EuMoBot: replicating euglenoid movement in a soft robot.

Swimming is employed as a form of locomotion by many organisms in nature across a wide range of scales. Varied strategies of shape change are employed to achieve fluidic propulsion at different scales due to changes in hydrodynamics. In the case of microorganisms, the small mass, low Reynolds number and dominance of viscous forces in the medium, requires a change in shape that is non-invariant under time reversal to achieve movement. The Euglena family of unicellular flagellates evolved a characteristic type of locomotion called euglenoid movement to overcome this challenge, wherein the body undergoes a giant change in shape. It is believed that these large deformations enable the organism to move through viscous fluids and tiny spaces. The ability to drastically change the shape of the body is particularly attractive in robots designed to move through constrained spaces and cluttered environments such as through the human body for invasive medical procedures or through collapsed rubble in search of survivors. Inspired by the euglenoids, we present the design of EuMoBot, a multi-segment soft robot that replicates large body deformations to achieve locomotion. Two robots have been fabricated at different sizes operating with a constant internal volume, which exploit hyperelasticity of fluid-filled elastomeric chambers to replicate the motion of euglenoids. The smaller robot moves at a speed of [Formula: see text] body lengths per cycle (20 mm min-1 or 2.2 cycles min-1) while the larger one attains a speed of [Formula: see text] body lengths per cycle (4.5 mm min-1 or 0.4 cycles min-1). We show the potential for biomimetic soft robots employing shape change to both replicate biological motion and act as a tool for studying it. In addition, we present a quantitative method based on elliptic Fourier descriptors to characterize and compare the shape of the robot with that of its biological counterpart. Our results show a similarity in shape of 85% and indicate that this method can be applied to understand the evolution of shape in other nonlinear, dynamic soft robots where a model for the shape does not exist.

Feeding and grazing impact by the bloom-forming euglenophyte Eutreptiella eupharyngea on marine eubacteria and cyanobacteria.

The phototrophic euglenophyte Eutreptiella eupharyngea often causes blooms in the coastal waters of many countries, but its mode of nutrition has not been assessed. This species has previously been considered as exclusively auxotrophic. To explore whether E. eupharyngea is a mixotrophic species, the protoplasm of E. eupharyngea cells were examined using light, epifluorescence, and transmission electron microscopy after eubacteria, the cyanobacterium Synechococcus sp., and diverse algal species were provided as potential prey. Furthermore, the ingestion rates of E. eupharyngea KR on eubacteria or Synechococcus sp. as a function of prey concentration were measured. In addition, grazing by natural populations of euglenophytes on natural populations of eubacteria in Masan Bay was investigated. This study is the first to report that E. eupharyngea is a mixotrophic species. Among the potential prey organisms offered, E. eupharyngea fed only on eubacteria and Synechococcus sp., and the maximum ingestion rates of these two organisms measured in the laboratory were 5.7 and 0.7 cells predator-1 h-1, respectively. During the field experiments, the maximum ingestion rates and grazing impacts of euglenophytes, including E. eupharyngea, on natural populations of eubacteria were 11.8 cells predator-1 h-1 and 1.228 d-1, respectively. Therefore, euglenophytes could potentially have a considerable grazing impact on marine bacterial populations.

Response of Epiphytic Algae to Heavy Pollution of Water Bodies.

The objective of the present work was to study the structure of epiphyton in water bodies significantly differing in the degree of their contamination. In the studied water bodies, the concentration of ammonia () accounted for 0.04-74.00 mg/L; nitrite (), 0.002-1.750 mg/L; nitrate (), 0.13-58.00 mg/L; inorganic compounds of phosphorus (Pinorg), 0.041-0.272 mg/L; chloride (Cl-), 34.5-560.5 mg/L; whereas the content of organic matter (DO) was 18.0-81.0 mg O/L. The numbers of algae were determined by a direct count. Biomass was calculated by the count cell volume method. The Generic Diatom Index, the Pantle-Buck saprobic index, and the Shannon index of species diversity were used to analyze algal community structure. This study found that epiphyton responded to heavy pollution with changes in its species richness, taxonomic structure, species composition, quantitative indices, numbers and biomass structure, dominant complex, and ecological spectrum.

Structure and dynamics of the phytoplankton community within a maturation pond in a semiarid region.

