• Journal of Ginseng Research
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• v.43 no.4
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• pp.645-653
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• 2019

Background: Cytochrome P450 enzymes catalyze a wide range of reactions in plant metabolism. Besides their physiological functions on primary and secondary metabolites, P450s are also involved in herbicide detoxification via hydroxylation or dealkylation. Ginseng as a perennial plant offers more sustainable solutions to herbicide resistance. Methods: Tissue-specific gene expression and differentially modulated transcripts were monitored by quantitative real-time polymerase chain reaction. As a tool to evaluate the function of PgCYP736A12, the 35S promoter was used to overexpress the gene in Arabidopsis. Protein localization was visualized using confocal microscopy by tagging the fluorescent protein. Tolerance to herbicides was analyzed by growing seeds and seedlings on Murashige and Skoog medium containing chlorotoluron. Results: The expression of PgCYP736A12 was three-fold more in leaves compared with other tissues from two-year-old ginseng plants. Transcript levels were similarly upregulated by treatment with abscisic acid, hydrogen peroxide, and NaCl, the highest being with salicylic acid. Jasmonic acid treatment did not alter the mRNA levels of PgCYP736A12. Transgenic lines displayed slightly reduced plant height and were able to tolerate the herbicide chlorotoluron. Reduced stem elongation might be correlated with increased expression of genes involved in bioconversion of gibberellin to inactive forms. PgCYP736A12 protein localized to the cytoplasm and nucleus. Conclusion: PgCYP736A12 does not respond to the well-known secondary metabolite elicitor jasmonic acid, which suggests that it may not function in ginsenoside biosynthesis. Heterologous overexpression of PgCYP736A12 reveals that this gene is actually involved in herbicide metabolism.

• Journal of Plant Biotechnology
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• v.46 no.3
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• pp.172-179
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• 2019

The aim of this study is to evaluate the effect of yeast extract (YE) and salicylic acid (SA) on the expression of curcuminoid-biosynthesis genes (CzDCS and CURS1-3), and accumulation of curcumin in Curcuma zedoaria cell cultures. The results showed that, in cells treated with YE or SA, the expression levels of curcuminoid genes were 1.14- to 3.64-fold higher than the control (untreated cells), in which the YE exhibited a stronger effect in comparison with SA. Curcumin accumulation also tended to be similar to gene expression, curcumin contents in YE- or SA-treated cells were 1.61- to 2.53-fold higher than the control. The SA treatment at the fifth day of culture stimulated the curcumin accumulation and expression in all four genes compared to that at the beginning. While the YE treatments gave different results, the CzCURS1 and CzCURS3 genes were expressed strongly in cells that were treated at the beginning. However, the CzDCS and CzCURS2 genes showed the opposite expression pattern, they were activated strongly in the treatments at day five of the culture. However, the content of curcumin reached its maximum value on the fifth day of culture in all investigations.

• Molecules and Cells
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• v.44 no.11
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• pp.830-842
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• 2021

When perceiving microbe-associated molecular patterns (MAMPs) or plant-derived damage-associated molecular patterns (DAMPs), plants alter their root growth and development by displaying a reduction in the root length and the formation of root hairs and lateral roots. The exogenous application of a MAMP peptide, flg22, was shown to affect root growth by suppressing meristem activity. In addition to MAMPs, the DAMP peptide PEP1 suppresses root growth while also promoting root hair formation. However, the question of whether and how these elicitor peptides affect the development of the vascular system in the root has not been explored. The cellular receptors of PEP1, PEPR1 and PEPR2 are highly expressed in the root vascular system, while the receptors of flg22 (FLS2) and elf18 (EFR) are not. Consistent with the expression patterns of PEP1 receptors, we found that exogenously applied PEP1 has a strong impact on the division of stele cells, leading to a reduction of these cells. We also observed the alteration in the number and organization of cells that differentiate into xylem vessels. These PEP1-mediated developmental changes appear to be linked to the blockage of symplastic connections triggered by PEP1. PEP1 dramatically disrupts the symplastic movement of free green fluorescence protein (GFP) from phloem sieve elements to neighboring cells in the root meristem, leading to the deposition of a high level of callose between cells. Taken together, our first survey of PEP1-mediated vascular tissue development provides new insights into the PEP1 function as a regulator of cellular reprogramming in the Arabidopsis root vascular system.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2022.09a
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• pp.99-99
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• 2022

