PHYLOGENY AND SYSTEMATICS OF THE MARINE ALGAL FAMILY GRACILARIACEAE (GRACILARIALES,RHODOPHYTA) BASED ON SMALL SUBUNIT rDNA AND ITS SEQUENCES OF ATLANTIC AND PACIFIC SPECIES1

We sequenced the small subunit rDNA and internal transcribed spacer region of Gracilariaceae from the tropical Atlantic and Pacific, with emphasis on flattened or compressed species. Sequence comparisons confirmed three main lineages of Gracilariaceae: Curdiea/Melanthalia, Gracilariopsis/Gracilariophila, and Gracilaria. The Curdiea/Melanthalia diverged early in the family. Gracilariopsis was paraphyletic, because at least one Gracilariophila species evolved from it. The Atlantic Gracilariopsis were monophyletic and separated from the Pacific lineages. The Gracilaria included all species referable to its own species and to Hydropuntia, which was paraphyletic, formed by distantly related lineages. The new combination Gracilaria pauciramosa (N. Rodríguez Ríos) Bellorin, M. C. Oliveira et E. C. Oliveira is proposed for Polycavernosa pauciramosa N. Rodríguez Ríos. Recognition of subgenera within Gracilaria, based on spermatangial arrangement, was not supported. Instead, infrageneric groups were delineated by geographic origins and combinations of reproductive characters. Most Pacific species with either “textorii” or “verrucosa” type spermatangia were deeply separated from Atlantic species. Within the Atlantic Gracilaria, a lineage encompassing mostly tropical cylindrical species with “henriquesiana” type spermatangia and distinctive cystocarp anatomy was recognized. A lineage was also retrieved for cold water stringy species with verrucosa type spermatangia. Several species from the western Atlantic are closely related to Gracilaria tikvahiae McLachlan with nearly identical morphology. On the other hand, most flattened species from the tropical Atlantic were closely related despite their diverse morphologies. The interpretation of our data in addition to the literature indicates that more populations from the Indo‐Pacific must be studied before a general picture of Gracilariaceae evolution can be framed.     

PHYLOGEOGRAPHY OF GRACILARIA TIKVAHIAE (GRACILARIACEAE,RHODOPHYTA): A STUDY OF GENETIC DISCONTINUITY IN A CONTINUOUSLY DISTRIBUTED SPECIES BASED ON MOLECULAR EVIDENCE1

Gracilaria tikvahiae, a highly morphologically variable red alga, is one of the most common species of Gracilariaceae inhabiting Atlantic estuarine environments and the Intracoastal Waterway of eastern North America. Populations of G. tikvahiae at the extremes of their geographic range (Canada and southern Mexico) are subjected to very different environmental regimes. In this study, we used two types of genetic markers, the chloroplast‐encoded rbcL and the nuclear internal transcribed spacer (ITS) region, to examine the genetic variability within G. tikvahiae, for inferring the taxonomic and phylogenetic relationships between geographically isolated populations, and to discuss its distributional information in a phylogeographic framework. Based on rbcL and ITS phylogenies, specimens from populations collected at the extreme distributional ranges reported for G. tikvahiae are indeed part of the same species; however, rbcL‐ but not ITS‐based phylogenies detected phylogenetic structure among the ten G. tikvahiae different haplotypes found in this study. The four distinct rbcL lineages were identified as 1) a Canadian–northeast U.S. lineage, 2) a southeast Florida lineage, 3) an eastern Gulf of Mexico lineage, and 4) a western Gulf of Mexico lineage. We found no evidence for the occurrence of G. tikvahiae in the Caribbean Sea. Observed phylogeographic patterns match patterns of genetic structures reported for marine animal taxa with continuous and quasicontinuous geographic distribution along the same geographic ranges.     