In northeastern Brazil, stabilization ponds are very suitable for wastewater treatment because of the relative great land availability and environmental conditions (e.g., high temperature) favorable for microorganism optimal development. However, blooms of potentially toxic cyanobacteria may affect the use of these treatment ponds due to resulting effluent poor quality. The objective of this study was to evaluate the dynamics of phytoplankton communities and the occurrence of cyanobacteria in a maturation pond located immediately after a series of two ponds. Temperature, dissolved oxygen, pH, BOD, N, and P were measured during a period of four months when samples were collected from the surface and the bottom of 7 sampling points distributed inside the pond. The phytoplankton of collected samples was also identified and classified using a conventional optical microscopy. Analysis of variance and Tukey test were used to evaluate the results. The three phytoplankton divisions found (Cyanophyta, Chlorophyta, and Euglenophyta) did not change considerably through surface and bottom. However, they changed greatly over the sampled months; great dominance of Cyanophyta was found at April and October, while Chlorophyta dominated the lagoon in September. Low superficial organic loads (between 78 and 109 kg BOD.ha-1.d-1) and N:P ≤ 10 were the determinant factors that favored the predominance of Cyanophyta. The presence of two potentially toxic species of Cyanophyta, Oscillatoria sp. and Microcystis aeruginosa, indicates that caution is required when considering the final destination of treated effluent and suggests a need to assess the risks and benefits associated with the use of the treatment technology.

Structure and dynamics of the phytoplankton community within a maturation pond in a semiarid region / Estrutura e dinâmica da comunidade fitoplanctônica de uma lagoa de maturação em região semiárida

Abstract In northeastern Brazil, stabilization ponds are very suitable for wastewater treatment because of the relative great land availability and environmental conditions (e.g., high temperature) favorable for microorganism optimal development. However, blooms of potentially toxic cyanobacteria may affect the use of these treatment ponds due to resulting effluent poor quality. The objective of this study was to evaluate the dynamics of phytoplankton communities and the occurrence of cyanobacteria in a maturation pond located immediately after a series of two ponds. Temperature, dissolved oxygen, pH, BOD, N, and P were measured during a period of four months when samples were collected from the surface and the bottom of 7 sampling points distributed inside the pond. The phytoplankton of collected samples was also identified and classified using a conventional optical microscopy. Analysis of variance and Tukey test were used to evaluate the results. The three phytoplankton divisions found (Cyanophyta, Chlorophyta, and Euglenophyta) did not change considerably through surface and bottom. However, they changed greatly over the sampled months; great dominance of Cyanophyta was found at April and October, while Chlorophyta dominated the lagoon in September. Low superficial organic loads (between 78 and 109 kg BOD.ha–1.d–1) and N:P ≤ 10 were the determinant factors that favored the predominance of Cyanophyta. The presence of two potentially toxic species of Cyanophyta, Oscillatoria sp. and Microcystis aeruginosa, indicates that caution is required when considering the final destination of treated effluent and suggests a need to assess the risks and benefits associated with the use of the treatment technology.

Resumo No nordeste do Brasil, as lagoas de estabilização são muito adequadas para o tratamento de águas residuárias por causa da disponibilidade relativamente grande de terra e das condições ambientais (por exemplo, altas temperaturas) favoráveis ao melhor desenvolvimento dos microorganismos. Entretanto, florações de cianobactérias potencialmente tóxicas podem afetar o uso dessas lagoas de tratamento, devido à consequente qualidade inferior do efluente. O objetivo deste estudo foi avaliar a dinâmica das comunidades de fitoplâncton e a ocorrência de cianobactérias em uma lagoa de maturação situada após duas lagoas em série. Temperatura, oxigênio dissolvido, pH, DBO, N e P foram medidos durante um período de quatro meses, durante o qual amostras foram coletadas na superfície e fundo em sete pontos de amostragem da lagoa. As comunidades de fitoplâncton das amostras coletadas foram também identificadas e classificadas utilizando-se um microscópio óptico convencional. Para avaliar os resultados utilizou-se a análise de variância e o teste de Tukey. Para as três divisões de fitoplâncton encontradas (Cyanophyta, Chlorophyta e Euglenophyta), não houve diferença significativa para as amostras de superfície e de fundo de um mesmo mês. Entretanto, ocorreu grande variação para as amostras dos diferentes meses; nos meses de abril e outubro houve uma predominância de Cyanophyta, ao passo que em setembro o predomínio na lagoa foi de Chlorophyta. Os fatores determinantes que favoreceram o predomínio de Cyanophyta foram a baixa carga orgânica superficial aplicada (entre 78 e 109 kg DBO.ha–1.d–1) e N:P ≤ 10. A presença de duas das espécies de Cyanophyta, Oscillatoria sp. e Microcystisaeruginosa, consideradas potencialmente tóxicas, indica que é necessária precaução quando se considera o destino final do efluente tratado e sugere a necessidade de avaliar os riscos e benefícios associados ao uso da tecnologia de tratamento.

