• Journal of The Korean Society of Grassland and Forage Science
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• v.41 no.4
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• pp.242-249
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• 2021

Forage crop management is severely challenged by global warming-induced climate changes representing diverse a/biotic stresses. Thus, screening of valuable genetic resources would be applied to develop stress-tolerant forage crops. We isolated two NAC (NAM, ATAF1, ATAF2, CUC2) transcription factors (ANAC032 and ANAC083) transcriptionally activated by multi-abiotic stresses (salt, drought, and cold stresses) from Arabidopsis by microarray analysis. The NAC family is one of the most prominent transcription factor families in plants and functions in various biological processes. The enhanced expressions of two ANACs by multi-abiotic stresses were validated by quantitative RT-PCR analysis. We also confirmed that both ANACs were localized in the nucleus, suggesting that ANAC032 and ANAC083 act as transcription factors to regulate the expression of downstream target genes. Promoter activities of ANAC032 and ANAC083 through histochemical GUS staining again suggested that various abiotic stresses strongly drive both ANACs expressions. Our data suggest that ANAC032 and ANAC083 would be valuable genetic candidates for breeding multi-abiotic stress-tolerant forage crops via the genetic modification of a single gene.

• Korean Journal of Plant Tissue Culture
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• v.24 no.2
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• pp.93-97
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• 1997

For the molecular genetic study of cold tolerance mechanism in plants, a cDNA encoding fatty acid desaturase (fad3), converting linoleic acid (18:2, $\omega$-6) to linolenic acid (18:3, $\omega$-3), was isolated from $\lambda$ZAPII Arabidopsis thaliana cDNA expression library by plaque hybridization using fad3 cDNA probe derived from Brassica napus. A 1.8 kb-EcoRI fragment from a lambda clone showing a strong positive hybridization signal was subcloned into pGEM7 and analyzed for its nucleotide sequence. From deduced amino acid sequences, the fad3 gene was revealed to have an open reading frame(ORF) consisting of 386 amino acids with a molecular mass of 44,075 Da. The fad3 gene was compared to chloroplast $\omega$-3 fatty acid desaturase (fad7) and endoplasmic reticulum Δ12 fatty acid desaturase (fad2) to show 70% and 58% amino acid sequence homology, respectively, Especially, amino acids of internal (82 to 151) and carboxy terminal (276 to 333) regions were highly conserved, implying their requisite role for enzymatic functioning of fatty acid desaturases. IPTG-induced fad3 cDNA expression in E. coli cells was suggested to be toxic to bacterial growth.

• Pediatric Infection and Vaccine
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• v.25 no.3
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• pp.113-122
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• 2018

Purpose: The aim of this study was to evaluate the clinical characteristics of children diagnosed as cryopyrin-associated periodic syndrome (CAPS) in Korea. Methods: Diagnosis was made based on clinical features and confirmed by a mutation in the cold-induced autoinflammatory syndrome 1 (CIAS1) gene. Especially, osteocartilaginous overgrowth in the patella or distal femur was so characteristic that its presence warranted a diagnosis of chronic infantile neurologic cutaneous and articular/NOMID. Results: We observed the clinical features of 9 Korean CAPS patients. All the patients suffered from an urticarial rash with recurrent fever. Among the 9 patients, 6 presented with rash and 4 with fever on the 1st or 2nd days of birth. Eight patients showed myalgia, and 7 patients showed arthralgia in the joints, and 6 patients showed radiologic findings of arthropathy including cupping of the metaphysis, excessive growth of the epiphysis, osteopenia or overgrowth of the cartilage. Four patients showed brain atrophy, enlarged ventricles or leptomeningeal enhancement on magnetic resonance imaging. Intellectual disability was observed in 1 patient. Five patients had eye involvement as conjunctivitis, uveitis, chorioretinitis, avascular area or papillary edema, and 3 patients showed progressive hearing loss. All 9 patients showed increased C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR). Conclusions: All the patients carried a mutation on exon 3 of the CIAS1 gene. After the anakinra (interleukin-1 receptor antagonist) therapy, the fever and rash immediately disappeared, and CRP and ESR were improved.

