosity ranging from 0.32 to 0.98 with no deviation from the Hardy–Weinberg equilibrium. In the test panel of 122 individuals from three populations, no null alleles, band-stuttering, large allele dropouts or linkage disequilibrium was detected.

Molecular phylogenetic studies on the evolution of the red algae indicate that this ancient division has many lineages that have recently undergone radiations. One such example is the cold-temperate family Palmariaceae. In this study, sequences from the ribosomal DNA internal transcribed spacer regions were compared among ten species in the Palmariaceae from both Atlantic and Pacific sites. Phylogenetic analyses of sequence data, in which Rhodophysema georgii Batters was used as outgroup and root, indicate a radiation into four clades, three of which contain species of ''Palmaria'' and the fourth species of Halosaccion. Palmaria palmata (L.) Kuntze, the type and only North Atlantic species in the genus, stands apart from all remaining species in the family and terminates the most basal branch in the rooted tree. The three more derived clades have radiated mainly in the North Pacific. Southern Ocean Palmaria and North Atlantic Devaleraea are hypothesized to have invaded from separate but closely related North Pacific ancestors. The ease with which sequences could be aligned combined with an unsaturated transition: transversion ratio and modest divergence involving predominantly point mutations suggests that the initial radiation is relatively recent (late Miocene-Pliocene) and that the Devaleraea-Palmaria clade is even more recent (late Pliocene-Pleistocene).

Genetic variation was examined within and among North Atlantic, North Sea and Baltic populations of the benthic red alga Phycodrys rubens using allozymes and random amplified polymorphic DNA (RAPD) markers. On western and eastern North Atlantic coasts distinct allozyme types were found, with the exception of western Newfoundland where East and West Atlantic types co-occur. Along the European coasts, two genetic groups were distinguished by fixed allelic differences: an outer oceanic group and a North Sea/Baltic group. The two genetic types co-occur in the Skagerrak and Kattegat region. Reproductive isolation between the two types is suggested by the lack of hybrids in the overlap zones, and they may therefore represent sibling species. Unexpectedly an analysis of RAPD variation was unable to recover the two cryptic species identified using allozymes. Within-population RAPD variation was similar to or greater than between-population variation. The lack of structure in the RAPD data cannot be attributed solely to technical artefacts of the method but appears to reflect real biological variability. Within-population genomic polymorphisms caused by frequent mutational events are discussed, as are high amounts of genetic drift and possible disruptive selection brought about by stressed habitats. Finally, Baltic and extra-Baltic salinity ecotypes are known to exist in P, rubens. However, no correlation between ecotypic variation and allozyme groups was detected.

In order to investigate how episodes of geological and climatic change have influenced the distribution and evolutionary diversification of Arctic to cold temperate-North Atlantic seaweed species, intraspecific genetic variation was analyzed among isolates of the sublittoral, benthic red alga Phycodrys rubens (collected between June 1992 and January 1994). Rooted phylogenetic analyses of nuclear ribosomal DNA internal transcribed spacer (ITS) sequences and the plastid encoded Rubisco spacer sequences suggest that P. rubens invaded the North Atlantic from the Pacific shortly after the opening of the Bering Strait (3 to 3.5 million years ago), colonizing both the western and eastern Atlantic coasts. Based on these data we further hypothesize that P. rubens survived along the European coasts during the more recent Pleistocene glaciations, while becoming locally extinct along the North American Atlantic coasts. Following retraction of the last ice sheet, the western Atlantic coast was colonized a second time from the Pacific. The presence of two distinct genetic types (based on ITS and Rubisco sequences) along the European coasts is postulated to be a result of isolation and subsequent differentiation. This is likely because ice-free areas are known to have existed in northern Scotland and Norway during the last glaciation. The presence of an East Atlantic genetic type along the West Atlantic coast is believed to be a recent introduction (caused by human activity) of P. rubens to Newfoundland.

