A molecular investigation of polymorphism in the North Atlantic red alga Chondrus crispus (Gigartinales)*

Seven samples of Chondrus crispus Stackhouse, representing widely contrasting forms from both sides of the North Atlantic, were compared by restriction digestion of their plastid DNA. The similar banding patterns confirmed that the seven forms were conspecific and distinct from Chondrus ocellatus Holmes f. ocellatus from Japan, used as an outgroup. Nucleotide sequences of the internal transcribed spacers (ITS1 and ITS21 and the intervening 5.8S rRNA gene of the nuclear rRNA operon were investigated as a potential indicator of genetic divergence among morphological variants of C. crispus. The combined ITS regions were relatively short in Chondrus (between 719 and 731 base pairs [bp] in C. crispus and 724 bp in C. ocellatus f. ocellatus), and the sequence of the 5.8s rDNA fragment (152 bp) was identical in both species. In the aligned ITS regions, there were 0–18 base pair differences (0–2.18% divergence) in pairwise comparisons of the seven forms of C. crispus but no consistent pattern of variation according to gross morphology or geographic origin. However, the ITS sequence differed at 41–54 sites (6.22-7.56%) between C. crispus and C. ocellatus f. ocellatus, again illustrating the genetic distinctiveness of the latter species.     

PHYLOGEOGRAPHIC PATTERNS INDICATE TRANSATLANTIC MIGRATION FROM EUROPE TO NORTH AMERICA IN THE RED SEAWEED CHONDRUS CRISPUS (GIGARTINALES,RHODOPHYTA)1

Inferring how the Pleistocene climate oscillations have repopulated the extant population structure of Chondrus crispus Stackh. in the North Atlantic Ocean is important both for our understanding of the glacial episode promoting diversification and for the conservation and development of marine organisms. C. crispus is an ecologically and commercially important red seaweed with broad distributions in the North Atlantic. Here, we employed both partial mtDNA Cox1 and nrDNA internal transcribed spacer region 2 (ITS2) sequences to explore the genetic structure of 17 C. crispus populations from this area. Twenty‐eight and 30 haplotypes were inferred from these two markers, respectively. Analysis of molecular variance (AMOVA) and of the population statistic Φ_ST not only revealed significant genetic structure within C. crispus populations but also detected significant levels of genetic subdivision among and within populations in the North Atlantic. On the basis of high haplotype diversity and the presence of endemic haplotypes, we postulate that C. crispus had survived in Pleistocene glacial refugia in the northeast Atlantic, such as the English Channel and the northwestern Iberian Peninsula. We also hypothesize that C. crispus from the English Channel refugium repopulated most of northeastern Europe and recolonized northeastern North America in the Late Pleistocene. The observed phylogeographic pattern of C. crispus populations is in agreement with a scenario in which severe Quaternary glaciations influenced the genetic structure of North Atlantic marine organisms with contiguous population expansion and locally restricted gene flow coupled with a transatlantic dispersal in the Late Pleistocene.     

Annual net primary production calculated from eastern Canadian Irish moss fishery data

Western Prince Edward Island Irish moss (Chondrus crispus) has been intensively dragraked since 1966. As well, most unattached fronds removed by wave surge, ice, etc. are brought to shore by waves and currents, where they are harvested eagerly. Accurate annual fishing yields were recorded between 1966 and 1981 inclusive. Given that herbivore densities are reduced, likely due to the intensive raking, and that mean annual bycatch (non-Irish moss seaweeds) (23.4 %) and commercial bed sizes (873 ha) were known, the fishing yields thus were considered a unique database from which to calculate net primary production (NPP). Factors used to convert from wet to dry wt (DW), and from dry wt to carbon were 0.22 an 0.31, respectively. Calculated mean annual NPP values were as follows: 2.101 ± 0.654 t (DW) ha^–1 y^–1; 210.1 g DW m^–2 yr^–1 and 63.0 g C m^–2 yr^–1. These values are much lower than those calculated for northwest Atlantic kelp and rockweed but similar to that determined for northeast Atlantic Gracilaria verrucosa. The interannual variability pattern for NPP was similar for both the wild Irish moss harvest and that of experimental Chondrus crispus outplants placed in one of the 14 commercial beds.     

