Kleptoplast distribution,photosynthetic efficiency and sequestration mechanisms in intertidal benthic foraminifera

Foraminifera are ubiquitously distributed in marine habitats, playing a major role in marine sediment carbon sequestration and the nitrogen cycle. They exhibit a wide diversity of feeding and behavioural strategies (heterotrophy, autotrophy and mixotrophy), including species with the ability of sequestering intact functional chloroplasts from their microalgal food source (kleptoplastidy), resulting in a mixotrophic lifestyle. The mechanisms by which kleptoplasts are integrated and kept functional inside foraminiferal cytosol are poorly known. In our study, we investigated relationships between feeding strategies, kleptoplast spatial distribution and photosynthetic functionality in two shallow-water benthic foraminifera (Haynesina germanica and Elphidium williamsoni), both species feeding on benthic diatoms. We used a combination of observations of foraminiferal feeding behaviour, test morphology, cytological TEM-based observations and HPLC pigment analysis, with non-destructive, single-cell level imaging of kleptoplast spatial distribution and PSII quantum efficiency. The two species showed different feeding strategies, with H. germanica removing diatom content at the foraminifer’s apertural region and E. williamsoni on the dorsal site. All E. williamsoni parameters showed that this species has higher autotrophic capacity albeit both feeding on benthic diatoms. This might represent two different stages in the evolutionary process of establishing a permanent symbiotic relationship, or may reflect different trophic strategies.Subject terms: Microbial ecology, Microbial ecology, Microbiology     

Molecular Identification of Algal Endosymbionts in Large Miliolid Foraminifera: 1. Chlorophytes

Large miliolid foraminifers bear various types of algal endosymbionts including chlorophytes, dinoflagellates, rhodophytes, and diatoms. Symbiosis plays a key role in the adaptation of large foraminifera to survival and growth in oligotrophic seas. The identity and diversity of foraminiferal symbionts, however, remain largely unknown. In the present work we use ribosomal DNA (rDNA) sequences to identify chlorophyte endosymbionts in large miliolid foraminifera of the superfamily Soritacea. Partial 18S and complete Internal Transcribed Spacer (ITS) rDNA sequences were obtained from symbionts of eight species representing all genera of extant chlorophyte-bearing Soritacea. Phylogenetic analysis of the sequences confirms the previous fine structure-based identification of these endosymbionts as belonging to the genus Chlamydomonas. All foraminiferal symbionts form a monophyletic group closely related to Chlamydomonas noctigama. The group is composed of seven types identified in this study, including one previously morphologically described species, Chlamydomonas hedleyi. Each of these types can be considered as a separate species, based on the comparison of genetic differences observed between other established Chlamydomonas species. Several foraminiferal species share the same symbiont type, but only one species, Archaias angulatus, was found to bear more than one type.     

A novel type of kleptoplastidy in Dinophysis (Dinophyceae): presence of haptophyte-type plastid in Dinophysis mitra

Red-fluorescent, non-phycobilin-containing plastids were found in the heterotrophic dinoflagellate, Dinophysis mitra. Transmission electron microscopy showed that they contained a three-layer thylakoid, the absence of girdle lamella, and an embedded pyrenoid with thylakoid intrusions. These characteristics all coincide with haptophyte plastids. Phylogenetic analysis of the plastid small-subunit ribosomal DNA (SSU rDNA) revealed that the Dinophysis mitra sequences are distantly related to those of phycobilin-containing Dinophysis species and are positioned within a lineage of haptophytes belonging to Prymnesiophyceae. Because the plastid SSU rDNA sequences of Dinophysis mitra showed significant heterogeneity, despite being derived from a single species, it is highly likely that they were not established as plastids through an evolutionary process but are "kleptoplastids" (temporally stolen plastids) from multiple sources of haptophytes in the environment. We deduced that Dinophysis mitra takes up haptophytes myzocytotically and selectively retains the plastid with surrounding plastidal membranes, whereas other haptophyte cell components are degraded. This represents another type of kleptoplastidy in the Dinophysis species, which mostly harbor cryptophyte plastids, and is the first evidence of kleptoplastidy originating from haptophytes.     

