A simple and effective cryopreservation protocol for the industrially important and model organism, Euglena gracilis

Abstract In order to obtain a model which allows to directly study Mg2+ influx into the cell, Mg2+ deficiency was induced in the unicellular photoautotrophic flagellate Euglena gracilis. Lack of Mg2+ in the culture medium leads to a number of morphological, biochemical, and physio logical changes in E uglena gracilis. The rate of cell division was reduced under Mg2+-free conditions. Subsequently an enlargement of the cells was observed and they changed from spindle-like to oval shape. The Mg2+-starved cells were well filled with paramylon granules, while their motility and vitality was not impaired. Concurrently with the larger cell size the protein-, carbohydrate-, and chlorophyll content of the cells increased. Further changes were observed in the surface carbohydrates. The proportion of cells with galactose, N-acetyl-galac-tosamine and mannose on the cell surface rose in the Mg2+-starved cultures, shown in a lectin-binding assay. Fucose was found on the pellicle of Mg2+-starved cells only. Cultivation of E uglena gracilis in Mg2+-free medium induced a drastic reduction of the intracellular Mg2+ concentration already after 24 h (from 233 nmol/106 cells to 82 nmol/106 cells). When Mg2+ was made available again, the Mg2+-starved cells took them up rapidly and the intracellular concentration of free Mg2+ rose. As Mg2+ depletion could be induced in Euglena gracilis easily by manipulating the culture conditions and as the cells remained viable, it was con cluded that this flagellate can be used as a model organism for studying the Mg2+ uptake of eukaryotic cells.

Download Full-text

Isolation and characterization of a motility-defective mutant of Euglena gracilis

PeerJ ◽

10.7717/peerj.10002 ◽

2020 ◽

Vol 8 ◽

pp. e10002

Author(s):

Shuki Muramatsu ◽

Kohei Atsuji ◽

Koji Yamada ◽

Kazunari Ozasa ◽

Hideyuki Suzuki ◽

...

Keyword(s):

Mutant Strain ◽

Euglena Gracilis ◽

Model Organism ◽

Cost Effective ◽

Industrial Applications ◽

Culture Conditions ◽

Wild Type ◽

Mass Cultivation ◽

Isolation And Characterization ◽

Defective Mutant

Euglena gracilis is a green photosynthetic microalga that swims using its flagellum. This species has been used as a model organism for over half a century to study its metabolism and the mechanisms of its behavior. The development of mass-cultivation technology has led to E. gracilis application as a feedstock in various products such as foods. Therefore, breeding of E. gracilis has been attempted to improve the productivity of this feedstock for potential industrial applications. For this purpose, a characteristic that preserves the microalgal energy e.g., reduces motility, should be added to the cultivars. The objective of this study was to verify our hypothesis that E. gracilis locomotion-defective mutants are suitable for industrial applications because they save the energy required for locomotion. To test this hypothesis, we screened for E. gracilis mutants from Fe-ion-irradiated cell suspensions and established a mutant strain, ${\mathrm{M}}_{3}^{-}$ZFeL, which shows defects in flagellum formation and locomotion. The mutant strain exhibits a growth rate comparable to that of the wild type when cultured under autotrophic conditions, but had a slightly slower growth under heterotrophic conditions. It also stores 1.6 times the amount of paramylon, a crystal of β-1,3-glucan, under autotrophic culture conditions, and shows a faster sedimentation compared with that of the wild type, because of the deficiency in mobility and probably the high amount of paramylon accumulation. Such characteristics make E. gracilis mutant cells suitable for cost-effective mass cultivation and harvesting.

Download Full-text

Organization of the Pellicle and the Muciferous Glands in Euglena Gracilis

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100067510 ◽

1970 ◽

Vol 28 ◽

pp. 104-105

Author(s):

Hilton H. Mollenhauer ◽

W. Evans

Keyword(s):

Plasma Membrane ◽

Euglena Gracilis ◽

Complex Forms ◽

Secondary Periodicity

The pellicular structure of Euglena gracilis consists of a series of relatively rigid strips (Fig. 1) composed of ridges and grooves which are helically oriented along the cell and which fuse together into a common junction at either end of the cell. The strips are predominantly protein and consist in part of a series of fibers about 50 Å in diameter spaced about 85 Å apart and with a secondary periodicity of about 450 Å. Microtubules are also present below each strip (Fig. 1) and are often considered as part of the pellicular complex. In addition, there may be another fibrous component near the base of the pellicle which has not yet been very well defined.The pellicular complex lies underneath the plasma membrane and entirely within the cell (Fig. 1). Each strip of the complex forms an overlapping junction with the adjacent strip along one side of each groove (Fig. 1), in such a way that a certain amount of sideways movement is possible between one strip and the next.