Prey or parasite? The first observations of live Euglenida in the intestine of Gastrotricha.

Euglenida are an important element in almost all aquatic ecosystems. They are known to parasitize animals such as copepods and flatworms, but have never been found in any other microscopic group. Gastrotrichs, a phylum of small microinvertebrates, are a constant and important element of marine and freshwater ecosystems. During our observation, 72 live gastrotrich specimens were collected from Europe (Poland, Germany) and South America (Brazil) containing active Euglenida of the genus Heteronema. Euglenida were found in the intestine of 10.4%, 52.6% and 1% of gastrotrichs, respectively. Taking into consideration the existing parasitism in euglenoids, it may be hypothesized that they either constitute fortuitous feed in the gastrotrich intestine or they are parasites or commensals. These observations suggest a new, so far unknown type of interaction between Gastrotricha and protists.

Ultrastructure and molecular phylogenetic position of Neometanema parovale sp. nov. (Neometanema gen. nov.), a Marine phagotrophic euglenid with skidding motility.

Heteronema is a commonly encountered genus of phagotrophic euglenids that contains very different morphotypes, including elongate gliding species and ovoid skidding forms. We report the first ultrastructural and sequence data from a culture of an ovoid skidding heteronemid, KM051. Cells were 8-23.5 µm long with 22 pellicular strips and a fibrous extracellular layer. The tubular extrusomes had dense centre sections. The feeding apparatus was barely visible by light microscopy, but included two microtubule-supported rods. The flagella had hollow, inflated transition zones, heteromorphic paraxonemal rods, and sheaths of flagellar hairs. The posterior flagellum bore a knob that, unusually, sat >2 µm distal to the flagellar base. No ultrastructural features were uniquely shared by KM051 and the elongate, gliding species Heteronema scaphurum. Conversely, the pellicular microtubule array resembles that in deep-branching primary osmotrophs (Aphagea). 18S ribosomal DNA (18S rDNA) phylogenies showed that KM051 is related to a recently obtained Heteronema c.f. exaratum sequence. These skidding heteronemids are not closely related to H. scaphurum, and instead are closely related to Dinema, Anisonema and specifically, Aphagea. The skidding species in Heteronema are transferred to Neometanema gen. nov. (along with most species of Metanema Klebs, 1893), with KM051 described as Neometanema parovale sp. nov.

The evolution of paralogous enzymes MAT and MATX within the Euglenida and beyond.

BACKGROUND: Methionine adenosyltransferase (MAT) is a ubiquitous essential enzyme that, in eukaryotes, occurs in two relatively divergent paralogues: MAT and MATX. MATX has a punctate distribution across the tree of eukaryotes and, except for a few cases, is mutually exclusive with MAT. This phylogenetic pattern could have arisen by either differential loss of old paralogues or the spread of one of these paralogues by horizontal gene transfer. Our aim was to map the distribution of MAT/MATX genes within the Euglenida in order to more comprehensively characterize the evolutionary history of MATX. RESULTS: We generated 26 new sequences from 23 different lineages of euglenids and one prasinophyte alga Pyramimonas parkeae. MATX was present only in photoautotrophic euglenids. The mixotroph Rapaza viridis and the prasinophyte alga Pyramimonas parkeae, which harbors chloroplasts that are most closely related to the chloroplasts in photoautotrophic euglenids, both possessed only the MAT paralogue. We found both the MAT and MATX paralogues in two photoautotrophic species (Phacus orbicularis and Monomorphina pyrum). The significant conflict between eukaryotic phylogenies inferred from MATX and SSU rDNA data represents strong evidence that MATX paralogues have undergone horizontal gene transfer across the tree of eukaryotes. CONCLUSIONS: Our results suggest that MATX entered the euglenid lineage in a single horizontal gene transfer event that took place after the secondary endosymbiotic origin of the euglenid chloroplast. The origin of the MATX paralogue is unclear, and it cannot be excluded that it arose by a gene duplication event before the most recent common ancestor of eukaryotes.