• Herbal Formula Science
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• v.20 no.2
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• pp.83-92
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• 2012

Objectives : The aim of this study was to evaluate the efficacy of Hwangryunhaedok-tang (HHT) and Gungangbuja-tang (GBT) on lipopolysaccharide (LPS)-induced mouse model of inflammation. HHT and GBT are one of the representative prescriptions of cold drug and one of the representative prescriptions of hot drug, respectively. For experimental evaluation of their efficacy, we investigated the anti-inflammatory effects of HHT and GBT on LPS-induced inflammation and the mechanisms of their action. Methods : ICR mice were given a HHT (50, 500 mg/kg), GBT (100, 1000 mg/kg) extract orally on three consecutive days. On the third day, they were administered LPS intraperitoneally (35 mg/kg), 1 h after the last sample administration. Blood and liver samples were taken 6 h after the LPS challenge. Cytokine expression and inflammation-related protein factor analyses were performed by Western blotting. Results : Oral administration of HHT significantly reduced pro-inflammatory cytokines, including interleukin (IL)-6, and interferon (IFN)-${\gamma}$ in the serum. While GBT inhibited an increase of IL-6, IFN-${\gamma}$ was not affected. Immunoblot analysis showed that LPS-induced NF-${\kappa}b$ activation was inhibited by GBT, meanwhile HHT only inhibited NF-${\kappa}b$ expression at high does (500 mg/kg). In addition, HHT and GBT inhibited LPS-induced phosphorylation of Erk1/2, Jnk and p38 MAPKs. GBT also significantly inhibited i-Nos and Cox-2 expression, and HHT inhibited only i-Nos expression. Conclusions : Both of HHT and GBT showed anti-inflammatory effects against LPS-induced endotoxemia. However, HHT significantly decreased inflammatory cytokine levels, such as IL-6 and IFN-${\gamma}$ more than GBT, while GBT significantly inhibited inflammatory proteins, including NF-${\kappa}b$, MAP Kinases, i-Nos and Cox-2, more than HHT. These results suggest that HHT and GBT regulate the different mechanisms of action and pathways, presumably by regulating cytokine levels (IL-6, IFN-${\gamma}$), NF-${\kappa}b$ activation, and several pro-inflammatory gene expression, although both of HHT and GBT have anti-inflammatory effects.

• Journal of Plant Biotechnology
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• v.42 no.1
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• pp.19-24
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• 2015

Glycine betaine (GB) is one of the compatible solutes that accumulate in the chloroplasts of certain halotolerant plants under salt or cold stress. The codA gene for choline oxidase, the enzyme that converts choline into GB, has been cloned from a soil bacterium Arthrobacter globiformis. We generated transgenic sweetpotato plants [Ipomoea batatas (L.) Lam] expressing codA gene in chloroplasts under the control of the SWPA2 promoter (referred to as SC plants) and evaluated SC plants under oxidative and drought stresses. SC plants showed enhanced tolerance to methyl viologen (MV)-mediated oxidative stress and drought stress due to induced expression of codA. At $5{\mu}M$ of MV treatment, all SC plants showed enhanced tolerance to MV-mediated oxidative stress through maintaining low ion leakage and increased GB levels compared to wild type plants. When plants were subjected to drought conditions, SC plants showed enhanced tolerance to drought stress through maintaining high relative water contents and increased codA expression compared to wild type plants. These results suggest that the SC plants generated in this study will be useful for enhanced biomass production on global marginal lands.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.1
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• pp.9-14
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• 2016

Objective: Autophagy contributes to the clearance and recycling of macromolecules and organelles in response to stress. We previously reported that vitrified mouse oocytes show acute increases in autophagy during warming. Herein, we investigate the potential role of Atg7 in oocyte vitrification by using an oocyte-specific deletion model of the Atg7 gene, a crucial upstream gene in the autophagic pathway. Methods: Oocyte-specific Atg7 deficient mice were generated by crossing Atg7 floxed mice and Zp3-Cre transgenic mice. The oocytes were vitrified-warmed and then subjected to in vitro fertilization and development. The rates of survival, fertilization, and development were assessed in the Atg7 deficient oocytes in comparison with the wildtype oocytes. Light chain 3 (LC3) immunofluorescence staining was performed to determine whether this method effectively evaluates the autophagy status of oocytes. Results: The survival rate of vitrified-warmed $Atg7^{f/f}$;Zp3-Cre ($Atg7^{d/d}$) metaphase II (MII) oocytes was not significantly different from that of the wildtype ($Atg7^{f/f}$) oocytes. Fertilization and development in the $Atg7^{d/d}$ oocytes were significantly lower than the $Atg7^{f/f}$ oocytes, comparable to the $Atg5^{d/d}$ oocytes previously described. Notably, the developmental rate improved slightly in vitrified-warmed $Atg7^{d/d}$ MII oocytes when compared to fresh $Atg7^{d/d}$ oocytes. LC3 immunofluorescence staining showed that this method can be reliably used to assess autophagic activation in oocytes. Conclusion: We confirmed that the LC3-positive signal is nearly absent in $Atg7^{d/d}$ oocytes. While autophagy is induced during the warming process after vitrification of MII oocytes, the Atg7 gene is not essential for survival of vitrified-warmed oocytes. Thus, induction of autophagy during warming of vitrified MII oocytes seems to be a natural response to manage cold or other cellular stresses.

