• Journal of Microbiology and Biotechnology
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• v.30 no.8
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• pp.1142-1148
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• 2020

Mitochondria play a vital role in iron uptake and metabolism in pathogenic fungi, and also influence virulence and drug tolerance. However, the regulation of iron transport within the mitochondria of Cryptococcus neoformans, a causative agent of fungal meningoencephalitis in immunocompromised individuals, remains largely uncharacterized. In this study, we identified and functionally characterized Mrs3/4, a homolog of the Saccharomyces cerevisiae mitochondrial iron transporter, in C. neoformans var. grubii. A strain expressing an Mrs3/4-GFP fusion protein was generated, and the mitochondrial localization of the fusion protein was confirmed. Moreover, a mutant lacking the MRS3/4 gene was constructed; this mutant displayed significantly reduced mitochondrial iron and cellular heme accumulation. In addition, impaired mitochondrial iron-sulfur cluster metabolism and altered expression of genes required for iron uptake at the plasma membrane were observed in the mrs3/4 mutant, suggesting that Mrs3/4 is involved in iron import and metabolism in the mitochondria of C. neoformans. Using a murine model of cryptococcosis, we demonstrated that an mrs3/4 mutant is defective in survival and virulence. Taken together, our study suggests that Mrs3/4 is responsible for iron import in mitochondria and reveals a link between mitochondrial iron metabolism and the virulence of C. neoformans.

• The Korean Journal of Physiology and Pharmacology
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• v.20 no.6
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• pp.581-593
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• 2016

The advantages of monounsaturated fatty acids (MUFAs) on insulin resistance and type 2 diabetes mellitus (T2DM) have been well established. However, the molecular mechanisms of the anti-diabetic action of MUFAs remain unclear. This study examined the anti-hyperglycemic effect and explored the molecular mechanisms involved in the actions of fish oil- rich in MUFAs that had been acquired from hybrid catfish (Pangasius larnaudii${\times}$Pangasianodon hypophthalmus) among experimental type 2 diabetic rats. Diabetic rats that were fed with fish oil (500 and 1,000 mg/kg BW) for 12 weeks significantly reduced the fasting plasma glucose levels without increasing the plasma insulin levels. The diminishing levels of plasma lipids and the muscle triglyceride accumulation as well as the plasma leptin levels were identified in T2DM rats, which had been administrated with fish oil. Notably, the plasma adiponectin levels increased among these rats. The fish oil supplementation also improved glucose tolerance, insulin sensitivity and pancreatic histological changes. Moreover, the supplementation of fish oil improved insulin signaling ($p-Akt^{Ser473}$ and p-PKC-${\zeta}/{\lambda}^{Thr410/403}$), $p-AMPK^{Thr172}$ and membrane GLUT4 protein expressions, whereas the protein expressions of pro-inflammatory cytokines (TNF-${\alpha}$ and nuclear NF-${\kappa}B$) as well as p-PKC-${\theta}^{Thr538}$ were down regulated in the skeletal muscle. These data indicate that the effects of fish oil-rich in MUFAs in these T2DM rats were partly due to the attenuation of insulin resistance and an improvement in the adipokine imbalance. The mechanisms of the anti-hyperglycemic effect are involved in the improvement of insulin signaling, AMPK activation, GLUT4 translocation and suppression of pro-inflammatory cytokine protein expressions.

• Journal of Applied Biological Chemistry
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• v.54 no.4
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• pp.244-251
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• 2011

Galactinol and rafinose accumulation in plants is associated with stressful environmental conditions such as cold, heat, or dehydration by the action of galactinols synthase (GolS) in the raffinose family of oligosaccharides biosynthetic pathway from UDP-galactose. Moreover, several reports mentioned that GolS transcription is up regulated by various environmental stresses like cold, heat, dehydration. Therefore, to determine whether MoGolS1 was induced with the abiotic stress we analyzed the expression pattern of the gene under various abiotic stresses like heat, cold, abscisic acid, sucrose and salt concentration in the lemon balm plants grown in standard MS medium. The MoGolS1 gene was 981-bp in length encoding 326 amino acids in its sequence and shared 77 and 76% sequence similarity with Arabidopsis thaliana galactinol synthase4 (AtGolS4) and AtGolS1 genes respectively. The MoGolS1 gene was strongly expressed by the abiotic stress induced by sucrose, ABA or heat shock. It was also expressed in responses to cold, Identification and Functional Characterization of the GALACTINOL SYNTHASgene induction with various stresses may be possible for itscrucial function in abiotic stress tolerance in plants, providing a good engineering target for genetic engineering.

