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Desmodesmus is a green microalgal genus that is frequently found in aquatic environments. Its high biomass productivity and potential as a source of lipids have attracted considerable attention. Although Desmodesmus is ubiquitous, it is difficult to identify; even within a small area, the diversity of the species and the fatty acids they produce are unknown. In this study, we performed scanning electron microscopy (SEM) and analyzed the genetic diversity of the internal transcribed spacer (ITS) region to accurately identify Desmodesmus in a local area of Japan (Saga City, Saga Pref.) and to assess its existence as a biological resource. In addition, we analyzed the fatty acid composition and content of the newly established strains. In total, 10 new strains were established, and 9 previously described species were identified. The presence of a cosmopolitan species indicated the global distribution of Desmodesmus. However, only regional species were found. One strain, dSgDes-b, did not form a clear clade with any described species in the phylogenetic analysis and had a characteristic ITS2 secondary structure. The cell wall of this strain exhibited a distinctive microstructure, and it produced docosahexaenoic acid (DHA); hence, the strain was described as a new species, Desmodesmus dohacommunis Demura sp. nov. Thus, useful information regarding the use of Desmodesmus as a bioresource was provided.

Separation of microalgae of various sizes and shapes is an important process that enables subsequent production of useful compounds. Herein, the separation of microalgae was accomplished using a magnetite-containing gel (42 μm) packed into a column. An algal suspension was injected into the top of the gel bed, after which water was passed through the column. The pressure generated during the process caused the lower domain of the gel bed to deform, resulting in narrowed gaps between the gel beads. When a suspension of Nannochloropsis sp. (0.0069-0.69 g L-1) was loaded and water was passed through the column at an applied pressure of 0.01-0.10 MPa, the majority of microalgae were captured within the upper domain of the gel bed, while only 20% were captured within the lower domain. The amount of Nannochloropsis sp. captured was expressed by an ordinary differential equation to determine the capture coefficient, K, and the maximum capture amount, Qmax. As pressure increased, gel gaps narrowed, K increased, and Qmax decreased because of a reduction in the number of effective capture sites upon compaction of the gel. When a mixed suspension of Anabaena sp., Monoraphidium sp., and Desmodesmus sp. (0.069 g L-1 each) was injected into the gel bed at an applied pressure of 0.01 MPa, only Anabaena sp. was captured at the bottom of the gel bed. This device can be applied for the separation of microalgae in rivers and the sea.

Pectic oligosaccharides have vital properties that are dependent on their molecular assemblies, such as the degree of esterification (DE) and branched side chains. The molecular structures are affected by the extraction solvents and conditions. In this study, we used pressurized carbon dioxide treatment (CT) as a novel method for pectin extraction from fresh and ripe orange peels and compared the total yield, DE, molecular weight, and monosaccharide compositions of the product obtained with those of products obtained using conventional extraction methods, such as heating with HCl or chelating with sodium hexametaphosphate (SHMP). CT provided a yield of approximately 3.8%, which was less than that of HCl (9.9%/dry weight). However, DE in the CT extract was approximately 94%, which was higher than that of HCl (75%) and SHMP (59%). The molecular weights of pectins in the CT extract appeared to be higher than that of HCl, except at a pressure of 2 MPa. β-Galacturonic acid was the major monosaccharide in pectin extracted by HCl and SHMP, whereas that was decreased in CT extract from fresh oranges. The results showed that extraction with CT contributed to a high DE pectin yield and diverse pectin molecular structures compared to conventional methods, although its application resulted in a lower overall pectin yield. These findings suggest the possibility of manufacturing oligosaccharides with novel properties.

