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A 96-h exposure experiment was conducted to elucidate the toxicity responses of the marine diatom Thalassiosira pseudonana upon exposure to different temperatures and copper (Cu) concentrations. Three Cu treatments (seawater control; 200 μg/L Cu, EC 50 for the yield at 25 °C; and 1000 μg/L Cu, EC 50 for growth inhibition at 25 °C) were conducted against four temperatures (10 °C, 15 °C, 25 °C and 30 °C). Growth rate and photosynthetic responses showed a significant interacting thermal-chemical effect with strong synergistic responses observed at 30 °C treatments. Expression of heat shock protein ( hsp ) was positively modulated by increasing temperatures. Hsp 90 , hsp90–2 and sit1 (related to silica shell formation) were highly expressed at 30 °C under 1000 μg/L Cu, while the genes encoding light harvesting proteins ( 3HfcpA and 3HfcpB ) and silaffin precursor sil3 were significantly up-regulated at 15 °C under 200 μg/L Cu. Our results indicated an increase Cu toxicity to T. pseudonana under high temperature and Cu dose. [ABSTRACT FROM AUTHOR]

Diatoms are one of the most successful marine eukaryotic algal groups, responsible for up to 20% of the annual global CO2 fixation. The evolution of a CO2-concentrating mechanism (CCM) allowed diatoms to overcome a number of serious constraints on photosynthesis in the marine environment, particularly low [CO2]aq in seawater relative to concentrations required by the CO2 fixing enzyme, ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO), which is partly due to the slow diffusion rate of CO2 in water and a limited CO2 formation rate from HCO-3 in seawater. Diatoms use two alternative strategies to take up dissolved inorganic carbon (DIC) from the environment: one primarily relies on the direct uptake of HCO-3 through plasma-membrane type solute carrier (SLC) 4 family HCO-3 transporters and the other is more reliant on passive diffusion of CO2 formed by an external carbonic anhydrase (CA). Bicarbonate taken up into the cytoplasm is most likely then actively transported into the chloroplast stroma by SLC4-type transporters on the chloroplast membrane system. Bicarbonate in the stroma is converted into CO2 only in close proximity to RubisCO preventing unnecessary CO2 leakage. CAs play significant roles in mobilizing DIC as it is progressively moved towards the site of fixation. However, the evolutionary types and subcellular locations of CAs are not conserved between different diatoms, strongly suggesting that this DIC mobilization strategy likely evolved multiple times with different origins. By contrast, the recent discovery of the thylakoid luminal θ-CA indicates that the strategy to supply CO2 to RubisCO in the pyrenoid may be very similar to that of green algae, and strongly suggests convergent coevolution in CCM function of the thylakoid lumen not only among diatoms but among eukaryotic algae in general. In this review, both experimental and corresponding theoretical models of the diatom CCMs are discussed. [ABSTRACT FROM AUTHOR]

The antibacterial potential of centric marine diatoms has been compared against the clinical pathogens and identified the potential diatom by rbcL gene sequencing. Totally, five diatoms namely Odontella aurita, Thalassiosira subtilis, Chaetoceros curvisetus, Skeletonema costatum and Coscinodiscus centralis were isolated from Cuddalore coastal waters. The diatoms were morphologically identified and isolated using micro capillary-pipette and serial dilution method. The isolated diatoms were cultured in Guillard's f/2 medium to get biomass for the antibacterial study. The dried biomass of the cultured diatoms was individually extracted with methanol, ethanol and hexane. All the obtained extracts were tested against Staphylococcus haemolyticus, Proteus vulgarius and Vibrio alginolyticus. The crude ethanol extract of O. aurita was exhibited highest zone of inhibition against all the test pathogens. The MIC of O. aurita was recorded as 50 μg/ml against both Staphylococcus haemolyticus and Proteus vulgarius whilst 75 μg/ml against Vibrio alginolyticus. This study indicates that O. aurita possesses antibacterial activities but the release of antibiotics depends on physical or chemical rupture of algal cells and extractive solvents. Based on the maximum antibacterial activity, O. aurita was further identified by rbcL gene sequencing. The rbcL gene could be an identical region for the species level identification of diatoms. [ABSTRACT FROM AUTHOR]

