Novel organization of the mitochondrial genome in the deep-sea coral, Madrepora oculata (Hexacorallia, Scleractinia, Oculinidae) and its taxonomic implications

Madrepora is one of the most ecologically important genera of reef-building scleractinians in the deep sea, occurring from tropical to high-latitude regions. Despite this, the taxonomic affinities and relationships within the genus Madrepora remain unclear. To clarify these issues, we sequenced the mitochondrial (mt) genome of the most widespread Madrepora species, M. oculata, and compared this with data for other scleractinians. The architecture of the M. oculata mt genome was very similar to that of other scleractinians, except for a novel gene rearrangement affecting only cox2 and cox3. This pattern of gene organization was common to four geographically distinct M. oculata individuals as well as the congeneric species M. minutiseptum, but was not shared by other genera that are closely related on the basis of cox1 sequence analysis nor other oculinids, suggesting that it might be unique to Madrepora.     

Trawling damage to Northeast Atlantic ancient coral reefs

This contribution documents widespread trawling damage to cold-water coral reefs at 840-1300 m depth along the West Ireland continental shelf break and at 200 m off West Norway. These reefs are spectacular but poorly known. By-catches from commercial trawls for deep-water fish off West Ireland included large pieces (up to 1 m(2)) of coral that had been broken from reefs and a diverse array of coral-associated benthos. Five azooxanthellate scleractinarian corals were identified in these by-catches, viz. Desmophyllum cristagalli, Enallopsammia rostrata, Lophelia pertusa, Madrepora oculata and Solenosmilia variabilis. Dating of carbonate skeletons using (14)C accelerator mass spectrometry showed that the trawled coral matrix was at least 4550 years old. Surveys by remotely operated vehicles in Norway showed extensive fishing damage to L. pertusa reefs. The urgent need for deep-water coral conservation measures is discussed in a Northeast Atlantic context.     

Calcification rates and the effect of ocean acidification on Mediterranean cold-water corals

Global environmental changes, including ocean acidification, have been identified as a major threat to scleractinian corals. General predictions are that ocean acidification will be detrimental to reef growth and that 40 to more than 80 per cent of present-day reefs will decline during the next 50 years. Cold-water corals (CWCs) are thought to be strongly affected by changes in ocean acidification owing to their distribution in deep and/or cold waters, which naturally exhibit a CaCO(3) saturation state lower than in shallow/warm waters. Calcification was measured in three species of Mediterranean cold-water scleractinian corals (Lophelia pertusa, Madrepora oculata and Desmophyllum dianthus) on-board research vessels and soon after collection. Incubations were performed in ambient sea water. The species M. oculata was additionally incubated in sea water reduced or enriched in CO(2). At ambient conditions, calcification rates ranged between -0.01 and 0.23% d(-1). Calcification rates of M. oculata under variable partial pressure of CO(2) (pCO(2)) were the same for ambient and elevated pCO(2) (404 and 867 μatm) with 0.06 ± 0.06% d(-1), while calcification was 0.12 ± 0.06% d(-1) when pCO(2) was reduced to its pre-industrial level (285 μatm). This suggests that present-day CWC calcification in the Mediterranean Sea has already drastically declined (by 50%) as a consequence of anthropogenic-induced ocean acidification.     

Phenotype-specific bacterial communities in the cold-water coral Lophelia pertusa (Scleractinia) and their implications for the coral's nutrition, health, and distribution

The pseudocolonial coral Lophelia pertusa (Scleractinia, Caryophylliidae) is a eurybathic, stenothermal cosmopolitan cold-water species. It occurs in two color varieties, white and red. L. pertusa builds vast cold-water coral reefs along the continental margins, which are among the most diverse deep-sea habitats. Microbiology of L. pertusa has been in scientific focus for only a few years, but the question of whether the coral holds a host-specific bacterial community has not been finally answered. Bacteria on coral samples from the Trondheimsfjord (Norway) were characterized by the culture-independent 16S rRNA gene-based techniques terminal restriction fragment length polymorphism and sequence analysis. L. pertusa revealed a high microbial richness. Clone sequences were dominated by members of the Alpha- and Gammaproteobacteria. Other abundant taxa were Bacteroidetes, Actinobacteria, Verrucomicrobia, Firmicutes, and Planctomycetes. The bacterial community of L. pertusa not only differed conspicuously from that of the environment but also varied with both the location and color variety of its host. Therefore, the microbial colonization cannot be termed "specific" sensu stricto. However, similarities to other coral-bacterium associations suggest the existence of "cold-water coral-specific" bacterial groups sensu lato. L. pertusa-associated bacteria appear to play a significant role in the nutrition of their host by degradation of sulfur compounds, cellulose, chitin, and end products of the coral's anaerobic metabolism. Some coral-associated microbes were regarded as opportunistic pathogens. Dominance of mixotrophic members of the Rhodobacteraceae in white L. pertusa could explain the wider dispersal of this phenotype by supplementary nutrition.     