The continuous decline in phosphorus (P) resources is a serious global issue. Therefore, it is important to develop methods to recover P from waste and wastewater. Most P ores are currently used in the phosphate form in the agriculture industry and in detergents, which results in a large release of phosphates into natural aquatic environments. Much attention has been given to measuring phosphate levels and monitoring water quality, survey, and control of algal phytoplankton dynamics. However, phosphite is oxidized from hypophosphite after plating and discharged as waste, so methods to recycle and reuse phosphite should also be developed. Currently, there is no evidence of phosphite utilization by photosynthetic eukaryotes, including eukaryotic algae. Thus, except for the possible utilization by some bacteria when phosphate is unavailable, the fate of the phosphite that is discharged is mostly unknown. Chlorella vulgaris (NIES-2170), Coccomyxa subellipsoidea (NIES-2166), Scenedesmus obliquus (NIES-2280), and Botryococcus braunii (BOT-22) were cultured in phosphite medium under conditions that prevented phosphate contamination and phosphite oxidation. As a result, the number of C. vulgaris and C. subellipsoidea increased in the phosphite medium, demonstrating the availability of phosphite for the growth of these strains. In particular, the growth of C. vulgaris increased as the phosphite concentration increased. After being cultured for 180 days in photosynthetic conditions, phosphite utilization rates were 32–38%. In contrast, S. obliquus and B. braunii strains did not grow in the phosphite medium. In conclusion, C. subellipsoidea and C. vulgaris utilize phosphite as a P resource, which is a novel finding in photosynthetic eukaryotes. The results of this study may have important implications for the phosphorus redox cycle.

Microalgae have potential as biomass energy sources with higher photosynthetic efficiency compared to terrestrial plants. The use of polyculture systems such as native microalgae communities for microalgae cultivation has several advantages, as well as challenges due to indeterminate species composition and growth rate variation between species. This paper presents an artificial neural network (ANN) model to estimate the growth of polyculture microalgae in a semi-continuous open raceway pond (ORP). The model was comprised of a multilayer backpropagation neural network with eight input parameters, one hidden layer, and one output parameter. The model was developed using datasets collected from the cultivation of polyculture microalgae in Minamisoma City, Fukushima Prefecture, Japan. The input parameters are as follows: initial algal concentration, harvesting period (between two and three days after the growth have begun), hydraulic retention time, addition of sodium acetate, average solar radiation (μmole m−2 s−1), average temperature (oC), pH condition, and nitrate ion ( NO 3 − ) concentration. The output variable is the microalgae concentration observed during the cultivation period. The output is represented using a single neuron. The result of the study showed that the designed three-layer ANN achieved a high prediction accuracy (R2 = 0.93) for all combinations of inputs.

Native polyculture microalgae is a promising scheme to produce microalgal biomass as biofuel feedstock in an open raceway pond. However, predicting biomass productivity of native polycultures microalgae is incredibly complicated. Therefore, developing polyculture growth model to forecast biomass yield is indispensable for commercial-scale production. This research aims to develop a polyculture growth model for native microalgal communities in the Minamisoma algae plant and to estimate biomass and biocrude oil productivity in a semi-continuous open raceway pond. The model was built based on monoculture growth of polyculture species and it is later formulated using species growth, polyculture factor (k value ), initial concentration, light intensity, and temperature. In order to calculate species growth, a simplified Monod model was applied. In the simulation, 115 samples of the 2014-2015 field dataset were used for model training, and 70 samples of the 2017 field dataset were used for model validation. The model simulation on biomass concentration showed that the polyculture growth model with k value had a root-mean-square error of 0.12, whereas model validation provided a better result with a root-mean-square error of 0.08. Biomass productivity forecast showed maximum productivity of 18.87 g/m2 /d in June with an annual average of 13.59 g/m2 /d. Biocrude oil yield forecast indicated that hydrothermal liquefaction process was more suitable with a maximum productivity of 0.59 g/m2 /d compared with solvent extraction which was only 0.19 g/m2 /d. With satisfactory root mean square errors less than 0.3, this polyculture growth model can be applied to forecast the productivity of native microalgae. This article is protected by copyright. All rights reserved.

Dewatering algal suspensions is an important step in the extraction of oil and other useful substances from algae. In this study, spherical Nannochloropsis sp. and ellipsoidal Monoraphidium sp. suspensions were dewatered in the presence of different amounts of 350-nm magnetite particles using a microfiltration membrane with 360-nm pores in cross-flow mode. Magnetite functions as a filter aid by reducing the deformation of the cake of filtered algae on the membrane and providing paths for water to flow through the filtration cake of algae. In the case of Nannochloropsis sp., the highest dewatering rate was obtained when the number ratio, defined based on the size and ideal density, between Nannochloropsis sp. and magnetite was 1:12.5, but the addition of magnetite had no observable effect on the filtration of ellipsoidal Monoraphidium sp. suspensions through the membrane. After dewatering, magnetite was effectively separated from the concentrated algal suspension by the application of a magnetic field in an open flow system. Magnetite has the potential to enhance dewatering performance using a cross-flow membrane system.

