Strain SAG Research Articles

Prasiolopsis wulf-kochii (Prasiolales, Trebouxiophyceae), a New Species Occurring in Hairs of the Sloth Bradypus tridactylus.

The monotypic genus Prasiolopsis has been known for a long time, but is often overlooked because of difficulties in identification and the morphological variability between uniseriate filaments and cell packages forming pseudoparenchymatic thalli depending on age. We investigated a strain (SAG 84.81) originally denoted as Trichophilus welckeri, which was isolated from the hairs of the sloth Bradypus tridactylus, and compared it with other available strains of Prasiolopsis and of the sister genus Pseudomarvania. Our investigations clearly showed that this strain differed in morphology, especially of the chloroplast, from those originally described for Trichophilus. Phylogenetic analyses of the SSU and ITS rDNA sequences revealed that the strain SAG 84.81 is sister to several strains of P. ramosa within the Prasiola clade (Trebouxiophyceae). Using the ITS-2/CBC approach, we clearly demonstrated that this strain represented a new species of Prasiolopsis, which we proposed here as P. wulf-kochii. In addition, we evaluated the ITS-2/CBC approach by comparing it with the two species of Pseudomarvania. All investigated strains showed CBCs and HCBCs, which support their species delimitation. The sequencing data of Trichophilus welckeri available in GenBank were phylogenetically re-evaluated by including all representatives of the Ulotrichales (Ulvophyceae). Our analyses showed that these sequences formed their own lineage within this order.

Solotvynia, a New Coccoid Lineage among the Ulvophyceae (Chlorophyta).

Coccoid Ulvophyceae are often overlooked despite their wide distribution. They occur as epiphytes on marine seaweeds or grow on stones or on shells of mussels and corals. Most of the species are not easy to identify based solely on morphology. However, they form two groups based on the flagellated cells during asexual reproduction. The biflagellated coccoids are monophyletic and represent the genus Sykidion (Sykidiales). In contrast, the quadriflagellated taxa are polyphyletic and belong to different genera and orders. The newly investigated strains NIES-1838 and NIES-1839, originally identified as Halochlorococcum, belong to the genus Chlorocystis (C. john-westii) among the order Chlorocystidales. The unidentified strain CCMP 1293 had almost an identical SSU and ITS-2 sequence to Symbiochlorum hainanense (Ignatiales) but showed morphological differences (single chloroplast, quadriflagellated zoospores) compared with the original description of this species (multiple chloroplasts, aplanospores). Surprisingly, the strain SAG 2662 (= ULVO-129), together with the published sequence of MBIC 10461, formed a new monophyletic lineage among the Ulvophyceae, which is highly supported in all of the bootstrap and Bayesian analyses and approximately unbiased tests of user-defined trees. This strain is characterized by a spherical morphology and also form quadriflagellated zoospores, have a unique ITS-2 barcode, and can tolerate a high variation of salinities. Considering our results, we emend the diagnosis of Symbiochlorum and propose the new genus Solotvynia among the new order Solotvyniales.

Complete mitochondrial genome of an oleaginous microalga Vischeria punctata (Eustigmatophyceae: Chlorobotryaceae) and phylogenetic analysis

Vischeria punctata, as first described by Vischer in 1945, is a member of the family Chlorobotryaceae, within the order Eustigmatales. This species is recognized for its potential as a source of biofuels and other high-value products. In the present investigation, the whole genome of V. punctata was sequenced utilizing the Illumina HiSeq 4000 platform, enabling the assembly and annotation of its complete mitochondrial genome. The resulting circular genome spans 41,528 base pairs (bp) with a guanine-cytosine (GC) content of 27.3%. This genome encompasses 36 protein-coding genes, alongside 28 transfer RNA (tRNA), and three ribosomal RNA (rRNA) genes. The evolutionary trajectory of V. punctata was further explored by constructing a phylogenetic tree derived from the mitochondrial 33 gene dataset of 16 Ochrophyta species. Comparative analysis reveals that V. punctata bears closer ties to Vischeria sp. CAUP Q202 than to Vischeria stellata strain SAG 33.83, suggesting shared evolutionary pathways and phenotypic traits. This investigation constitutes the inaugural study into the mitochondrial evolution and phylogenetic patterning of the mitogenome in V. punctata. The outcomes from this research bolster our understanding of the genetic diversity and evolutionary processes within the class Eustigmatophyceae. In particular, the mitochondrial genome of V. punctata serves as a valuable resource in elucidating these aspects.

