Isolation and characterization of a single‐stranded RNA virus that infects the marine planktonic diatom Chaetoceros sp. (SS08‐C03)

Diatoms are the major primary producers in the world's aquatic environment; hence, their dynamics are an important focus in current studies. Viruses, along with other physical, chemical, and biological factors, have recently been recognized as potential factors of diatom mortality. We isolated and characterized a new diatom virus (Csp03RNAV) that causes lysis of the marine planktonic diatom Chaetoceros sp. strain SS08‐C03 isolated from Hiroshima Bay, Japan. Here, we present the physiology, morphology, and genome characteristics of this virus. Csp03RNAV was isolated from surface waters of Yatsushiro Sea, Japan. Virions were icosahedral and 32 nm in diameter, and accumulated in the cytoplasm of the host cells. The latent period was estimated to be <48 h. Csp03RNAV harbors a single‐stranded RNA genome, which has 9417 bases encoding two open reading frames that code for putative replication‐related proteins and putative structural proteins, respectively. The monophyly of Csp03RNAV and the other known diatom‐infecting single‐stranded RNA viruses (genus Bacillarnavirus), Rhizosolenia setigera RNA virus, Chaetoceros socialis f. radians RNA virus, and Chaetoceros tenuissimus RNA virus was strongly supported by phylogenetic analysis based on the amino acid sequence of the RNA‐dependent RNA polymerase domain. On the basis of these results, Csp03RNAV is considered to be a new member of the genus Bacillarnavirus.     

Vallodiscus gen. nov., a new fossil resting spore morpho-genus related to the marine diatom genus Chaetoceros (Bacillariophyceae)

A new fossil marine diatom resting spore morphogenus, Vallodiscus Suto gen. nov., is described using samples from Deep Sea Drilling Project Site 338 in the Norwegian Sea, Sites 436 and 438 in the north‐west Pacific Ocean and the onland Newport Beach Section, California. Vallodiscus is characterized by a single ring of veins along the epivalve margin and a hypovalve covered with circular depressions of several sizes with gentle elevation. The morpho‐genus bears three new species and one new combination: Vallodiscus simplexus Suto sp. nov., Vallodiscus complexus Suto sp. nov., Vallodiscus lanceolatus Suto sp. nov. and Vallodiscus chinchae (Mereschkowsky) Suto comb. nov.     

Global diversity of two widespread,colony‐forming diatoms of the marine plankton,Chaetoceros socialis (syn. C. radians) and Chaetoceros gelidus sp. nov.

Marine phytoplankton samples containing diatoms of the Chaetoceros socialis group were collected from Thailand, China, Denmark, and Greenland, and cells were isolated into culture for light and electron microscopy and DNA sequencing of D1–D3 of the LSU rDNA. Species of this lineage are characterized by three short and one long setae extending from each cell, the long setae from several cells joining into a common center to form large colonies, which are sometimes visible with the naked eye. Phylogenetic analyses including sequences from other parts of the world revealed segregation into three groups. Most sequences fell into two large clades, one comprising material from cold waters, whereas the other contained material from warmer waters. Strain CCMP 172 from the Strait of Georgia, Washington State, USA, formed a separate group. The warm‐water species included Chinese and Thai material and therefore probably also material from the type locality of C. socialis, Hong Kong. It is characterized by all setae being covered by spines and the setae extending from the valve at some distance from the margin. In the resting spores, both valves are ornamented with spines. The cold‐water material is characterized by three spiny and one mostly smooth long setae, and the setae extend from the valve near the margin. Both valves of the resting spore are smooth. This material is described as C. gelidus sp. nov. C. radians, described from the Baltic in 1894, is considered a synonym of C. socialis. CCMP172 is in many ways intermediate and probably constitutes a separate species. The published evidence on this globally distributed and sometimes bloom‐forming group of species indicates higher species diversity than presently thought.     

Proprietes antifongiques d'une diatomee Chaetoceros lauderi ralfs C C

Résumé Une souche de la diatomée marine Chaetoceros lauderi a montré une activité antifongique très spécifique vis-à-vis des Dermatophytes. Dans l'état actuel des travaux de purification, cette activité semble liée à l'activité antibactérienne à large spectre mise en évidence précédemment chez cette même espèce.

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In fungicidal action researches from phytoplanctonic algae strains in culture an significant activity against pathogenic fungi of man was found for substances from the marine diatom Chaetoceros lauderi.