Diatoms operate a CO2-concentrating mechanism (CCM) that drives upwards of 20% of annual global primary production. Recent progress in CCM research in the marine pennate diatom Phaeodactylum tricornutum revealed that this diatom directly takes up HCO3- from seawater through low-CO2-inducible plasma membrane HCO3- transporters, which belong to the solute carrier (SLC) 4 family. Apart from this, studies of carbonic anhydrases (CAs) in diatoms have revealed considerable diversity in classes and localization among species. This strongly suggests that the CA systems, which control permeability and flux of dissolved inorganic carbon (DIC) by catalysing reversible CO2 hydration, have evolved from diverse origins. Of particular interest is the occurrence of low-CO2-inducible external CAs in the centric marine diatom Thalassiosira pseudonana, offering a strategy of CA-catalysed initial CO2 entry via passive diffusion, contrasting with active DIC transport in P. tricornutum. Molecular mechanisms to transport DIC across chloroplast envelopes are likely also through specific HCO3- transporters, although details have yet to be elucidated. Furthermore, recent discovery of a luminal θ-CA in the diatom thylakoid implied a common strategy in the mechanism to supply CO2 to RubisCO in the pyrenoid, which is conserved among green algae and some heterokontophytes. These results strongly suggest an occurrence of convergent coevolution between the pyrenoid and thylakoid membrane in aquatic photosynthesis. [ABSTRACT FROM AUTHOR]

Phosphorus (P) is a limiting macronutrient for diatom growth and productivity in the ocean. Much effort has been devoted to the physiological response of marine diatoms to ambient P change, however, the whole-genome molecular mechanisms are poorly understood. Here, we utilized RNA-Seq to compare the global gene expression patterns of a marine diatom Skeletonema costatum grown in inorganic P-replete, P-deficient, and inorganic- and organic-P resupplied conditions. In total 34,942 unique genes were assembled and 20.8% of them altered significantly in abundance under different P conditions. Genes encoding key enzymes/proteins involved in P utilization, nucleotide metabolism, photosynthesis, glycolysis and cell cycle regulation were significantly up-regulated in P-deficient cells. Genes participating in circadian rhythm regulation, such as circadian clock associated 1, were also up-regulated in P-deficient cells. The response of S. costatum to ambient P deficiency shows several similarities to the well-described responses of other marine diatom species, but also has its unique features. S. costatum has evolved the ability to re-program its circadian clock and intracellular biological processes in response to ambient P deficiency. This study provides new insights into the adaptive mechanisms to ambient P deficiency in marine diatoms.

An unprecedented phosphatidylmonogalactosyldiacylglycerol pool (PGDG, 1) rich in polyunsaturated fatty acids was isolated from the marine diatoms Thalassiosira weissflogii. Here we report for the first time the NMR characterization of this rare lipid from marine organisms along with a synthetic strategy for the preparation of a PGDG analog (2). PGDG 1 exhibited immunostimulatory activity in human dendritic cells (DCs) and the synthetic PGDG 2 was prepared to explore its mechanism of action. A Toll-like receptor-4 (TLR-4) agonistic activity was evidenced in human and murine DCs underlying the antigen-specific T-cell activation of this class of molecules.

ABSTRACT To describe potential intraspecific variation in phosphorus incorporation in two strains of Phaeodactylum tricornutum (Bohlin), Ub3 and Ub7, alkaline phosphatase (AP) activity was evaluated via enzyme-labeled fluorescence assay. Analysis using the probe ELF-97(r) provides individual evaluation, and therefore can determine the nutritional status of inorganic phosphorus in phytoplanktonic cells. Bioassays compared the control treatment to both phosphate-enriched and phosphate-depleted treatments by varying only the phosphate concentration in the media. The P. tricornutum strains exhibited differences in their development when incubated in the phosphate-enriched media. The development of the Ub7 strain differed by exhibiting "luxury uptake" and utilization of organic phosphorus, and the alkaline phosphatase analysis indicated limitations of this clone under such conditions. The Ub7 strain showed higher AP activity, when compared to Ub3, in the P-enriched condition. P. tricornutum presented increases in AP activity and low variation in Surface/Volume ratio, by increasing biovolume and its maximum linear dimension, as strategies for phosphate incorporation. Our results highlight intraspecific differences in alkaline phosphatase activity, and hence differences in the incorporation of organic phosphorus, as the tested species regulated enzymatic activity under different external phosphate concentrations.