Amoeboid swimming: a generic self-propulsion of cells in fluids by means of membrane deformations.

Microorganisms, such as bacteria, algae, or spermatozoa, are able to propel themselves forward thanks to flagella or cilia activity. By contrast, other organisms employ pronounced changes of the membrane shape to achieve propulsion, a prototypical example being the Eutreptiella gymnastica. Cells of the immune system as well as dictyostelium amoebas, traditionally believed to crawl on a substratum, can also swim in a similar way. We develop a model for these organisms: the swimmer is mimicked by a closed incompressible membrane with force density distribution (with zero total force and torque). It is shown that fast propulsion can be achieved with adequate shape adaptations. This swimming is found to consist of an entangled pusher-puller state. The autopropulsion distance over one cycle is a universal linear function of a simple geometrical dimensionless quantity A/V(2/3) (V and A are the cell volume and its membrane area). This study captures the peculiar motion of Eutreptiella gymnastica with simple force distribution.

Ultrastructure and molecular phylogenetic position of Heteronema scaphurum: a eukaryovorous euglenid with a cytoproct.

Euglenids comprise a distinct clade of flagellates with diverse modes of nutrition, including phagotrophy, osmotrophy and phototrophy. Much of the previous research on euglenids has focused on phototrophic species because of their ecological abundance and significance as indicators for the health of aquatic ecosystems. Although largely understudied, phagotrophic species probably represent the majority of euglenid diversity. Phagotrophic euglenids tend to be either bacterivorous or eukaryovorous and use an elaborate feeding apparatus for capturing prey cells. We characterized the ultrastructure and molecular phylogenetic position of Heteronema scaphurum, a eukaryovorous euglenid collected in freshwater. This species was equipped with a distinct cytoproct through which waste products were eliminated in the form of faecal pellets; a cytoproct has not been reported in any other member of the Euglenida. Heteronema scaphurum also had a novel predatory mode of feeding. The euglenid ensnared and corralled several green algal prey cells (i.e. Chlamydomonas) with hook-like flagella covered in mucous before engulfing the bundle of prey cells whole. Molecular phylogenetic analyses inferred from small subunit rDNA sequences placed this species with other eukaryovorous euglenids, which was consistent with ultrastructural features associated with the feeding apparatus, flagellar apparatus, extrusomes, and pellicle.

Reverse engineering the euglenoid movement.

Euglenids exhibit an unconventional motility strategy amongst unicellular eukaryotes, consisting of large-amplitude highly concerted deformations of the entire body (euglenoid movement or metaboly). A plastic cell envelope called pellicle mediates these deformations. Unlike ciliary or flagellar motility, the biophysics of this mode is not well understood, including its efficiency and molecular machinery. We quantitatively examine video recordings of four euglenids executing such motions with statistical learning methods. This analysis reveals strokes of high uniformity in shape and pace. We then interpret the observations in the light of a theory for the pellicle kinematics, providing a precise understanding of the link between local actuation by pellicle shear and shape control. We systematically understand common observations, such as the helical conformations of the pellicle, and identify previously unnoticed features of metaboly. While two of our euglenids execute their stroke at constant body volume, the other two exhibit deviations of about 20% from their average volume, challenging current models of low Reynolds number locomotion. We find that the active pellicle shear deformations causing shape changes can reach 340%, and estimate the velocity of the molecular motors. Moreover, we find that metaboly accomplishes locomotion at hydrodynamic efficiencies comparable to those of ciliates and flagellates. Our results suggest new quantitative experiments, provide insight into the evolutionary history of euglenids, and suggest that the pellicle may serve as a model for engineered active surfaces with applications in microfluidics.

[Persistent properties of microorganisms inhabiting highly mineralized water bodies].