     

Research note: High efficiency transformation of the diatom Phaeodactylum tricornutum with a promoter from the diatom Cylindrotheca fusiformis

A high efficiency transformation system was established for the pennate diatom Phaeodactylum tricornutum Bohlin using a plasmid containing fucoxanthin chlorophyll a/c binding protein ( fcp ) promoter/terminator and nitrate reductase ( NR ) promoter/terminator that are derived from the pennate diatom Cylindrotheca fusiformis . The plasmid that contains the zeocin resistance gene ( ble ) with the fcp promoter and enhanced green fluorescent protein gene ( egfp ) with the NR promoter was introduced into P. tricornutum using microparticle bombardment. Transformants (650 ± 58 per 10⁸ cells) were obtained. The yield of transformants was between 1.5 and 130 times higher than previously reported P. tricornutum transformation systems. Four to seven copies of the ble gene were integrated into genomic DNA of the transformants. This high efficiency transformation system of P. tricornutum is expected to provide a powerful tool for high-throughput analysis of gene function using homologous recombination or RNAi.     

Characterization of the growth and lipid content of the diatom Chaetoceros muelleri

Chaetoceros muelleri (Schütt) was cultured on a thermal gradient plate, subjected to two media types with a range of specific conductances, and evaluated for growth and neutral lipid accumulation. Growth was measured directly by daily changes in cell numbers and indirectly by changes in optical density at 750 nm. C. muelleri exhibited a growth rate of at least two doublings day-1 over broad temperature (20 to 35 °C) and conductance ranges (10 to over 60 mS cm-1) and the optimum growth rate approached 4.0 doublings day-1 at 30 °C and a conductance of 25 mS cm-1. Intracellular neutral lipid storage was evaluated with fluorometry and epifluorescent microscopy using the fluorochrome Nile Red. Gravimetric analysis revealed a total lipid content in nitrogen-depleted cultures of C. muelleri of over 400 mg L-1, five to seven times that observed in nitrogen-replete cultures. Based on its high growth rate, tolerance to a broad range of temperatures and specific conductances, and large quantity of intracellular lipid, C. muelleri may have potential for exploitation as a renewable precursor to liquid fuels or as a lipid source. This revised version was published online in August 2006 with corrections to the Cover Date.     

角毛藻光合作用对连续强光照射的动态响应

利用调制荧光技术研究了角毛藻(Chaetocerossp.)光合作用对连续强光照射的动态响应。用1000μmolm-2s-1的强光处理12h内,角毛藻的快速光曲线和荧光参数呈动态变化。处理30min内,最大光合速率P(m即最大相对电子传递速率)和光系统Ⅱ的最大量子产量Fv/Fm均逐渐下降至最低值,表明角毛藻受到了光抑制;初始斜率α的降低和非光化学淬灭NPQ的升高,表明角毛藻可以通过降低捕光色素对光能的吸收和增加热耗散来保护光合器官免受过度损伤。此后,α开始增加,但始终不能恢复到初始值,说明角毛藻逐渐适应了强光照状态,捕光能力逐渐增加;Pm逐渐恢复并超过初始值,最终(8h后)达初始值的2倍,表明经过光系统Ⅱ的电子传递明显加快;而Fv/Fm只能恢复到初始值的一半左右,说明强光下光系统Ⅱ的光能转换效率明显降低;NPQ达最大值(1h)后逐渐下降至低于初始值,表明热耗散在光保护中不再起主要作用。随处理时间的延长,出现光化光关闭后的短期荧光上升现象,且上升速度逐渐加快,说明围绕光系统Ⅰ的环式电子传递速度逐渐加快,可能在光保护中发挥重要作用。     

Concerted response of xanthophyll-cycle pigments in a marine diatom, Chaetoceros gracilis, to shifts in light condition

The interconversion rate of diadinoxanthin (DD) cycle under high irradiance and subsequent darkness was analyzed using the cultivated centric marine diatom, Chaetoceros gracilis Schütt. A prompt de‐epoxidation from diadinoxanthin to diatoxanthin (DT) occurred immediately after the onset of higher irradiance. The fist‐order rate constant, k, for this de‐epoxidation was 0.1–0.2 min‐¹ irrespective of the irradiance. The difference in photon fluence rate lead to the difference of the final amount of DT, leaving the rate constant at almost the same value. After turning off the light, epoxidation from DT to DD occurred. The first‐order constant of epoxidation was much slower than that of de‐epoxidation: 0.005 – 0.009 min‐¹. Independent of this epoxidation process, de novo synthesis of DD‐cycle pigments was also observed under the subsequent darkness. Based on these findings, a common nature of the DD‐cycle as a protection mechanism for photo‐systems was demonstrated for C. gracilis.     