GENETIC DATA HINT AT A COMMON DONOR REGION FOR INVASIVE ATLANTIC AND PACIFIC POPULATIONS OF GRACILARIA VERMICULOPHYLLA (GRACILARIALES,RHODOPHYTA)1

Gracilaria vermiculophylla (Ohmi) Papenf., an agar‐producing red alga introduced from northeast Asia to Europe and North America, is often highly abundant in invaded areas. To assay its genetic diversity and identify the putative source of invasive populations, we analyzed the mitochondrial cytochrome c oxidase subunit I (cox1) gene from 312 individuals of G. vermiculophylla collected in 37 native and 32 introduced locations. A total of 19 haplotypes were detected: 17 in northeast Asia and three in Europe and eastern and western North America, with only one shared among all regions. The shared haplotype was present in all introduced populations and in ～99% of individuals in the introduced areas. This haplotype was also found at three native locations in east Korea, west Japan, and eastern Russia. Both haplotype and nucleotide diversities were extremely low in Europe and North America compared to northeast Asia. Our study indicates that the East Sea/Sea of Japan is a likely donor region of the invasive populations of G. vermiculophylla in the east and west Atlantic and the east Pacific.     

MOLECULAR DELINEATION OF SPECIES AND SPECIES RELATIONSHIPS IN THE RED ALGAL AGAROPHYTES GRACILARIOPSIS AND GRACILARIA (GRACILARIALES)1

Delineation of species in the economically important agarophyte genera Gracilaria and Gracilariopsis has proven extremely difficult using available morphological characteristics. In this study, we examine the usefulness of two transcribed spacers for molecular systematic studies of these genera. The polymerase chain reaction was used to amplify the internal transcribed spacers (ITSs) and the intervening 5.8S ribosomal DNA of the nuclear ribosomal repeat region. In addition, a plastid spacer region and flanking regions of coding genes were amplified from the RUBISCO operon. Both regions were sequenced for individuals and populations of Gracilariopsis lemaneiformis (Bory) Dawson, Acleto, et Foldvik to determine the usefulness of these spacers in delimiting populations. These studies reveal that there is as much variation among individuals of a population as there is between individuals of geographically separate populations. In addition, the ITS spacer regions were compared between different species of Gracilariopsis and Gracilaria. The nuclear ITS spacer region is conserved at a species level in both genera and provides phylogenetically informative characters that can be used to examine species interrelationships among relatively closely related taxa. However, because of the difficulties of aligning this entire region among species from the two genera, the ITS region is not useful for examining intergenera relationships. ITS interspecies sequence comparisons indicate that Gracilariopsis lemaneiformis from California is significantly different from G. lemaneiformis from China and that a species of Gracilariopsis from Peru is more closely related to G. lemaneiformis from North Carolina than it is to the other Gracilariopsis species examined. In addition, these studies indicate that Gracilaria chilensis Bird, McLachlan, et Oliveira from New Zealand and Gracilaria tenuistipitata Chang et Xia from southeast Asia are as closely related as are Gracilaria verrucosa (Hudson) Papenfuss, G. pacifica Abbott, and Gracilaria robusta Kylin. Phylogenetic analysis of aligned plastid spacer sequences from Gracilaria and Gracilariopsis taxa provide similar conclusions about species relationships.     

GENETIC DIVERSITY AND INTROGRESSION IN TWO CULTIVATED SPECIES (PORPHYRA YEZOENSIS AND PORPHYRA TENERA) AND CLOSELY RELATED WILD SPECIES OF PORPHYRA (BANGIALES,RHODOPHYTA)1

We investigated the genetic variations of the samples that were tentatively identified as two cultivated Porphyra species (Porphyra yezoensis Ueda and Porphyra tenera Kjellm.) from various natural populations in Japan using molecular analyses of plastid and nuclear DNA. From PCR‐RFLP analyses using nuclear internal transcribed spacer (ITS) rDNA and plastid RUBISCO spacer regions and phylogenetic analyses using plastid rbcL and nuclear ITS‐1 rDNA sequences, our samples from natural populations of P. yezoensis and P. tenera showed remarkably higher genetic variations than found in strains that are currently used for cultivation. In addition, it is inferred that our samples contain four wild Porphyra species, and that three of the four species, containing Porphyra kinositae, are closely related to cultivated Porphyra species. Furthermore, our PCR‐RFLP and molecular phylogenetic analyses using both the nuclear and plastid DNA demonstrated the occurrence of plastid introgression from P. yezoensis to P. tenera and suggested the possibility of plastid introgression from cultivated P. yezoensis to wild P. yezoensis. These results imply the importance of collecting and establishing more strains of cultivated Porphyra species and related wild species from natural populations as genetic resources for further improvement of cultivated Porphyra strains.     