Mitogenome polymorphism in a single branch sample revealed by SOLiD deep sequencing of the Lophelia pertusa coral genome

We present an initial genomic analysis of the non-symbiotic scleractinian coral Lophelia pertusa, the dominant cold-water reef-building coral species in the North Atlantic Ocean. A significant fraction of the deep sequencing reads was of mitochondrial and microbial origins. SOLiD deep sequencing reads from fragment library experiments of total DNA and PCR amplified mitogenome generated about 21,000 times and 136,000 times coverage, respectively, of the 16,150bp mitogenome. Five polymorphic sites that include two non-synonymous sites in the NADH dehydrogenase subunit 5 genes were detected in both experiments. This observation is surprising since anthozoans in general exhibit very low mtDNA sequence variation at intraspecific level compared to nuclear sequences. More than fifty bacterial species associated with the coral isolate were also sequence detected, representing at least ten complete genomes. Most reads, however, were predicted to originate from the Lophelia nuclear genome.     

Genetic structure of the deep-sea coral Lophelia pertusa in the northeast Atlantic revealed by microsatellites and internal transcribed spacer sequences

The azooxanthellate scleractinian coral Lophelia pertusa has a near-cosmopolitan distribution, with a main depth distribution between 200 and 1000 m. In the northeast Atlantic it is the main framework-building species, forming deep-sea reefs in the bathyal zone on the continental margin, offshore banks and in Scandinavian fjords. Recent studies have shown that deep-sea reefs are associated with a highly diverse fauna. Such deep-sea communities are subject to increasing impact from deep-water fisheries, against a background of poor knowledge concerning these ecosystems, including the biology and population structure of L. pertusa. To resolve the population structure and to assess the dispersal potential of this deep-sea coral, specific microsatellites markers and ribosomal internal transcribed spacer (ITS) sequences ITS1 and ITS2 were used to investigate 10 different sampling sites, distributed along the European margin and in Scandinavian fjords. Both microsatellite and gene sequence data showed that L. pertusa should not be considered as one panmictic population in the northeast Atlantic but instead forms distinct, offshore and fjord populations. Results also suggest that, if some gene flow is occurring along the continental slope, the recruitment of sexually produced larvae is likely to be strongly local. The microsatellites showed significant levels of inbreeding and revealed that the level of genetic diversity and the contribution of asexual reproduction to the maintenance of the subpopulations were highly variable from site to site. These results are of major importance in the generation of a sustainable management strategy for these diversity-rich deep-sea ecosystems.     

Plectroninia celtica n. sp. (Calcarea, Minchinellidae), a new species of “pharetronid” sponge from bathyal depths in the northern Porcupine Seabight, NE Atlantic

Recent pharetronid sponges were regarded as relict species in tropical and subtropical waters, inhabiting cryptic habitats on coral reefs and in caves. More recent findings of a new species of the genus Plectroninia off northern Norway, with an inner fused skeleton have changed that view. Recent investigations on the sponge fauna of the “Propeller Mound”, northern Porcupine Seabight, focusing on sponges growing on the azooxanthellate cold-water coral Lophelia pertusa (Linné 1758) and Madrepora oculata Linné 1758, established the presence of a species of Plectroninia new to science. Its status as a common species within this deep-water coral habitat and the general status of the genus Plectroninia are discussed.     

Molecular studies of photobionts of selected lichens from the coastal vegetation of Brazil

A light microscopic and molecular analysis of photobionts in Ramalina and Cladonia from coastal habitats of Brazil is presented. A Bayesian phylogenetic analysis of ITS rDNA sequences suggests a Trebouxia lineage which is preferentially tropical in geographic distribution. This highly diverse clade also includes the morphological similar species Trebouxia higginsiae and galapagensis. Within the predominantly tropical clade of Trebouxia we distinguish several subclades, three of which are represented in our samples of Ramalina species. Since sexuality has not been recognized in coccal lichenised photobionts until recently, we cannot apply a biological species concept, but when compared with the sequence diversity between known species we conclude that several new species need to be described in this clade. The mutually exclusive presence of other Trebouxia lineages in temperate samples of Ramalina suggests an evolution towards higher selectivity in this genus. A strictly tropical lineage is not conspicuous in the photobionts of the genus Asterochloris sampled from Cladonia so far.     