Mutant swarms of a totivirus‐like entities are present in the red macroalga Chondrus crispus and have been partially transferred to the nuclear genome

Chondrus crispus Stackhouse (Gigartinales) is a red seaweed found on North Atlantic rocky shores. Electrophoresis of RNA extracts showed a prominent band with a size of around 6,000 bp. Sequencing of the band revealed several sequences with similarity to totiviruses, double‐stranded RNA viruses that normally infect fungi. This virus‐like entity was named C. crispus virus (CcV). It should probably be regarded as an extreme viral quasispecies or a mutant swarm since low identity (<65%) was found between sequences. Totiviruses typically code for two genes: one capsid gene (gag) and one RNA‐dependent RNA polymerase gene (pol) with a pseudoknot structure between the genes. Both the genes and the intergenic structures were found in the CcV sequences. A nonidentical gag gene was also found in the nuclear genome of C. crispus, with associated expressed sequence tags (EST) and upstream regulatory features. The gene was presumably horizontally transferred from the virus to the alga. Similar dsRNA bands were seen in extracts from different life cycle stages of C. crispus and from all geographic locations tested. In addition, similar bands were also observed in RNA extractions from other red algae; however, the significance of this apparently widespread phenomenon is unknown. Neither phenotype caused by the infection nor any virus particles or capsid proteins were identified; thus, the presence of viral particles has not been validated. These findings increase the known host range of totiviruses to include marine red algae.     

Open water Chondrus crispus Stackhouse cultivation

Selected strains of Chondrus crispus were acclimated to relativelywarm Pacific waters off the Baja California peninsula, Mexico, and werecultured in semi-exposed oceanic conditions. Specific techniques weredeveloped for culturing this species in semi-exposed oceanic environment,in an effort to establish the basis for its commercial farming. This is the firsttime that Chondrus crispus has been cultured in the ocean and outsideits natural Atlantic environment. The information generated in this studyis being used to feed an economic model to test the feasibility of thecommercial culturing of Chondrus crispus.     

广东省水库微囊藻的产毒特征和ITS序列的遗传多样性分析

为了解广东省水库微囊藻的产毒特征和ITS 序列的遗传多样性,从广东省供水水库中分离得到28 株微囊藻(Microcystisspp.),对它们的产毒特征和15 株微囊藻的ITS 序列进行了分析.高效液相色谱(HPLC)和微囊藻毒素合成酶基因mcyE 的检测结果表明,广东省水库中的微囊藻以产毒藻株占优势,微囊藻毒素的主要类型为MC-RR.广东省15 株藻株的ITS 序列相似性大于93.2%,在用相邻法(NJ)构建的系统树上,不同形态的种和不同地理区域的藻株没有区分开,产毒和非产毒藻株没有形成独立分支.这说明微囊藻ITS 序列的遗传多样性较低,ITS 序列和mcyE 存在没有相关性,表型不能够反映藻株的进化关系.因此,有必要将藻类传统分类方法与分子方法结合起来对蓝藻进行重新分类.     

Evolution of nuclear rDNA its sequences in the Cladophora albida/sericea clade (Chlorophyta)

Ribosomal DNA ITS sequences were compared among 13 different species and biogeographic isolates from the monophyletic albida/sericea clade in the green algal genus Cladophora. Six distinct ITS sequence types were found, characterized by multiple insertions and deletions and high levels of nucleotide substitution. Conserved domains within the ITS regions indicate the presence of ITS secondary structure. Low transition/transversion ratios among the six types and nearly symmetrical tree-length frequency distributions indicate some saturation, and low phylogenetic signal. Although branching order among five of the six ITS sequence types could not be resolved, estimates of ITS sequence divergence as compared with 18S divergence in a subset of the taxa suggests that the origin of the different ITS types is probably in the mid-Miocene (12 Ma ago) but that biogeographic isolates within a single ITS type (including both Pacific and Atlantic representatives) have probably dispersed on a time scale of thousands rather than millions of years.Correspondence to: J.J. Olsen     

Trends in abundance of rocky intertidal seaweeds and filter feeders across gradients of elevation,wave exposure,and ice scour in eastern Canada