DISSECTION OF TWO DISTINCT DEFENSE‐RELATED RESPONSES TO AGAR OLIGOSACCHARIDES IN GRACILARIA CHILENSIS (RHODOPHYTA) AND GRACILARIA CONFERTA (RHODOPHYTA)1

The two agar‐producing red algae, Gracilaria chilensis C. J. Bird, McLachlan & E. C. Oliveira and Gracilaria conferta (Schousboe ex Montagne) Montagne, responded with hydrogen peroxide (H₂O₂) release when agar oligosaccharides were added to the medium. In G. conferta, a transient release was observed, followed by a refractory state of 6 h. This response was sensitive to chemical inhibitors of NADPH oxidase, protein kinases, protein phosphatases, and calcium translocation in the cell, whereas it was insensitive to inhibitors of metalloenzymes. Transmission electron microscopic observations of the H₂O₂‐dependent formation of cerium peroxide from cerium chloride indicated oxygen activation at the plasma membrane of G. conferta. A putative system, consisting of a receptor specific to agar oligosaccharides and a plasma membrane‐located NADPH oxidase, appears to be responsible for the release of H₂O₂ in G. conferta. Subcellular examination of G. chilensis showed that the H₂O₂ release was located in the cell wall. It was sensitive to inhibitors of metalloenzymes and flavoenzymes, and no refractory state was observed. The release was correlated with accumulation of an aldehyde in the algal medium, suggesting that an agar oligosaccharide oxidase is present in the apoplast of G. chilensis. The presence of this enzyme could also be demonstrated by polyacrylamide electrophoresis under nondenaturating conditions and proven to be variable. Cultivation of G. chilensis at 16 to 17°C resulted in significantly stronger expression of agar oligosaccharide oxidase than cultivation at 12°C, which indicates that the enzyme is used under conditions that generally favor microbial agar macerating activity.     

GENETIC VARIABILITY AND POTENTIAL SOURCES OF GRATELOUPIA DORYPHORA (HALYMENIACEAE,RHODOPHYTA), AN INVASIVE SPECIES IN RHODE ISLAND WATERS (USA)1

Grateloupia doryphora (Montagne) M.Howe is an invasive foliose alga that was reported for the first time in Rhode Island, USA in 1997. The population has since increased in size and expanded in range. In this study, the genetic variation and potential sources of the Rhode Island G. doryphora population were examined using three types of molecular markers: randomly amplified polymorphic DNA (RAPD), nuclear internal transcribed spacer (ITS) sequences, and mitochondrial cox2–cox3 intergenic spacer (COX) sequences. No variation was detected in ITS or COX sequences among Rhode Island G. doryphora individuals. RAPDs, however, did reveal genetic variation, although banding patterns were similar, with RAPD genetic distances between individuals ranging from 0.00 to 0.17. The low level of genetic diversity observed within the Rhode Island population may be due to a small founder population or a founder population derived from a genetically uniform source. To identify possible sources of the Rhode Island invasion, individuals from nine geographically diverse populations of foliose Grateloupia were compared. Phylogenetic trees inferred from RAPD distances and ITS and COX sequences had similar topologies; thus there was phylogenetic congruence among these independent loci. The Rhode Island G. doryphora specimens were genetically similar to specimens from G. doryphora populations located in Portsmouth, England; Tholen Island, The Netherlands; and Brittany and Hérault, France. Interestingly, the G. doryphora population in each of these locations is itself due to an introduction event within the past 40 years.     

