• Molecules and Cells
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• v.43 no.1
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• pp.48-57
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• 2020

The microalga Chlamydomonas reinhardtii accumulates triacylglycerols (TAGs) in lipid droplets under stress conditions, such as nitrogen starvation. TAG biosynthesis occurs mainly at the endoplasmic reticulum (ER) and requires fatty acid (FA) substrates supplied from chloroplasts. How FAs are transferred from chloroplast to ER in microalgae was unknown. We previously reported that an Arabidopsis thaliana ATP-binding cassette (ABC) transporter, AtABCA9, facilitates FA transport at the ER during seed development. Here we identified a gene homologous to AtABCA9 in the C. reinhardtii genome, which we named CrABCA2. Under nitrogen deprivation conditions, CrABCA2 expression was upregulated, and the CrABCA2 protein level also increased. CrABCA2 knockdown lines accumulated less TAGs and CrABCA2 overexpression lines accumulated more TAGs than their untransformed parental lines. Transmission electron microscopy showed that CrABCA2 was localized in swollen ER. These results suggest that CrABCA2 transports substrates for TAG biosynthesis to the ER during nitrogen starvation. Our study provides a potential tool for increasing lipid production in microalgae.

• The Plant Pathology Journal
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• v.37 no.5
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• pp.437-445
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• 2021

Tilletia laevis Kuhn (syn. Tilletia foetida (Wallr.) Liro.) causes wheat common bunt, which is one of the most devastating plant diseases in the world. Common bunt can result in a reduction of 80% or even a total loss of wheat production. In this study, the characteristics of T. laevis infection in compatible wheat plants were defined based on the combination of scanning electron microscopy, transmission electron microscopy and laser scanning confocal microscopy. We found T. laevis could lead to the abnormal growth of wheat tissues and cells, such as leakage of chloroplasts, deformities, disordered arrangements of mesophyll cells and also thickening of the cell wall of mesophyll cells in leaf tissue. What's more, T. laevis teliospores were found in the roots, stems, flag leaves, and glumes of infected wheat plants instead of just in the ovaries, as previously reported. The abnormal characteristics caused by T. laevis may be used for early detection of this pathogen instead of molecular markers in addition to providing theoretical insights into T. laevis and wheat interactions for breeding of common bunt resistance.

• ALGAE
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• v.38 no.4
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• pp.241-252
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• 2023

The genera Carolibrandtia and Chlorella have been described as small green algae with spherical cell shapes that inhabit various environments. Species of these genera are often difficult to identify because of their simple morphology and high phenotypic plasticity. We investigated two small coccoid strains from Antarctica based on morphology, molecular phylogeny by two alignment methods which have been applied to previous phylogenetic studies of the genus Chlorella, and comparison of the secondary structures of nuclear small subunit (SSU) and internal transcribed spacer (ITS) rDNA sequences. Light microscopy of two strains revealed spherical cells containing chloroplasts with pyrenoids, and the morphological characteristics of the strains were nearly identical to those of other Chlorella species. However, based on the phylogenetic analyses of nuclear SSU and ITS rDNA sequences, it was determined that the Antarctic microalgal strains belonged to two genera, as the Chlorella and Carolibrandtia. In addition, the secondary structures of the SSU and ITS2 sequences were analyzed to detect compensatory base changes (CBCs) that were used to identify and describe the two strains. A unique CBC in the SSU rDNA gene was decisive for distinguishing strain CCAP 211/45. The ITS2 rDNA sequences for each strain were compared to those obtained previously from other closely related species. Following the comparison of morphological and molecular characteristics, we propose KSF0092 as a new species, Chlorella terrestris sp. nov., and the reassignment of the strain Chlorella antarctica CCAP 211/45 into Carolibrandtia antarctica comb. nov.

• Journal of Plant Biotechnology
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• v.33 no.1
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• pp.1-9
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• 2006

