Production of a thermal stress resistant mutant <i>Euglena gracilis</i> strain using Fe-ion beam irradiation

Yamada, Koji; Kazama, Yusuke; Mitra, Sharbanee; Marukawa, Yuka; Arashida, Ryo; Abe, Tomoko; Ishikawa, Takahiro; Suzuki, Kengo

doi:10.1080/09168451.2016.1171702

Cited by 15 publications

(22 citation statements)

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“…Reports suggest that E. gracilis exhibits polyploidy (Schiff and Epstein 1965;Hill et al 1966), which might be responsible for the observed difficulty in obtaining the nuclear genome mutants of E. gracilis (Schiff et al 1980). Although it is difficult to breed mutated E. gracilis, in the recent years, the use of heavy ion beams for inducing mutations has resulted in the appearance of mutants that exhibit previously unknown phenotypes such as tolerance to high temperature (Yamada et al 2016a) and higher lipid content (Yamada et al 2016b). Mutants generated by heavy ion irradiation are recognized as natural mutants.…”

Section: Breeding Potential Of Euglenamentioning

confidence: 99%

Large-Scale Cultivation of Euglena

Suzuki¹

2017

Advances in Experimental Medicine and Biology

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From the middle of the twentieth century, microalgae have been exploited as a candidate biomass source of food and other products. One such candidate source is the fast-proliferating microalga Euglena gracilis. The commercial cultivation of E. gracilis began in 2007, after the success of its outdoor mass cultivation and improvement of the harvesting and drying methods suitable for Euglena cells. The commercialization of Euglena production is based on the strategy of "5Fs of Biomass," which refers to the development and production of commercial products including food, fiber, feed, fertilizer, and fuel from biomass." Although room for improvement remains in the productivity of Euglena biomass, the product with the highest value-food-is already profitable. By enhancing the productivity of its biomass, other Euglena products, including fiber, feed, fertilizer, and fuel, can be commercialized. Breeding and recombinant DNA technology studies are being conducted to accomplish more extensive application of Euglena. In addition, the search for a better place for outdoor mass cultivation of Euglena is ongoing.

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Section: Breeding Potential Of Euglenamentioning

confidence: 99%

Large-Scale Cultivation of Euglena

Suzuki¹

2017

Advances in Experimental Medicine and Biology

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Section: Introductionmentioning

confidence: 99%

“…To improve the value of E. gracilis for industrial application, breeding methods for this species are established (Yamada et al, 2016a;Yamada et al, 2016b). Fe-ion beam irradiation efficiently induces random mutations and produces a population including various phenotypes of mutants (Yamada et al, 2016b). In addition, efficient selective breeding of oil-rich E. gracilis strain is demonstrated by isolating the desired cells from the mutant pool by using fluorescence-activated cell sorting (Yamada et al, 2016a).…”

Section: Introductionmentioning

confidence: 99%

Characterization of the eyespot and hematochrome-like granules of Euglena gracilis by scan-free absorbance spectral imaging A(x, y, λ) for quantification of carotenoids within the live cells

Yagi

Isono

Nishiyama

et al. 2018

Preprint

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Euglena gracilis is an edible photosynthetic single-cell alga that can synthesize carotenoids. It is highly demanded to establish the technology to select and grow individual cells capable of synthesizing more carotenoids because it contributes to safe and inexpensive production of carotenoids. In the cells of E. gracilis, carotenoids are mainly contained in chloroplasts and eyespots, and typical carotenoids have a characteristic absorption maximum in common. E. gracilis also has an organelle resembling hematochrome, which has an appearance similar to the eyespot and the absorption band spectrally overlapping that of the carotenoid although reportedly it does not contain carotenoids. To discriminate the eyespot and hematochrome-like granules and to investigate the intracellular distribution of carotenoids, scan-free, non-invasive, absorbance spectral imaging A(x, y, λ) microscopy of single live cells was applied. It was demonstrated that this technique is a powerful tool not only for basic research on intracellular structural analysis but also for identifying difference in carotenoid content in individual cells applicable to screening of carotenoid-rich cells. By this technique, it was confirmed that carotenoids exist in chloroplasts and eyespots, and a number of characteristic absorption spectra of pigments observed specific to the eyespot or hematochrome-like granules were identified. In addition, it was found that hematochromelike granules have a characteristic absorption peak at 620 nm as well as at 676 nm, suggesting that its origin is a component of chloroplast including Chlorophyll a.