Phylogenetic relationships in the Arctic-Antarctic disjunct seaweed species Acrosiphonia arcta (Dillwyn) J. G. Agardh (Acrosiphoniales, Chlorophyta) were examined using restriction fragment-length polymorphism analysis of the fast-evolving nuclear ribosomal intergenic spacer (IGS) region and random amplified polymorphic DNA (RAPD) markers. Twenty-two isolates collected from 10 different locations in both hemispheres were compared. Five IGS length classes were identified among the 10 locations. Throughout the North Atlantic, IGS regions were found to be extremely homogeneous whereas RAPD patterns revealed subdivided populations that suggest founder effects. Acrosiphonia arcta populations found in the Arctic and North Atlantic oceans are hypothesized to be of Pacific origin. Extensive differences found between Arctic Greenland populations and those in the North Atlantic suggest that colonization of Arctic Greenland occurred as an independent event. Recolonization of the Antarctic peninsula from Southern Chile is favored, whereas the directionality of transequatorial passage along the western coast of the Americas could be in either direction.

Fucus serratus and F. evanescens commonly occur on Northern European shores. Nine microsatellite loci were developed for F. serratus (8-22 alleles, observed heterozygosities = 0.367-0.850) and one for F. evanescens (seven alleles, observed heterozygosity = 0.804). Cross-amplification was apparent, as five F. serratus loci were polymorphic in F. evanescens and 2-5 were polymorphic in F. vesiculosus, F. distichus, and F. spiralis.

Two examples of the most extreme biogeographic disjunctions in benthic marine algae are found in Acrosiphonia arcta (Chlorophyta) and Desmarestia viridis/willii (Phaeophyta). Both species are members of the Arctic and Antarctic boreal and subboreal marine floras. Although both genera have temperate species, neither genus has subtropical or tropical representatives. Comparisons of the fast-evolving ribosomal DNA internal transcribed spacer regions among isolates in each of the two species collected from both hemispheres showed an unexpected near sequence identity suggesting that these biogeographic disjunctions are of recent origin, possibly as recent as the last Pleistocene glacial maximum (18000 yr ago). Paleoclimatic explanations that rely on a much earlier transequatorial passage of cold-adapted species through a narrowed and cooler tropical belt during the Oligocene/Miocene (38 to 7 Ma ago) are unlikely. We hypothesize that despite the separated evolutionary histories of the northern and southern hemisphere cold-water marine floras, deep-water dispersal of microthalli has occurred and probably occurs on a regular basis.

Nucleotide sequences of the nuclear ribosomal DNA internal transcribed spacers (ITS1 and ITS2), the 5.8S, and short stretches of the adjacent 18S and 26S coding regions were determined in isolates from four disjunct Cladophora albida (Huds.) Kutz. populations (NE-America, W-Europe, Japan, and W-Australia). The two Pacific isolates share nearly identical ITS sequences as do the two Atlantic isolates. In contrast, interoceanic comparisons exhibit a 21% sequence difference. Variation within ITS regions is useful for identification of population groups on a regional or oceanic scale. However, both spacers are characterized by numerous repeat motifs as well as point mutations, which result in alignment problems at the interspecific level within Cladophora.

Analysis of the reassociation kinetics of the DNA from Cladophora pellucida (Huds.) Kutz. indicates that the genome of this benthic alga is comprised of approximately 75% repetitive sequences. Single-copy sequences reassociated with a rate constant of 1.8 x 10(-3) M-1.s-1, which corresponds to a haploid genome size of 4.7 x 10(8) bp. Genotypic relationships between members of the form section Longiarticulatae were determined by the method of DNA-DNA hybridization. No significant divergence was observed between the single-copy sequences of C. pellucida isolates from the East Atlantic coast and Mediterranean Sea. Cladophora feredayi Harv. and C. att. ad pellucida from Australia and C. pellucidoidea van den Hoek from the West Atlantic coast were highly and about equally divergent from C. pellucida. The data support the hypothesis that the West Atlantic-West Pacific divergence reflects the middle Miocene closure of the Mediterranean-Indo-Pacific seaways, and the hypothesis that the Northwest Atlantic-Northeast Atlantic divergence reflects the middle Miocene thermal separation of these coasts.