Intra-genomic ribosomal RNA polymorphism and morphological variation in Elphidium macellum suggests inter-specific hybridization in foraminifera

Elphidium macellum is a benthic foraminifer commonly found in the Patagonian fjords. To test whether its highly variable morphotypes are ecophenotypes or different genotypes, we analysed 70 sequences of the SSU rRNA gene from 25 specimens. Unexpectedly, we identified 11 distinct ribotypes, with up to 5 ribotypes co-occurring within the same specimen. The ribotypes differ by varying blocks of sequence located at the end of stem-loop motifs in the three expansion segments specific to foraminifera. These changes, distinct from typical SNPs and indels, directly affect the structure of the expansion segments. Their mosaic distribution suggests that ribotypes originated by recombination of two or more clusters of ribosomal genes. We propose that this expansion segment polymorphism (ESP) could originate from hybridization of morphologically different populations of Patagonian Elphidium. We speculate that the complex geological history of Patagonia enhanced divergence of coastal foraminiferal species and contributed to increasing genetic and morphological variation.     

UK intertidal foraminiferal distributions: implications for sea-level studies

Foraminiferal assemblages have been collected from ten intertidal study areas situated on the east, south and west coasts of the UK. The assemblages display a vertical zonation which indicates that the distribution of foraminifera in these intertidal environments is usually the direct function of altitude with the duration and frequency of intertidal exposure the most important factors. Multivariate analyses separate foraminiferal assemblages into two faunal zones: a high- and middle marsh zone consisting of differing abundances of Jadammina macrescens, Trochammina inflata and Miliammina fusca; and a low-marsh and tidal flat zone dominated by calcareous foraminiferal species, notably Elphidium williamsoni, Haynesina germanica and Quinqueloculina spp. These faunal zones are similar to those in other mid-latitude, cool temperate intertidal environments although there are spatial and temporal variations between areas. The altitudinal ranges of the faunal zones are employed to identify the vertical relationship of the local environment in which the assemblage accumulated to a reference tide level.     

PHYLOGENETIC POSITION OF ATTHEYA LONGICORNIS AND ATTHEYA SEPTENTRIONALIS (BACILLARIOPHYTA)1

The phylogenetic position of diatoms belonging to the genus Attheya is presently under debate. Species belonging to this genus have been placed in the subclasses Chaetocerotophycidae and Biddulphiophycidae, but published phylogenetic trees based on 18S rDNA, morphology, and sexual reproduction indicate that this group of diatoms may be a sister group of the pennates. To clarify the position of Attheya, we studied the morphology, 18S rDNA, 16S rDNA of the chloroplasts, the rbcL large subunit (LSU) sequences of the chloroplasts, and the sterol composition of three different strains of Attheya septentrionalis (Østrup) R. M. Crawford and one strain of Attheya longicornis R. M. Crawford et C. Gardner. These data were compared with data from more than 100 other diatom species, covering the whole phylogenetic tree, with special emphasis on species belonging to the genera that have been suggested to be related to the genus Attheya. All data suggest that the investigated Attheya species form a separate group of diatoms, and there is no indication that they belong to either the Chaetocerotophycidae or the Biddulphiophycidae. Despite applying these various approaches, we were unable to determine the exact phylogenetic position of the investigated Attheya species within the diatoms.     

Benthic foraminifera contribution to fjord modern carbon pools: A seasonal study in Adventfjorden,Spitsbergen

The aim of this study was to determine the amount of organic and inorganic carbon in foraminifera specimens and to provide quantitative data on the contribution of foraminifera to the sedimentary carbon pool in Adventfjorden. The investigation was based on three calcareous species that occur commonly in Svalbard fjords: Cassidulina reniforme, Elphidium excavatum and Nonionellina labradorica. Our results show that the species investigated did not contribute substantially to the organic carbon pool in Adventfjorden, because they represented only 0.37% of the organic carbon in the sediment. However, foraminiferal biomass could have been underestimated as it did not include arenaceous or monothalamous taxa. Foraminiferal carbonate constituted up to 38% of the inorganic carbon in the sediment, which supports the assumption that in fjords where non‐calcifying organisms dominate the benthic fauna foraminifera are among the major producers of calcium carbonate and that they play crucial roles in the carbon burial process. The results presented in this study contribute to estimations of changes in foraminiferal carbon levels in contemporary environments and could be an important reference for palaeoceanographic studies.     