Coastal areas in the Tohoku region were severely damaged by the tsunami that followed the March 2011 earthquake. Consequently, many lands have become non-arable. To assess the use of these non-arable lands for the production of algal biomass, we carried out experimental cultivations of native algal communities in Minamisoma city, Fukushima Prefecture, located in the cool temperate zone of Japan. Productivity of the native algal community was measured using 1-m2 raceway ponds (120 L) and open vessel-type bioreactors (50 L–500 L), with different hydraulic retention times (HRTs) and in the presence or absence of sodium acetate. Maximum productivity in 1-m2 raceway ponds (10-cm water depth) was observed with an HRT of 4 days and sodium acetate supplementation in June 2015 (13.2 g/m2/day); average productivity throughout the year was 10.6 g/m2/day. In the case of deep-water cultivation (80-cm water depth) in vessel-type cultures, the highest productivity (> 29 g/m2/day) was obtained between January and September 2015. The dominant native microalgal species in the cultivations were Desmodesmus sp. and Scenedesmus acuminatus. Heterotrophic organisms, native bacteria, and protista, such as ciliates and amoeba, were also observed. Species composition and abundance varied depending on the medium content and seasonal conditions.

In this study, we report a two-dimensional (2D) hexagonal disk obtained by carbonization of Botryococcus braunii (B. braunii) residues. Carbonization at 700℃followed by naturally cooling down to room temperature under a non-inert gas flow atmosphere affords to yield this unique structure. The 2D hexagonal disks consist of more than 52% carbon and more than 25% oxygen. Slight amount of Fe, silicon and magnesium would be the trigger of the formation of hexagonal structure. Treatment of biomass residue is a challenge in the near future accompanied by the achievement of new energy technology in the industrial level. This research points out that efficient use of discharged biomass residue could create a new avenue for material science. The morphology of obtained crystals carbonized in different conditions, especially with the existence of argon flow, was also investigated.

The biosequestration of CO2 using microalgae has emerged as a promising means of recycling CO2 into biomass via photosynthesis, which could be used to produce biofuels as an attractive approach to CO2 mitigation. We investigated the CO2 fixation capability of the native nonaxenic microalgal culture using a 2 L photobioreactor operated in batch mode. The cultivation was carried out at varying concentrations of total dissolved CO2 (Tco2) in the bulk media ranging from 200 to 1000 mg L−1, and the temperature and light intensities were kept constant. A maximum CO2 fixation rate was observed at 400 mg L−1 of Tco2. Characteristic growth parameters such as biomass productivity, specific growth rate, maximum biomass yield, and biochemical parameters such as carbohydrate, protein, and lipids were determined and discussed. We observed that the effect of CO2 concentration on growth and biochemical composition was quite significant. The maximum biomass productivity was 22.10 ± 0.70 mg L−1 day−1, and the rate of CO2 fixation was 28.85 ± 3.00 mg L−1 day−1 at 400 mg L−1 of Tco2. The maximum carbohydrate (8.17 ± 0.49% dry cell weight) and protein (30.41 ± 0.65%) contents were observed at 400 mg L−1, whereas the lipid content (56.00 ± 0.82% dry cell weight) was the maximum at 800 mg L−1 of Tco2 in the bulk medium.

Cryopreservation is useful for long-term maintenance of living strains in microbial culture collections. We applied this technique to environmental specimens from two monitoring sites at Sendai Bay, Japan and compared the microbial diversity of photosynthetic picoeukaryotes in samples before and after cryopreservation. Flow cytometry (FCM) showed no considerable differences between specimens. We used 2500 cells sorted with FCM for next-generation sequencing of 18S rRNA gene amplicons and after removing low-quality sequences obtained 10,088-37,454 reads. Cluster analysis and comparative correlation analysis of observed high-level operational taxonomic units indicated similarity between specimens before and after cryopreservation. The effects of cryopreservation on cells were assessed with representative culture strains, including fragile cryptophyte cells. We confirmed the usefulness of cryopreservation for genetic studies on environmental specimens, and found that small changes in FCM cytograms after cryopreservation may affect biodiversity estimation.