Characterization of 13 microsatellite loci for the deep-sea coral, Lophelia pertusa (Linnaeus 1758), from the western North Atlantic Ocean and Gulf of Mexico

A suite of 13 polymorphic tri- and tetranucleotide microsatellite loci were isolated from the ahermatypic deep-sea coral, Lophelia pertusa. Among 51 individuals collected from three disjunct oceanic regions, allelic diversity ranged from six to 38 alleles and averaged 9.1 alleles per locus. Observed heterozygosity ranged from 9.1 to 96.8% and averaged 62.3% in the Gulf of Mexico population. For some loci, amplification success varied among collections, suggesting regional variation in priming site sequences. Four loci showed departures from Hardy-Weinberg equilibrium in certain collections which may reflect nonrandom mating.     

Solenosmilia variabilis-bearing cold-water coral mounds off Brazil

Coral Reefs - Cold-water corals (CWC), dominantly Desmophyllum pertusum (previously Lophelia pertusa), and their mounds have been in the focus of marine research during the last two decades;...     

Phylogenetic Analysis of Mediterranean Mugilids by Allozymes and 16S mt-rRNA Genes Investigation: Are the Mediterranean Species of Liza Monophyletic?

The family Mugilidae (Pisces, Mugiliformes) includes species which are present in all tropical and temperate regions. Six species, Chelon labrosus, Mugil cephalus, Liza aurata, L. ramada, L. saliens, Oedalechilus labeo, are commonly found in the Mediterranean. These species have been widely studied through morphological, biochemical, and molecular markers. However, their phylogenetic relationships, and therefore the assumed monophyly of Liza species, still remain unclear: To further investigate this topic, gene-enzyme systems and sequences of the partial 16S rRNA mitochondrial gene were analyzed in Italian samples of all six Mediterranean species. The phylogenetic reconstructions indicated M. cephalus as being the most divergent species and the existence of a main cluster including all the Mediterranean species of Liza and C. labrosus. The parametric bootstrap approach adopted to test alternative phylogenetic hypotheses indicated that the Mediterranean species of Liza do not form a monophyletic group exclusive of Chelon.     

孔石莼(Ulva pertusa)质体蓝素基因的克隆及基因特征分析(英文)

采用cDNA末端快速扩增的办法,从孔石莼(Ulva pertusa)中克隆获得质体蓝素基因。该基因完整的cDNA为787bp,包括40 bp 5’端非编码区和327 bp的3’端非编码区,以及一个420 bp的开放阅读框架,编码139个氨基酸的蛋白质。该基因编码质体蓝素的前体肽,其N端41个氨基酸残基为信号肽,后面为98个氨基酸残基的成熟肽。从Genbank中选择了13个质体蓝素的前体肽基因进行序列比对分析和构建进化树。孔石莼质体蓝素基因与其它质体蓝素基因的同源性为48.2%至78.8%。该进化树将来源于6种藻类植物的7个质体蓝素基因聚类在一起,显示出它们较近的进化关系。同样,也表现出11种生物的分子进化关系。序列比对结果显示,在质体蓝素的基因序列中存在两个高度保守的基序,它编码质体蓝素蛋白的铜结合活性位点。     

The reproductive biology of two deep-water,reef-building scleractinians from the NE Atlantic Ocean

The reproductive ecology of colonies of Lophelia pertusa (Linné 1758) and Madrepora oculata Linné (1758) from the Porcupine Seabight (Thérèse Mound and South Porcupine Seabight site) and the Darwin Mounds (NE Rockall Trough—L. pertusa only) was investigated using histological techniques. Samples of L. pertusa exhibited seasonal reproduction, whereas the evidence for M. oculata is equivocal but suggests multiple cohorts of gamete production. L. pertusa produces a single cohort of around 3,000 oocytes, whereas M. oculata produces two cohorts, with a total fecundity of around 60 oocytes. The maximum observed oocyte size in L. pertusa was 140 μm and in M. oculata was 405 μm. From these oocyte sizes and the timing of reproduction, a lecithotrophic larva is expected, though not observed. This seasonality of reproduction fits with the phytodetrital food fall occurring around July in the Seabight area. L. pertusa was found to be non-reproductive at the Darwin Mound site. Though unable to be specifically tested, this may suggest that the increased trawling activity in this area might be keeping colonies below sexually viable sizes, as seen in numerous shallow water situations. All areas in the NE Atlantic are coming under threat from increased fishing and commercial exploration practices. This study shows that these highly seasonal reproducers could be sensitive to these fishing operations and care must be taken so as not to repeat the destruction that has occurred on shallower reefs.     