Dinophyte chloroplasts and phylogeny - A review

Dinophytes acquired chloroplasts obviously early in evolution and later lost them multiple times. Most families and genera contain both photosynthetic and heterotrophic species. Chloroplasts enveloped by three membranes with thylakoids in stacks of three, containing peridinin as the main pigment, are regarded as the original dinophyte plastids. Pyrenoids are generally present. Stigmata, if present, are usually parts of the chloroplast or are modified original plastids. The form II type RUBISCO found in the dinophytes is unique for eukaryotes, otherwise known only in some anaerobic bacteria. It is disputed whether the original dinophyte chloroplasts are derived from a prokaryotic or an eukaryotic endosymbiosis. Various dinoflagellates contain aberrant chloroplasts. Glenodinium foliaceum and Peridinium balticum have a single complete endosymbiont, originally a pcnnate diatom. Podolampas bipes houses several dictyophycean symbiont cells. The “symbionts” of Lepidodiniurn viride and Gymnodinium chlorophorum are highly reduced prasinophyte cells. The chloroplasts of Gymnodinium mikimotoi have aberrant pigments (fucoxanthin derivatives, no peridinin) and fine structure. The dinoflagellate hosts do not seem to contain any parts of the former endosymbiont except the chloroplasts. Photosynthetic Dinophysis species have cryptophycean-like chloroplasts, whereas symbiotic cyanobacteria are found in other members of the Dinophysiales, e.g., Ornithocercus. Various dinophytes, e.g. Gymnodinium aeruginosum, use kleptochloroplasts from ingested cryptophytes transiently for photosynthesis. Original or secondarily acquired chloroplasts can only be used for phylogenetic considerations in exceptionally cases: it seems unlikely that the Prorocentrales have evolved from the Dinophysiales because all Prorocentrales possess original dinoflagellate chloroplasts, whereas no member of the Dinophysiales has such chloroplasts.     

Breaching biogeographic barriers: the invasion of <Emphasis Type="Italic">Haynesina germanica</Emphasis> (Foraminifera,Protista) in the Bahía Blanca estuary,Argentina

Large populations of the living benthic foraminifera Haynesina germanica (Ehrenberg 1840) are reported for the first time from marsh and intertidal mud flat sediments of the Bahía Blanca estuary (Argentina). Maximum abundance of living specimens was recorded in shallow intertidal environments. The species was previously recorded from many European and North American shallow-water coastal settings, but has not been documented from Argentina. Comparative faunal assemblage analysis from dated core sample material from within the Bahía Blanca estuary shows that the species has not been present for at least the last 8,200 years. This supports the hypothesis that the species has been accidentally introduced outside its natural range as a probable result of ballast water and/or shipping activities. This study is the first report of a successful invasion of non-indigenous benthic foraminifera to the South Atlantic coast of Argentina. The introduction of nonnative foraminiferal species provides new evidence that human-mediated breaching of biogeographic barriers, will ultimately result in the biotic homogenization of foraminiferal intertidal faunal assemblages.     

Molecular Identification of Algal Endosymbionts in Large Miliolid Foraminifera: 2. Dinoflagellates

