• Journal of Plant Biotechnology
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• v.31 no.2
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• pp.175-178
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• 2004

Transgenic tobacco (Nicotiana tabacum L. cv Bright Yellow-2) cell lines expressing a human histone H1.5 (referred to as hH1.5), which suppress collagen-induced rheumatoid arthritis, were developed under the oxidative stress-inducible peroxidase (SWPA2) promoter. Tobacco BY-2 cells were transformed by Agrobacterium-mediated method. The kanamycin-resistant calli were selected on the modified MS medium containing 150mg/L kanamycin and 300mg/L claforan. Transgenic cell lines were confirmed by PCR and northern blot analysis. Recombinant hH1.5 (rhH1.5) protein (42 kDa) was also detected by Western blot analysis, showing a different molecular weight of human hH1.5 (32 kDa). These results suggested that a hH1.5 gene was properly introduced in tobacco cultured cells under the control of SWPA2 promoter. The further characterization of rhH1.5 protein remains to be studied.

• Journal of Plant Biology
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• v.36 no.1
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• pp.113-120
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• 1993

Effects of various NaCl concentrations on salt-tolerant callus of tobacco were investigated. Selection of NaCl-tolerant (S) callus was conducted by subculturing Nicotiana tabacum cv. BY 4 callus in 200 mM NaCl-containing MS medium for more than 18 months. In spite of the long subculture period, characteristics of salt tolerance were maintained very stably. Significant differences were found in ion contents of each callus which was subcultured with treatment of various NaCl concentrations: Na+ and Cl- became higher but Mg2+, Ca2+ and K+ became lower with the increasing external salt contents. Therefore, the ratios of Na+/Ca2+ and Na+/K+ also increased resulting close to those of halophytic property. The contents of chlorophylls and carotenoids in S callus were estimated to 3.1 and 2.9 times more, respectively. than those of non-selected (NS) callus (control). The higher content of external NaCl tended to increase the amount of water soluble proteins and to decrease the amounts of the total sugars, reducing sugars and free amino acids. The activity of peroxidase was increased with higher contents of external NaCl in S callus, but it was maintained at a higher level than S callus at lower NaCl, followed by a subsequent decrease above 80 mM NaCl in NS callus. These results suggest that S callus may have a biological system converting energy source to efficient growth leading to reduction of the growth inhibition under stress environment.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2002.04b
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• pp.49-58
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• 2002

Oxidative stress derived from reactive oxygen species (ROS) is one of the major damaging factors in plants exposed to environmental stress. In order to develop the platform technology to solve the global food and environmental problems in the 21st century, we focus on the understanding of the antioxidative mechanism in plant cells, the development of oxidative stress-inducible antioxidant genes, and the development of transgenic plants with enhanced tolerance to stress. In this report, we describe our recent results on industrial transgenic plants by the gene manipulation of antioxidant enzymes. Transgenic tobacco plants expressing both superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts were developed and were evaluated their protection effects against stresses, suggesting that simultaneous overexpression of both SOD and APX in chloroplasts has synergistic effects to overcome the oxidative stress under unfavorable environments. Transgenic tobacco plants expressing a human dehydroascorbate reductase gene in chloroplasts were showed the protection against the oxidative stress in plants. Transgenic cucumber plants expressing high level of SOD in fruits were successfully generated to use the functional cosmetic purpose as a plant bioreactor. In addition, we developed a strong oxidative stress-inducible peroxidase promoter, SWPA2 from sweetpotato (Ipomoea batatas). We anticipate that SWPA2 promoter will be biotechnologically useful for the development of transgenic plants with enhanced tolerance to environmental stress and particularly transgenic cell lines engineered to produce key pharmaceutical proteins.

