• Korean Journal of Weed Science
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• v.15 no.2
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• pp.160-165
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• 1995

Anthocyanin formation in callus cultures using Populus alba ${\times}$ Populus glandulosa was evaluated on basal MS medium supplemented with various levels of growth regulators, sucrose and nitrate concentrations. The highest yield of anthocyanin from cultured cells was produced under 5% sucrose, 1/8 strength of nitrate(12.5% of basic concentration) and combination of 1.0 mg/l IAA with 2 mg/l BAP, respectively. The high anthocyanin producing cell line no. 11 was selected among 15 cell lines, showing over 80% cells contained anthocyanin producing cells. From these cells, the highly productive red protoplast was isolated and the highest protoplast yield, $6.7{\times}10^6$ was obtained in enzyme combination IV which is composed of 2.0% cellulase, 0.5% macerozyme and 0.1% pectolyase.

• Korean Journal of Breeding Science
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• v.49 no.3
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• pp.170-179
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• 2017

Plant molecular farming has attracted a lot of attention lately in the field of mass production of industrially valuable materials by extending application of the plant as a kind of factory concept. Among them, protein expression system using rice(Oryza sativa L.) callus is a technology capable of mass culture and industrialization because of a high expression rate of a target protein. This study was carried out to develop an Agrobacterium-mediated transformation system to increase the utilization of rice callus. The transformation efficiency was improved by using the hand when seeds were de-husked for callus induction. Furthermore, we were possible induction of callus from 6 years old seed smoothly. Selection of the callus contained the target gene was required a cultivation period of at least 3 weeks, and the most efficient selection period was after 6 weeks of culture including one passage. This selection was confirmed that the gene was stably inserted into the genomic DNA of the plant cell by the southern blot analysis and progeny test. Such an efficient selection system of rice callus that can be cultured in the long term will be contribute to the industrialization of useful recombinant proteins using rice.

• KSBB Journal
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• v.31 no.4
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• pp.277-283
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• 2016

In this system, rice cells were genetically modified to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) using RAmy3D promoter induced by sugar depletion. Even though the target protein fused with signal sequence peptide, plant cell wall can be a barrier against secretion of recombinant proteins. Therefore, hCTLA4Ig can be trapped inside cell wall or remained in intracellular space. In this study, to enhance the secretion of hCTLA4Ig from cytoplasm and cell walls into the medium, permeabilizing agents, such as dimethyl sulfoxide (DMSO), Triton X-100 and Tween 20, were applied in transgenic rice cell cultures. When 0.5% (v/v) of DMSO was added in sugar-free medium, intracellullar hCTLA4Ig was increased, on the other hand, the secreted extracellular hCTLA4Ig was lower than that of control. DMSO did not give permeable effects on transgenic rice cell cultures. And Triton X-100 was toxic to rice cells and also did not give enhancing permeability of cells. When 0.05% (v/v) Tween 20 was added in rice cell cultures, however, intracellular hCTLA4Ig was lower than that of control cultures. And the maximum 44.76 mg/L hCTLA4Ig was produced for 10 days after induction, which was 1.4-fold increase compared to that of control cultures. Especially, Tween 20 at 0.05% (v/v) showed the positive effect on the secretion of hCTLA4Ig though the decrease of intracellular hCTLA4Ig. Also, Tween 20 as a non-toxic surfactant did not affect the cell growth, cell viability and protease activity. In conclusion, secretion of hCTLA4Ig could be increased by enhancing permeability of cells regardless of the cell growth, cell viability and protease activity.

• The Journal of the Convergence on Culture Technology
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• v.7 no.4
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• pp.721-726
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• 2021

Plant cells have a totipotencial capacity, the ability of each cell to produce a new complete individual through development. By applying this, several technologies are being developed for widespread application of somatic embryogenesis by processing hormones in vitro as a method of propagation of plants. In order to use this technology, in Kalanchoe pinnata, a plant capable of asexual reproduction with more regular cell division, kinetin belonging to cytokinin and picloram among hormones belonging to auxin were added in combination and treated for 8 weeks, and then the typical performance was evaluated. As a result of our experiment, the rooting effect in leaf slices showed a 70% incidence rate at a picloram concentration of 0.1 mg/L. It has been proven that a concentration difference of 1:5-1:10 in the ratio of kinetin and picloram is effective. It is the experimental result that the effect of auxin is essential for the development of Kalanchoe roots. As for the effect of shooting, the incidence rate was 60% at the picloram concentration of 0.5 mg/L. The kinetin concentration from 0.5 and 1.0 mg/L and has a significant effect on development. It has been proven that the ratio of kinetin to picloram is effective with a concentration difference of 1:1-1:2. These results show that the combination of cytokinin and auxin is crucially important for shooting. It is thought that it can be the basis of a technology for inducing mass proliferation in vitro by inducing direct organogenesis with a combination of hormones.