Tempo and mode of concerted evolution in the L1 repeat family of mice

A 300-bp DNA sequence has been determined for 30 (10 from each of three species of mice) random isolates of a subset of the long interspersed repeat family L1. From these data we conclude that members of the L1 family are evolving in concert at the DNA sequence level in Mus domesticus, Mus caroli, and Mus platythrix. The mechanism responsible for this phenomenon may be either duplicative transposition, gene conversion, or a combination of the two. The amount of intraspecies divergence averages 4.4%, although between species base substitutions accumulate at the rate of approximately 0.85%/Myr to a maximum divergence of 9.1% between M. platythrix and both M. domesticus and M. caroli. Parsimony analysis reveals that the M. platythrix L1 family has evolved into a distinct clade in the 10-12 Myr since M. platythrix last shared a common ancestor with M. domesticus and M. caroli. The parsimony tree also provides a means to derive the average half-life of L1 sequences in the genome. The rates of gain and loss of individual copies of L1 were estimated to be approximately equal, such that approximately one-half of them turn over every 3.3 Myr.      

Evolution of the ascidian anural larva: evidence from embryos and molecules.

Most ascidians pass through a tadpole (urodele) larval stage, although some species have derived a tailless (anural) larva. New insights into the evolution of anural larvae in the Roscovita clade of molgulid ascidians were obtained from studing embryonic development of the transitional anural species Molgula bleizi and from phylogenetic analysis based on muscle and cytoskeletal actin gene sequences. By observing in vitro fertilized eggs, we found that M. bleizi, previously described as a typical anural developer, actually forms a short immotile tail during embryogenesis. The short tail contains notochord lineage cells, which undergo abbreviated morphogenetic movements but eventually arrest in development. Molgula bleizi tail muscle lineage cells produce the muscle enzyme acetylcholinesterase (AChE) but do not express muscle actin genes. The presence of a short tail, a vestigial notochord, and AChE-positive muscle cells suggest that M. bleizi is a recently derived anural species. An M. bleizi larval muscle actin gene (MbMA1) was isolated, sequenced, and shown to be a pseudogene based on critical deletions in its coding region that would result in a nonfunctional actin protein. The mutations in MbMA1 are distinct from and have evolved independent of the larval muscle actin pseudogenes MoccMA1a and MoccMA1b in Molgula occulta, another anural developer in the Roscovita clade. Pseudogene formation explains the absence of muscle actin mRNA in M. bleizi embryos. The 3' untranslated region of an M. bleizi cytoskeletal actin gene was also isolated and sequenced. Phylogenetic trees reconstructed using muscle and cytoskeletal actin sequences suggest that the anural developer M. bleizi evolved prior to the divergence of the urodele developer Molgula oculata and the anural developer M. occulta in the Roscovita clade. Since M. bleizi lives attached to hard substrata in the tidal zone, whereas M. oculata and M. occulta live buried in subtidal sand flats, our results suggest that the anural larva evolved at least twice in the Roscovita clade of molgulid ascidians as an adaptation to different habitats.     

Tracing paternal ancestry in mice, using the Y-linked, sex-determining locus, Sry

The molecular evolution of mammalian Y-linked DNA sequences is of special interest because of their unique mode of inheritance: most Y- linked sequences are clonally inherited from father to son. Here we investigate the use of Y-linked sequences for phylogenetic inference. We describe a comparative analysis of a 515-bp region from the male sex- determining locus, Sry, in 22 murine rodents (subfamily Murinae, family Muridae), including representatives from nine species of Mus, and from two additional murine genera--Mastomys and Hylomyscus. Percent sequence divergence was < 0.01% for comparisons between populations within a species and was 0.19%-8.16% for comparisons between species. Our phylogenetic analysis of 12 murine taxa resulted in a single most- parsimonius tree that is highly concordant with phylogenies based on mitochondrial DNA and allozymes. A total evidence tree based on the combined data from Sry, mitochondrial DNA, and allozymes supports (1) the monophyly of the subgenus Mus, (2) its division into a Palearctic group (M. musculus, M. domesticus, M. spicilegus, M. Macedonicus, and M. spretus) and an Oriental group (M. cookii++, M. cervicolor, and M. caroli), and (3) sister-group relationships between M. spicilegus and M. macedonicus and between M. cookii and M. cervicolor. We argue that Y- chromosome DNA sequences represent a valuable new source of characters for phylogenetic inference.      