AIM: Study prevalence and intensity of persistent properties in bacteria inhabiting highly min eralized water bodies and determine their role in interaction with halophilous heterotrophic protozoa. MATERIALS AND METHODS: 300 bacteria strains and 3 cultures of heterotrophic protozo isolated from water bodies with mineralization of 2-350 g/l were studied. Antilysozyme (ALA) antihistone (AHA) activity of bacteria, protozoa lysozyme were evaluated by dish and photometric methods. Protozoa histones were evaluated cytochemically. Interaction of protozoa and Escherichia coli was evaluated by experimental co-cultivation. RESULTS: Presence of lysozyme an histones was shown in halophilous heterotrophic protozoa. Prevalence of ALA and AHA in bacteria was shown to increase as water body mineralization decreases. Intensity of E. colielimination from brine was determined to depend on the bacteria ALA level and phagocytic activity of protozoa. Participation of halotolerant protozoa in formation of heterogeneity of bacterial population by ALA was shown. CONCLUSION: In biocenoses of highly mineralized water bodies functioning of lysozyme-antilysozyme, histone-antihistone systems was shown. Bacteria with high persistent potential may impair sanitary parameters of highly mineralized water bodies, process of self-purification of which depends directly on phagocytic activity of protozoa.

Morphostasis in a novel eukaryote illuminates the evolutionary transition from phagotrophy to phototrophy: description of Rapaza viridis n. gen. et sp. (Euglenozoa, Euglenida).

BACKGROUND: Morphostasis of traits in different species is necessary for reconstructing the evolutionary history of complex characters. Studies that place these species into a molecular phylogenetic context test hypotheses about the transitional stages that link divergent character states. For instance, the transition from a phagotrophic mode of nutrition to a phototrophic lifestyle has occurred several times independently across the tree of eukaryotes; one of these events took place within the Euglenida, a large group of flagellates with diverse modes of nutrition. Phototrophic euglenids form a clade that is nested within lineages of phagotrophic euglenids and that originated through a secondary endosymbiosis with green algae. Although it is clear that phototrophic euglenids evolved from phagotrophic ancestors, the morphological disparity between species representing these different nutritional modes remains substantial. RESULTS: We cultivated a novel marine euglenid, Rapaza viridis n. gen. et sp. ("green grasper"), and a green alga, Tetraselmis sp., from the same environment. Cells of R. viridis were comprehensively characterized with light microscopy, SEM, TEM, and molecular phylogenetic analysis of small subunit rDNA sequences. Ultrastructural and behavioral observations demonstrated that this isolate habitually consumes a specific strain of Tetraselmis prey cells and possesses a functional chloroplast that is homologous with other phototrophic euglenids. A novel feeding apparatus consisting of a reduced rod of microtubules facilitated this first and only example of mixotrophy among euglenids. R. viridis also possessed a robust photoreception apparatus, two flagella of unequal length, euglenoid movement, and a pellicle consisting of 16 strips and one (square-shaped) whorl of posterior strip reduction. The molecular phylogenetic data demonstrated that R. viridis branches as the nearest sister lineage to phototrophic euglenids. CONCLUSIONS: The unusual combination of features in R. viridis combined with its molecular phylogenetic position completely conforms to the expected transitional stage that occurred during the early evolution of phototrophic euglenids from phagotrophic ancestors. The marine mixotrophic mode of nutrition, the preference for green algal prey cells, the structure of the feeding apparatus, and the organization of the pellicle are outstanding examples of morphostasis that clarify pivotal stages in the evolutionary history of this diverse group of microbial eukaryotes.

Eukaryote-to-eukaryote gene transfer gives rise to genome mosaicism in euglenids.