This study investigates the potential of microalgae oil production as an alternative renewable energy source, in a pilot project located at Minamisoma City in the Fukushima Prefecture of Japan. The algal communities used in this research were the locally mixed species, which were mainly composed of Desmodesmus collected from the Minamisoma pilot project. The microalgae oil-production processes in Minamisoma consisted of three stages: cultivation, dewatering, and extraction. The estimated theoretical input-energy requirement for extracting oil was 137.25 MJ to process 50 m3 of microalgae, which was divided into cultivation 15.40 MJ, centrifuge 13.39 MJ, drum filter 14.17 MJ, and hydrothermal liquefaction (HTL) 94.29 MJ. The energy profit ratio (EPR) was 1.41. The total energy requirement was highest in the HTL process (68%) followed by cultivation (11%) and the drum filter (10%). The EPR value increased along with the yield in the cultivation process. Using HTL, the microalgae biomass could be converted to bio-crude oil to increase the oil yield in the extraction process. Therefore, in the long run, the HTL process could help lower production costs, due to the lack of chemical additions, for extracting oil in the downstream estimation of the energy requirements for microalgae oil production.

Microalgae is a promising next-generational energy. In this research, we focus on oil-extracted Botryococcus braunii residues collected by adding polysilicato-iron (PSI) as a flocculant followed by carbonization under argon atmosphere. We conducted carbonization at various temperatures as a first attempt to reveal the fundamental properties of the carbonization process of the microbes. The carbons thus obtained by heat treatment at 900 °C present a unique magnetic behavior due to reduced magnetite (Fe3O4) inclusion, which is produced from polysilicato iron (Fe2O3) during the heating process. Experimental results suggest that this carbonic material can be applied as a heavy metal-capturing carbon and magnetic porous substrate catalyst. The effective use of the waste may open a new avenue for an energy-microbiology-materials system.

In this study, the effects of an ethanolic extract of Aurantiochytrium mangrovei 18W-13a strain (AM18W-13a) on lipopolysaccharide (LPS)-induced inflammatory responses in RAW264 murine macrophages were studied. Pre-treatment with the AM18W-13a extract significantly suppressed the LPS-induced production of nitric oxide and pro-inflammatory cytokines. RAW264 cells treated with the AM18W-13a extract for 1 and 24 h were subjected to DNA microarray analyses for detecting the differentially expressed genes. The treatment of RAW264 cells with the AM18W-13a extract for 24 h significantly suppressed the expression of several genes associated with inflammation or chemotaxis. Furthermore, treatment with the AM18W-13a extract for 1 h suppressed the expression of Pde4b, but induced the expression of Egr2 and Egr3 in RAW264 cells. Additionally, the AM18W-13a extract significantly enhanced the expression of certain anti-inflammatory mediators. This study is the first report of the anti-inflammatory effects of the AM18W-13a extract and its mechanism of action in LPS-stimulated murine macrophages.

Algae have been recognized as important resources providing functional components due to their capacity to exert beneficial effects on health. Therefore, there is increasing interest in investigating the biological activity of algae. In this study, we evaluated the antidepressant-like effect of the administration of 100 mg/kg/day of the ethanol extract of colonial green alga Botryococcus braunii (EEB) for 14 consecutive days in the forced swimming test (FST)-induced depression in imprinting control region (ICR) mice. Imipramine, a commercial antidepressant drug, was used as a positive control. In addition, we investigated the molecular mechanisms underlying the effect of EEB by measuring ATP production and by assessing any change in gene expression at the end of the treatment using real-time polymerase chain reaction (PCR) and microarray assays. We showed that the immobility time in the water-administered control (FST stress) group gradually increased from day 1 to day 14. However, treatment with EEB caused a significant decrease of immobility time in the FST compared with that in the FST stress group. Microarray and real-time PCR results revealed that EEB treatment induced variation in the expression of several genes associated with neurogenesis, energy metabolism, and dopamine synthesis. Interestingly, we revealed that only EEB treatment enhanced the promotion of energy production, while treatment with imipramine was ineffective. Our study provides the first evidence that B. braunii enhances energy production, which may contribute to the modulation of neurogenesis and to the enhancement of dopaminergic function, in turn potentially underlying the antistress- and antidepressant-like effects that we observed.