Large miliolid foraminifers of the subfamily Soritinae bear symbiotic dinoflagellates morphologically similar to the species of the "Symbiodinium" complex, commonly found in corals and other marine invertebrates. Soritid foraminifers are abundant in coral reefs and it has been proposed that they share their symbionts with other dinoflagellate-bearing reef dwellers. In order to test this hypothesis, we have analysed partial large subunit ribosomal DNA sequences from dinoflagellates symbionts obtained from 28 foraminiferal specimens, and compared them to the corresponding sequences of Symbiodinium-like endosymbionts from various groups of invertebrates. Phylogenetic analysis of our data shows that all soritid symbionts belong to the "Symbiodinium" species complex, within which they form seven different molecular types (Frl-Fr7). Only one of these types (Fr1) branches within a group of invertebrate symbionts, previously described as type C. The remaining six types form sister groups to coral symbionts previously designed as types B, C, and D. Our data indicate a high genetic diversity and specificity of Symbiodinium-like symbionts in soritids. Except for type C, we have found no evidence for the transmission of symbionts between foraminifers and other symbiont-bearing invertebrates from the same localities. However, exchanges must have occurred frequently between the different species of Soritinae, as suggested by the lack of host specificity and some biogeographical patterns observed in symbiont distribution. Our data suggest that members of the subfamily Soritinae acquired their symbionts at least three times during their history, each acquisition being followed by a rapid diversification and independent radiation of symbionts within the foraminiferal hosts.     

The roles of elevation and salinity as primary controls on living foraminiferal distributions: Cowpen Marsh, Tees Estuary, UK

Benthic foraminiferal assemblages in subrecent deposits are commonly used to reconstruct past sea level. Interpretations are generally made by comparison with either modern dead or total (live plus dead) assemblages. In both cases there will have been post-mortem changes that have differentially affected preservation. It is therefore important to establish the primary ecological controls by analysis of the living assemblages. We have determined the spatial and temporal variability of intertidal benthic foraminifera in the surface (0–1 cm) sediments from a time series survey of 31 sampling stations at Cowpen Marsh, for a period of 12 months. We counted 112,067 live foraminifera assigned to 28 species. The fauna was dominated by two agglutinated species (Jadammina macrescens and Trochammina inflata) on the high and middle marshes, and three calcareous species (Elphidium williamsoni, Haynesina germanica and Quinqueloculina spp.) on the low marsh and tidal flat.The standing crop of the whole intertidal zone, including the high, middle, low marsh and tidal flat habitats, and the individual species varied both temporally and spatially. The standing crop of the intertidal zone as a whole was greatest in the summer months and showed a positive correlation with elevation. The standing crops of the high and middle marshes showed similar temporal variation with peaks in summer and autumn and a trough in winter. The low marsh showed numerous peaks and troughs of standing crop during the year, whereas the tidal flat showed a single peak in summer. The standing crops of Jadammina macrescens and Trochammina inflata on the high and middle marshes peaked from April to May and August to October with troughs in winter. These agglutinated species showed a strong correlation with elevation. Haynesina germanica peaked in May to August and November to January on the low marsh, whereas on the tidal flat there was a single peak in July. The standing crops of E. williamsoni on the low marsh and tidal flat were relatively high in June and May, and July, respectively. Quinqueloculina spp. peaked in May to July on the low marsh and July on the tidal flat. The species was also found in the middle marsh from July to May and high marsh from September to November. Haynesina germanica showed a strong negative correlation with elevation, whereas the other two dominant calcareous species demonstrated weak negative correlations with both elevation and salinity.Reconstructing former sea level depends primarily on the recognition of high and middle marsh assemblages and in this study these are shown to be strongly controlled by elevation rather than salinity. Caution may be needed in interpreting low marsh and tidal flat data as salinity plays a more important role here.     

Synthetic analogues of a transit peptide inhibit binding or translocation of chloroplastic precursor proteins

Although amino-terminal transit peptides of chloroplastic precursor proteins are known to be necessary and sufficient for import into chloroplasts, the mechanism by which they mediate this process is not understood. Another important question is whether different precursors share a common transport apparatus. We used 20-residue synthetic peptides corresponding to regions of the transit peptide of the precursor to the small subunit of ribulose bisphosphate carboxylase (prSS) as competitive inhibitors for the binding and translocation of precursor proteins into chloroplasts. Synthetic peptides with sequences corresponding to either end of the transit peptide had little to no effect on binding of prSS to chloroplasts, but significantly inhibited its translocation. Synthetic peptides corresponding to the central region of the transit peptide inhibited binding of prSS to chloroplasts. Each of the peptides inhibited binding or translocation of precursors to light-harvesting chlorophyll a/b protein, ferredoxin, and plastocyanin in the same manner and to a similar extent as prSS transport was inhibited. The results presented in this paper suggest that the central regions of the transit peptide of prSS mediate binding to the chloroplastic surface, whereas the ends of this transit peptide are more important for translocation across the envelope. Furthermore, all of the precursors tested appear to share components in the transport apparatus even though they are sorted to different chloroplastic compartments.     