Metabolic fingerprinting reveals differences between northern and southern strains of the cryptic diatom Chaetoceros socialis

Morphology and molecular phylogeny constitute the structural elements of diatom taxonomy. These approaches do not, however, give information on the functioning of taxa. Additional methods to serve a more integrated and wide-ranging taxonomy have therefore been called for. Metabolic fingerprinting is one approach used within the field of metabolomics, often applied in classification of samples. Here we apply metabolic fingerprinting in a taxonomic study of a cryptic diatom species. Strains of the cosmopolitan diatom Chaetoceros socialis from two geographical areas; the north-east Atlantic and Arctic and the Gulf of Naples, were cultivated at three different temperatures; 2.5, 8 and 13°C. The strains from the two different geographical areas exhibited different growth rates as well as different photosynthetic efficiencies. Algal extracts, collected at the end of the growth experiments, were analysed by Ultra-Performance Liquid Chromatography High Resolution Mass Spectrometry. The two groups of strains were separated by principal component analysis of their metabolic fingerprints. Analysis of the data revealed both qualitative and quantitative differences in metabolite markers. These phenotypic differences reinforce differences also found for morphology, phylogenetic markers and growth rates, and point at different adaptive characteristics in organisms living under different temperature regimes.     

Observations on Chaetoceros salsugineus (Chaetocerotales,Bacillariophyceae): first record of this bloom-forming diatom in a European estuary

Examination of material obtained from the Urdaibai estuary (North Spain) during a 2 year phytoplankton sampling programme revealed a rare species of the genus Chaetoceros, C. salsugineus Takano. The morphology of this small (4·0×5·5?µm) member of the subgenus Hyalochaetae has been described using both light and electron microscopy. Regarding the original material, two new features have been observed: longer chains up to 24 cells and a new type of aperture, bisected by the fusion of central protuberances of adjacent valves. This bloom-forming diatom reached high densities in the poly-euhaline zone of the estuary closely related with high temperatures, denoting its neritic and summer preferences. Maximal cell densities (10⁶–10⁷ cells l^?1) were reached in August and September when the water column was completely mixed and the salinity and temperature were about 31 psu and 20°C, respectively. This is the first report of C. salsugineus in Europe and contributes to the knowledge of the morphological and ecological features of this species. Relationships with other small species of the subgenus Hyalochaetae are also discussed.     

Stephanodiscus kusuensis, sp. nov a new Pleistocene diatom from Southern Japan

A new fossil diatom, Stephanodiscus kusuensis sp. nov., is described from a diatomite deposit from the Kusu Basin, Oita Prefecture Japan. Radiometric age estimates of the strata in which the taxon occurs are 400–500 kyr. The new species is distinct from all other known species of Stephanodiscus because it possesses a highly pronounced double concentric undulation and two distinct arrangements of central strutted processes. Cladistic analysis is used to identify the phylogenetic position of this taxon with respect to other Stephanodiscus species.     

The role of external carbonic anhydrase in photosynthesis during growth of the marine diatom Chaetoceros muelleri

Carbon dioxide concentrating mechanisms (CCMs) act to improve the supply of CO₂ at the active site of ribulose‐1,5‐bisphosphate carboxylase/oxygenase. There is substantial evidence that in some microalgal species CCMs involve an external carbonic anhydrase (CA_ext) and that CA_ext activity is induced by low CO₂ concentrations in the growth medium. However, much of this work has been conducted on cells adapted to air‐equilibrium concentrations of CO₂, rather than to changing CO₂ conditions caused by growing microalgal populations. We investigated the role of CA_ext in inorganic carbon (C_i) acquisition and photosynthesis at three sampling points during the growth cycle of the cosmopolitan marine diatom Chaetoceros muelleri. We observed that CA_ext activity increased with decreasing C_i, particularly CO₂, concentration, supporting the idea that CA_ext is modulated by external CO₂ concentration. Additionally, we found that the contribution of CA_ext activity to carbon acquisition for photosynthesis varies over time, increasing between the first and second sampling points before decreasing at the last sampling point, where external pH was high. Lastly, decreases in maximum quantum yield of photosystem II (F_v/F_m), chlorophyll, maximum relative electron transport rate, light harvesting efficiency (α) and maximum rates of C_i‐ saturated photosynthesis (V_max) were observed over time. Despite this decrease in photosynthetic capacity an up‐regulation of CCM activity, indicated by a decreasing half‐saturation constant for CO₂ (K_0.5CO₂), occurred over time. The flexibility of the CCM during the course of growth in C. muelleri may contribute to the reported dominance and persistence of this species in phytoplankton blooms.     

Continuous culture with deep seawater of a benthic food diatom Nitzschia sp.