Hydrocytium expands the phylogenetic, morphological, and genomic diversity of the poorly known green algal order Chaetopeltidales.

Chaetopeltidales is a small, understudied order of the green algal class Chlorophyceae, that is slowly expanding with the occasional discoveries of novel algae. Here we demonstrate that hitherto unrecognized chaetopeltidaleans also exist among previously described but neglected and misclassified species. Strain SAG 40.91 of Characium acuminatum, shown by previous preliminary evidence to have affinities with the orders Oedogoniales, Chaetophorales, and Chaetopeltidales (together constituting the OCC clade), was investigated with light and electron microscopy to characterize its morphology and ultrastructure. Sequence assemblies of the organellar and nuclear genomes were obtained and utilized in bioinformatic and phylogenetic analyses to address the phylogenetic position of the alga and its salient genomic features. The characterization of strain SAG 40.91 and a critical literature review led us to reinstate the forgotten genus Hydrocytium A.Braun 1855, with SAG 40.91 representing its type species, Hydrocytium acuminatum. Independent molecular markers converged on placing H. acuminatum as a deeply diverged lineage of the order Chaetopeltidales, formalized as the new family Hydrocytiaceae. Both chloroplast and mitochondrial genomes shared characteristics with other members of Chaetopeltidales and were bloated by repetitive sequences. Notably, the mitochondrial cox2a gene was transferred into the nuclear genome in the H. acuminatum lineage, independently of the same event in Volvocales. The nuclear genome data from H. acuminatum and from another chaetopeltidalean that was reported by others revealed endogenized viral sequences corresponding to novel members of the phylum Nucleocytoviricota. The resurrected genus Hydrocytium expands the known diversity of chaetopeltidalean algae and provides the first glimpse into their virosphere.

Comparing three different Phaeodactylum tricornutum strains for the production of chrysolaminarin in flat panel airlift photobioreactors

In recent years, various applications for algae-based ß-1,3-glucans have been postulated, including animal feed and human nutrition. Chrysolaminarin is a ß-1,3–1,6-glucan produced by diatoms such as Phaeodactylum tricornutum for energy storage. It is accumulated under nutrient-depleted cultivation conditions. In this study, the production of chrysolaminarin in artificially illuminated scalable flat panel airlift photobioreactors (FPA) was investigated by using P. tricornutum in a two-stage production process. In the growth stage primarily biomass is produced, and the subsequent nitrogen-depleted stage induces the accumulation of chrysolaminarin. Three P. tricornutum strains (SAG 1090-1a, SAG 1090-1b, SAG 1090–6) were cultured at laboratory scale in 6 L-FPA reactors under controlled light conditions to characterize the process and identify a production strain. The chrysolaminarin content of the algae strains was analysed and additionally their contents of eicosapentaenoic acid and fucoxanthin, both of which could be involved in a possible co-production. Strain SAG 1090-1b exhibited the highest biomass productivity and chrysolaminarin content (317 ± 9 mg gDW−1) after nitrogen depletion, and thus stood out as the most suitable for chrysolaminarin production in a two-stage process. A co-production of the three compounds is possible. However, during nitrogen depletion there occurred trade-offs between the compounds. As chrysolaminarin was produced, the amount of fucoxanthin in the culture stagnated or even decreased depending on the selected strain.

Removal of sugars in wastewater from food production through heterotrophic growth of Galdieria sulphuraria.

The unicellular extremophilic red alga Galdieria sulphuraria is capable of chemoheterotrophy and its growth has been investigated on some defined and undefined substrates. In this study, the removal of sugars in wastewater from fruit‐salad production with G. sulphuraria strain SAG 21.92 was analyzed. Growth and sugar consumption were determined under variation of temperature, pH‐value and concentration of a model substrate, containing sucrose, glucose and fructose. In shake flask cultivation maximum specific growth rate and specific substrate consumption rate of 1.53±0.09 day−1 and 2.41±0.14 gSub·gDW −1·day−1 were measured at pH 2 and 42°C. A scale‐up of this process was conducted in a 3 L stirred tank reactor (STR). Wastewater from fruit‐salad production was diluted to 15 g·L−1 total sugar concentration, supplemented with micronutrients and ammonia and pH was set to 3. Determined growth rate and substrate consumption were 1.21 day−1 and 1.88 gSub·gDW −1·day−1, respectively. It was demonstrated, that high sugar concentrations in wastewater streams from food production processes can be significantly reduced with G. sulphuraria SAG 21.92. This strain could achieve substrate consumption rates in wastewater, equal to the more common strain 074G, but at higher pH values. Generated biomass can be used for production of phycocyanin, a valuable nutraceutical.