On NW Atlantic rocky shores, the main basal organisms in intertidal communities are seaweeds (Ascophyllum nodosum, Fucus spp. and Chondrus crispus) and filter feeders (barnacles, Semibalanus balanoides, and mussels, Mytilus spp.). Their ecology has been extensively studied in New England (United States), but knowledge gaps exist for northern shores, which are subjected to stronger environmental stress. Therefore, we studied the above organisms on Canadian shores. We quantified the summer abundance of these seaweeds and filter feeders across full vertical (intertidal elevation) and horizontal (wave exposure and winter ice scour) environmental gradients on the Gulf of St. Lawrence and open Atlantic coasts of Nova Scotia. At the regional scale along the open Atlantic coast, seaweeds showed similar abundances in Nova Scotia than values reported for New England. However, both filter feeders were considerably less abundant in Nova Scotia. At the local scale in Nova Scotia, intense winter ice scour (which only occurs on the Gulf of St. Lawrence coast) was associated with a very low abundance of all species except barnacles. Spatial trends in Nova Scotia were similar to patterns known for certain species elsewhere, such as A. nodosum being almost restricted to sheltered habitats, regardless of elevation, and C. crispus being almost restricted to low elevations, regardless of exposure. Other trends were, however, characteristic of Nova Scotia, such as C. crispus being frequent at low elevations in exposed habitats, unlike in New England, where mussels often predominate there because of competitive advantages. In Nova Scotia, mussels were always restricted to cracks and crevices, unlike in New England, where they form extensive intertidal beds on exposed shores. The direct effects of increased environmental stress and indirect effects through altered interspecific interactions might explain the regional differences in local species distribution, which will require experimental validation. Handling editor: K. Martens     

Blitzkrieg in a Marine Invasion: Caulerpa racemosa var. cylindracea (Bryopsidales, Chlorophyta) Reaches the Canary Islands (North-East Atlantic)

On the basis of morphological and genetic studies (rDNA ITS1, 5.8S, ITS2, and a 18S rDNA intron), we confirm here that Caulerpa racemosa var. cylindracea (Sonder) Verlaque, Huisman et Boudouresque, a southwestern Australian taxon recently introduced into the Mediterranean Sea also occurs in the Canary Islands. This is the first report of C. racemosa var. cylindracea in the Atlantic. It was observed for the first time in the Canary Archipelago in 1997–1998. The speed and regional scale of expansion (north Atlantic Ocean and the Mediterranean Sea) of this invasive species appear to be among the most dramatic ever recorded. The possible outcome of this introduction in the Atlantic is discussed.     

Recent changes in the seaweed community of Western Prince Edward Island: implications for the seaweed industry

Prior to 1980, seaweed beds of western Prince Edward Island, Canada, were dominated by Chondrus crispus Stackhouse, associated with 37 other species of common algae. Furcellaria lumbricalis (Huds.) Lamour. occurred in 4.6% of the samples from the Pleasant View bed but never reached measurable levels of cover. By 1991 F. lumbricalis was reported in 67% of the samples from this bed. Furcellaria lumbricalis biomass was negatively correlated with C. crispus biomass and positively correlated with Phyllophora Grev. spp. biomass. F. lumbricalis biomass and cover peaked in subtidal areas below 4 m at 359 ± 299 SD fresh g 0.25 m^–2. Harvesters avoid concentrations of F. lumbricalis, resulting in a 5% content of this species in the commercial harvest. Systematic harvesting located concentrations of F. lumbricalis in seven of 10 commercial beds. Proposed causes for change in species dominance from C. crispus to F. lumbricalis range from general environmental change, natural succession to excessive human perturbation.     

The 18S rRNA gene sequences of four commercially important seaweeds

18S rRNA gene sequences are presented forAhnfeltia plicata, Chondrus crispus, Furcellaria lumbricalis andPalmaria palmata, commercially important marine algae of the North Atlantic. The sequences range from 1765 to 1777 nucleotides in length, with guanine + cytosine content of 50.1% to 52.4%. Sequence divergence between species in different orders was 11.3–12.3%, whereas the variation betweenC. crispus andF. lumbricalis, both from the Gigartinales, was only 3.6%. Based on limited experience with other groups of Rhodophyta, these sequences obtained from single populations are likely to be representative of the species as a whole, with little variation expected among conspecifics regardless of morphological aberration or apparent genetic isolation.NRCC 34824.     

Ecological impact of marine plant harvesting in the northwest Atlantic: a review

The ecological impact of marine plant harvesting is related to the intensity of exploitation, the harvesting technique, and the vulnerability of the species or habitat to perturbation. In eastern Canada information was available on four levels of impact: long-term changes in the target species and direct loss or damage to non-target species, direct or indirect impact on the habitat or community, indirect effects of changes in habitat or community structure, and trophic level impact. Near monoculture stands of Chondrus crispus have associated with them up to 36 animals species and 19 major species of algae that are vulnerable to removal as by-catch. Indirect effects of changes in macrophyte cover were not observed in fish species utilization of Ascophyllum nodosum beds on rising tides. Subtidal areas devoid of all macrophyte cover had lower levels of the preferred foods for Homarus americanus than kelp-covered areas; however, barren grounds are not created by macrophyte exploitation rates of 20% to 80% in eastern Canada. Long-term harvesting has altered the population structure and population ecology of C. crispus and A. nodosum in some areas. In general both target species and associated communities are resistant to perturbation.     