Seven Indo-West Pacific Prochloron Lewin samples and one Caribbean cyanobacterial sample were compared by DNA-DNA reassociation experiments. The quality and quantity of extracted DNA were affected by several cell preservation methods and DNA extraction and purification procedures. We conclude that Prochloron samples from six different hosts and three different geographical locations spanning ca. 9000 km belong to a single species but are not closely related to the strain of cyanobacteria used. The intraspecific genotypic variation within Prochloron is discussed with respect to host specificity and geographical distribution (...)

The nuclear small-subunit ribosomal DNA sequence from the freshwater red alga Boldia erythrosiphon Herndon emend Howard et Parker was determined. Phylogenetic analysis confirms the positioning of this species within the bangiophycidean order of the Compsopogonales. The results strongly suggest that occupation of freshwater habitats by marine red algae has happened at least twice during red algal evolution.

The phylogeography of thornback rays (Raja clavata) was assessed from European waters, using five nuclear microsatellite loci and mitochondrial cytochome b sequences. Strong regional differentiation was found between the Mediterranean basin, the Azores and the European continental shelf. Allelic and haplotype diversities were high in Portuguese populations, consistent with the existence of a refugium along the Iberian Peninsula. Unexpectedly, high diversity was also found in the English Channel/North Sea area. The lowest genetic diversity was found in the Black Sea. Populations sampled from the Mediterranean, Adriatic and Black Seas were characterized by a single mitochondrial haplotype. This haplotype was also the most ancestral and widespread outside of the Mediterranean basin except for the Azores. Populations from the Azores were dominated by a second ancestral haplotype which was shared with British populations. Results from multidimensional scaling, amova and nested clade analysis indicate that British waters are a secondary contact zone recolonized from at least two refugia--one around the Iberian Peninsula and one possibly in the Azores. Links to a potential refugium known as the Hurd Deep, between Cornwall and Brittany, are discussed. Finally, a historical demographic analysis indicates that thornback ray populations started to expand between 580,000 and 362,000 years ago, which suggests that the Last Glacial Maximum (20,000 years ago) had mainly affected the distribution of populations rather than population size.

As the most widespread seagrass in temperate waters of the Northern Hemisphere, Zostera marina provides a unique opportunity to investigate the extent to which the historical legacy of the last glacial maximum (LGM18 000-10 000 years bp) is detectable in modern population genetic structure. We used sequences from the nuclear rDNA-internal transcribed spacer (ITS) and chloroplast matK-intron, and nine microsatellite loci to survey 49 populations (2000 individuals) from throughout the species' range. Minimal sequence variation between Pacific and Atlantic populations combined with biogeographical groupings derived from the microsatellite data, suggest that the trans-Arctic connection is currently open. The east Pacific and west Atlantic are more connected than either is to the east Atlantic. Allelic richness was almost two-fold higher in the Pacific. Populations from putative Atlantic refugia now represent the southern edges of the distribution and are not genetically diverse. Unexpectedly, the highest allelic diversity was observed in the North Sea-Wadden Sea-southwest Baltic region. Except for the Mediterranean and Black Seas, significant isolation-by-distance was found from ~150 to 5000 km. A transition from weak to strong isolation-by-distance occurred at ~150 km among northern European populations suggesting this scale as the natural limit for dispersal within the metapopulation. Links between historical and contemporary processes are discussed in terms of the projected effects of climate change on coastal marine plants. The identification of a high genetic diversity hotspot in Northern Europe provides a basis for restoration decisions.