GRACILARIA HUMMII SP. NOV. (GRACILARIALES,RHODOPHYTA), A NEW NAME FOR THE AGAROPHYTE “GRACILARIA CONFERVOIDES” HARVESTED IN NORTH CAROLINA DURING WORLD WAR II1

Gracilaria hummii Hommers. et Freshwater is proposed as a new name for the inshore cylindrical species found in North Carolina that was treated as Gracilaria confervoides (L.) Grev. during World War II, and more recently as G. verrucosa (Huds.) Papenf. Molecular evidence places G. hummii in the Gracilis‐group in Gracilaria together with G. gracilis (Stackh.) Steentoft, L. M. Irvine et Farnham, the name currently applied to specimens formerly identified as G. confervoides and G. verrucosa. G. hummii differs from G. gracilis in possessing shallower male conceptacles in which the spermatangial filament originates from a surface cortical cell rather than from a subcortical cell. The cystocarps are similar, except that the gonimoblasts of G. hummii are attached to the base of the pericarp by numerous, prominent thickened terminal tubular cells and because terminal tubular cells are absent above the midregion of the cystocarp or in the vicinity of the ostiole. The gonimoblasts are subtended by a bundle of longitudinally oriented, thick‐walled secondary filaments of a type that has not been described before in Gracilaria but that may be characteristic of some other species. G. hummii occupies a basal position in the Gracilis‐group and is distinct from all other cylindrical North Carolina Gracilaria species, according to the molecular and morphological evidence.     

GRACILARIA VERMICULOPHYLLA (RHODOPHYTA,GRACILARIALES) IN THE VIRGINIA COASTAL BAYS,USA: COX1 ANALYSIS REVEALS HIGH GENETIC RICHNESS OF AN INTRODUCED MACROALGA

Gracilaria vermiculophylla (Ohmi) Papenfuss is an invasive alga that is native to Southeast Asia and has invaded many estuaries in North America and Europe. It is difficult to differentiate G. vermiculophylla from native forms using morphology and therefore molecular techniques are needed. In this study, we used three molecular markers (rbcL, cox2‐cox3 spacer, cox1) to identify G. vermiculophylla at several locations in the western Atlantic. RbcL and cox2‐cox3 spacer markers confirmed the presence of G. vermiculophylla on the east coast of the USA from Massachusetts to South Carolina. We used a 507 base pair region of cox1 mtDNA to (i) verify the widespread distribution of G. vermiculophylla in the Virginia (VA) coastal bays and (ii) determine the intraspecific diversity of these algae. Cox1 haplotype richness in the VA coastal bays was much higher than that previously found in other invaded locations, as well as some native locations. This difference is likely attributed to the more intensive sampling design used in this study, which was able to detect richness created by multiple, diverse introductions. On the basis of our results, we recommend that future studies take differences in sampling design into account when comparing haplotype richness and diversity between native and non‐native studies in the literature.     

GENETIC POPULATION STRUCTURE AND MATING SYSTEM IN CHONDRUS CRISPUS (RHODOPHYTA)1

Chondrus crispus Stackh. has been intensely studied, yet no study to date has elucidated its population structure or mating system despite many populations in which there was a haploid bias and lack of male gametophytes. Therefore, 12 nuclear microsatellite loci were identified in this red alga. Microsatellite markers were developed and tested against a panel of specimens collected from two shore levels at two sites in Brittany, France: Pointe de Primel and Pointe de la Jument, Concarneau. Single locus genetic determinism was verified at eight polymorphic loci, as only one band was observed for haploid genotypes, whereas one or two bands were observed for diploids. These markers enabled the detection of unique genotypes within sampled populations, indicating that very few fronds shared the same multilocus genotype. This finding suggests that asexual reproduction was not the prevailing mode of reproduction. In addition, we explored the hierarchical population structure showing that gene flow is restricted at small spatial scales (<50 m) between upper and lower Chondrus‐range populations within a shore. Sexual reproduction predominated in the populations of C. crispus studied, but probably due to fine‐scale spatial substructuring, inbreeding was also significant. In conclusion, this study reveals that fine‐scale genetic variation is of major importance in C. crispus, suggesting that differences between microhabitats should be essential in understanding evolutionary processes in this species.     