An assessment of the AFLP method for investigating population structure in the red alga Chondrus crispus Stackhouse (Gigartinales, Florideophyceae)

The appropriateness of the Amplified Fragment Length Polymorphism (AFLP) technique for investigating Chondrus crispus Stackhouse populations in the Maritime Provinces of Canada was assessed. The AFLP procedure was first subjected to reproducibility testing and three shortcomings were noted: 1) failure to reproduce band intensity between replicate runs for the same individual and primer pair; 2) failure of some bands to replicate; 3) lack of reproducibility for complete replicate runs for some individuals and primer pairs. In the last-mentioned case, the lack of reproducibility resulted in characteristic electropherograms indicative of weak reactions. These weak runs can be attributed to poor restriction digest/ligation reactions and/or substandard PCR, these failures ultimately resulting from low and inconsistent DNA quality. We recommend that reproducibility testing should be completed routinely in studies using the AFLP technique. In the current work, only fragments and individuals that gave reproducible results were used in subsequent analyses. The AFLP method resulted in highly variable markers within and between the populations of C. crispus included in this investigation, which prevented successful resolution of population structure. This situation could result from a lack of suitability for AFLP markers in population genetic studies, and/or too extensive genetic variation for C. crispus populations to be discerned by the AFLP technique. These two possible explanations are discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

基于ITS序列分析钩苞大丁草九个居群的亲缘关系

该研究对我国西南地区钩苞大丁草(Gerbera delavayi)9个居群rDNA ITS序列进行PCR的扩增和检测序列,并以非洲菊(G.jamesonii)的ITS序列作为外类群,比较了序列之间的差异,同时分析了钩苞大丁草不同居群在地理距离与遗传距离之间的关系,构建了NJ系统发育树。结果表明:(1)钩苞大丁草9个居群的ITS序列全长介于600~700 bp之间,平均长度约为657 bp,其中,ITS1长度为243~246 bp,(G+C)含量为45.67%~46.80%之间,5.8S长度191~193 bp,(G+C)含量为58.60%~58.61%之间,ITS2长度为220~221 bp,(G+C)含量为57.00%~57.45%之间;ITS序列共有22个变异位点,ITS1序列(17个)、5.8S序列(2个)以及ITS2序列(3个)上均有变异。(2)地理距离与遗传距离有正相关(r2=0.652),序列间遗传分化距离为0.001 1~0.024 3,其中普洱居群与其他居群间遗传距离最大。(3)钩苞大丁草9个居群分成三个分支,普洱居群单独成支,丽江和洱源居群聚为一支,富源、武定、德昌、石林、新平和开远6个居群聚为一支。rDNA ITS序列可以用于钩苞大丁草群体遗传研究的分析,该研究结果为其保护性开发提供了参考依据。     

Laboratory biology and host range ofHydrellia balciunasi [Diptera: Ephydridae]

Hydrellia balciunasi Bock, a native of Australia, was evaluated in quarantine in Florida, USA, for its potential as a biocontrol agent of the submersed aquatic weed,Hydrilla verticillata (L.f) Royle. Larvae are leafminers. Mean total development time at 27°C was 22.8 days. Mean duration of the egg stage was 3.0 days, larval was 11.5 days, and puparial was 8.3 days. Mean fecundity was 35.5 eggs. Mean female longevity was 19.7 days, and mean male longevity was 15.6 days. The sex ratio was 1.1∶1 (male: female). Fourteen plant species closely related to hydrilla in 4 families plus rice were tested in no-choice larval development tests and an additional 27 plant species in 16 families were tested in multi-choice tests. Larvae mined in 2 test plant species,Potamogeton pusillus L. andP. crispus L., but developed (1%) only on the introduced weedP. crispus L. Females oviposited on most test plants. Permission to release this fly in the United States was received from federal and state (Florida) officials, and it was released from quarantine on 24 May 1989.      