Artemisia fukudo is a biennial plant and has been reported to have anticancer, anti-melanogenesis, and anti-inflammatory effects. However, it is difficult to produce biomass from A. fukudo, so it is not used as a material for cosmetics or pharmaceuticals. In vitro culture can stably produce biomass throughout the year. In this study, the culture system for producing the highest biomass and bioactive substances was compared. Ex vitro plants were collected in Pyoseon-eup, Jeju island in May 2021, and in vitro culture was harvested after culturing for 8 weeks (plantlet) and 4 weeks (adventitious roots), respectively. After harvest, total polyphenol content (TPC), total flavonoid content (TFC), and DPPH scavenging activity were analyzed. In biomass production, adventitious roots (FW: 5.1 g·100 ml^-1, DW: 0.6 g·100 ml^-1) were about 4 times higher than that of plantlets (FW: 1.8 g·200 ml^-1, DW: 0.3 g·200 ml^-1). Both TPC and TFC were highest in ex vitro plants (9.2 ㎍·mL^-1, 31.6 ㎍·mL^-1), and were 3.0 times and 1.8 times higher than those of plantlets (3.0 ㎍·mL^-1, 17.8 ㎍·mL^-1), respectively. The IC₅₀ value of DPPH scavenging activity was also the best in ex vitro plants (69.8 ㎍·mL^-1), followed by root root (184.4 ㎍·mL^-1) and plants (325.3 ㎍·mL^-1) in that order. Through additional elicitor treatment, scale-up, and advanced compounds analysis such as HPLC, it can be used as an industrial material.

• Mycobiology
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• v.51 no.4
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• pp.230-238
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• 2023

Glabridin is a well-known active isoflavone found in the root of licorice (Glycyrrhiza glabra L.) that possess a wide range of biological activity. Plant cells, hairy roots, and fungal endophytes cultures are the most important alternative methods for plant resources conservation and sustainable production of natural compounds, which has received much attention in recent decades. In the present study, an efficient culture condition was optimized for the biomass accumulation and glabridin production from fungal endophyte Aspergillus eucalypticola SBU-11AE isolated from licorice root. Type of culture medium, range of pH, and licorice root extract (as an elicitor) were tested. The results showed that the highest and lowest biomass production was observed on PCB medium (6.43 ± 0.32 g/l) and peptone malt (5.85 + 0.11 g/l), respectively. The medium culture PCB was produced the highest level of glabridin (7.26 ± 0.44 mg/l), while the lowest level (4.47 ± 0.02 mg/l) was obtained from the medium peptone malt. The highest biomass (8.51 ± 0.43 g/l) and glabridin (8.30 ± 0.51 mg/l) production were observed from the PCB medium adjusted with pH = 6, while the lowest value of both traits was obtained from the same medium with pH = 7. The highest production of total glabridin (10.85 ± 0.84 mg/l) was also obtained from the culture medium treated with 100 mg/l of the plant root extract. This information can be interestingly used for the commercialization of glabridin production for further industrial applications.

• KSBB Journal
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• v.19 no.6 s.89
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• pp.471-477
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• 2004

Oldenlandia diffusa is a Chinese medicinal herb with antitumor activity capable of suppressing the growth of some cancer cell lines. Oleanolic acid and ursolic acid are triterpenoid compounds that exist in Oldenlandia diffusa. Recently, these have been noted for anti-inflammatory, anti-cancer, and hepato-protective effects. Application of both plant growth regulators, 2,4-D and kinetin, was found to be essential for the initiation of callus and suspension cells. Leaf blades of Oldenlandia diffusa was transformed into callus on Schenk and Hildebrandt medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L kinetin, while optimum initiation condition for suspension cells of Oldenlandia diffusa was determined to be 0.75 mg/L 2,4-D and 0.1 mg/L kinetin. Chromatographic separation of oleanolic acid from its derivatives was achieved using Rexchrom S5-100-ODS column. Analytical conditions for oleanolic acid were determined as follows: flow rate at 1.0 mL/min, UV length at 200 nm and mobile phase of $80\%$ acetonitrile and $20\%$ water. Production of secondary metabolites was found to be increased by the treatment with elicitors or signal transducers. The maximum production of oleanolic acid was 99.6 mg/L in cultures with 0.5 mM salicylic acid. It is 1.74 times higher than that of control.

• KSBB Journal
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• v.11 no.4
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• pp.411-419
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• 1996

In this research, an extractive production system for alkaloids, where production and some degree of separation occur simultaneously, was developed in a way that the fast removal of alkaloid produced from the suspension cultures was done by capturing alkaloid with cyclodextrins. The alkaloid production was substantially enhanced up to 40 fold when the solid cultures of E. califonica cells treated with ${\beta}$-cyclodextrin compared to the control. The enhancement of alkaloid production was also observed in the suspension cultures. Interestingly, the production pattern seemed to change when the cultures were treated with ${\beta}$-cyclodextrin so that the major part of the alkaloids in the treated cultures was present in the medium, while the non-treated cultures produced the alkaloids intracellularly. ${\beta}$-cyclodextrin was the most effective one in terms of the alkaloid production among the cyclodextrilns(${\alpha}$-cylodextrin, ${\beta}$-cyclodextrin and ${\gamma}$-cyclodextrin) tested in the suspension cultures. ${\beta}$-cyclodextrin showed no adverse effect on the cell growth. The most effective concentration of ${\beta}$-cyclodextrin was observed around 1.5% (w/v) in the suspension cultures. The formation of the inclusion complex of the alkaloids with ${\beta}$-cyclodextrin in the suspension cultures was confirmed by detecting the shift of UV absorbance from 274 nm to 282 nm with a UV spectrophotometer.

