• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.189-189
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• 2022

Soil salinity negatively affects plant growth, productivity, and metabolism. Rice is known to have more sensitive phenotypes than other cereal crops, such as wheat, sorghum, and barley. We characterized the molecular function of rice C3HC4 as a really interesting new gene (RING). Oryza sativa RING finger protein H2-3 (OsRFPH2-3) was highly expressed in 100 mM NaCl. To identify the localization of OsRFPH2-3, we fused vectors that include C-terminal GFP protein (35S;;OsRFPH2-3-GFP). OsRFPH2-3 was expressed in the nucleus in rice protoplasts. An in vitro ubiquitin assay demonstrated that OsRFPH2-3 possessed E3-ubiquitin ligase activity. However, the mutated OsRFPH2-3 were not possessed any E3-ubiquitin ligase activity. Under normal conditions, there is no significant phenotypic difference between transgenic plants and WT plants. However, OsRFPH2-3-overexpressing plants exhibited higher fresh weight and length under saline conditions. Also, transgenic plants maintain higher chlorophyll, proline, and soluble sugar contents and lower H₂O₂ and MDA contents than the wild type; these results support transgenic plants with enhanced salinity tolerance phenotypes.

• Proceedings of the Korean Society of Crop Science Conference
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• 2023.04a
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• pp.156-156
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• 2023

Salinity is a major abiotic stress that limits rice productivity in many regions of the world. In order to develop salt stress tolerant rice plants, genetic engineering is a promising approach. We characterized the molecular function of rice C3H2C3 as a really interesting new gene (RING). Oryza sativa RING finger protein H2-3 (OsRFPH2-3) was highly expressed in 100 mM NaCl. To identify the localization of OsRFPH2-3, we fused vectors that include C-terminal GFP protein (35S;;OsRFPH2-3-GFP). OsRFPH2-3 was expressed in the nucleus in rice protoplasts. An in vitro ubiquitin assay demonstrated that OsRFPH2-3 possessed E3-ubiquitin ligase activity. However, the mutated OsRFPH2-3 were not possessed any E3-ubiquitin ligase activity. Under salinity conditions, OsRFPH2-3-overexpressing plants exhibited higher chlorophyll, proline, SOD, POD, CAT, and soluble sugar contents and lower H₂O₂ accumulation than wild-type plants, supporting transgenic plants with enhanced salinity tolerance phenotypes. OsRFPH2-3-overexpressing plants exhibited low Na+ accumulation and Na+/K+ ratios in their roots. Theses results suggest that overexpression of OsRFPH2-3 can make plant insensitivity about salinity conditions.

• Journal of Plant Biotechnology
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• v.31 no.3
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• pp.185-190
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• 2004

We established a transient gene expression system in chili pepper leaves based on Agrobacterium-mediated transformation of GUS gene. For the best GUS transient expression, two step culture system was adopted. When the Agrobacterium tumefaciens cell density of pre-culture was $A_{600nm}$ 0.3, the cells were harvested and diluted to $A_{600nm}$ 0.8 with virulence induction medium after cell harvested. The addition of acetosyringone (200 $\mu$M) in virulence induction step was a key factor for successful transient expression. Additionally, Younger leaves showed more effective transient expression than older leaves. Temporally, the strongest intensity of GUS expression was detected at 2 days after infiltration. These results demonstrate that Agrobacterium-mediated transient expression can be used for a simple in vivo assays of plant promoters, transcription factors and furthermore provide efficient protocol for chili pepper transformation.

• Korean Journal of Plant Resources
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• v.28 no.5
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• pp.582-590
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• 2015

In the present study, we investigated the effect of 70% EtOH extract from Hippophae Rhamnoides L. leaves (HRL) on the anti-obesity effect in 3T3-L1 cells. The effects of HRL on lipid accumulation in 3T3-L1 cells were examined using Oil Red O staining. In addition, we examined the gene expression levels by using RT-PCR and western blot. The results of this analysis showed that 100 ㎍/㎖ HRL significantly increased the inhibition of lipid accumulation by 82.25%; significantly decreased the mRNA expression of sterol regulatory element binding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding proteins α (C/EBPα), and fatty acid synthase (FAS) in 3T3-L1 cells as well as the stimulated protein expression of AMP-activated protein kinase (AMPK); and suppressed the expression level of PPARγ. These results suggest that HRL can prevent adipogenesis through activation of AMPKα and inhibition of adipogenesis transcription factors.