Multifaceted changes in marine environments as a result of anthropogenic activities are likely to have a compounding impact on the physiology of marine phytoplankton. Most studies on the combined effects of rising p CO 2 , sea surface temperature, and UVB radiation on marine phytoplankton were only conducted in the short-term, which does not allow to test the adaptive capacity of phytoplankton and associated potential trade-offs. Here, we investigated populations of the diatom Phaeodactylum tricornutum that were long-term (∼3.5 years, ∼3000 generations) adapted to elevated CO 2 and/or elevated temperatures, and their physiological responses to short-term (∼2 weeks) exposure of two levels of ultraviolet-B (UVB) radiation. Our results showed that while elevated UVB radiation showed predominantly negative effects on the physiological performance of P. tricornutum regardless of adaptation regimes. Elevated temperature alleviated these effects on most of the measured physiological parameters (e.g., photosynthesis). We also found that elevated CO 2 can modulate these antagonistic interactions, and conclude that long-term adaptation to sea surface warming and rising CO 2 may alter this diatom's sensitivity to elevated UVB radiation in the environment. Our study provides new insights into marine phytoplankton's long-term responses to the interplay of multiple environmental changes driven by climate change. • Elevated UVB showed predominantly negative effects on P. tricornutum. • Elevated temperature alleviated these negative effects. • Elevated CO 2 modulated these antagonistic interactions between temperature and UVB. [ABSTRACT FROM AUTHOR]

Biosilica with a surface area of 143 m 2 g −1 derived from marine diatoms was prepared using an easy two-step method involving washing with dilute acid and baking. The extracted biosilica was used to remove divalent lead ions, i.e., Pb(II), from aqueous solution. The effects on Pb(II) adsorption of initial pH, shaking speed, and adsorbent loading were investigated. The adsorption of Pb(II) in the presence of other ions was also investigated. The biosilica showed a high adsorption capacity with high selectivity for Pb(II). The experimental maximum adsorption capacity was 108.2–120.4 mg g −1 at an adsorbent loading of 1 g L −1 . The adsorption process was best described by the Langmuir model. The adsorbent selectively adsorbed Pb(II) from binary ion systems in the presence of Cu(II), Cd(II), Ni(II), and Ag(I). The results of this study show that biosilica extracted from fresh marine diatoms is a more efficient and selective adsorbent for Pb(II) than other inorganic adsorbents. [ABSTRACT FROM AUTHOR]

Integrated Ocean Drilling Program (IODP) Expedition 337 drilled a new core in 2012 that extended the coring depth of Hole C9001D at Site C9001 below the 647 meters below seafloor (mbsf) that was penetrated in 2006 during the Chikyu Shakedown Cruise CK06-06. Drilling operations at Site C0020 (formerly Site C9001), located 80 km off the Shimokita Peninsula of northeastern Honshu, Japan, penetrated Hole C0020A to a total depth of 2466 mbsf. IODP Exp. 337 sought to explore a deep hydrocarbon system and coalbeds that, prior to drilling, were estimated to be of Eocene age. Combined shipboard diatom and palynological analyses, however, revealed unexpectedly thick sections of Pliestocene and Miocene sediments, an approximately 12–15 million year hiatus at 1086.5 mud depth below sea floor (m MSF), and coalbeds younger than originally predicted. New biostratigraphic results from Hole C0020A indicate the target coalbed is Early Miocene. The age of the youngest sediments obtained in Hole C0020A is revised from Late Pliocene to Early Pleistocene, and the basal age of Hole C9001C is confirmed as Early Pleistocene. The age of the deepest sediments obtained at Site C0020 is Late Oligocene-Early Miocene. All new data are integrated with the overlying interval obtained from Holes C9001C and C9001D to produce a composite chronostratigraphic framework for Site C0020. [ABSTRACT FROM AUTHOR]