BACKGROUND: Euglenophytes are a group of photosynthetic flagellates possessing a plastid derived from a green algal endosymbiont, which was incorporated into an ancestral host cell via secondary endosymbiosis. However, the impact of endosymbiosis on the euglenophyte nuclear genome is not fully understood due to its complex nature as a 'hybrid' of a non-photosynthetic host cell and a secondary endosymbiont. RESULTS: We analyzed an EST dataset of the model euglenophyte Euglena gracilis using a gene mining program designed to detect laterally transferred genes. We found E. gracilis genes showing affinity not only with green algae, from which the secondary plastid in euglenophytes evolved, but also red algae and/or secondary algae containing red algal-derived plastids. Phylogenetic analyses of these 'red lineage' genes suggest that E. gracilis acquired at least 14 genes via eukaryote-to-eukaryote lateral gene transfer from algal sources other than the green algal endosymbiont that gave rise to its current plastid. We constructed an EST library of the aplastidic euglenid Peranema trichophorum, which is a eukaryovorous relative of euglenophytes, and also identified 'red lineage' genes in its genome. CONCLUSIONS: Our data show genome mosaicism in E. gracilis and P. trichophorum. One possible explanation for the presence of these genes in these organisms is that some or all of them were independently acquired by lateral gene transfer and contributed to the successful integration and functioning of the green algal endosymbiont as a secondary plastid. Alternative hypotheses include the presence of a phagocytosed alga as the single source of those genes, or a cryptic tertiary endosymbiont harboring secondary plastid of red algal origin, which the eukaryovorous ancestor of euglenophytes had acquired prior to the secondary endosymbiosis of a green alga.

Complex array of endobionts in Petalomonas sphagnophila, a large heterotrophic euglenid protist from Sphagnum-dominated peatlands.

Petalomonas sphagnophila is a poorly studied plastid-lacking euglenid flagellate living in Sphagnum-dominated peatlands. Here we present a broad-ranging microscopic, molecular and microspectrophotometric analysis of uncultured P. sphagnophila collected from four field locations in Nova Scotia, Canada. Consistent with its morphological characteristics, 18S ribosomal DNA (rDNA) phylogenies indicate that P. sphagnophila is specifically related to Petalomonas cantuscygni, the only other Petalomonas species sequenced to date. One of the peculiar characteristics of P. sphagnophila is the presence of several green-pigmented particles approximately 5 mum in diameter in its cytoplasm, which a previously published study suggested to be cyanobacterial endosymbionts. New data presented here, however, suggest that the green intracellular body may not be a cyanobacterium but rather an uncharacterized prokaryote yet to be identified by molecular sequencing. 16S rDNA library sequencing and fluorescence in situ hybridizations show that P. sphagnophila also harbors several other endobionts, including bacteria that represent five novel genus-level groups (one firmicute and four different proteobacteria). 16S rDNA phylogenies suggest that three of these endobionts are related to obligate intracellular bacteria such as Rickettsiales and Coxiella, while the others are related to the Daphnia pathogen Spirobacillus cienkowskii or belong to the Thermoactinomycetaceae. TEM, 16S rDNA library sequencing and a battery of PCR experiments show that the presence of the five P. sphagnophila endobionts varies markedly among the four geographic collections and even among individuals collected from the same location but at different time points. Our study adds significantly to the growing evidence for complex and dynamic protist-bacterial associations in nature.

Cascades of convergent evolution: the corresponding evolutionary histories of euglenozoans and dinoflagellates.

The majority of eukaryotic diversity is hidden in protists, yet our current knowledge of processes and structures in the eukaryotic cell is almost exclusively derived from multicellular organisms. The increasing sensitivity of molecular methods and growing interest in microeukaryotes has only recently demonstrated that many features so far considered to be universal for eukaryotes actually exist in strikingly different versions. In other words, during their long evolutionary histories, protists have solved general biological problems in many more ways than previously appreciated. Interestingly, some groups have broken more rules than others, and the Euglenozoa and the Alveolata stand out in this respect. A review of the numerous odd features in these 2 groups allows us to draw attention to the high level of convergent evolution in protists, which perhaps reflects the limits that certain features can be altered. Moreover, the appearance of one deviation in an ancestor can constrain the set of possible downstream deviations in its descendents, so features that might be independent functionally, can still be evolutionarily linked. What functional advantage may be conferred by the excessive complexity of euglenozoan and alveolate gene expression, organellar genome structure, and RNA editing and processing has been thoroughly debated, but we suggest these are more likely the products of constructive neutral evolution, and as such do not necessarily confer any selective advantage at all.

[Spatial structure of communities of heterotrophic flagellates from a sphagnum bog].