Philopatry to spawning grounds combined with well-known migratory patterns in the flatfish Pleuronectes platessa (plaice) has led to the hypothesis that regional populations may reflect relatively discrete, genetic stocks. Using six microsatellite loci we genotyped 240 adult individuals collected from locations in Norway, the Faeroe plateau, the Irish Sea, the Femer Baelt, Denmark, and the southern North Sea, and 240 0-class juveniles collected from five nursery-ground locations in Iceland, northwest Scotland, two sites in the Wadden Sea, and the Bay of Vilaine in Southern Brittany. The mean number of alloles/locus ranged from 5.3 to 20.4, with a mean of 13.9. Expected heterozygosity was uniformly high across all locations (multilocus H-exp = 0.744+/-0.02). Pairwise comparisons of theta among all 11 locations revealed significant differentiation between Iceland and all other locations (theta = 0.0290*** to 0.0456***), which is consistent with the deep-water barrier to dispersal in plaice. In contrast, no significant differentiation was found among any of the remaining continental-shelf sampling locations. This suggests that regional stocks are themselves composed of several genetic stocks under a model of panmixia which persists even to the spawning grounds. The presence of significant heterozygote deficiencies at all locations (not due to null alleles) suggests a temporal Wahlund effect yet the absence of significant population differentiation among continental shelf localities makes this explanation alone difficult to. reconcile. Sampling of eggs at the spawning grounds will be required to resolve this issue. Causes of the mismatch between genetic and geographical stocks is discussed in the context of high gene flow.

Axenic cultures of the tropical green seaweed Cladophoropsis membranacea (C. Agardh) Boergesen were obtained by cutting 2-mm apical tips from fast-growing unialgal filaments and incubating them in 1/2PES containing 100-mu-g.mL-1 cefotaxime. After 1 week, apical tips were cut from the newly grown plantlets, washed through a series of sterile drops of seawater, and incubated in sterile 1/2PES. Plantlets were screened for the presence of bacteria by incubating droplets of the culture medium on peptone agar plates and by examining DAPI-stained filaments. Ninety to one hundred percent of the plantlets obtained with this treatment were axenic. Cefotaxime was also effective against cyanobacteria but of only limited value in rhodophytes.

Seven microsatellite loci were developed for Pleuronectes platessa, a commercially important flatfish found throughout northern European waters. The number of alleles per locus ranged from 11 to 33 and heterozygosities from 0.74 to 0.96. Cross-reactivity was tested against six other flatfish species. Significant amplification was found in all species from one to all seven of the loci.

Ascophyllum nodosum L. dominates rocky intertidal shores throughout the temperate North Atlantic. Six microsatellite loci were developed for A. nodosum using enriched libraries. The number of alleles ranged from 9 to 24 and heterozygosities from 0.2213 to 0.7785. Ascophyllum is monotypic. There was no cross-reactivity observed with Fucus serratus, F. vesiculosus or F. evanescens.

Ascophyllum nodosum is dominant seaweed along many rocky intertidal shores throughout the North Atlantic. Next to the kelps, fucalean taxa such as Ascophyllum are the largest macrophytes and provide important habitat for invertebrates. Understanding the underlying genetic structure of natural populations over a range of spatial scales can reveal how the causes of structure may change with scales. Separating population history from population structure may also be elucidated. The analysis is based on six polymorphic microsatellite loci and> 1000 individuals. Strong genetic structure at small spatial scale was found and is consistent with demographic models based on long-lived individuals, low recruitment and many sib matings. At large spatial scales only weak population differentiation was found. This is consistent with recent recolonization of the North Atlantic following the last glacial maximum.

DNA-DNA hybridizations between single-copy nuclear DNA from Laminaria digitata and total DNA from L. saccharina, L. Hyperborea, L. rodriguezii, L. ochroleuca and Chorda filum, respectively, show that these species of Laminaria are genotypically closely related.Chorda filum is only distantly related with L. digitata. Based on the thermal elution patterns of the DNA hybrids, as quantified by ΔTm(e) values, it is hypothesized that all five species of Laminaria evolved at about the same time from their most recent common ancestor some 15–19 Ma ago. This phylogenetic hypothesis is discussed in relation to the history of modern laminarialean distribution patterns.