浙江钱塘江冰后期有孔虫化石记录及其古环境意义

本研究对浙江萧山SE2钻孔近50m沉积物岩心进行了详细的有孔虫定量统计分析,并结合测年资料重建了冰后期以来钱塘江沉积河谷的环境演化过程。有孔虫群落特征反映的古环境变化与沉积相分析结果一致。有孔虫资料显示:大约1万年以来该地区受到海水的影响、以胶结质壳Textularia-Reophax组合为特征,并在中全新世海水深度到达最大、以近岸浅水组合Ammonia beccarii-Elphidium magellanicum为代表,而晚全新世以来在全球海平面下降的影响下、该地区海水变浅的同时,却由于狭窄的特殊河口地形,水动力相对较强,显示强潮型河口地区特有的近岸分子Ammonia beccarii,Elphidium advenum和相对较深水分子Ammonia compressiuscula共存的底栖有孔虫群落特征。     

Comprehensive comparisons of three pennate diatoms, Diatoma tenuae, Fragilaria vaucheriae, and Navicula pelliculosa, isolated from summer Arctic reservoirs (Svalbard 79°N), by fine-scale morphology and nuclear 18S ribosomal DNA

Here we report morphological and molecular characteristics of dominant freshwater diatoms in summer Arctic reservoirs of Svalbard (Norway), using four culture isolates, when we collected the samples in the field on 15 August 2005. Analyses of morphology and BLAST searches with 18S rDNA sequences identified them to Diatoma tenue (HYNP006, HYNP013), Navicula pelliculosa (HYNP021), and Fragilaria vaucheriae (HYNP022), respectively. Comparative studies of morphology revealed that the body shapes of the three polar diatoms were nearly identical to the known morphology of each species; however, they were considerably shorter in body length than previously described identical species from other locations. The 18S rDNA sequences of the diatoms were nearly identical to the same species from temperate and other regions. Phylogenetic analysis showed that the polar diatoms each formed a clade with their identical species and genera according to their taxonomic positions. This suggests that the polar diatoms may possess little or no genetic or morphological variation compared to more temperate strains.     

How can organellar protein N-terminal sequences be dual targeting signals? In silico analysis and mutagenesis approach

Organellar nuclear-encoded proteins can be mitochondrial, chloroplastic or localized in both mitochondria and chloroplasts. Most of the determinants for organellar targeting are localized in the N-terminal part of the proteins, which were therefore analyzed in Arabidopsis thaliana. The mitochondrial, chloroplastic and dual N-terminal sequences have an overall similar composition. However, Arg is rare in the first 20 residues of chloroplastic and dual sequences, and Ala is more frequent at position 2 of these two types of sequence as compared to mitochondrial sequences. According to these observations, mutations were performed in three dual targeted proteins and analyzed by in vitro import into isolated mitochondria and chloroplasts. First, experiments performed with wild-type proteins suggest that the binding of precursor proteins to mitochondria is highly efficient, whereas the import and processing steps are more efficient in chloroplasts. Moreover, different processing sites are recognized by the mitochondrial and chloroplastic processing peptidases. Second, the mutagenesis approach shows the positive role of Arg residues for enhancing mitochondrial import or processing, as expected by the in silico analysis. By contrast, mutations at position 2 have dramatic and unpredicted effects, either enhancing or completely abolishing import. This suggests that the nature of the second amino acid residue of the N-terminal sequence is essential for the import of dual targeted sequences.     