The algal pyrenoid is a large plastid body, where the majority of the CO2-fixing enzyme, ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) resides, and it is proposed to be the hub of the algal CO2-concentrating mechanism (CCM) and CO2 fixation. The thylakoid membrane is often in close proximity to or penetrates the pyrenoid itself, implying there is a functional cooperation between the pyrenoid and thylakoid. Here, GFP tagging and immunolocalization analyses revealed that a previously unidentified protein, Pt43233, is targeted to the lumen of the pyrenoid-penetrating thylakoid in the marine diatom Phaeodactylum tricornutum. The recombinant Pt43233 produced in Escherichia coli cells had both carbonic anhydrase (CA) and esterase activities. Furthermore, a Pt43233:GFP-fusion protein immunoprecipitated from P. tricornutum cells displayed a greater specific CA activity than detected for the purified recombinant protein. In an RNAi-generated Pt43233 knockdown mutant grown in atmospheric CO2 levels, photosynthetic dissolved inorganic carbon (DIC) affinity was decreased and growth was constantly retarded; in contrast, overexpression of Pt43233:GFP yielded a slightly greater photosynthetic DIC affinity. The discovery of a θ-type CA localized to the thylakoid lumen, with an essential role in photosynthetic efficiency and growth, strongly suggests the existence of a common role for the thylakoid-luminal CA with respect to the function of diverse algal pyrenoids. [ABSTRACT FROM AUTHOR]

The lipid content, composition and productivity of the marine benthic diatom Halamphora coffeaeformis were studied in order to evaluate its potential as feedstock for biodiesel production. Cultures were carried out in two stages: I) in photobioreactors (PBRs) to increase the biomass as inoculum for larger volumes, and II) in raceway ponds to increase naturally the triacylglycerol (TAG) content during the stationary growth phase. Biomass concentrations of 0.64 g L −1 and 0.23 g L −1 were reached in the PBR and the raceway pond, respectively. Total lipid content was 54.4 (±11.6) % ash free dry weight (AFDW) in the raceway pond on day 19 (harvest day), with a neutral lipid content of 34% AFDW. The TAG productivity in the raceway pond was 1.2 mg L −1 d −1 . The indicators of biodiesel, calculated from fatty acid profile composition, showed that H. coffeaeformis oil was of good quality, according to international standards. Some hypothetical aspects are proposed in order to improve lipid productivity and net energy ratio in processes at larger scales. [ABSTRACT FROM AUTHOR]

Diatoms are phytoplanktonic organisms that grow successfully in the ocean where light conditions are highly variable. Studies of the molecular mechanisms of light acclimation in the marine diatom Phaeodactylum tricornutum show that carotenoid de-epoxidation enzymes and LHCX1, a member of the light-harvesting protein family, both contribute to dissipate excess light energy through non-photochemical quenching (NPQ). In this study, we investigate the role of the other members of the LHCX family in diatom stress responses. Our analysis of available genomic data shows that the presence of multiple LHCX genes is a conserved feature of diatom species living in different ecological niches. Moreover, an analysis of the levels of four P. tricornutum LHCX transcripts in relation to protein expression and photosynthetic activity indicates that LHCXs are differentially regulated under different light intensities and nutrient starvation, mostly modulating NPQ capacity. We conclude that multiple abiotic stress signals converge to regulate the LHCX content of cells, providing a way to fine-tune light harvesting and photoprotection. Moreover, our data indicate that the expansion of the LHCX gene family reflects functional diversification of its members which could benefit cells responding to highly variable ocean environments. [ABSTRACT FROM AUTHOR]