• Journal of Microbiology and Biotechnology
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• v.26 no.7
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• pp.1311-1319
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• 2016

Alternaria brassicicola (Schwein.) invades Brassicaceae and causes black spot disease, significantly lowering productivity. Mitogen-activated protein kinases (MAPKs) and their upstream kinases, including MAPK kinases (MAPKKs) and MAPKK kinases (MAPKKK), comprise one of the most important signaling pathways determining the pathogenicity of diverse plant pathogens. The AbSte7 gene in the genome of A. brassicicola was predicted to be a homolog of yeast Ste7, a MAPKK; therefore, the function was characterized by generating null mutant strains with a gene replacement method. AbSte7 replacement mutants (RMs) had a slower growth rate and altered colony morphology compared with the wild-type strain. Disruption of the AbSte7 gene resulted in defects in conidiation and melanin accumulation. AbSte7 was also involved in the resistance pathways in salt and oxidative stress, working to negatively regulate salt tolerance and positively regulate oxidative stress. Pathogenicity assays revealed that AbSte7 RMs could not infect intact cabbage leaves, but only formed very small lesions in wounded leaves, whereas typical lesions appeared on both intact and wounded leaves inoculated with the wild-type strain. As the first studied MAPKK in A. brassicicola, these data strongly suggest that the AbSte7 gene is an essential element for the growth, development, and pathogenicity of A. brassicicola.

• Korean Journal of Plant Resources
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• v.23 no.6
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• pp.526-534
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• 2010

Genomes of clusters of related eukaryotes are now being sequenced at an increasing rate. In this paper, we developed an accurate, low-cost method for annotation of gene prediction and exon-intron structure. The gene prediction was adapted for delta 1-pyrroline-5-carboxylate-synthetase (p5cs) gene from China wild-type of the halophytic Leymus chinensis (Trin.), naturally adapted to highly-alkali soils. Due to complex adaptive mechanisms in halophytes, more attentions are being paid on the regulatory elements of stress adaptation in halophytes. P5CS encodes delta 1-pyrroline-5-carboxylate-synthetase, a key regulatory enzyme involved in the biosynthesis of proline, that has direct correlation with proline accumulation in vivo and positive relationship with stress tolerance. Using analysis of reverse transcription-polymerase chain reaction (RT-PCR) and PCR, and direct sequencing, 1076 base pairs (bp) of cDNA in length and 2396 bp of genomic DNA in length were obtained from direct sequencing results. Through gene prediction and exon-intron structure verification, the full-length of cDNA sequence was divided into eight parts, with seven parts of intron insertion. The average lengths of determinated coding regions and non-coding regions were 154.17 bp and 188.57 bp, respectively. Nearly all splice sites displayed GT as the donor sites at the 5' end of intron region, and 71.43% displayed AG as the acceptor sites at the 3' end of intron region. We conclude that this method is a cost-effective way for obtaining an experimentally verified genome annotation.

• Journal of Environmental Science International
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• v.21 no.12
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• pp.1435-1447
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• 2012

The present study was conducted to determine the effect of hexaconazole (HEX), a triazole fungicide, on the growth, yield, photosynthetic response and antioxidant potential in cucumber (Cucumis sativus L.) plants subjected to UV-B stress. UV-B radiation and HEX were applied separately or in combination to cucumber seedlings. The growth parameters were significantly reduced under UV-B treatment, however, this growth inhibition was less in HEX treated plants. HEX caused noticeable changes in plant morphology such as reduced shoot length and leaf area, and increased leaf thickness. HEX was quite persistent in inhibiting shoot growth by causing a reduction in shoot fresh and dry weight. HEX noticeably recovered the UV-B induced inhibition of biomass production. Significant accumutation in anthocyanin and flavonoid pigments in the leaves occurred as a result of HEX or UV-B treatments. HEX permitted the survival of more green leaf tissue preventing chlorophyll content reduction and higher quantum yield for photosystemII under UV-B exposure. HEX treatment induced a transient rise in ABA levels in the leaves, and combined application of HEX and UV-B showed a significant enhancement of ABA content which activates $H_2O_2$ generation. UV-B exposure induced accumulation of $H_2O_2$ in the leaves, while HEX prevented UV-B induced increase in $H_2O_2$, indicating that HEX serves as an antioxidant agent able to scavenge $H_2O$ to protect cells from oxidative damage. An increase in the ascorbic acid was observed in the HEX treated cucumber leaves affecting many enzyme activities by removing $H_2O_2$ during photosynthetic processes. The activities of antioxidant enzymes including catalase(CAT), ascorbate peroxidase(APX), superoxide dismutase(SOD) and peroxidase(POD) in the leaves in the presence of HEX under UV-B stress were higher than those under UV-B stress alone. These findings suggest that HEX may participate in the enhanced tolerance to oxidative stress. From these results it can be concluded that HEX moderately ameliolate the effect of UV-B stress in cucumber by improving the components of antioxidant defense system.