Peroxiredoxins (Prxs) are ubiquitously distributed and play important functions in diverse cellular signaling systems. The proteins are largely classified into three groups, such as typical 2-Cys Prx, atypical 2-Cys Prx, and 1-Cys Prx, that are distinguished by their catalytic mechanisms and number of Cys residues. From the three classes of Prxs, the typical 2-Cys Prx containing the two-conserved Cys residues at its N-terminus and C-terminus catalyzes $H_2O_2$ with the use of thioredoxin (Trx) as an electron donor. During the catalytic cycle, the N-terminal Cys residue undergoes a peroxide-dependent oxidation to sulfenic acid, which can be further oxidized to sulfinic acid at the presence of high concentrations of $H_2O_2$ and a Trx system containing Trx, Trx reductase, and NADPH. The sulfinic acid form of 2-Cys Prx is reduced by the action of sulfiredoxin which requires ATP as an energy source. Under the strong oxidative or heat shock stress conditions, 2-Cys Prx in eukaryotes rapidly switches its protein structure from low-molecular-weight species to high-molecular-weight protein structures. In accordance with its structural changes, the protein concomitantly triggers functional switching from a peroxidase to a molecular chaperone, which can protect its substrate denaturation from external stress. In addition to its N-terminal active site, the C-terminal domain including 'YF-motif' of 2-Cys Prx plays a critical role in the structural changes. Therefore, the C-terminal truncated 2-Cys Prxs are not able to regulate their protein structures and highly resistant to $H_2O_2$-dependent hyperoxidation, suggesting that the reaction is guided by the peroxidatic Cys residue. Based on the results, it may be concluded that the peroxidatic Cys of 2-Cys Prx acts as an '$H_2O_2$-sensor' in the cells. The oxidative stress-dependent regulation of 2-Cys Prx provides a means of defense systems in cells to adapt stress conditions by activating intracellular defense signaling pathways. Particularly, 2-Cys Prxs in plants are localized in chloroplasts with a dynamic protein structure. The protein undergoes conformational changes again oxidative stress. Depending on a redox-potential of the chloroplasts, the plant 2-Cys Prx forms super-molecular weight protein structures, which attach to the thylakoid membranes in a reversible manner.

• Horticultural Science & Technology
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• v.19 no.4
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• pp.501-504
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• 2001

The effect of soil EC on tissue morphology of leaf and shoot tip in cucumber (Cucumis sativus L. cv. Euinchim-baekdadagi) seedlings was investigated. Number of trichomes on leaf upper epidermis increased with the increase in soil EC from 1.0 to $3.0dS{\cdot}m^{-1}$, but the shape and number of stomata on lower epidermis remained unchanged. Epidermal cells of cucumbers grown in EC $1.5dS{\cdot}m^{-1}$ soil was occupied mostly by large vacuole whereas those grown in EC $3.0dS{\cdot}m^{-1}$ soil were filled with a nucleus, mitochondria, chloroplast and other micro-organelles. Sponge parenchima cells were also larger and contained fewer chloroplasts at EC $1.5dS{\cdot}m^{-1}$ than those grown at EC $3.0dS{\cdot}m^{-1}$. Leaf thickness decreased at high EC and the color of epidermal cells became significantly darker on the photograph of optical microscope. Normal tissue differentiation was greatly suppressed in plants grown in soils with $3.5dS{\cdot}m^{-1}$ or higher EC.

• Reproductive and Developmental Biology
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• v.41 no.4
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• pp.65-70
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• 2017

Alpha-linolenic acid (ALA) is one of n-3 polyunsaturated fatty acids and found mainly in the chloroplasts. Many studies have been reported that intracellular reactive oxygen species (ROS) in mammalian oocytes were reduced by supplementation of ALA in in vitro maturation (IVM) medium. Based on these reports, we expected that ALA acts as an antioxidant during IVM of porcine oocytes. Therefore, the objective of this study was to investigate the antioxidant effect of ALA supplementation during IVM in porcine oocytes. The cumulus-oocyte complexes (COCs) were incubated in IVM medium containing $200{\mu}m$ $H_2O_2$ or $H_2O_2$ with $50{\mu}m$ ALA for 44 h. Nuclear maturation stage of oocytes was evaluated using aceto-orcein method. For measurement of oxidative stress state, intracellular ROS and glutathione (GSH) levels were measured using carboxy-DCFDA and cell tracker red, respectively. In results, oocytes in metaphase-II (MII) stage development was significantly reduced in $H_2O_2$ group compared to non-treated control group $61.84{\pm}1.42%$ and 80.00%, respectively; p<0.05) and it was slightly recovered by treatment of ALA ($69.76{\pm}1.67%$; p<0.05). The intracellular GSH levels was decreased in $H_2O_2$ groups compared with control groups, but it was enhanced by ALA treatment (p<0.05). On the contrary, $H_2O_2$ treatment increased intracellular ROS level in oocytes and $H_2O_2$-induced ROS was decreased by treatment of ALA (p<0.05). Our findings suggested that ALA treatment under oxidative stress condition improve oocyte maturation via elevated GSH and reduced ROS levels in oocytes. Therefore, these results suggest that ALA have an antioxidative ability and it could be used as antioxidant in in vitro production system of porcine embryo.