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.69-77
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• 2002

Oxidative stress derived from reactive oxygen species (ROS) is one of the major damaging factors in plants exposed to environmental stress. In order to develop the platform technology to solve the global food and environmental problems in the 21st century, we focus on the understanding of the antioxidative mechanism in plant cells, the development of oxidative stress-inducible antioxidant genes, and the development of transgenic plants with enhanced tolerance to stress. In this report, we describe our recent results on industrial transgenic plants by the gene manipulation of antioxidant enzymes. Transgenic tobacco plants expressing both superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts were developed and were evaluated their protection effects against stresses, suggesting that simultaneous overexpression of both SOD and APX in chloroplasts has synergistic effects to overcome the oxidative stress under unfavorable environments. Transgenic tobacco plants expressing a human dehydroascorbate reductase gene in chloroplasts were showed the protection against the oxidative stress in plants. Transgenic cucumber plants expressing high level of SOD in fruits were successfully generated to use the functional cosmetic purpose as a plant bioreactor. In addition, we developed a strong oxidative stress-inducible peroxidase promoter, SWPA2 from sweetpotato (lpomoea batatas). We anticipate that SWPA2 promoter will be biotechnologically useful for the development of transgenic plants with enhanced tolerance to environmental stress and particularly transgenic cell lines engineered to produce key pharmaceutical proteins.

• Korean Journal Plant Pathology
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• v.14 no.3
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• pp.278-285
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• 1998

Effects of chitosan on growth of tomato plant, and suppression of Fusaruim wilt caused by Fusarium oxysporum f. sp. lycopersici and late blight casued by Phytophthora infestans, were examined. Both late blight and fusarium wilt were suppressed by spray and irrigation of chitosan, respectively. Inhibition of mycelial growth was not greatly affected by molecular size of chitosan but, concentration dependent effects was observed. Ninty percent of P. infestans and 80% of F. oxysporum f. sp. lycopersici of mycelial growth was inhibited by 1,000 ppm of chitosan (MW 30,000~50,000) when amended in plate media. Induction of defense-related gene expression in plant by chitosan treatments were observed when chitosan treated tobacco and tomato RNA samples were hybridized with several defense-related genes as probes. The results revealed that $\beta$-1,3-glucanase and chitinase genes were strongly induced, while pathogenesis-related protein-1, 3-hydroxy-3-methylglutaryl coenzyme A reductase, anionic peroxidase, phenylalanine ammonia lyase genes were weakly induced by chitosan treatment. These results suggest that chitosan have dual effects on these host-pathogen interactions. Possible roles of chitosan in suppression of tomato diseases by inhibition of mycelial growth and activation of plant defense responses are discussed.

• Proceedings of the Botanical Society of Korea Conference
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• 1999.07a
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• pp.79-84
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• 1999

Oxidative stress is one of the major environmental stresses to plants. Reactive oxygen species (ROS) generated during metabolic processes damage cellular functions and consequently lead to cell death. Fortunately plants have in vivo defense system by which the ROS is scavenged by enzymes such as superoxide dismutase (SOD) and ascorbate peroxidase (APX). In attempts to understand the protection mechanism of plant against oxidative stress, we developed transgenic tobacco (Nicotiana tabacum cv. Xanthi) plansts thet expressed both SOD and APX in chloroplast using Agrobacterum-mediated transformation and evaluated their protection capabilities against methyl viologen (MV, paraquat) -mediated oxidative damage. Three double transformants (CAI, CA2, and CA3) expressed the chimeric CuZnSOD and chimeric APX in chloroplast, and one transformant (AM) expressed the chimeric APX and chimeric MnSOD in chloroplast. In addition, we obtained three lines of transformants (C/Al, C/A2, and A/C) that expressed the APX and SOD than control plants, and more resistant to oxidative stress caused by MV. TRansformants (C/A and A/C) overexpressing MnSOD, CuZnSOD and APX at the same time showed the highest resistance to MV-mediated oxidative stress among the transformants.

• BMB Reports
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• v.32 no.1
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• pp.51-59
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• 1999