Effect of UV radiation on photoinhibition of marine macrophytes in culture systems

The present study examined the effect of UV andphotosynthetically active radiation (PAR) onphotoinhibition and recovery in the Phaeophyte Macrocystis pyrifera, the Rhodophyte Chondruscrispus and the Chlorophyte Ulva lactuca underoutdoor culture conditions. There was an increase inphotoinhibition as a consequence of high exposure toUV-B radiation in M. pyrifera, however, highlevels of PAR accounted for most of thephotoinhibition in C. crispus and U.lactuca. Photodamage by UV-A, UV-B and PAR wascompletely repaired within 5 h and effective quantumyield reached pretreatment values in the three speciesstudied. Species were less susceptible tophotoinhibition after being incubated for 5 d underhigh exposures of natural irradiance suggesting aphotoadaptive process. The recovery of the effectivequantum yield was impaired by long exposure to highlevels of UV-B in C. crispus and UV-A, UV-B andPAR in M. pyrifera. This suggests a differentkind of damage by UV-A and PAR radiation, one to thephotosynthetic apparatus and another which affects therepair mechanism of some species. There was anincrease in UV-absorption ( 330 nm) in M. pyrifera and C. crispus within four days ofthe initiation of the experiment suggesting that thesespecies photoprotect their photosynthetic system whenexposed to elevated UV and PAR levels.     

Effects of sediment anoxia and light on turion germination and early growth of Potamogeton crispus

Potamogeton crispus is a cosmopolitan aquatic species and is widely used as a pioneer species for vegetation restoration of eutrophic lakes. However, many restoration projects applying P. crispus turions have not been successful. Earlier studies focused on effects of light and temperature on turion germination. The purpose of this study was to determine whether sediment anoxia and light interactively affected the turion germination and early growth of P. crispus. Anoxic conditions in the experiment were produced by adding sucrose to the sediment. The germination rate of the turions was 68–73% lower in the highly anoxic condition treatment than in the control. Medium light intensity (10% of natural light at the water surface) was more favorable for germination under slightly anoxic conditions than either low or high light intensity. The growth of newly-formed sprouts was also significantly inhibited by sediment anoxia. Photosynthesis and shoot biomass were reduced under sediment anoxia, whereas total chlorophyll content, root biomass, and soluble protein content were highest in the low anoxic condition treatment. Medium light improved net photosynthesis and biomass production of the sprouts. We conclude that turion germination and sprout growth can be significantly inhibited by sediment anoxia. Medium light intensity may alleviate this inhibition by anoxia, but light has little effect when sediment anoxia is severe. For the purposes of vegetation restoration, more attention should be paid to the role of sediment anoxia, and it is necessary to improve sediment and light conditions for turion germination and early growth of P. crispus in eutrophic lakes. These results will contribute to a more complete understanding of turion germination dynamics of P. crispus and will be useful for future restoration programs. Handling editor: S. M. Thomaz     

Seasonal acclimatization of thallus proline contents of Mastocarpus stellatus and Chondrus crispus: intertidal rhodophytes that differ in freezing tolerance

Mastocarpus stellatus and Chondrus crispus often co‐occur in the lower intertidal of Northern Atlantic rocky shorelines. At our field site along the Maine coast (USA), Mastocarpus stellatus thalli possessed greater contents of proline when compared with thalli of Chondrus crispus. In addition, M. stellatus thalli acclimated to colder growth conditions in winter/early spring by increasing proline content several fold; no seasonal acclimation in proline content was observed in C. crispus. Proline accumulates in the tissues of a broad diversity of freezing‐tolerant organisms and is among the most common cryoprotectant molecules. Thus, our observations provide a basis for the previously well‐documented greater freezing tolerance of Mastocarpus stellatus when compared with Chondrus crispus.     

The Freshwater Animal Diversity Assessment: an overview of the results

The geographic genetic structure of two common encrusting sponges, Hymeniacidon sinapium and Hymeniacidon flavia (family Halichondriidae), was investigated using two DNA markers, Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA and NADH dehydrogenase subunit 5 (nad5) of mitochondrial DNA. In the ITS analyses, multiple sequence types were identified within each species. Geographic distribution patterns of sequence types showed higher diversity in the western than eastern areas in both species. However, intraspecific genetic diversity of the two species in Japan differed markedly. Hymeniacidon flavia had far more diverse sequence types, and several genetic differentiations between localities were detected. In contrast, H. sinapium had only four sequence types in Japan, and two Atlantic Hymeniacidon species had sequence types similar to this species. In comparison to ITS, nad5 showed very low genetic diversity in both species, with two haplotypes identified in each species. In H. flavia, frequency of haplotype changed gradually from north to south. In H. sinapium, one haplotype was predominant in most regions, and another haplotype was minor and distributed only in the Korean and Tsushima populations. Based on the unique distribution patterns of sequence types around Shikoku and Kyushu, geographical history and ocean currents were assumed to affect the generation of genetic structure. The geographic genetic structure of H. flavia suggests low dispersal ability of pelagic larvae, whereas higher larval dispersal ability and a far broader distribution range are suggested in H. sinapium. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. Handling editor: C. Sturmbauer