GRACILARIA VERMICULOPHYLLA (RHODOPHYTA,GRACILARIALES) IN HOG ISLAND BAY,VIRGINIA: A CRYPTIC ALIEN AND INVASIVE MACROALGA AND TAXONOMIC CORRECTION1

Gracilaria in Virginia, USA, is abundant and composed of thalli either having relatively flat or cylindrical branches. These two morphologies were referred to previously as G. foliifera (Forsskål) Bøgesen and G. verrucosa (Hudson) Papenfuss. However, G. verrucosa is regarded an invalid name, and the flat specimens are now referred to as G. tikvahiae McLachlan. This has created confusion about the nomenclature of Gracilaria from this region. Here we document that the cylindrical form that dominates Hog Island Bay, Virginia, is G. vermiculophylla (Ohmi) Papenfuss, an alien macroalga from the West Pacific. Most of the ecological studies performed at the Long Term Ecological Research (LTER) site in Hog Island Bay used this cylindrical species. The present study clarifies the taxonomical status of this species, and we identify attributes that make this alien successful in turbid coastal lagoons.     

HIGH LEVELS OF GENETIC DIVERSITY AND LOW LEVELS OF GENETIC DIFFERENTIATION IN NORTH SEA PSEUDO‐NITZSCHIA PUNGENS (BACILLARIOPHYCEAE) POPULATIONS1

Six microsatellite markers were used to investigate the genetic structure of North Sea Pseudo‐nitzschia pungens (Grunow ex P. T. Cleve) Hasle populations. Isolates were collected on 42 separate occasions from waters surrounding the German islands of Helgoland and Sylt over the course of three sampling periods: spring 2002, spring 2003, and autumn 2003. In total, 464 isolates were genotyped, of which 453 were different (i.e. clonal diversity was 98%). The numbers of alleles per locus ranged from 6 to 24 and the observed heterozygosities from 0.59 to 0.87 (mean H_o and H_e were 0.73); there were no significant departures from Hardy‐Weinberg equilibrium at any of the six loci. Sexual reproduction therefore appears to be important in the production of genetic variation. Over the temporal and spatial scales sampled (18 months and 100 km), weak genetic differentiation was detected both within and between sampling periods (significant F_ST values ranged from 0.0018 to 0.0389), suggesting that the German North Sea supports a single largely unstructured population of P. pungens.     

GENETIC VARIABILITY AND POPULATION DIFFERENTIATION INFERRED FROM DNA FINGERPRINTING IN SILVEREYES (AVES: ZOSTEROPIDAE)

This study evaluated DNA fingerprinting as a tool for estimating population genetic diversity and differentiation by comparing minisatellite variation in island and mainland populations of silvereyes (Aves: Zosterops lateralis). Three populations with different recent histories were compared: (1) Heron Island and neighboring islands, colonized 3000 to 4000 yr ago; (2) Lady Elliot Island, colonized within the past two decades; and (3) an adjacent mainland population, which presumably has existed for thousands of years. The degree of genetic variability within the three populations reflected both their size and the time since their colonization. Minisatellite diversity was highest in the mainland population, intermediate in the Capricorn Island group (which was shown to represent a single admixture), and lowest in the Lady Elliot Island population, possibly because of a recent population bottleneck during colonization. Mean band sharing between any two populations was less than the mean within either of those populations, and four fingerprint bands common to island birds were rare or absent in the fingerprints of mainland birds. In the absence of significant gene flow between the mainland and the islands, the populations have apparently become distinct at minisatellite loci, as evidenced by differences in both allelic diversity and in the frequencies of specific fragments. Within the Heron Island population, cohort analyses demonstrated the temporal stability of the fingerprint profile over 6 yr. This study demonstrates that length polymorphisms at minisatellite loci may be stable enough over time to retain information about recent historical and demographic effects on the relative genetic variability and differentiation of small, closely related populations.     

GENETIC VARIABILITY AND MOLECULAR PHYLOGENY OF DINOPHYSIS SPECIES (DINOPHYCEAE) FROM NORWEGIAN WATERS INFERRED FROM SINGLE CELL ANALYSES OF rDNA1