Spatial distribution pattern of heterotrophic flagellates within a macroscopically homogenous sphagnum parcel of a transitional bog in the southern taiga was studied. Under investigation was horizontal pattern at different scales (1 cm, 10 cm, 1 m, 10 m) and the vertical heterogeneity of the community in the sphagnum quagmire. 105 species and forms of heterotrophic flagellates were revealed. Predominating were euglenids, less abundant are kynetoplastids and cercomonads. The most numerous appeared to be Cryptomonas sp., Heteromita minima, Goniomonas truncata, Protaspis simplex, Bodo designis, B. saltans, Phyllomitus apiculatus, Paraphysomonas sp., Petalomonas minuta. More abundant species were characterized by less patchy distribution than less abundant. At a smaller scale, the community was formed by the species with different degree of patchiness while at larger scales, all the species possess nearly the same distribution pattern. The same number of samples of equal sizes revealed nearly the same species numbers independently of distances between the sample sites, as the samples at each scale differ from each other nearly at the same magnitude. An averaged size of the species aggregations in the community is as large as several centimeters. Such a scale is probably a characteristic size (minimum area) of the community of the sphagnum dwelling heterotrophic flagellates. Rather low environmental heterogeneity within the sphagnum quagmire leads to significant homogeneity of the community at larger scales. Vertical differentiation of the heterotrophic flagellate community within that quagmire appeared to be very unstable with the time. The same species are characterized by different preferences to the depths at different spatial-temporal loci. Specific vertical distributions and community patterns are formed under different local conditions.

The complete chloroplast genome of the chlorarachniophyte Bigelowiella natans: evidence for independent origins of chlorarachniophyte and euglenid secondary endosymbionts.

Chlorarachniophytes are amoeboflagellate cercozoans that acquired a plastid by secondary endosymbiosis. Chlorarachniophytes are the last major group of algae for which there is no completely sequenced plastid genome. Here we describe the 69.2-kbp chloroplast genome of the model chlorarachniophyte Bigelowiella natans. The genome is highly reduced in size compared with plastids of other photosynthetic algae and is closer in size to genomes of several nonphotosynthetic plastids. Unlike nonphotosynthetic plastids, however, the B. natans chloroplast genome has not sustained a massive loss of genes, and it retains nearly all of the functional photosynthesis-related genes represented in the genomes of other green algae. Instead, the genome is highly compacted and gene dense. The genes are organized with a strong strand bias, and several unusual rearrangements and inversions also characterize the genome; notably, an inversion in the small-subunit rRNA gene, a translocation of 3 genes in the major ribosomal protein operon, and the fragmentation of the cluster encoding the large photosystem proteins PsaA and PsaB. The chloroplast endosymbiont is known to be a green alga, but its evolutionary origin and relationship to other primary and secondary green plastids has been much debated. A recent hypothesis proposes that the endosymbionts of chlorarachniophytes and euglenids share a common origin (the Cabozoa hypothesis). We inferred phylogenies using individual and concatenated gene sequences for all genes in the genome. Concatenated gene phylogenies show a relationship between the B. natans plastid and the ulvophyte-trebouxiophyte-chlorophyte clade of green algae to the exclusion of Euglena. The B. natans plastid is thus not closely related to that of Euglena, which suggests that plastids originated independently in these 2 groups and the Cabozoa hypothesis is false.

Photoreceptor for curling behavior in Peranema trichophorum and evolution of eukaryotic rhodopsins.

When it is gliding, the unicellular euglenoid Peranema trichophorum uses activation of the photoreceptor rhodopsin to control the probability of its curling behavior. From the curled state, the cell takes off in a new direction. In a similar manner, archaea such as Halobacterium use light activation of bacterio- and sensory rhodopsins to control the probability of reversal of the rotation direction of flagella. Each reversal causes the cell to change its direction. In neither case does the cell track light, as known for the rhodopsin-dependent eukaryotic phototaxis of fungi, green algae, cryptomonads, dinoflagellates, and animal larvae. Rhodopsin was identified in Peranema by its native action spectrum (peak at 2.43 eV or 510 nm) and by the shifted spectrum (peak at 3.73 eV or 332 nm) upon replacement of the native chromophore with the retinal analog n-hexenal. The in vivo physiological activity of n-hexenal incorporated to become a chromophore also demonstrates that charge redistribution of a short asymmetric chromophore is sufficient for receptor activation and that the following isomerization step is probably not required when the rest of the native chromophore is missing. This property seems universal among the Euglenozoa, Plant, and Fungus kingdom rhodopsins. The rhodopsins of animals have yet to be studied in this respect. The photoresponse appears to be mediated by Ca2+ influx.