A continuous culture system for a benthic food diatom Nitzschia sp. wasestablished by using properties of high nutrient and clean of deep seawater(DSW). DSW collected from 320 m depth in Muroto City, Japan, was introducedinto a glass-pipe bioreactor (14 cm length, 3 cm diam.) containing glassbeads of 0.5 cm diam. as substrata for the alga, and it was incubated at18°C · 80Em^–2sec^–1 · L:D=14:10. The chlorophyll a yield of benthicdiatoms in a reactor as a unit of surface area of the substratum was only0.001–0.003 g cm^–2 when the flow rate of DSW was 0(batch culture conditions). However, when DSW was supplied continuously to areactor, the yield increased to 1.4 g-chl.a cm^–2 alongwith the increase in flow rate of DSW. Moreover, amounts of chl.a washed outof the system were negligible, 0.0014 to 0.0045%, even though theflow rate of DSW was as much as 25 times h^–1, suggesting thatsloughing of benthic diatoms from the substratum was minimized. Although theyield of diatoms fluctuated significantly at the time that the DSW wascollected, the variation could be minimized by increasing the flow rate ofDSW. These results indicate that the continuous culturing system with DSWsupports the stable and effective mass culture of benthic food diatom.     

Continuous production of diatom Entomoneis sp. in mechanically stirred tank and flat-panel airlift photobioreactors

Continuous production of diatom Entomonies sp. was performed in mechanically stirred tank and flat-panel airlift photobioreactors (FPAP). The maximum specific growth rate of diatom from the batch experiment was 0.98 d^?1. A series of dilution rate and macronutrient concentration adjustments were performed in a stirred tank photobioreactor and found that the dilution rate ranged from 0.7 to 0.8 d^?1 and modified F/2 growth media containing nitrate at 3.09?mg N/L, phosphate at 2.24?mg P/L, and silicate at 11.91?mg Si/L yielded the maximum cell number density. Finally, the continuous cultivation of Entomonies sp. was conducted in FPAP using the optimal conditions determined earlier, resulting in the maximum cell number density of 19.69?×?10⁴ cells/mL, which was approximately 47 and 73% increase from the result using the stirred tank photobioreactor fed with modified and standard F/2 growth media, respectively.     

Differential responses in EPA and fucoxanthin production by the marine diatom Stauroneis sp. under varying cultivation conditions

There has been an increasing drive toward better valorising raw biological materials in the context of the sustainability of bio-based industries and the circular economy. As such, microalgae hold the ability to biosynthesise valuable metabolites, which are sought after within the bioenergy, pharmaceuticals, cosmetics or nutrition sectors. Owing to their bioactivities, the xanthophyll pigment fucoxanthin and the omega-3 polyunsaturated fatty acid eicosapentaenoic acid (EPA) have fostered increasing interests in terms of sustainably refining them from natural sources, such as microalgae. Together with the suitability of individual species to industrial cultivation, a key challenge resides in optimizing the yields of these compounds within the microalgal biomass they are retrieved from. The marine diatom Stauroneis sp. LACW24 was batch cultivated into its stationary phase of growth prior to being subjected at high cell density (1 × 10⁶ cells mL⁻¹) to seven different regimes of light exposure in replenished medium and under nutritional limitation (silica and nitrate) for 12 days. The highest EPA proportions and yields were obtained under blue LED in f/2 medium (16.5% and 4.8 mg g⁻¹, respectively), double the values obtained under red LED illumination. The fucoxanthin yield was the highest when cells were subjected to blue LEDs (5.9 mg g⁻¹), a fourfold increase compared to the nitrogen-limited treatment under white LEDs. These results indicate that a two-stage approach to the batch cultivation of this diatom can be used for enhancing the production of the high-value metabolites fucoxanthin and EPA post-stationary phase.     

镧—氨基酸络合物对牟氏角毛藻的影响

研究了镧 脯氨酸和镧 苯丙氨酸络合物对牟氏角毛藻 (Chaetocerosmulleri)的影响 ,结果表明 ,两种络合物对牟氏角毛藻细胞繁殖和叶绿素的合成有明显的促进作用 ,浓度为 1～ 1 5mg·L- 1 时 ,至第 6天细胞数分别提高 1 3.2～ 1 9.2 %、1 2 .6～ 2 1 .5% (P <0 .0 5) ;叶绿素浓度分别提高 2 5.2～ 33.9%、2 6.2～ 45.1 % (P <0 .0 5) ;其中以浓度为 5mg·L- 1 的实验组最高 .镧的氨基酸络合物对牟氏角毛藻的作用效果与镧相比是等同的 ,但最佳浓度明显降低 .