• Korean Journal of Plant Resources
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• v.26 no.4
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• pp.411-416
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• 2013

This study was conducted not only to analyze ${\alpha}$-glucosidase inhibition activity with fronds and rhizomes of nine Pteridophyte species, but also to select the plant materials suitable for natural ${\alpha}$-glucosidase inhibitor. Harvested rhizomes and fronds were washed, freeze-dried and grinded. After conducting ultrasonification extraction for 30 minutes in ultrasonic water tank with 100% methanol solvent, and vacuum filtration, ${\alpha}$-glucosidase inhibition activity was measured. Acarbose was used as the positive control. After mixing $100{\mu}L$ of 0.7 unit ${\alpha}$-glucosidase enzyme solution into $50{\mu}L$ of extract and reacting them at $37^{\circ}C$ for 10 minutes, $50{\mu}L$ of 1.5 mM ${\rho}$-NPG solution was taken and reacted at $37^{\circ}C$ for 20 minutes. The reaction was stopped with 1 mL of 1 M $Na_2CO_3$ and absorbance was measured in 405 nm. With the regression analysis, the content of solubility solids (the value of $IC_{50}$) which can inhibit 50% of 0.7 unit ${\alpha}$-glucosidase solution's activity was investigated. The frond ($IC_{50}=14.00{\sim}913.33{\mu}g{\cdot}mL^{-1}$) and rhizome extracts ($IC_{50}=12.93{\sim}205.84{\mu}g{\cdot}mL^{-1}$) of nine Pteridophyte species showed higher ${\alpha}$-glucosidase inhibition activity in comparison with acarbose ($IC_{50}=1413.70{\mu}g{\cdot}mL^{-1}$). The extracts of fronds and rhizomes showed higher value than acarbose by 1.55~100.98 and 6.87~109.33 times each. Especially, ${\alpha}$-glucosidase inhibition activities of Pyrrosia lingua in fronds and Osmunda cinnamomea var. fokiensis in rhizomes were the highest. The necessary biomass of fronds and rhizomes for inhibiting 50% of ${\alpha}$-glucosidase activity showed the lowest value, 0.35, 0.27 mg each, in O. cinnamomea var. fokiensis. $IC_{50}$ value of P. lingua was the highest among fronds of nine Pteridophyte species, but content of soluble solids was 2.4 times less than O. cinnamomea var. fokiensis. So frond of O. cinnamomea var. fokiensis is more economic in comparison with P. lingua. As the result of this study, O. cinnamomea var. fokiensis showed high ${\alpha}$-glucosidase inhibition activity even with small biomass. Therefore it was considered to be high-valued economic material as natural ${\alpha}$-glucosidase inhibitor.

• Korean Journal of Plant Tissue Culture
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• v.25 no.4
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• pp.277-282
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• 1998

Whereas cationic extracellular peroxidases (PODs) were observed in the suspension cultures of rose (Rosa sp. L. cv Pual's scarlet) grown under normal conditions, new anionic isozymes were induced within 24 hr by the treatment of low host-specific elicitor (10 mg glucan/L media) prepared from yeast cell wall. Prominent anionic (pI 6.1) and cationic POD (pI 8.4) were purified and characterized to understand the physiological role of the enzymes. Both enzymes were purified (ca.200 fold) by the ammonium sulfate precipitation, ion exchange chromate-graphy and gel filtration chromatography. The Km values of the purified anionic POD for ferulic acid and $\textrm{H}_2\textrm{O}_2$ were 4.64 mM and 0.72 mM, whereas those of the cationic POD were 1.38 mM and 0.48 mM, respetively. The activity of the anionic POD as NADH oxidase was twice higher than that of cationic POD. The NADH oxidation in the anionic POD fraction was inhibited by 60% on the addition of 0.1 mM coniferyl alcohol, while that in the cationic fraction was inhibited by 15%.

• Korean Journal of Soil Science and Fertilizer
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• v.43 no.5
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• pp.616-623
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• 2010

Plant samples from 49 sites for Chinese cabbage, 28 sites for radish, 16 sites for cabbage, 8 sites for head lettuce, 20 sites for onion from farmers' and experimental fields in highland of Korea were collected and analyzed to find out the uptake patterns of nitrogen (N), phosphorus (P) and potassium (K) by altitude. Dry weight and uptake of N, P and K were increased at higher altitude in most vegetable crops. Nutrition uptake by Chinese cabbage was 163 ~ 283 kg $ha^{-1}$ for N, 42 ~ 69 kg $ha^{-1}$ for $P_2O_5$ and 146 ~ 270 kg $ha^{-1}$ for $K_2O$ according to altitude. Nutrient uptake by radish according to altitude was 153~159 kg $ha^{-1}$ for N, 38 ~ 46 kg $ha^{-1}$ for $P_2O_5$, and 151 ~ 185 kg $ha^{-1}$ for $K_2O$. In case of cabbage, the plant uptakes of N, P, and K were increased at altitudes of 600 ~ 1,000 m. Nutrient uptake of cabbage was 280 ~ 348 kg $ha^{-1}$ for N, 34 ~ 87 kg $ha^{-1}$ for $P_2O_5$, and 209 ~ 290 kg $ha^{-1}$ for $K_2O$ according to altitude. Uptakes of N-$P_2O_5-K_2O$ by head lettuce at an altitude of 800 ~ 850 m were 93-26-126 kg $ha^{-1}$, respectively. Uptakes of N-$P_2O_5-K_2O$ by onions at an altitude of 600 ~ 800 m were 313-140-234 kg $ha^{-1}$, respectively, but there was no tendency in nutrition uptake patterns by altitude. Small cultivation areas used for leaf vegetable crops do not have fertilizer recommendation standards in alpine regions. It might be preferable to use a correction factor equivalent to the index of available nutrient uptake for the determination of N, P and K fertilizer application rates.