In traditional chlorophytan systems the organizational level was the primary character for the distinction of main groups (classes and orders). For instance, in Fott (1971), the flagellate level corresponds with the Volvocales, the coccoid level with the Chlorococcales, the filamentous level with the Ulotrichales, the siphonocladous level with the Siphonocladales, and the siphonous level with the Bryopsidales. The new system presented here is an elaboration and emendation of recently proposed taxonomies and their underlying phylogenetic hypotheses, and it is mainly based on ultrastructural features which have become available over the last 15 years. The following criteria are used for the distinction of classes and orders: (1) architecture of the flagellate cell (flagellate cells are considered as the depositories of primitive characters); (2) type of mitosis-cytokinesis; (3) place of meiosis in the life history and, consequently, the sexual life history type; (4) organizational level and thallus architecture; (5) habitat type (marine versus feshwater and terrestrial); (6) chloroplast type. The following classes are presented: Prasinophyceae, Chlamydophyceae, Ulvophyceae (orders Codiolales, Ulvales, Cladophorales, Bryopsidales, Dasycladales), Pleurastrophyceae (?), Chlorophyceae s.s. (orders Cylindrocapsales, Oedogoniales, Chaetophorales), Zygnematophyceae, Trentepohliophyceae, Charophyceae (orders Klebsormidiales, Coleochaetales, Charales). The new system no longer reflects the traditional hypothesis of a stepwise evolutionary progression of organizational levels in which the flagellate level represents the most primitive lineage, the coccoid and sarcinoid levels lineages of intermediate derivation, and the filamentous, siphonocladous and siphonous levels the most derived lineages. Instead, it is now hypothesized that these levels have arisen over and over again in different chlorophytan lineages which are primarily characterized by their type of flagellate cell. The flagellate green algal classes Prasinophyceae (with organic body scales) and Chlamydophyceae probably represent bundles of highly conservative lineages that diverged very long ago. Consequently, extant genera and species in these classes can be expected to have emerged long ago. Fossil evidence points to a minimum age of 600 Ma of certain extant Prasinophycean genera, and molecular evidence to a minimum age of 400–500 Ma of a fewChlamydomonas species. On the contrary, the most derived “green algal” lineage, the Angiosperms, can be expected to consist of, on average, much younger genera and species. Fossil evidence points to a minimum age of genera of 5–60 Ma. Lineages of intermediate evolutionary derivation (Ulvophyceae, Chlorophyceae, Charophyceae) can be expected to encompass genera and species of intermediate age. Fossil and (limited) molecular evidence point to a minimum age of 230–70 Ma of extant genera in Bryopsidales, Dasycladales and Cladophorales (Ulvophyceae) and of 250–80 Ma of extant genera in Charales (Charophyceae).

Genomic relationships between North Atlantic, Australian and Japanese isolates of the benthic seaweed Cladophora albida (Huds.) Kutz. were examined in 1987 by means of DNA-DNA hybridization. The data indicate that C. albida can be divided into a North Atlantic and an Indo-West Pacific group with an intergroup hybridization response of 25 to 30% and 5.5° to 6.0°C for hybridization percentage and ΔT me the melting temperature reduction of hybridized sequences, respectively. This level of genome divergence is considerably higher than that observed in most other eukaryotes. The separation between the two C. albida groups presumably dates back to the closure of the Asian part of the Tethys Ocean, about 12 million years ago. The data also indicate that transatlantic C. albida populations have a greater genetic inter-relatedness than have Japanese and Australian populations. In C. albida there is no clear correlation between molecular evolution and the evolution of morphological traits. C. albida and C. rupestris (L.) Kutz have hardly any DNA sequences in common.

Genotypic relationships between seven Prochloron samples isolated from different didemnid ascidian hosts collected at the Palau archipelago and Munda (Solomon Islands) and one cyanobacterial (Synechocystis) strain were determined by DNA-DNA reassociations. Thermal stability values of DNA-DNA hybrids indicate that all Prochloron samples involved are mutually very closely related and only slightly related with the Synechocystis strain. It is concluded that the Prochloron samples tested are representatives of one and the same species.