In this paper, we present an evaluation of genotoxic responses in marine diatom, Chaetoceros tenuissimus, isolated from Kandla Creek (lat 23.03° N, long 70.22° E), Gujarat, India, in terms of impairment of DNA integrity as a function of their exposure to elevated levels of mercury (Hg) under laboratory conditions. DNA integrity in C. tenuissimus was determined by partial alkaline unwinding assay. To our knowledge, this is the first such genotoxicity study to be conducted on marine diatom cultures towards understanding the relationship between Hg toxicity and DNA damage. Furthermore, we studied the impact of Hg on the growth of C. tenuissimus as a function of their exposure to enhanced levels of Hg in terms of decreasing chlorophyll a (chl a) concentrations. The data show the genotoxic effect of Hg on the growth of C. tenuissimus as well as DNA integrity to a great extent. Based on the results of our investigations, it is suggested that C. tenuissimus can be used as sentinel species for bio-monitoring of pollution due to genotoxic contaminants. [ABSTRACT FROM AUTHOR]

In this paper we determine whether a realistic mixture of hydrophobic chemicals affects the growth dynamics of a marine diatom and how this effect compares to the effect of temperature, light regime and nutrient conditions. To do so, we examine the specific growth rate of Phaeodactylum tricornutum in a 72 h algal growth inhibition test using a full factorial design with three nutrient regimes, two test temperatures, three light intensities and three chemical exposures. Passive samplers were used to achieve exposure to realistic mixtures of organic chemicals close to ambient concentrations. Nutrient regime, temperature and time interval (24, 48 and 72 h) explained 85% of the observed variability in the experimental data. The variability explained by chemical exposure was about 1%. Overall, ambient concentrations of hydrophobic compounds present in Belgian coastal waters, and for which the passive samplers have affinity, are too low to affect the intrinsic growth rate of P. tricornutum. [ABSTRACT FROM AUTHOR]

This study presents a new approach in aquatic toxicity testing combining passive sampling and passive dosing. Polydimethylsiloxane sheets were used to sample contaminant mixtures in the marine environment. These sheets were subsequently transferred to ecotoxicological test medium in which the sampled contaminant mixtures were released through passive dosing. 4 out of 17 of these mixtures caused severe effects in a growth inhibition assay with a marine diatom. These effects could not be explained by the presence of compounds detected in the sampling area and were most likely attributable to unmeasured compounds absorbed to the passive samplers during field deployment. The findings of this study indicate that linking passive sampling in the field to passive dosing in laboratory ecotoxicity tests provides a practical and complimentary approach for assessing the toxicity of hydrophobic contaminant mixtures that mimics realistic environmental exposures. Limitations and opportunities for future improvements are presented. [ABSTRACT FROM AUTHOR]

Cette étude consiste à évaluer le biotransfert du tributylétain (TBT) par le filtreur bivalve Mya arenaria, à partir d'une culture semi-continue de la diatomée Phaeodactylum tricornutum exposée à un niveau de contamination n'arrêtant pas sa croissance à long terme. La dose du contaminant employée (100 ng I-1) est de l'ordre de grandeur de celles décelées dans certaines régions contaminées, telles les marinas et les régions portuaires. Dans ces conditions, la culture de P tricornutum a maintenu son équilibre de croissance durant plus de 50 jours. Sa productivité était légèrement inférieure (de 4 %) à celle de la culture témoin (sans contaminant). L'ingestion des diatomées contaminées, sur une période de 90 jours, a entraîné une bioaccumulation de TBT et DBT (dibutylétain) dans la chair du mollusque, atteignant des concentrations respectives de 133 et 128 ng g-1 (poids humide). L'étude suggère que le biotransfert du TBT et du DBT associés à des micro-algues en croissance continue ou semi-continue pourrait constituer un facteur de bioamplification important du biocide en milieu naturel.