Choricystis and Lewiniosphaera gen. nov. (Trebouxiophyceae Chlorophyta), two different green algal endosymbionts in freshwater sponges

Associations of freshwater sponges with coccoid green algae have been known for a long time. Two types of coccoid green algae, which are commonly assigned as zoochlorellae, are recognized by morphology: small coccoids (< 3 μm) without pyrenoids and larger Chlorella-like algae (4–6 μm) with pyrenoids. Despite their wide distribution in some freshwater sponges, these green algae were never studied using a combined analysis of morphology and molecular phylogeny. We investigated several endosymbiotic strains isolated from different Spongilla species, which were available in culture collections. Phylogenetic analyses of SSU and ITS rDNA sequences revealed that the strain SAG 211-40a is a member of the Chlorellaceae and represents a new species of the newly erected genus Lewiniosphaera, L symbiontica. The phylogenetic position was confirmed by morphology and ITS-2 barcode. The endosymbionts without pyrenoid were identified as Choricystis parasitica by morphology and phylogenetic analyses. The comparison with free-living strains revealed the recognition of two new Choricystis species, C. krienitzii and C. limnetica, which were confirmed by molecular signatures in V9 region of SSU rDNA and ITS-2 barcode.

The Draft Genome of Coelastrum proboscideum (Sphaeropleales, Chlorophyta)

Coelastrum proboscideum Bohlin, 1896 (Sphaeropleales, Scenedesmaceae, Chlorophyta) is a coenobial species with cosmopolitan distribution in diverse freshwater habitats. Coelastrum spp. are widely tested for biotechnological applications such as carotenoid and lipid production, and in bioremediation of wastewater. Here, we report the draft genome of C. proboscideum var. dilatatum strain SAG 217-2. The final assembly comprised 125,935,854 bp with over 8357 scaffolds. The whole-genome data is publicly available in the Nucleotide Sequence Archive (CNSA) of China National GeneBank (CNGB) (https://db.cngb.org/cnsa/) under the accession number CNA0014153.

Overlooked diversity with terrestrial lifestyle in the predominantly freshwater and snow phylogroup Chloromonadinia (Volvocales, Chlorophyceae)

ABSTRACTThe phylogroup Chloromonadinia (Volvocales, Chlorophyceae) encompasses ecologically versatile green microalgae with a predominantly freshwater and snow lifestyle. Although a few species have been described from soils, the evolutionary history of terrestrial volvocalean flagellates is poorly known due to undersampling. Here we provide new insights into the phylogenetic position of terrestrial flagellates within the Chloromonadinia and investigate six unresolved strains from terrestrial and freshwater habitats in both hemispheres. The terrestrial strains differed in cell morphology, ultrastructure and lifestyle from the freshwater ones, therefore, we expected them to represent distinct evolutionary lineages (species or genera). While the terrestrial strain SAG 25.87 (Chloromonas gracillima comb. & stat. nov.) from Czech mountains clustered within the psychrotolerant core Chloromonas clade, the other terrestrial strains formed a phylogenetically separate lineage which we proposed as Ostravamonas gen. nov. The new genus differs from other closely related Chloromonadinia genera by chloroplast, eyespot, papilla and cell-wall morphology. Two terrestrial strains, CAUP G 1401 (coal spoil heap; Czechia) and SAG 12.72 (acidic soil; New Zealand), were designated as Ostravamonas chlorostellata comb. nov. and represent the first Chloromonadinia microalgae discovered in acidic terrestrial environments. Closely related ‘Chlamydomonas’ strains SAG 75.81 (ditch; France) and CCAP 11/108 (origin unknown) were reassigned to Ostravamonas meslinii comb. nov. and Ostravamonas tenuiincisa sp. nov., respectively. Our study shows that the traditional ‘Chlamydomonas’ still hides an overlooked diversity of species with a terrestrial lifestyle that are nested phylogenetically within the Chloromonadinia phylogroup.