Algae have been recognized as important resources providing functional components due to their capacity to exert beneficial effects on health. Therefore, there is increasing interest in investigating the biological activity of algae. In this study, we evaluated the antidepressant-like effect of the administration of 100 mg/kg/day of the ethanol extract of colonial green alga Botryococcus braunii (EEB) for 14 consecutive days in the forced swimming test (FST)-induced depression in imprinting control region (ICR) mice. Imipramine, a commercial antidepressant drug, was used as a positive control. In addition, we investigated the molecular mechanisms underlying the effect of EEB by measuring ATP production and by assessing any change in gene expression at the end of the treatment using real-time polymerase chain reaction (PCR) and microarray assays. We showed that the immobility time in the water-administered control (FST stress) group gradually increased from day 1 to day 14. However, treatment with EEB caused a significant decrease of immobility time in the FST compared with that in the FST stress group. Microarray and real-time PCR results revealed that EEB treatment induced variation in the expression of several genes associated with neurogenesis, energy metabolism, and dopamine synthesis. Interestingly, we revealed that only EEB treatment enhanced the promotion of energy production, while treatment with imipramine was ineffective. Our study provides the first evidence that B. braunii enhances energy production, which may contribute to the modulation of neurogenesis and to the enhancement of dopaminergic function, in turn potentially underlying the antistress- and antidepressant-like effects that we observed.

Summary Chattonella marina var. antiqua and C. marina var. marina (Raphidophyceae) are red tide-forming, harmful phytoplankton species. We investigated the genetic diversity and genetic relationship among the populations using microsatellite markers to identify putative sources of C. marina var. antiqua and C. marina var. marina in Japanese coastal populations. A positive correlation between genetic divergence and geographical distance (isolation by distance) was recognized for C. marina var. antiqua. The C. marina var. antiqua populations were established throughout a geological time scale, and genetic divergence had progressed in each population with gene flow depending on geographic distances. In contrast, isolation by distance was not observed for C. marina var. marina populations, and the genetic divergence among populations was extremely high. The Tokyo Bay population of C. marina var. marina, which was first recognized in 2008, had many private alleles but was related to the Kagoshima Bay population. The Tokyo Bay population may have been established by several invasions from the Kagoshima Bay population and other regions.

The relationship between hydrocarbons and the molecular phylogeny of Botryococcus braunii was investigated using 31 axenic strains isolated in Japan. The resulting 18S rRNA tree, which includes additional sequences from the gene bank, revealed the existence of three major pylogenetic clades (I, II, and III) and four subclades (II1, II2, III1, and III2) with high bootstrap values. By gas chromatography/mass spectrometry analysis, nine hydrocarbon species were detected and categorized as 4 types: the three known races A, B, and L and the tentatively named race S comprising epoxy-n-alkane and saturated n-alkane chains with carbon numbers 18 and 20, respectively. Strains of clade I were coincident with race A. Clade II contained race B, and clade III contained race L and its derivatives, and race S. Clade III was further categorized into subclade III1 containing the typical race L strains and subclade III2 containing race S with high bootstrap values. The phylogenetic relationship of Botryococcus strains appeared to be in considerable agreement with unique hydrocarbon synthesis pathways. Maximum divergence values of 18S rRNA indicated large speciation among the clades at almost the species level, suggesting the utility of biochemical properties, e.g., hydrocarbon types, in determining the taxonomy of Botryococcus .