• Korean Journal of Medicinal Crop Science
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• v.26 no.3
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• pp.227-232
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• 2018

Background: Angelica gigas Nakai is a perennial herb belonging to the family Umbelliferae. Its roots are utilized in traditional medicine. The aim of this study was to increase the yield of and the content of two indicator components (decursin and decursinol angelate) in A. gigas Nakai. Methods and Results: The roots of A. gigas Nakai were harvested 4-times from late August to late October in 2017. Two agents (trace element-TE, and plant hormone-HM) were applied 4 times at intervals of 2 weeks. The content of the two indicator components were analyzed by high performance liquid chromatography. The HM treatment showed the greatest increase in underground part yield and root diameter. The content of the two indicator components in the control (non-treatment) group was the highest in the underground part, but was higher in the aerial parts in the agent treatment group. After treatment with the agents, the content of the indicator components tended to decrease in the underground part. However, the total content of the indicator components in the two agent treatment groups exceeded the level of 6% set by the Korean Pharmacopoeia. Conclusions: The highest underground part yield was found in the HM treatment group, while the highest content of decursin and decursinol angelate were found in the control group. This study provides basic information for yield improvement in A. gigas.

• Journal of Bio-Environment Control
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• v.28 no.2
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• pp.158-165
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• 2019

The effects of microbubbles on glucosinolate accumulation and growth of watercress (Nasturtium officinale R. Br.) were investigated. Watercress plant at the 4th mature leaf stage (2 weeks old) were exposed to microbubbles or non-microbubbles generated in an Otsuka-house nutrient solution for 3 weeks in a controlled environment culture room. Stem length of the watercress grown under the microbubbles was 41% shorter than that of the non-microbubbles, showing significantly different. However, shoot fresh and dry weights, root length, leaf length, leaf width, SPAD, and quentum yield of the watercress were not significantly different between treatments. Glucoiberin, glucobrassicin, gluconapin, gluconasturtiin of the watercress grown under microbubbles, excepted for 4-methoxyglucobrassicin, were significantly higher than those of the watercress grown in non-microbubbles. In addition, watercress grown under microbubbles for 3 weeks contained 85% (${\mu}mol/g$ DW) and 65% (${\mu}mol/plant$) more total glucosinolate, respectively. Results indicated that microbubbles generated in a deep flow technique hydroponics system could increase the accumulation of glucosinolate without growth reduction.

• Journal of Bio-Environment Control
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• v.27 no.1
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• pp.94-101
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• 2018

Mugwort (Artemisia princeps) is a medicinal plant that has a substance called euphatilin, which is effective for cell damage and gastritis recovery. The objectives of this study were to investigate the annual growth characteristics of Artemisia princeps in greenhouse and to increase the eupatiline content by environmental stresses. Growth and eupatilin content of the plants were compared after 6 weeks of seedling and subsequent 8 weeks of greenhouse cultivation. Photosynthesis of mugwort plants did not saturate even at a relatively high light intensity of $1,200{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$. Growth rate of the plants reached its highest at two weeks after transplanting and began to decrease since 8 weeks after transplanting. The plants showed typical characteristics of a perennial herbaceous plant as they were sensitive to seasonal changes. In particular, the plants showed high growth and eupatilin content in spring and summer as vegetative growth periods, but flowering and wintering caused considerable decreases in growth and eupatilin content in fall and winter. Therefore, application of night interruption is essential for year-round cultivationof the plant. Two stresses and a elicitor were treated: drought stresses by stopping irrigation at 5, 6, 7, and 8 days before harvest; salt stresses with nutrient solution concentrations of 2, 4, 6, 8, and $10dS{\cdot}m^{-1}$ by adding sodium chloride at 3 days before harvest; and foliar applications of methyl jasmonates of 12.5, 25, 50, and $100{\mu}M$ at 3 days before harvest. Significant increase in eupatilin content was observed at drought stresses of 7- and 8-days of irrigation stop and foliar application of $25{\mu}M$ methyl jasmonate, while no significant increase observed at salt stresses. From the results, it was confirmed that the environmental treatments can improve the productivity and quality of Artemisia princeps as a phamaceutical raw material.