The Draft Genome of the Small, Spineless Green Alga Desmodesmus costato-granulatus (Sphaeropleales, Chlorophyta)

Desmodesmus costato-granulatus (Skuja) Hegewald 2000 (Sphaeropleales, Chlorophyta) is a small, spineless green alga that is abundant in the freshwater phytoplankton of oligo- to eutrophic waters worldwide. It has a high lipid content and is considered for sustainable production of diverse compounds, including biofuels. Here, we report the draft whole-genome shotgun sequencing of D. costato-granulatus strain SAG 18.81. The final assembly comprises 48,879,637bp with over 4,141 scaffolds. This whole-genome project is publicly available in the CNSA (https://db.cngb.org/cnsa/) of CNGBdb under the accession number CNP0000701.

The Draft Genome of Hariotina reticulata (Sphaeropleales, Chlorophyta) Provides Insight into the Evolution of Scenedesmaceae

Hariotina reticulata P. A. Dangeard 1889 (Sphaeropleales, Chlorophyta) is a common member of the summer phytoplankton of meso- to highly eutrophic water bodies with a worldwide distribution. Here, we report the draft whole-genome shotgun sequencing of H. reticulata strain SAG 8.81. The final assembly comprises 107,596,510bp with over 15,219 scaffolds (>100bp). This whole-genome project is publicly available in the CNSA (https://db.cngb.org/cnsa/) of CNGBdb under the accession number CNP0000705.

High density long-term cultivation of Chlorella vulgaris SAG 211-12 in a novel microgravity-capable membrane raceway photobioreactor for future bioregenerative life support in SPACE

Hybrid life support systems are of great interest for future far-distant space exploration missions to planetary surfaces, e.g. Mars, planned until 2050. By synergistically combining physicochemical and biotechnological algae-based subsystems, an essential step towards the closure of the carbon loop in environmental control and life support systems (ECLSS) shall be accomplished, offering a wide beneficial potential for ECLSS through the utilization of oxygenic photosynthesis: O2 and potential human food can be formed in-situ from CO2 and water. The wild type green alga Chlorella vulgaris strain SAG 211-12 was selected as model microorganism due to its photoautotrophic growth, high biomass yield, cultivation flexibility and long-term cultivation robustness. The current study presents for the first time a stable xenic long-term processing of microalgae in a novel microgravity capable membrane raceway photobioreactor for 188 days with the focus on algal growth kinetics and gas evolution. In particular, culture homogeneity and viability were monitored and evaluated during the whole cultivation process due to their putative crucial impact on long-term functionality and efficiency of a closed cultivation system. Based on a specially designed cyclic batch cultivation process for SAG 211-12, a successive biomass growth up to a maximum of 12.2 g l−1 with a max. global volumetric productivity of 1.3 g l−1 d−1 was reached within the closed loop system. The photosynthetic capacity was assessed to a global molar photosynthetic quotient of 0.31. Furthermore, cultivation parameters for a change from batch to continuous processing at high biomass densities and proliferation rates are introduced. The presented µgPBR miniature plant and the developed high throughput cultivation process are planned to be tested under real space conditions within the PBR@LSR project (microgravity and cosmic radiation) aboard the International Space Station with an operation period of up to 180 days to investigate the impact on long-term system stability.

Complete mitogenome of the streptophyte green alga Coleochaete scutata (Coleochaetophyceae)

We have sequenced the mitogenome of Coleochaete scutata strain SAG 110.80M. This mitogenome is the largest among the streptophyte green algae examined to date. At 242,024 bp, it is 4.3-fold larger than the mitogenome of Chaetosphaeridium globosum, the only other mitogenome available for the Coleochaetophyceae. This size difference is mainly explained by differences in the abundance of introns and in the length of intergenic regions containing vestiges of coding sequences thought to be of foreign origin. With 31 group I and 26 group II introns, the C. scutata mitogenome is the most intron-rich organelle genome known among streptophyte algae.