Summary Red tides of Chattonella spp. have caused continuous damage to Japanese aquaculture, however, the life cycle of this organism remains incompletely understood. To further investigate this matter, we assessed genotypes at 14 microsatellite markers in three varieties of Chattonella marina, viz., C. marina var. antiqua, C. marina var. marina, and C. marina var. ovata, to establish whether Chattonella undergoes asexual diploidization or sexual reproduction. After genotyping 287 strains of C. marina, all but one of these strains was shown to be heterozygous for at least some loci, and thus, in the diploid state, suggesting that Chattonella strains undergo sexual reproduction. In addition, we performed single-cell amplification on ‘small cells’ that are derived from vegetative cells under dark and low-nutrient conditions. The results indicated the existence of two types of small cells. The ‘Small cell Type 1’ was found to be heterozygous, genotypically equivalent to the vegetative cells, and is therefore diploid. These small cells may change to resting cells (cysts) directly. The ‘Small cell Type 2’ was homozygous at all analyzed loci, suggesting that these small cells are haploid and may be derived by meiosis. As fusion between small cells has previously been observed, the ‘Small cell Type 2’ may be the gamete of Chattonella. We present a construct of the full life cycle of Chattonella marina based on our own and previous results.

Botryococcus braunii is a green colonial alga that produces large volumes of liquid hydrocarbon. Therefore, B. braunii is expected to be useful as an alternative fuel resource. Natural blooms of B. braunii have been recorded in several lakes and reservoirs. Elucidation of natural B. braunii blooming would provide important information for the development of an open-pond cultivation system. In this study, we periodically assessed the genetic diversity and colony density of B. braunii populations, along with several environmental parameters, in two Japanese reservoirs (provisionally called “N” and “S”) from December 2008 to December 2009. Reservoir N had low numbers of B. braunii colonies whereas Reservoir S was characterized by periodic density increases that occurred in December 2008, and in March, September, and December 2009. Population genetics analysis using specific environmental sequences (PGA-SES method) was conducted for B. braunii populations for the first time. Among the B. braunii -dominated samples of Reservoir S, high levels of genetic diversity were observed in December 2008 and March 2009, whereas the diversity levels in September and December 2009 were low. The results suggest that B. braunii periodicity can be categorized into a high genetic diversity type and a low genetic diversity type. The high genetic diversity type may be caused by simultaneous growth of many genotypes, whereas the low genetic diversity type seems to be explained by increases in the cell density of only a few adapted genotypes.

Metagenomics approaches have been developing rapidly in marine sciences. However, the application of these approaches to marine eukaryotes, and in particular to the smallest ones, is challenging because marine microbial communities are dominated by prokaryotes. One way to circumvent this problem is to separate eukaryotic cells using techniques such as single-cell pipetting or flow cytometry sorting. However, the number of cells that can be recovered by such techniques remains low and genetic material needs to be amplified before metagenomic sequencing can be undertaken. In this methodological study, we tested the application of whole-genome amplification (WGA) to photosynthetic eukaryotes. We performed various optimization steps both on a mixture of known microalgal strains and on natural photosynthetic eukaryote populations sorted by flow cytometry. rRNA genes were used as markers for assessing the efficiency of different protocols. Our data indicate that WGA is suitable for the amplification of photosynthetic eukaryote genomes, but that biases are induced, reducing the diversity of the initial population. Nonetheless, this approach appears to be suitable for obtaining metagenomics data on microbial eukaryotic communities.

This article presents an engineering study of an integrated system to produce bio-oil from microalgae biomass. The analysis is based on a pilot plant located at Minami-soma Fukushima, Japan, which further simulates 1 ha based-cultivation. Municipal wastewater and flue gases were utilized as nutrient sources for the microalgae culture of the proposed design. A flow sheet diagram of the integrated plant was synthesized by process engineering software to allow simulation of a continuous system. The design and sizing of the process equipment were performed to obtain a realistic estimation of possible production cost. The results demonstrated that nutrient savings was achieved by wastewater and CO2 utilization to the polyculture of native microalgae. Process simulation gave an estimated CO2 sequestration of 82.77 to 140.58 tons ha&minus, 1year&minus, 1 with 63 to 107 tons ha&minus, 1 of potential biomass production. The integrated process significantly improved the energy balance and economics of biofuel production and also the wastewater treatment plant (WWTP). The economic analysis confirmed that higher biomass production and technology improvement were required to achieve operational feasibility and profitability of the current microalgae-based bio-oil production.