The Korean radish cationic peroxidase (KRCP) promoter, comprising nucleotides -471 to +704 relative to the transcriptional initiation site, was fused to the GUS gene and transformed to tobacco BY-2 cells. We examined how auxin (2,4-dichlorophenoxyacetic acid, 2,4-D), cytokinin (6-benzylaminopurine, BAP), gibberellic acid ($GA_3$), abscisic acid (ABA), methyl jasmonate (MeJA), and phosphatidic acid (PA) affect the GUS expression in the presence or absence of 2,4-D in a modified LS medium. Exogenous 2,4-D or BAP greatly decreased the GUS expression regulated by the KRCP promoter in a modified LS medium containing 0.2 mg/l 2,4-D. $GA_3$ increased the GUS expression and ABA completely reduced the inductive effect of $GA_3$. The GUS expression was also increased dose-dependently by plant defense regulators, MeJA and PA. In contrast to the above results, auxin deprivation from the modified LS medium increased the GUS expression after treatment with exogenous 2,4-D whereas BAP still greatly decreased the GUS expression dose-dependently. $GA_3$ or MeJA slightly decreased the GUS expression. The data suggest that auxin deprivation changes the sensitivity of the suspension cells to exogenous chemicals and that the regulation of the KRCP promoter by 2,4-D, $GA_3$, and MeJA is dependent on auxin, whereas the regulation by BAP is not. This study will be valuable for understanding the function and expression mode of the Korean radish cationic peroxidase in Korean radish.

• Korean Journal of Plant Tissue Culture
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• v.28 no.2
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• pp.69-74
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• 2001

Calli were induced on MS medium supplemented with 0.5 mg/L 2,4-D by using the leaf explants of haploid which were derived from the diploid and haploid of Nicotiana tabacum cv BY4. These calli were subcultured on MS medium with the combination of 2.0 mg/L 2,4-D, 1.0 mg/L kinetin and 0.1 mg/L BAP. Cell propagation of diploid plants were good in a combination of 2.0 mg/L 2,4-D, 0.1mg/L BAP in vitro conditions, suspension cultures were conducted in equal condition. Homogenized suspension cultured cells were smeared 2.0 mL each on MS medium with 0~100 $\mu$M PFP, to select the resistant colony to PFP, and were examined after 10d, 20d and 30d. Measurment of fresh weight of cells after 30d of culture shows that with more concentration of PFP in medium the fresh weight of the cells decreased. In case of diploid, selected callus was the highest in vitro treated with 5 $\mu$M PFP. It was higher than control until 100 $\mu$M PFP. The active degree of catalase was the highest in vitro with 5 $\mu$M PFP but the lowest in vitro with 10 $\mu$M PFP on the other hand, in case of haploid plant, the active degree of peroxidase and catalase was the highest in vitro treated with 50 $\mu$M PFP. It's sure that enzyme active degree of between diploid and haploid had big differences.

• Korean Journal of Environmental Biology
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• v.19 no.3
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• pp.211-217
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• 2001

Salicylate is involved in the induction of pathogen-related proteins and plant defense response. The effects of salicylate on the activity isoperoxidase $A_3$ from tobacco callus (Nicotiana tabacum L.) and the protection against the enzyme inactivation by salicylate in the presence of $Fe^{2+}$ were examined. About 20% and 85% activity losses of peroxidase occurred at 0.48 mM and 0.6 mM salicylate, respectively, showing that isoperoxidase $A_3$ was inactivated by salicylate. The inactivation occurred depending on pH and showed noncompetitive inhibition mode. Moreover, inactivation of the enzyme by salicylate was completely protected in the presence of $Fe^{2+}$. Apoperoxidase without heme moiety was constructed and the effects of various metal ions on the recovery of enzyme activities were investigated. More than 80% of the activity was reconstituted by the addition of $Fe^{2+}$ or hemin. However, the enzyme activity was not recovered by $Cu^{2+},\;Zn^{2+},\;Co^{2+},\;or\;Mn^{2+}$.

• Journal of Plant Biotechnology
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• v.45 no.4
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• pp.364-368
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• 2018

The R2R3-type protein IbMYB1 transcription factor is a key regulator for anthocyanin biosynthesis in the storage roots of sweet potatoes. It was previously demonstrated that the IbMYB1 expression stimulates anthocyanin pigmentation in tobacco leaves, arabidopsis and storage roots of sweet potatoes. In this study, we generated the transgenic potato plants that express the IbMYB1 genes, which accumulated high levels of anthocyanins under the control of either the tuber-specific patatin (PAT) promoter or oxidative stress-inducible peroxidase anionic 2 (SWPA2) promoter. The PAT-MYB1 transgenic lines exhibited higher anthocyanin levels in the tuber than the empty vector control (EV) or SWPA2-MYB1 plants. When combined, our results indicated that overexpression of the IbMYB1 is a highly promising strategy for the generation of transgenic plants with enhanced tissue specific anthocyanin production.