• The Korean Journal of Ecology
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• v.4 no.3_4
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• pp.114-123
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• 1981

Diurnal acid fluctuations, stomatal resistance, and morphology and anatomy were investigated in leaves and stems P. oleracea L. growing under the natural environmental condition. A CAM-like pattern of acid fluctuation was exhibited not in leaves of the young purslane but in its stems. Defoliated stems showed a decreased in diurnal changes of total acidity as compared with normal stems. Excised stems stored in continuous darkness exhibited diurnal acid rhythms, and they showed light deacidifications for three days. Kranz-type arrangement was observed in leaves, but not in stems. Micrography of cross sections of stems showed cells with relatively large vaculoles and a few chloroplasts. The number of stomata was 3,275cm-2 in leaves, while the stomata could not be observed in stems. Stomatal resistance was high at night and low in daytime in leaves of the young purslane, and the range of its value was 5~40 sec.$\textrm{cm}^{-1}$. But stomatal resistance in leaves of the water-stressed plant was comparratively high in day time, and its value was 30 sec.$\textrm{cm}^{-1}$. The result of these studies showed the possibility that the stem of P.oleracea L. possesses CAM under certain stressed conditions.

• ALGAE
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• v.35 no.3
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• pp.253-262
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• 2020

Haematococcus pluvialis is a commercial microalga that can produce high quantities of astaxanthin. Under induced conditions, some important changes in the subcellular structures related to astaxanthin accumulation were observable. For example, a large number of astaxanthin granules, oil structures and starch granules appeared in the thick-walled cells; Astaxanthin granules gradually dissolved into the oil structures and spread throughout the entire cell with the fusion and diffusion process of oil structures during the middle and late stages of induction; The plastoglobules were closed to the newly formed structures, and some plastoglobules would abnormally increase in size under stress. Based on observations of cell damage, the degradation of membrane structures, such as chloroplasts, was found to be the primary form of damage during the early stage of induction. During the middle stage of induction, some transparent holes were exposed in the dissolving astaxanthin granules in the cytoplasm. In thick-walled cells, these transparent holes were covered by oil substances dissolving astaxanthin, thereby avoiding further damage to cells. Given the relatively few oil structures, in non-thick-walled cells, the transparent holes expanded to form multiple transparent areas, eventually resulting in the rupture and death of cells. These results suggested that the high level of synthesis and the wide range diffusion of oil explained the expansion of astaxanthin in H. pluvialis.

• Journal of Life Science
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• v.24 no.10
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• pp.1055-1062
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• 2014

We investigated the expression of cucumber genes involved in lipid mobilization and metabolism during cotyledon development to compare gene activity and to study the direction of carbon (acetyl unit) transport between glyoxysomes and mitochondria. The core metabolic pathway involving 10 genes was examined in four intracellular compartments: glyoxysomes (peroxisomes), mitochondria, chloroplasts, and cytosol. Additionally, we tested the early germination response of dark-grown seedlings and the immediate light response for a further 3 days. According to the reverse transcription polymerase chain reaction (RT-PCR), 3-L-ketoacyl-CoA thiolase 2 (Thio2), isocitrate lyase (ICL), and malate synthase (MS), the genes involved in storage lipid mobilization showed a similar and consistent pattern of gene expression in seedling development. Furthermore, coordinate expression of the A BOUT DE SOUFFLE (BOU) gene with ICL and MS during seedling emergence pointed to a possible secondary route of acetyl unit (acetyl-CoA) transport between peroxisomes and mitochondria in cucumber. The expression of the BOU gene was light dependent, as shown by BOU activity in Arabidopsis, suggesting that the dark condition also results in weak membrane biogenesis. In addition, several genes were active throughout the development of the green cotyledon, even during senescence. In conclusion, this study summarizes oil-seed germination and gene expression during cucumber cotyledon development and proposes an additional route for acetyl unit transport.

• Korean Journal of Ecology and Environment
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• v.34 no.3 s.95
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• pp.185-191
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• 2001

The morphological variation and density of the Euglena viridis cells and environmental factors of urban waterways of Daejoncheon , Jeonjucheon, Kwangjucheon, Kumhogang, Mihocheon,and Musimcheon, Korea were studied from 25December, 1995 to 5 January, 1997 in order toelucidate possible relationships among the bio-logical and abiological factors. All E. viridis cells were same in having single star-cluster of chlo-roplast lobes and included two morphotypes based on other detailed morphology. The morphotype I cells agreed well with the typical form off. viridrs and commonly occurred in most of waters and bloomed with $5386\;cells\;{\cdot}\;mL^{-1}$ in Kwangjucheon. The density of the morphotype Ipositively correlated with ammonium (r = 0.80)and nitrite (r = 0.68), while negatively with nit-rate concentration. The morphotype II cells were characterized by having randomly scattered cytoplasmic granules beneath pellicle and unevenmargined lobes of chloroplasts. The density of the morphotype II positively correlated with nitrate (r = 0.98), while negatively correlated with ammonium and nitrite. However, the density of each morphotype was not significantly related with inorganic phosphate, temperature and pH of surface water. These results indicate that E. viridis includes two morphotypes in urban waterways in Korea, that coexist in the same period and station as a response of allocation of nitrogenous nutrients.