• Journal of Korean Society of Forest Science
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• v.103 no.2
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• pp.189-195
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• 2014

Basic leucine zipper (bZIP) protein is a regulatory transcription factor that plays crucial roles in growth, development and stress response of plant. In this study, we isolated a PagbZIP1 gene that belonged to Group SE3 of bZIP from Populus alba ${\times}$ P. glandulosa, and investigated its expressional characteristics. The PagbZIP1 is 844 base pairs long and encodes a putative 144-amino-acid protein with an expected molecular mass of 16.6 kDa. The PagbZIP1 has two conserved domains including the basic and leucine zipper portions. Southern blot analysis revealed that two copies of the gene are presented in the poplar genome. PagbZIP1 was specifically expressed in the root and suspension cells. Moreover, the expression of PagbZIP1 was induced by drought, salt, cold and ABA. Therefore, our results indicated that PagbZIP1 might be expressed in response to abiotic stress through the ABA-mediated signaling pathway in poplar.

• Journal of Plant Biotechnology
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• v.36 no.1
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• pp.23-29
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• 2009

We have previously isolated a CCCH type zinc-finger protein gene, OsZF2 (Oryza sativa Zinc Finger 2), from the cold-treated rice cDNA library. To investigate the potential role of OsZF2, transgenic rice lines over-expressing OsZF2 under the control of CaMV 35S promoter have been developed through Agrobacterium-mediated transformation. Elevated level of OsZF2 transcripts was confirmed by RNA gel blot analysis in transgenic rice. Under the 100 mM NaCl condition, the transgenic rice showed significantly enhanced growth rate in terms of shoot length and fresh weight, implicating that OsZF2 is likely to be involved in salt response of rice. In the field condition, however, the transgenic rice showed a dwarf phenotype and flowering time was delayed. Genome expression profiling analysis of transgenic plants using the 20K NSF rice oligonucleotide array revealed many up-regulated genes related to stress responses and signaling pathways such as chaperone protein dnaJ 72, salt stress-induced protein, PR protein, disease resistance proteins RPM1 and Cf2/Cf5 disease resistance protein, carbohydrate/ sugar transporter, OsWAK kinase, brassinosteroid LRR receptor kinase, and jasmonate O-methyltransferase. These data suggest that the CCCH type zinc-finger protein OsZF2 is a upstream transcriptional factor regulating growth and stress responsiveness of rice.

• Weed & Turfgrass Science
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• v.2 no.4
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• pp.368-375
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• 2013

Whole genome transcriptomes from Miscanthus species were sequenced and analyzed, which provided 50 different types of transcription factor (TF) involving various developmental processes or environmental stresses. Among the explored TF, WRKY gene family was the major type and one of the WRKY genes, MSIR7180_WRKY4, induced under low temperature environment was selected to investigate how the Miscanthus-originated MSIR7180_WRKY4 TF responds when exposed to low temperature in four warm-season turfgrasses (Z. matrella 'Semil', bermudagrass, St. Augustinegrass, and seashore paspalum). The MSIR7180_WRKY4 was expressed higher during low temperature period in Bermudagrass, but the expression was enhanced in St. Augustinegrass. In contrast, the gene in 'Semil' cultivar was barely expressed and relatively less expressed, but repressed gradually in seashore paspalum, which seems to allow two turfgrasses stay-green longer in the fall season. The results indicate that bermudagrass and St. Augustinegrass adapt to low temperature quickly, but relative tolerance to low or cold temperature at the molecular level needs to be further investigated at different physiological stages and the corresponding genes systematically.

• Journal of Plant Biotechnology
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• v.44 no.3
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• pp.296-302
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• 2017

Rice (Oryza sativa) cultivars show an impairment of growth and development in response to abiotic stresses such as drought, salinity, heat and cold at the early seedling stage. The tolerance to heat stress in plants has been genetically modulated by the overexpression of heat shock transcription factor genes or proteins. In addition to a high temperature-tolerance that has also been altered by elevating levels of osmolytes, increasing levels of cell detoxification enzymes and through altering membrane fluidity. To examine the heat tolerance in transgenic rice plants, three OsOPT10 overexpressing lines were characterized through a physiological analysis, which examined factors such as the electrolyte leakage (EL), soluble sugar and proline contents. We further functionally characterized the OsOPT10 gene and found that heat induced the expression of OsOPT10 and P5CS gene related proline biosynthesis. It has been suggested that the expression of OsOPT10 led to elevated heat tolerance in transgenic lines.