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“…For the development of these technologies, noninvasive and safe cell viability assay 77 plays an important role as a basic technology. 78 For example, the production of nuclear mutants of E. gracilis requires a physically and 79 chemically significant load on cells due to the robustness of their genome, so it is necessary to 80 confirm whether the cells are viable or dead at screening (Yamada et al, 2016a). Moreover, 81 when the alga is mass cultured for edible use, it is necessary to confirm the state of growth.…”

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Peer Review #2 of "Noninvasive and safe cell viability assay for Euglena gracilis using natural food pigment (v0.1)"

2019

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Noninvasive and safe cell viability assay is required in many fields such as regenerative medicine, genetic engineering, single-cell analysis, and microbial food culture. In this case, a safe and inexpensive method which is a small load on cells and the environment is preferable without requiring expensive and space-consuming equipment and a technician to operate. We examined 8 typical natural food dyes to find Monascus (MP) or anthocyanin pigment (AP) works as a good viability indicator of dye exclusion test (DET) for Euglena gracilis which is an edible photosynthetic green microalga. This is the first report using natural food dyes as cell viability assay. Euglena gracilis stained by MP or AP can be visually judged with a bright field microscope. This was spectrally confirmed by scan-free, non-invasive absorbance spectral imaging A(x, y, λ) microscopy of single live cells and principal component analysis (PCA) . To confirm the ability of staining dead cells and examine the load on the cells, these two natural pigments were compared with trypan blue (TB) and methylene blue (MP), which are synthetic dyes conventionally used for DET. As a result, MP and AP had as good ability of staining dead cells treated with microwave as TB and MB and showed faster and more uniform staining for dead cells in benzalkonium chloride than them. The growth curve and the ratio of dead cells in the culture showed that the synthetic dyes inhibit the growth of E. gracilis, but the natural pigments do not. As the cell density increased, however, AP increased the ratio of stained cells, which was prevented by the addition of glucose. MP can stain dead cells in a shorter time than AP, while AP is more stable in color against long-term irradiation of intense light than MP. Due to the low toxicity of these pigments, viability of cells in culture can be monitored with them over a long period. PeerJ reviewing PDF | (Manuscript to be reviewed 38 39 40 Abstract 41Noninvasive and safe cell viability assay is required in many fields such as regenerative 42 medicine, genetic engineering, single-cell analysis, and microbial food culture. In this case, a 43 safe and inexpensive method which is a small load on cells and the environment is preferable 44 without requiring expensive and space-consuming equipment and a technician to operate. We 45 examined 8 typical natural food dyes to find Monascus (MP) or anthocyanin pigment (AP) works 46 as a good viability indicator of dye exclusion test (DET) for Euglena gracilis which is an edible 47 photosynthetic green microalga. This is the first report using natural food dyes as cell viability 48 assay. Euglena gracilis stained by MP or AP can be visually judged with a bright field 49 microscope. This was spectrally confirmed by scan-free, non-invasive absorbance spectral 50 imaging A(x, y, λ) microscopy of single live cells and principal component analysis (PCA). To 51 confirm the ability of staining dead cells and examine the load on the cells, these two natural 52 pigments were compared with trypan blue (TB)...

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Production of a thermal stress resistant mutant Euglena gracilis strain using Fe-ion beam irradiation

Cited by 15 publications

References 30 publications

Large-Scale Cultivation of Euglena

Large-Scale Cultivation of Euglena

Characterization of the eyespot and hematochrome-like granules of Euglena gracilis by scan-free absorbance spectral imaging A(x, y, λ) for quantification of carotenoids within the live cells

Peer Review #2 of "Noninvasive and safe cell viability assay for Euglena gracilis using natural food pigment (v0.1)"

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