Antifreeze proteins are a group of proteins that allow organisms to survive in subzero environments. These proteins possess thermal hysteresis and ice recrystallization inhibition activities. In the present study, we demonstrated the efficiency of a recombinant antifreeze protein from the Arctic yeast Leucosporidium sp. AY30, LeIBP, in cryopreservation of the marine diatom Phaeodactylum tricornutum, which is one of the classical model diatoms and has most widely been studied with regard to its ecology, physiology, biochemistry, and molecular biology. P. tricornutum cells were frozen by either a fast or two-step freezing method in freezing medium containing 10 % dimethyl sulfoxide, glycerol, propylene glycol, and ethylene glycol, respectively, with or without LeIBP supplement. When cells were frozen using the two-step freezing method, cell survival was significantly increased and statistically the same as that of unfrozen native cells in the presence of 0.1 mg/ml LeIBP in 10 % propylene glycol or 10 % ethylene glycol at day 11 of post-thaw culture. In the presence of LeIBP, the concentration of chlorophyll a was dramatically increased to 14-, 48-, 1.6-, and 8.8-fold when cells were frozen in freezing medium containing dimethyl sulfoxide (DMSO), glycerol, propylene glycol (PG), and ethylene glycol (EG), respectively. Scanning electron microscopy observations demonstrated that the cells were also successfully preserved and epitheca or hypotheca were not deformed. These results demonstrate that LeIBP was successfully applied to improve cryopreservation of the marine diatom P. tricornutum. [ABSTRACT FROM AUTHOR]

Fistulifera sp. strain JPCC DA0580, a marine pennate diatom, contains extremely high levels of intracellular triglyceride and has been suggested as a promising source of feedstock for biodiesel fuels. JPCC DA0580 was isolated from a mangrove swamp located in Sumiyo Bay below the mouths of the Sumiyo and Yakugachi Rivers in Amami- Ohshima, Kagoshima, Japan. In this study, the morphology and the 18S rDNA sequence of JPCC DA0580 were compared with those of other Fistulifera strains. JPCC DA0580 possesses morphological characters of the genus Fistulifera, namely lightly silicified frustules, a distinct median costa (raphe sternum), and a wart-like central pore (fistula). Morphometric analysis revealed that JPCC DA0580 differs from other Fistulifera species by the presence of a valve with coarser striation and coarser areolation. On the basis of 18S rDNA phylogeny, JPCC DA0580 formed a well-supported clade with other members of the Fistulifera species complex, although the number of nucleotide substitutions was highest in JPCC DA0580. Our results led us to propose the taxonomic name Fistulifera solaris sp. nov. for JPCC DA0580. [ABSTRACT FROM AUTHOR]

In the genome of the marine diatom- Thalassiosira pseudonana, there are several putative genes encoding enzymes potentially constitute a classical C type biochemical CO-concentrating mechanism. Two genes encode a carboxylation enzyme phosphoenolpyruvate carboxylase (PEPC)1 and PEPC2; and another two encode decarboxylation enzymes, NAD-dependent malic enzyme (NAD-ME) and phosphoenolpyruvate carboxykinase (PEPCK). These genes were tagged by the enhanced-green fluorescence protein, egfp, ligated in the transformation vector, and transformed into the cells of T. pseudonana for localization of GFP fusion products. The PEPC1:GFP fusion was localized at the matrix of the periplastidal compartment, while the PEPC2:GFP fusion was localized at the mitochondria. The NAD-ME:GFP fusion was localized in the cytosol and the PEPCK:GFP fusion at the mitochondria. The transcripts level of NAD-ME was extremely low, and PEPCK transcript was significantly induced under the dark, suggesting that PEPCK is involved in the dark metabolism such as respiration and amino acid metabolism in the mitochondria. Treatments of low-COgrown T. pseudonana cells with inhibitors for PEPCK and PEPC efficiently dissipated the maximum rate of photosynthesis while these treatments did not affect high-affinity photosynthesis. These data strongly suggest that classical C enzymes play little role in the CCM in T. pseudonana. [ABSTRACT FROM AUTHOR]