• Journal of Life Science
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• v.22 no.6
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• pp.788-793
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• 2012

High concentration of antibiotics has been used to treat the outbreak of edwardsiellosis caused by Edwardsiella tarda in aquaculture. However, not all of the bacteria have been killed with high concentrations of antibiotics treatment by the formation of persister cells with a dormant state. The main objective of this study was to kill persister cell using antibiotics with the addition of natural plant compounds. Antibiotics used in this study consist of 100 mg/ml florfenicol and 100 mg/ml amoxicillin. Ten natural plant compounds with persister cell inhibitor activity to E. coli were obtained from Protein Engineering and Systems Biology Lab. of Sungkyunkwan University. The persister cell inhibition activities of those natural plant compounds were evaluated in test tube. Concentrations of the antibiotics were in the ranges of 25~200 ${\mu}g/ml$. The persister cell formation was observed after 16 hours of culture. Persister cells were killed by antibiotics with natural plant compounds. Among ten natural plant compounds, Gynostemma pentaphyllum, Mallotus japonicus, and Orixa japonica showed persister cell formation inhibition activities. The optimal concentrations of G. pentaphyllum, M. japonicus, and O. japonica for the inhibitor of persister cell formation were 100 ${\mu}g/ml$, 100 ${\mu}g/ml$, and 200 ${\mu}g/ml$, respectively. In vivo study was carried out to evaluate the effect of the antibiotics with natural plant compounds using aquacultural fish, olive flounder, as test animals. G. pentaphyllum, M. japonicus, and O. japonica of 30 ${\mu}g/ml$, 10 ${\mu}g/ml$, and 10 ${\mu}g/ml$ with antibiotics reduced cumulative mortalities, showing the effectiveness of persister cell inhibition.

• Journal of Microbiology and Biotechnology
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• v.10 no.5
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• pp.728-732
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• 2000

Propagation of Anagrapha falcifera nuclear polyhedrosis virus(AfNPV) was investigated using well-plates and split-flow air-lift bioreactors. In well-plate experiments, the effects of pH, cell density at a point of infection, serum concentration, DEAE-dextran, and lipid on virus propagation were all closely examined. The AfNPV titer in well-plates was optimal at pH 6.8 and $3{\times}10^6$ cells/$cm^2$. The virus titer was not dramatically affected when the fetal bovine serum concentration was reduced from 10% to 5%. The addition of cholesterol at AfNPV infection of Sf21 cells enhanced the virus titer, whereas the addition of DEAE-dextran did not improve the titer. The AfNPV titer ($3.8{\times}10^7$ $TCID_{50}/ml$) at optimized conditions for well-plate experiments was 2.5-fold higher than for the control. In bioreactor experiments, the AfNPV titer showed its maximum level at air flow rates of 20-40 ml/min. In a split-flow air-lift bioreactor, AfNPV titer ($2.3{\times}10^7\;TCID_{50}/ml$) was 1.5-fold higher than the control when the culture was at pH 6.8 and supplemented with 0.34 mM cholesterol.

• Archives of Pharmacal Research
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• v.28 no.5
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• pp.541-545
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• 2005

Flowers of Erigeron annuus L. were extracted with 80% aqueous MeOH, and the concentrated extract was partitioned with EtOAc, n-BuOH, and H$_2$O. Repeated silica gel and OD S column chromatography of the EtOAc fraction led to the isolation of a sterol, through activityguided fractionation, using ACAT inhibitory activity measurements. From the physico-chemical data, including NMR, MS, and IR, the chemical structure of the compound was determined to be an ergosterol peroxide (1), which has been isolated for the first time from this plant. This compound exhibited hACAT-1 and Lp-PLA$_2$ inhibitory effects, with inhibitory values of 51.6 ${\pm}$ 0.9 and 51 .7 ${\pm}$ 1.2%, at a treatment concentration of 0.23 mM.