Early evolutionary origin of the planktic foraminifera inferred from small subunit rDNA sequence comparisons

Phylogenetic analysis of five partial planktic foraminiferal small subunit (SSU) ribosomal (r) DNA sequences with representatives of a diverse range of eukaryote, archaebacterial, and eubacterial taxa has revealed that the evolutionary origin of the foraminiferal lineage precedes the rapid eukaryote diversification represented by the crown of the eukaryotic tree and probably represents one of the earliest splits among extant free-living aerobic eukaryotes. The foraminiferal rDNA sequences could be clearly separated from known symbionts, commensals, and food organisms. All five species formed a single monophyletic group distinguished from the crown group by unique foraminiferal specific insertions as well as considerable nucleotide distance in aligned regions.     

Limitations in chloroplast multiplication inAcetabularia mediterranea

Summary The incorporation of thymidine into chloroplastic DNA inAcetabularia, examined two times after enucleation, has been compared with known data on the evolution of the DNA content of these organelles after enucleation.Division of the chloroplasts has been examined in cytoplasts obtained from intact and anucleate algae and compared with known data on chloroplast division in algae enucleated since various periods of time (up to three weeks) and with corresponding electron micrographs.The distribution curves of the chloroplast size in intact algae, anucleate fragments, and cytoplasts were established.On the basis of the experimental evidence, it is proposed that both replication of chloroplastic DNA and division of the chloroplasts are under nuclear control but that information or factors involved in the former have a longer life time than those involved in the latter.In addition, some factors involved in division seem to be located in the peripheral part of the cytoplasm ofAcetabularia. Alternatively, they could be lost by the cytoplasts.     

Growth responses of the mixotrophic dinoflagellates, Cryptoperidiniopsis sp. and Pfiesteria piscicida, to light under prey-saturated conditions

Recent research emphasis on the ecology of Pfiesteria spp. (Dinophyceae) has led to recognition of several morphologically similar heterotrophic dinoflagellates that often co-occur with Pfiesteria spp. in estuaries along the United States Atlantic coast. These include cryptoperidiniopsoid dinoflagellates, which resemble Pfiesteria spp. in having complex life cycles that include zoospores capable of kleptoplastidy. To examine and compare the role of kleptoplastidy in Cryptoperidiniopsis sp. and Pfiesteria piscicida, we tested the effects of irradiance on growth under prey-saturated (Storeatula major, Cryptophyceae) conditions. Growth of Cryptoperidiniopsis was strongly influenced by light intensity while no major effects were observed in P. piscicida. In Cryptoperidiniopsis, highest cell numbers and specific growth rates, but lowest specific cryptophyte consumption rates, were found at the highest light intensity tested (100 μmol photons m⁻² s⁻¹). A growth model was developed and used to estimate that the average half-life of chloroplasts ingested by Cryptoperidiniopsis decreased 3.4-fold from 12.6 h at high light to 3.7 h in the dark. These results show that light strongly enhances specific growth rate and growth efficiency of Cryptoperidiniopsis feeding on cryptophytes, and suggest that retained kleptochloroplasts may play a quantitatively significant role in carbon and energy metabolism of this organism. Differences in the effects of light between Cryptoperidiniopsis and P. piscicida may reflect different nutritional strategies, and allow these closely related dinoflagellates to occupy different niches and co-exist.     

南海北部琼东南盆地BD-2井中新世有孔虫生物地层及沉积环境

南海北部琼东南盆地BD-2井中新世地层沉积连续,是琼东南盆地中新统较典型的钻井剖面之一。该井中新世地层中含丰富的有孔虫化石,共鉴定有孔虫62属98种,其中浮游有孔虫13属41种,底栖有孔虫49属57种。根据有孔虫标志种及螺旋浮游有孔虫旋向优势度的变化,对该井的有孔虫生物地层进行了详细划分,从早中新世至晚中新世共识别出11个有孔虫化石带或联合化石带。探讨了下中新统与中中新统,中中新统与上中新统界线的有孔虫划分标志。依据有孔虫丰度、分异度及组合特征,讨论了BD-2井从早中新世至晚中新世沉积环境演化特征。