• Journal of Plant Biotechnology
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• v.1 no.1
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• pp.61-68
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• 1999

Rapid accumulation of reactive oxygen species (ROS) and their toxic reaction products with lipids and proteins significantly contributes to the damage of crop plants under biotic and abiotic stresses. We have identified several stress activated alfalfa genes, including the gene of the alfalfa ferritin and a novel NADPH-dependent aldose/aldehyde reductase enzyme. Transgenic tobacco plants that synthesize alfalfa ferritin in vegetative tissues-either in its processed form in chloroplast or in the cytoplasmic non-processed form-retained photosynthetic function upon free radical toxicity generated by paraquat treatment and exhibited tolerance to necrotic damage caused by viral and fungal infections. We propose that by sequestering intracellular iron involved in generation of the very reactive hydroxyl radicals through a Fenton reaction, ferritin protects plant cells from oxidative damage. Our preliminary results with the other stress-inducable alfalfa gene (a NADPH-dependent aldo-keto reductase) indicate, that the encoded enzyme may play role in the stress response of the plant cells. These studies reveal new pathways in plants that can contribute to the increased stress resistance with a potential use in crop improvement.

• Biomolecules & Therapeutics
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• v.21 no.6
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• pp.442-446
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• 2013

Here in this study, we isolated 1,2,3,4,6-penta-O-galloyl-${\beta}$-D-glucose (PGG) from Curcuma longa L. and elucidated the lifespan-extending effect of PGG using Caenorhabditis elegans model system. In the present study, PGG demonstrated potent lifespan extension of worms under normal culture condition. Then, we determined the protective effects of PGG on the stress conditions such as thermal and oxidative stress. In the case of heat stress, PGG-treated worms exhibited enhanced survival rate, compared to control worms. In addition, PGG-fed worms lived longer than control worms under oxidative stress induced by paraquat. To verify the possible mechanism of PGG-mediated increased lifespan and stress resistance of worms, we investigated whether PGG might alter superoxide dismutase (SOD) activities and intracellular ROS levels. Our results showed that PGG was able to elevate SOD activities of worms and reduce intracellular ROS accumulation in a dose-dependent manner.

• Journal of Plant Biotechnology
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• v.34 no.4
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• pp.293-298
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• 2007

Robinia pseudoacacia var. umbraculifera, commonly called umbrella black locust were regenerated after co-cultivation of internode segments with Agrobacterium tumefaciens which included yeast cadmium factor 1 (YCF 1) gene. The tolerance to cadmium and lead for plants can be increased by the YCF1 gene expression. Moreover, the recent studies have shown that YCF1 gene transgenic plants increase the accumulation of cadmium and lead into plant vacuoles. The effect of plant growth regulator such as 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}$-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and thidiazuron (TDZ) were studied to evaluate the propagation of plants through internode explants. The efficient induction of multiple adventitious shoots and callus were observed on a medium supplemented with 0.1 mg/L TDZ + 0.2 mg/L BA. To induce shoot elongation and rooting, regenerated shoots were transferred into basal MS medium without any plant growth regulator. Successful Agrobacterium tumefaciens mediated transformation was obtained by 20 min vacuum-infiltration with $50{\mu}M$ acetosyringone on the optimal multiple shoot induction medium with 30 mg/L hygromycin and 300 mg/L cefotaxime. To confirm the integration and expression of transgene, Polymerase Chain Reaction (PCR) and Reverse Transcriptase PCR (RT-PCR) were performed with specific primers. The frequency of transformation was approximately 18.94%. This study can be used to genetic engineering of phytoremediator.

• Journal of The Korean Society of Grassland and Forage Science
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• v.21 no.4
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• pp.259-264
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• 2001

To investigate the physiological responses to naturally occurring winter freezing stress in creeping bentgrass, changes in carbohydrates were monitored during winter period. Turf quality and leaf growth was nearly parallel with temperature fluctuation. The concentration of glucose, fructose and sucrose in both shoot and root gradually increased from November to January, and then sharply decreased until April. Sucrose was the largest pool of soluble sugars. Fructan also slightly accumulated in both shoot and roots from November to February. Fructan hydrolysis in both organs was found to be much active between February to April. Shoot contained largely higher carbohydrate content in all compounds examined than roots did. Fructan was found to be a main carbohydrate storage form, showing the highest concentration (176.7 and 126.7 mg g-' DW for shoot and root in February). The depolymerization of fructan from February coincided with the high declines in mono- and disaccharide. These results suggest that the accumulation of non-structural carbohydrate until January could be associated with freezing tolerance, and the active decrease from February with shoot regrowth.