Download Full-text

Immunocytochemical studies on the behavior of RuBisCO and LHCP II during the cell cycle of synchronized Euglena gracilis

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100160868 ◽

1990 ◽

Vol 48 (3) ◽

pp. 662-663

Author(s):

Tetsuaki Osafune ◽

Shuji Sumida ◽

Tomoko Ehara ◽

Eiji Hase ◽

Jerome A. Schiff

Keyword(s):

Cell Cycle ◽

Immunoelectron Microscopy ◽

Growth Phase ◽

Euglena Gracilis ◽

Dark Period ◽

Light Period ◽

Carboxylase Activity ◽

Immunocytochemical Studies ◽

Lhcp Ii

Changes in the morphology of pyrenoid and the distribution of RuBisCO in the chloroplast of Euglena gracilis were followed by immunoelectron microscopy during the cell cycle in a light (14 h)- dark (10 h) synchronized culture under photoautotrophic conditions. The imrnunoreactive proteins wereconcentrated in the pyrenoid, and less densely distributed in the stroma during the light period (growth phase, Fig. 1-2), but the pyrenoid disappeared during the dark period (division phase), and RuBisCO was dispersed throughout the stroma. Toward the end of the division phase, the pyrenoid began to form in the center of the stroma, and RuBisCO is again concentrated in that pyrenoid region. From a comparison of photosynthetic CO2-fixation with the total carboxylase activity of RuBisCO extracted from Euglena cells in the growth phase, it is suggested that the carboxylase in the pyrenoid functions in CO2-fixation in photosynthesis.

Download Full-text

Three-dimensional distribution of ribulose-1, 5-bisphosphate carboxylase/oxygemase during the early development of proplastids in dark-grown Euglena cells transferred to an inorganic medium

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100162090 ◽

1990 ◽

Vol 48 (3) ◽

pp. 906-907

Author(s):

Tomoko Ehara ◽

Shuji Sumida ◽

Tetsuaki Osafune ◽

Eiji Hase

Keyword(s):

Cell Suspension ◽

Euglena Gracilis ◽

Carbon Materials ◽

Three Dimensional ◽

Peripheral Region ◽

Plastid Development ◽

Bisphosphate Carboxylase ◽

Inorganic Medium ◽

Ribulose 1 ◽

Dimensional Distribution

As shown previously, Euglena cells grown in Hutner’s medium in the dark without agitation accumulate wax as well as paramylum, and contain proplastids showing no internal structure except for a single prothylakoid existing close to the envelope. When the cells are transferred to an inorganic medium containing ammonium salt and the cell suspension is aerated in the dark, the wax was oxidatively metabolized, providing carbon materials and energy 23 for some dark processes of plastid development. Under these conditions, pyrenoid-like structures (called “pro-pyrenoids”) are formed at the sites adjacent to the prolamel larbodies (PLB) localized in the peripheral region of the proplastid. The single prothylakoid becomes paired with a newly formed prothylakoid, and a part of the paired prothylakoids is extended, with foldings, in to the “propyrenoid”. In this study, we observed a concentration of RuBisCO in the “propyrenoid” of Euglena gracilis strain Z using immunoelectron microscopy.