• Microbiology and Biotechnology Letters
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• v.21 no.4
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• pp.329-335
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• 1993

Ascorbate oxidase activity in various cucumber tissue extracts was highest in young fruit peeling. Cucumber callus was induced from young fruit peeling and callus cell lines were selected for more than 7 months, which porduced high levels of ascorbate oxidase and had a high growth rate. Induction of callus was optimized with Linsmaier-Skoog(LS) medium at 25$^{\circ}C$ in dark phase. Ascorbate oxidase activity reached a maximum at 5 days after transfer to LS basal liquid-medium ant then declined. The enzyme activity in callus cells was stimulated by addition of 10${\mu}$M $CuSO_4$ in the early logarithmic phase of growth. And also, adding 10${\mu}$M $CuSO_4$ at 3rd day 7th day of culture period, ascorbate oxidase activity in callus cells was maintained to high level. Maximum yield of ascorbate oxidase was found at the 25th day by flask shaking culture, but three-fold of ascorbate oxidase activity was obtained at the 16th day by jar fermentation.

• Journal of Microbiology and Biotechnology
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• v.5 no.5
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• pp.280-284
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• 1995

Aspergi11us niger F-580, a potent producer of aminopeptidase M inhibitor, was isolated from the brown spots of plant leaves with a pathological trait. The inhibitory activity was found only in the fungal mat formed by surface culture of Aspergi11us niger F-580, but not in the culture supernatant or cell pellet. The inhibitor was purified from the hot water extract of this fungal mat by using chromatographies on Diaion HP-20, DEAE-cellulose, Sephadex G-l0 and YMC-ODS-AQ columns. The purified inhibitor was analyzed by UV, mass, and NMR spectroscopies, and identified as OF494911, which had been isolated as an aminopeptidase B inhibitor from Penicillium rugulosum OF4949

• Journal of Environmental Science International
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• v.18 no.4
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• pp.435-443
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• 2009

A bacterial strain SY-97 that showed algicidal activity against Cochlodinium polykrikoides was isolated from coastal water of Uljin (eastern coast of Korea) in August, 2005. The isolated strain was identified as Brachybacterium sp. by morphological and biological tests, and analysis of 16S rDNA sequence. The optimal culture conditions for the growth of strain SY-97 were $30^{\circ}C$, initial pH 7.0, and salinity 2.0%. From the result of cell culture insert experiment, Brachybacterium sp. SY-97 is assumed to produce secondary metabolites which have algicidal activity. When 10% culture filtrate of this strain was applied to C. polykrikoides ($1.2{\times}10^4\;cells/m{\ell}$) cultures, 100% of C. polykrikoides cells was destroyed within 15 hours. The released algicides were heat-tolerant to $100^{\circ}C$ and stable in pH $6.0{\sim}10.0$. These results suggest that Brachybacterium sp. SY-97 is potentially useful for controlling outbreaks of C. polykrikoides.

• Korean Journal of Plant Tissue Culture
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• v.21 no.6
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• pp.341-345
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• 1994

The effect of phytohormones on in vitro production of ubiquinone 10 from the callus cultures of Nicotiana tabacum cv Xanthi was investigated. The growth of callus cultures of Xanthi was in proved by addition of NAA and 2,4-D, especially NAA 0.5 mg/L alone, at the light condition. Ubiquinone 10 was detected by HPLC, and confirmed from Xanthi callus cultured on the all of uppermedia. The ubiquinone 10 content in Xanthi tobacco callus cultured on the medium with NAA 0.5 mg/L only was higher than that of other mixed medium with NAA and 2,4-D. However addition of IBA 1 mg/L and NAA 0.5 mg/L to the medium was more effective in promoting ubiquinone 10 formation than that of NAA 0.5 mg/L only As the callus growth of Xanthi was considerabley restrained at concentration of kinetin, Content and production of ubiquinone low as the highest at kinetin 0.5mg/L and 2,4-D 0.5mg/L in the light.

• 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.

• Journal of Life Science
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• v.28 no.8
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• pp.938-944
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• 2018

This study evaluated tyrosinase, elastase inhibitory, and antioxidant activities of fermented Rhododendron micranthum Turcz. extract using a lactic acid bacterium, Lactobacillus rhamnosus. The optimum conditions for fermentation of R. micranthum Turcz. extract were $37^{\circ}C$ and 3% R. micranthum Turcz. extract for 3 days based on the bacterial cell number, total phenolic compounds, DPPH radical scavenging activity, and tyrosinase and elastase inhibitory activity. After culture for 3 days using 3% R. micranthum Turcz. extract, the cell mass of L. rhamnosus reached $5.7{\times}10^9CFU/ml$. The results indicated that R. micranthum Turcz. extract can be used for industrial lactic acid bacteria culture. After fermentation under optimum conditions, the total content of phenolic compounds of the fermented R. micranthum Turcz. extract was 157 GAE mg/ml, and the $IC_{50s}$ of DPPH radical scavenging activity, ABTS radical scavenging activity, and tyrosinase and elastase inhibitory effects were 78.8, 79.8, 329.1, and $449.5{\mu}g/ml$, respectively. The fermented R. micranthum Turcz. extract exhibited 1.2-, 1.3-, 1.5-, 2.4-, and 5.6-fold higher total content of phenolic compounds, DPPH radical scavenging activity, ABTS radical scavenging activity, and tyrosinase and elastase inhibitory effects than the nonfermented R. micranthum Turcz. extract. These results indicated that fermented R. micranthum Turcz. extract using L. rhamnosus can be used for developing new natural functional ingredients for the health food or cosmetic industry.