New record of the rare genus Crinalium Crow (Oscillatoriales, Cyanobacteria) from sand dunes of the Baltic Sea, Germany: epitypification and emendation of Crinalium magnum Fritsch et John based on an integrative approach

Representatives of the Gomontiellaceae (Oscillatoriales) are rare and hence unstudied cyanobacteria with unusual morphology, distributed in terrestrial and aquatic habitats all over the world. Investigation of the group based on an integrative approach is only beginning, and to understand the actual biodiversity and ecology, a greater number of cultivated strains is necessary. However, some ecological traits of these cyanobacteria (e.g. low population densities, the absence of conspicuous growth in nature) led to methodological difficulties during isolation in culture. One species in the family Gomontiellaceae, Crinalium magnum Fritsch et John, is characterized by prominent wide and flattened trichomes, and represented by the non-authentic strain SAG 34.87. Detailed previous investigation of this strain clearly showed its morphological discrepancy with the original description of C. magnum and the genus Crinalium in general. The new isolate from maritime sand dunes of the Baltic Sea coast (Germany), however, revealed morphological characters completely corresponding with the diagnosis of C. magnum. Phylogenetic analysis based on 16S rRNA sequences indicated a position of the new strain inside Gomontiellaceae. Both morphology and ultrastructure of the strain are congruous with characters of the family. Epitypification and emendation of C. magnum are proposed since the ecology and habitat of the original strain are congruent with the type locality of this rare species (sand, Irish Sea coast, North Wales, UK). We expanded the description of C. magnum by details of the filament development and specified dimensional ranges for trichomes and cells, as well as by new data about the transversely striated structure of mucilaginous sheath.

The complete mitochondrial genome of an oleaginous microalga vischeria stellata strain SAG 33.83 (Eustigmatophyceae)

The complete mitochondrial genome of an oleaginous microalga Vischeria stellata strain SAG 33.83 was sequenced and annotated. The circular genome was 42,608 bp long with 26.7% GC content. The mitochondrial genome of V. stellata strain SAG 33.83 contained 36 protein-coding genes (PCGs), 27 transfer RNA genes (tRNAs), three ribosomal RNA genes (rRNAs). Furthermore, the phylogenetic tree was constructed to validate the evolutionary relationship based on the complete mitogenomes of V. stellata strain SAG 33.83 through combining with seven species of Eustigmatophyceae, two species of Phaeophyceae and one species of Synurophyceae algae. Phylogenetic analysis revealed that Eustigmatophyceae were more closely related to Synurophyceae than Phaeophyceae and V. stellata strain SAG 33.83 had close genetic relationship with Vischeria sp. CAUP Q 202.

Transient genetic transformation of Mougeotia scalaris (Zygnematophyceae) mediated by the endogenous α-tubulin1 promoter.

Mougeotia scalaris is a filamentous streptophyte alga renowned for light-inducible plastid rotation and microtubule-dependent polarity establishment. As a first step toward transgenic approaches we determined the 5,825 base pair genomic sequence encoding the α-tubulin1 gene (MsTUA1) of M.scalaris (strain SAG 164.80). The subcloned MsTUA1 promoter facilitated strong transgene expression in M.scalaris and tobacco leaf cells, as shown by particle bombardment and the subsequent visualization of expressed fluorescent protein markers. Our results provide a route for the genetic transformation of the filamentous streptophyte alga M.scalaris based on the endogenous TUA1 promoter.

Microglena redcarensis sp. nov. (Volvocales, Chlorophyceae), a brackish water chlamydomonad with contractile vacuoles

SUMMARYIn 1960, Butcher isolated a strain of unicellular green alga, ‘Chlamydomonas subehrenbergii’ SAG 18.89, from a brackish water locality in England, facing the North Sea. Although C. subehrenbergii Butcher was previously described by the same author in 1959, the strain SAG 18.89 is in fact morphologically different, possessing shorter flagella, a different‐shaped papilla, and different‐sized vegetative cells. Light and fluorescence microscopy, molecular phylogenetic analyses, and ITS2 comparisons revealed that this strain represents an undescribed species of the genus Microglena (Volvocales, Chlorophyceae), described here as M. redcarensis sp. nov. The presence of contractile vacuoles, which are osmoregulatory organelles, suggests potential inhabitation in freshwater. Contractile vacuole activity and growth of M. redcarensis were therefore examined in media of different salinity.