从rbcL序列探讨单室茱萸属的系统位置

为了探讨单室茱萸属Mastixia的系统位置,本文对八蕊单室萸（Mastixia euony moides),云南单室茱萸（M.pentandra subsp,chinensis)及近缘物种云南紫树（Nyssa yunnanensis)和毛八角枫（Alangium kurzii)的cpDNA中编码1,5－二磷酸核酸糖羧化酶大亚基的rbcL基因序列进行了双脱氧测定。通过对这些序列与广义山茱萸科Cornaceae和Mastixia曾被归入的科的类群的rbcL序列进行PAUP（Phylogenetic Analysis Using Parsimony)分析,得到24棵最简约的分支树图。PAUP分析表明单室茱萸属与马蹄参属Diplopanax,紫树属Nyssa,喜树属Camptotheca和珙桐属Davidia的亲缘关系最近,并由分支图支持可以将Mastixia提升为山茱萸目Cornales中的一个科－单室萸科Masstixiaceae。     

The structural genes coding for the L and M subunits of Rhodospirillum rubrum photoreaction center

In Rhodospirillum rubrum, pufL, and pufM, the structural genes coding for the photoreaction center L and M polypeptides, are comprised respectively of 831 and 921 nucleotides. They are separated by a stretch of 12 nucleotides between the TAA stop codon of pufL and the first base of the ATG initiation codon of pufM. The predicted amino acid sequence of the L and M polypeptides, respectively, contain 275 and 305 residues with corresponding molecular weights of 30,473 and 33,978. Their sequences are highly homologous to those of Rhodobacter sphaeroides, Rhodobacter capsulatus, and Rhodopseudomonas viridis. As can be deduced from the crystallographic structure of other photoreaction centers, the regions of greatest similarity are the binding sites of the cofactors involved in the photochemical reaction rather than the protein secondary structure. L and M contain, at conserved positions of their sequences, three main clusters of positively charged residues on the cytoplasmic side of the membrane. This arrangement may be involved in protein orientation during membrane assembly. Evolutionary distance of pufL and pufM, as assessed by substitution frequency analysis, confirms the closeness of the two Rhodobacter species, the other two species being equidistant from one another. Interspecies evolutionary distance is greater for pufL than for pufM.     

Molecular evolution of a Y-chromosomal repetitive sequence family in the genus Mus

A 522-base-long Y-chromosomal sequence was isolated from a BALB/c genomic library and was designated "BF046." It is repeated about 200 times in the male genome, and a difference was detected between the Mus musculus musculus and the M. m. domesticus type Y chromosomes. BF046- related sequences were present over the entire length of the Y chromosome as visualized by in situ hybridization. Southern blot analysis against DNAs isolated from eight species in the genus Mus showed that BF046-related sequences were amplified in the Y chromosomes of three closely related species: M. musculus, M. spicilegus, and M. spretus. To gain insight into the stability of the BF046 sequence family, we isolated 18 additional clones from these three mouse species and compared their sequences. The M. musculus sequences differed from the M. spicilegus and M. spretus sequences by two indels. The remaining parts of the sequences were very similar, but both parsimony and distance-based analytical methods divided the sequences into the same four subgroups, with each species having its own subgroup(s). Thus, the Y chromosomes of M. musculus, M. spicilegus, and M. spretus can be distinguished from one another.      

Shallow-buried Pleistocene Madrepora-dominated coral mounds on a muddy continental slope, Tuscan Archipelago, NE Tyrrhenian Sea

Subfossil azoxanthellate deep-sea coral mounds occur at 355–410 m on the continental slope of the NE Tyrrhenian Sea between Gorgona and Capraia islands, Tuscan Archipelago. These low-relief patch reefs are at present buried by a thin muddy drape. Their age is latest Pleistocene. The colonial scleractinian Madrepora oculata is the major frame builder, in association with the solitary coral Desmophyllum dianthus and the colonial coral Lophelia pertusa. These NE Tyrrhenian Madrepora-dominated coral mounds represent one of the few known Mediterranean examples of deep-coral colonization of a muddy, low-gradient continental slope.