Demura M., Noel M.-H., Kasai F., Watanabe M.M. and Kawachi M. 2009. Taxonomic revision of Chattonella antiqua, C. marina and C. ovata (Raphidophyceae) based on their morphological characteristics and genetic diversity. Phycologia 48: 518–535. DOI: 10.2216/08-98.1. Three raphidophyte species, Chattonella antiqua, C. marina and C. ovata, are red tide–forming harmful phytoplankton species, causing mass mortality at fish farms. These species were originally distinguished from each other solely based on their morphological characters. Recent genetic diversity analyses showed that the three species were extremely similar. It was therefore necessary to re-examine these three species to determine if they were independent single species. We compared 104 strains of the three species using morphological characters, nuclear ITS rDNA regions, the chloroplast rbcL gene, the mitochondrion COI gene and selected microsatellite regions. The morphological characters for the three species formed a continuum of varia...

We have developed 11 microsatellite markers that are specific to Chattonella antiqua, C. marina, and C. ovata, the red tide-forming harmful phytoplanktons. The 11 loci were amplified in the three species. The number of alleles per locus ranged from 5 to 16. The three species shared most microsatellite regions, although the genetic differences in specific loci were detected among them. These markers of the Chattonella species will be beneficial for biogeographical, detailed taxonomic, studies.

Botryococcus braunii Kützing, a green colonial microalga, occurs worldwide in both freshwater and brackish water environments. Despite considerable attention to B. braunii as a potential source of renewable fuel, many ecophysiological properties of this alga remain unknown. Here, we examined the desiccation and temperature tolerances of B. braunii using two newly isolated strains BOD-NG17 and BOD-GJ2. Both strains survived through 6- and 8-month desiccation treatments but not through a 12-month treatment. Interestingly, the desiccation-treated cells of B. braunii gained tolerance to extreme temperature shifts, i.e., high temperature (40 °C) and freezing (−20 °C). Both strains survived for at least 4 and 10 days at 40 and −20 °C, respectively, while the untreated cells barely survived at these temperatures. These traits would enable long-distance dispersal of B. braunii cells and may account for the worldwide distribution of this algal species. Extracellular substances such as polysaccharides and hydrocarbons seem to confer the desiccation tolerance.

In the cytochrome c oxidase subunit I (cox1) gene of four raphidophycean flagellates Chattonella antiqua, C. marina, C. ovata, and C. minima we found two group II introns described here as Chattonella cox1-i1 and Chattonella cox1-i2 encoding an open reading frame (ORF) comprised of three domains: reverse transcriptase (RT), RNA maturase (Ma) and zinc finger (H-N-H) endonuclease domains. The secondary structures show both Chattonella cox1-i1 and Chattonella cox1-i2 belong to group IIA1, albeit the former possesses a group IIB-like secondary structural character in the epsilon' region of arm I. Our phylogenetic analysis inferred from RT domain sequences of the intronic ORF, comparison of the insertion sites, and the secondary structures of the introns suggests that Chattonella cox1-i1 likely shares an evolutionary origin with the group II introns inserted in cox1 genes of five phylogenetically diverged eukaryotes. In contrast, Chattonella cox1-i2 was suggested to bear a close evolutionary affinity to the group II introns found in diatom cox1 genes. The RT domain-based phylogeny shows a tree topology in which Chattonella cox1-i2 is nested in the diatom sequences suggesting that a diatom-to-Chattonella intron transfer has taken place. Finally, we found no intron in cox1 genes from deeper-branching raphidophyceans. Based on parsimonious discussion, Chattonella cox1-i1 and Chattonella cox1-i2 have invaded into the cox1 gene of an ancestral Chattonella cell after diverging from C. subsalsa.

Ships carry ballast water for better stability and to control trim. However, the discharge of ballast water near ports is known to transport invasive species from one coastal area to another. The exchange of ballast water on the high seas is supposed to reduce such invasions of exotic species. In this study, we used denaturing gradient gel electrophoresis (DGGE) to analyze the composition of the bacterial community in ballast water before and after such a mid-ocean exchange, and we also measured total bacterial counts. Our findings confirmed that the ballast water was replaced by the mid-ocean exchange, as indicated by the marked change in the composition of the bacterial community. There was also a significant decrease in bacterial abundance after the mid-ocean exchange. Finally, our findings support the incubation hypothesis, because the composition of the bacterial communities changed over time within the same ballast water.