Download Full-text

TK NIH funding for model organism databases

Nature ◽

10.1038/nature.2016.20133 ◽

2016 ◽

Keyword(s):

Model Organism ◽

Nih Funding ◽

Model Organism Databases

Download Full-text

Zebrafish as model organism for craniosynostosis

Bone Abstracts ◽

10.1530/boneabs.6.p019 ◽

2017 ◽

Author(s):

Rabea Bluemel ◽

Eva Klopocki ◽

Daniel Liedtke

Keyword(s):

Model Organism

Download Full-text

Effect of K2Cr2O7 on the photosynthetic activity and mobility of Euglena gracilis Klebs cells

International Journal on Algae ◽

10.1615/interjalgae.v9.i3.30 ◽

2007 ◽

Vol 9 (3) ◽

pp. 222-235

Author(s):

I. P. Novikova ◽

T. V. Parshikova ◽

V. V. Vlasenko ◽

I. B. Zubenko

Keyword(s):

Euglena Gracilis ◽

Photosynthetic Activity

Download Full-text

Genomic insights from Monoglobus pectinilyticus: a pectin-degrading specialist bacterium in the human colon

10.26686/wgtn.12585650.v1 ◽

2020 ◽

Author(s):

CC Kim ◽

GR Healey ◽

WJ Kelly ◽

ML Patchett ◽

Z Jordens ◽

...

Keyword(s):

Cell Surface ◽

Degradation Products ◽

Model Organism ◽

Human Colon ◽

Surface Proteins ◽

Human Gut ◽

Pectate Lyases ◽

Unusual Distribution ◽

Degrading Bacteria ◽

Acetyl Esterases

© 2019, International Society for Microbial Ecology. Pectin is abundant in modern day diets, as it comprises the middle lamellae and one-third of the dry carbohydrate weight of fruit and vegetable cell walls. Currently there is no specialized model organism for studying pectin fermentation in the human colon, as our collective understanding is informed by versatile glycan-degrading bacteria rather than by specialist pectin degraders. Here we show that the genome of Monoglobus pectinilyticus possesses a highly specialized glycobiome for pectin degradation, unique amongst Firmicutes known to be in the human gut. Its genome encodes a simple set of metabolic pathways relevant to pectin sugar utilization, and its predicted glycobiome comprises an unusual distribution of carbohydrate-active enzymes (CAZymes) with numerous extracellular methyl/acetyl esterases and pectate lyases. We predict the M. pectinilyticus degradative process is facilitated by cell-surface S-layer homology (SLH) domain-containing proteins, which proteomics analysis shows are differentially expressed in response to pectin. Some of these abundant cell surface proteins of M. pectinilyticus share unique modular organizations rarely observed in human gut bacteria, featuring pectin-specific CAZyme domains and the cell wall-anchoring SLH motifs. We observed M. pectinilyticus degrades various pectins, RG-I, and galactan to produce polysaccharide degradation products (PDPs) which are presumably shared with other inhabitants of the human gut microbiome (HGM). This strain occupies a new ecological niche for a primary degrader specialized in foraging a habitually consumed plant glycan, thereby enriching our understanding of the diverse community profile of the HGM.

Download Full-text

The Mechanisms Behind the Biological Activity of Flavonoids

Current Medicinal Chemistry ◽

10.2174/0929867325666180706104829 ◽

2019 ◽

Vol 26 (39) ◽

pp. 6976-6990 ◽

Cited By ~ 12

Author(s):

Ana María González-Paramás ◽

Begoña Ayuda-Durán ◽

Sofía Martínez ◽

Susana González-Manzano ◽

Celestino Santos-Buelga

Keyword(s):

Gene Expression ◽

Biological Activity ◽

Phenolic Compounds ◽

Intracellular Signaling ◽

Molecular Mechanisms ◽

Radical Scavenging ◽

Model Organism ◽

Stress Theory ◽

Radical Scavenging Properties

: Flavonoids are phenolic compounds widely distributed in the human diet. Their intake has been associated with a decreased risk of different diseases such as cancer, immune dysfunction or coronary heart disease. However, the knowledge about the mechanisms behind their in vivo activity is limited and still under discussion. For years, their bioactivity was associated with the direct antioxidant and radical scavenging properties of phenolic compounds, but nowadays this assumption is unlikely to explain their putative health effects, or at least to be the only explanation for them. New hypotheses about possible mechanisms have been postulated, including the influence of the interaction of polyphenols and gut microbiota and also the possibility that flavonoids or their metabolites could modify gene expression or act as potential modulators of intracellular signaling cascades. This paper reviews all these topics, from the classical view as antioxidants in the context of the Oxidative Stress theory to the most recent tendencies related with the modulation of redox signaling pathways, modification of gene expression or interactions with the intestinal microbiota. The use of C. elegans as a model organism for the study of the molecular mechanisms involved in biological activity of flavonoids is also discussed.

Download Full-text