The objectives of this study were to determine rDNA sequences of the most common Dinophysis species in Scandinavian waters and to resolve their phylogenetic relationships within the genus and to other dinoflagellates. A third aim was to examine the intraspecific variation in D. acuminata and D. norvegica, because these two species are highly variable in both morphology and toxicity. We obtained nucleotide sequences of coding (small subunit [SSU], partial large subunit [LSU], 5.8S) and noncoding (internal transcribed spacer [ITS]1, ITS2) parts of the rRNA operon by PCR amplification of one or two Dinophysis cells isolated from natural water samples. The three photosynthetic species D. acuminata, D. acuta, and D. norvegica differed in only 5 to 8 of 1802 base pairs (bp) within the SSU rRNA gene. The nonphotosynthetic D. rotundata (synonym Phalacroma rotundatum[Claparède et Lachmann] Kofoid et Michener), however, differed in approximately 55 bp compared with the three photosynthetic species. In the D1 and D2 domains of LSU rDNA, the phototrophic species differed among themselves by 3 to 12 of 733 bp, whereas they differed from D. rotundata by more than 100 bp. This supports the distinction between Dinophysis and Phalacroma. In the phylogenetic analyses based on SSU rDNA, all Dinophysis species were grouped into a common clade in which D. rotundata diverged first. The results indicate an early divergence of Dinophysis within the Dinophyta. The LSU phylogenetic analyses, including 4 new and 11 Dinophysis sequences from EMBL, identified two major clades within the phototrophic species. Little or no intraspecific genetic variation was found in the ITS1–ITS2 region of single cells of D. norvegica and D. acuminata from Norway, but the delineation between these two species was not always clear.     

LOW GENETIC VARIABILITY OF SARGASSUM MUTICUM (PHAEOPHYCEAE) REVEALED BY A GLOBAL ANALYSIS OF NATIVE AND INTRODUCED POPULATIONS1

Sargassum muticum (Yendo) Fensholt is one of the most well‐known invasive species in the world. There have, however, been few genetic investigations on both its introduced and native populations. There are also some questions about the taxonomic status of this species. This study is the first to assess the genetic diversity of S. muticum on a global scale, by utilizing one marker each from the extranuclear genomes, namely, plastidial RUBISCO and mitochondrial TrnW_I spacers, as well as the nuclear internal transcribed spacer 2 (ITS2). Based on the markers investigated, both the invasive as well as the native populations of this species appeared very homogenous, when compared with other invasive and brown macroalgae. No variation in ITS2 and RUBISCO spacer was revealed in S. muticum populations, including those from its native ranges in Asia and the introduced ranges in Europe and North America. Two TrnW_I spacer haplotypes with a fixed two‐nucleotide difference were found between the populations of eastern Japan and the other 15 populations examined. This study confirms that there is no cryptic diversity in the introduced range of this species. All the materials collected globally are indeed S. muticum. Results depicting the distribution range of the two TrnW_I spacer haplotypes also support the earlier suggestion that the source of the introduced S. muticum populations is most likely western and central Japan (Seto Inland Sea), where the germlings of S. muticum were likely to have been transported with the Pacific oysters previously introduced for farming in Canada, UK, and France in earlier years.     

GENETIC STRUCTURE OF NATURAL POPULATIONS IN THE RED ALGAE GELIDIUM CANARIENSE (GELIDIALES,RHODOPHYTA) INVESTIGATED BY RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD) MARKERS1

Random amplified polymorphic DNA (RAPD) marker variation was analyzed in female gametophytes in natural populations of Gelidium canariense (Grunow) Seoane‐Camba ex Haroun, Gil‐Rodríguez, Diaz de Castro et Prud'Homme van Reine from the Canary Islands to estimate the degree and distribution of genetic variability and differentiation. A total of 190 haploid individuals were analyzed with 60 polymorphic RAPDs bands which produced 190 distinct multilocus genotypes. A high level of polymorphism was detected in all populations analyzed. Within‐population gene diversity ranged from 0.156 to 0.264. The populations on the island of Gran Canaria showed higher genetic variation than the other populations analyzed. The partitioning of molecular variance by analysis of molecular variance showed that most genetic variation resides within populations (68.85%). These results suggest that sexual reproduction is the predominant mode of reproduction for G. canariense gametophytic populations, and the main determinant in reaching high levels of genetic diversity. The Neighbor‐Joining tree and FCA analysis displayed two subclusters that correspond to the populations from the western islands (Tenerife, La Palma, Gomera) and the eastern island (Gran Canaria). In addition, we have detected a significant relationship between F_ST/(1?F_ST) and geographical distance consistent with data on water circulation and age of islands. The results obtained agree with an isolation by distance model, with gene flow from eastern to the western islands, and a high level of genetic differentiation between populations (F_ST=0.311, P<0.001).     