Effects of Daphnia exudates and sodium octyl sulphates on filament morphology and cell wall thickness of Aphanizomenon gracile (Nostocales), Cylindrospermopsis raciborskii (Nostocales) and Planktothrix agardhii (Oscillatoriales)

ABSTRACTGrazing is recognized as one of the selective factors shaping the morphology and physiology of cyanobacteria. A recent study has shown that the filamentous cyanobacterium Aphanizomenon gracile strain SAG 31.79 thickened in the presence of Daphnia (Cladocera) and its exudates. The aims of our study were: (1) to determine whether this type of response to Daphnia cues is common for other strains of A. gracile, and other species of filamentous cyanobacteria, (2) to test whether the response is due to nutrients recycled by Daphnia, or kairomone induced, and (3) whether it is related to toxin production. Prior to the experiment, cyanobacterial strains were inspected using chromatographic methods for the presence of two toxins, cylindrospermopsin (CYN) and three homologues of microcystin (MC-RR, MC-YR, MC-LR). HPLC analyses showed that all strains were free of cylindrospermopsin, whereas microcystins were detected only in one strain (Planktothrix agardhii). We then tested whether Daphnia exudates can cause thickening of cyanobacterial filaments, which would suggest the morphological changes in cyanobacterial filaments are caused by recycled nutrients. Cyanobacteria were also exposed to sodium octyl sulphate (a commercially available Daphnia kairomone). Transmission electron microscopy (TEM) was used to check whether Daphnia exudates and sodium octyl sulphate trigger thickening of cyanobacterial cell walls, which would be a defence mechanism against grazing. The TEM analysis revealed no significant effect of either Daphnia exudates or kairomone (sodium octyl sulphate) on the cell wall thickness of cyanobacteria. However, our study showed that Daphnia exudates triggered filament thickening in nostocalean cyanobacteria, while filaments of the oscillatorialean strain P. agardhii did not show this response. It was also demonstrated that sodium octyl sulphate alone can also cause filament thickening, which suggests that this might be a specific defence response to the presence of grazers.

Cytological study of Сhloromonas typhlos (Chlamydomonadaceae, Chlorophyta) from North-Western Russia

The strain of biflagellated green monad, selected from the specimen gathered in the Novgorod Region was studied by light and electron microscopy. The species was identified on the base of coincidence of the majority of specific light and ultrastructural signs revealed in both the strain SAG 26.86 of Chloromonas typhlos (Gerloff) Matsuzaki et al. and in studied strain. On LM level the cells are ellipsoid, some asymmetric, 12–17 μm long, 8–13 μm wide, with an apical, hardly distinguished hemispherical papilla flattened on the top. Chloroplast parietal, cup-shaped, thickened on one side, with big lobes, irregular incisions and fissures on the surface. These and other features are characteristic of the type and the studied strain of C. typhlos, excluding stigma observed in the studied strain only. On the ultrastructural level, large thylakoid packs, similar with chloroplast thylakoids, were observed in pyrenoids of both strains of C. typhlos. The detected pyrenoid type was never discovered in chlamydomonads earlier. It is the first record of C. typhlos for the Novgorod Region and the whole Russia, and adds the list of «bloom-forming» species. The strain was deposited in the collection of microalgae strains in the Institute of Biology of Komi Scientific Centre under the name SYKOA Ch-063-17.

Differences in cell wall of thin and thick filaments of cyanobacterium Aphanizomenon gracile SAG 31.79 and their implications for different resistance to Daphnia grazing

&lt;p&gt;Recent studies have shown that the filamentous cyanobacterium &lt;em&gt;Aphanizomenon gracile&lt;/em&gt; Lemmermann, strain SAG 31.79, consists of two types of filaments that differ in thickness. These two types are known to vary in resistance to &lt;em&gt;Daphnia&lt;/em&gt; &lt;em&gt;magna&lt;/em&gt; grazing: thin filaments (&amp;lt;2.5 µm) are more vulnerable to grazing than the thick ones (&amp;gt;2.5 µm). In this study, we investigated whether the difference in the vulnerability to grazing of thin and thick filaments is a result of different thickness of their cell walls, a filament stiffness determinant. We expected thick filaments to have thicker cell walls than the thin ones. Additionally, we analysed whether cell wall thickness correlates with filament thickness regardless of the filament type. A morphometric analysis of cell walls was performed using transmission electron micrographs of ultra-thin sections of the batch-cultured cyanobacterial material. Our study revealed that the thin type of filaments had thinner cell walls than the thick filaments. Moreover, cell wall thickness was positively correlated with filament thickness. TEM (transmission electron microscopy) observations also revealed that the thin type of filaments was often at different stages of autocatalytic cell destruction, which was mainly manifested in the increase in cell vacuolization and degradation of the cytoplasm content. Based on our findings, we assume that previously reported higher resistance of thick filaments to &lt;em&gt;Daphnia&lt;/em&gt; grazing results from greater stiffness and excellent physiological conditions of thick filaments. &lt;/p&gt;