Thirteen putative carbonic anhydrase (CA) genes have been identified in the marine multipolar centric diatom, Thalassiosira pseudonana, and two of these CAs have been localized previously. The first, an alpha CA (TpαCA1), was localized in the chloroplast stroma; the second, a zeta-type CA (TpζCA1), was localized to the periplasmic space. In the present study, cloning and localization of the remaining CAs were carried out. TpγCA2, TpγCA3, TpγCA4, TpγCA5, TpδCA1, TpδCA2, TpδCA3, and TpζCA1 were responsive to CO availability at the transcriptional level, being significantly reduced in cells grown at 0.4 % CO, whereas TpαCA1, TpαCA2, TpαCA3, TpγCA1, and TpδCA4 transcript levels were constitutive with respect to CO concentration. Full-length cDNAs for TpγCA1, TpγCA2, TpγCA3, TpγCA4, TpδCA1, and TpδCA2 were isolated and fused with the enhanced- green fluorescent gene at their 3′ termini. These GFP-fusion constructs were transformed into T. pseudonana, and the resulting GFP fusion products were localized using fluorescence microscopy. The δ-type TpδCA1 was localized on the periphery of the cell, strongly suggesting localization to the periplasmic space or the frustule. The δ-type TpδCA3 and the γ-type TpγCA2 were, respectively, localized in a periplastidal compartment and the cytosol. The δ-type TpδCA2, and the γ-types TpγCA1, 3, and 4 were localized in the mitochondria. The distribution of CAs in T. pseudonana contrasts notably with that of the raphid pennate diatom P. tricornutum, with likely consequences for CCM function including modes of CO acquisition, regions in which DIC is accumulated, and needs for minimizing CO leakage from the chloroplast. [ABSTRACT FROM AUTHOR]

It is well established that iron is one of the major constraints of primary productivity of marine diatoms in world oceans. In the present study, changes in the transcript levels of the 20 iron related genes were profiled in the marine diatom Phaeodactylum tricornutum during an early stage of acclimation from iron replete to iron-limited conditions. The results clearly showed that the profiles differ depending on genes, suggesting the occurrence of several modes of iron-responsive regulation at the transcriptional level. Upstream DNA sequences of iron starvation induced protein1 ( Isi1 ), ferrichrome binding protein1 ( FBP1 ), and flavodoxin ( Fld ) genes were isolated, fused with the GUS reporter gene, uidA , and transformed into P. tricornutum . Obtained transformants were subjected to the GUS reporter assay and the result clearly revealed that the GUS activity of all transformants was significantly increased upon iron limitation. Iron responsive Cis -elements in each promoter region were determined by the promoter truncation technique, demonstrating the occurrence of the critical iron-responsive regulatory regions of about 30 bp in the promoter regions of three genes, Isi1 , FBP1 , and Fld . Interestingly, these sequences were similar with each other revealing two conserved motifs, A; A(A/C)G(G/C)C(G/-)C(A/G)TG; and B; CACGTG(T/C)C, which are homologous to the iron responsive Cis -element in the green alga, Chlamydomonas reinhardtii . The impairment of the motif B in the Isi1 promoter resulted in the loss of iron response and the core regulatory region of the FBP1 promoter conferred an iron response on the constitutive cytomegalovirus promoter, P CMV , indicating that these conserved promoter sequences are iron-responsive elements. Finally, the inductive regulation of these promoters under iron-limited conditions was dissipated specifically by 5% CO 2 , strongly suggesting the participation of CO 2 in the transcriptional regulation of the iron-related gene promoters. [ABSTRACT FROM AUTHOR]

Among the proteins localized on the cell wall (frustule) of diatoms (frustule-associated proteins), several proteins tightly associated with the cell wall have been implicated in frustule formation. These proteins include diatom-specific unique serine- and lysine-rich sequences represented by silaffins. Taking advantage of available genome information, we used a recently described bioinformatics approach to screen silaffin-like proteins rich in serine and lysine from the genome of the marine pennate diatom Fistulifera sp. strain JPCC DA0580 and identified 7 proteins. All of the proteins shared a sequence motif called the XGXG domain, which was also confirmed in a silaffin-like protein identified in other diatoms. In vivo localization analysis revealed that one of the identified proteins, G7408, occurs throughout the frustule with a slightly uneven distribution. This novel frustule-associated protein could be a useful tool to elucidate the mechanism of biosilica formation in diatoms and to functionalize this strain for future biotechnological applications. [ABSTRACT FROM AUTHOR]