REDUCED GENETIC DIVERSITY AND INCREASED POPULATION DIFFERENTIATION IN PERIPHERAL AND OVERHARVESTED POPULATIONS OF GIGARTINA SKOTTSBERGII (RHODOPHYTA,GIGARTINALES) IN SOUTHERN CHILE1

This study assesses two hypotheses on the genetic diversity of populations of Gigartina skottsbergii Setchell et Gardner (Rhodophyta, Gigartinales) at the border of the species distribution: 1) peripheral populations display a reduced genetic diversity compared with central populations, and 2) genetic differentiation is higher among peripheral than among central populations. Two peripheral and four central populations were sampled along the Chilean coast and 113 haploid individuals were analyzed using 17 random amplification of polymorphic DNA loci. The genetic diversity was estimated by allele diversity (H_e), allele richness (Â), and the mean pair‐wise differences among multilocus genotypes. All three estimates consistently and significantly indicated a lower genetic diversity within the peripheral than within the central populations. Genetic differentiation between the two peripheral populations was stronger (F_ST=0.35) than between central populations at similar spatial scales (F_ST ranging from 0 to 0.25). In addition, it appeared from the distribution of pair‐wise differences that peripheral populations are in demographic expansion after a recent bottleneck. The results are discussed in the specific context of potential overharvesting of these wild populations.     

AN EXAMINATION OF PACHYMENIA AND AEODES (HALYMENIACEAE,RHODOPHYTA) IN NEW ZEALAND AND THE TRANSFER OF TWO SPECIES OF AEODES IN SOUTH AFRICA TO PACHYMENIA1

A phylogenetic study was conducted of species of Halymeniaceae from New Zealand presently placed in Aeodes or Pachymenia, based on maximum‐likelihood (ML), maximum‐parsimony (MP), and Bayesian analyses of rbcL and nuclear internal transcribed spacer (ITS) rDNA sequences. We used molecular and morphological data in combination with exhaustive sampling of herbarium collections to clarify the taxonomy and distributions of New Zealand members of Pachymenia and Aeodes. Our study confirms the presence of three erect species of Pachymenia on the New Zealand mainland, and we resurrect the name Pachymenia dichotoma J. Agardh for the widely distributed, southernmost species. Species of Aeodes from South Africa are shown to be closely related to Pachymenia carnosa (J. Agardh) J. Agardh, the type species of Pachymenia, and are accordingly transferred to Pachymenia.     

CONSTRUCTION OF A GENETIC LINKAGE MAP FOR PORPHYRA HAITANENSIS (BANGIALES,RHODOPHYTA) BASED ON SEQUENCE‐RELATED AMPLIFIED POLYMORPHISM AND SIMPLE SEQUENCE REPEAT MARKERS1

Molecular markers and molecular genetic maps are prerequisites for molecular breeding in any plant species. A comprehensive genetic linkage map for cultivated Porphyra haitanensis T. J. Chang et B. F. Zheng has not yet been developed. In this study, 157 double haploid (DH) lines [derived from a YSIII (wildtype) × RTPM (red‐type artificial pigmentation mutant) cross] were used as a mapping population in P. haitanensis. A total of 60 pairs of sequence‐related amplified polymorphism (SRAP) primers and 39 pairs of simple sequence repeat (SSR) primers were used to detect polymorphisms between the two parents. Fifteen SRAP and 16 SSR polymorphic primer pairs were selected to analyze the DH population. A linkage genetic map comprising 67 SRAP markers and 20 SSR markers in five linkage groups, with a total length of 830.6 cM and an average of 10.13 cM between markers, was constructed. The markers were distributed evenly in all linkage groups without clustering. The linkage groups comprised 12–23 markers ranging in length from 134.2 to 197.3 cM. The estimated genome length of P. haitanensis was 942.4 cM, with 88.1% coverage. This is the first report of a comprehensive genetic map in P. haitanensis. The map presented here will provide a basis for the development of high‐density genetic linkage maps and lay the foundation for molecular breeding work in P. haitanensis.