• Korean Journal of Plant Tissue Culture
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• v.27 no.2
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• pp.133-136
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• 2000

To evaluate the biological effects of dykellic acid, a novel apoptosis inhibitor, isolated from microorganism on the plant cells, the cell growth, protein contents, and superoxide dismutase (SOD) activity were investigated in suspension cultures of tobacco photomixotrophic cultured (PM) cells on 12 days after different concentration of chemical treatment. The cells were cultured in MS medium containing 0.7 mg/L 2,4-D, 0.3 mg/L kinetin, 30 g/L sucrose and 200 mM NaCl at $25^{\circ}C$ in the light (100 rpm). Dykellic acid strongly inhibited the cell growth by evaluating the cell fresh wt and the ion conductivity in the medium ($IC_{50}$/, about 20 $\mu$M). The results as inhibition of cell growth and cell wall damage were same. The compound significantly increased the protein contents and the SOD specific activity in proportion with the dosage. The results suggested that dykellic acid may have biological activity in plant cells and tobacco PM cells may be suitable biomaterials for in vitro evaluation of the biological activity of natural products.

• Journal of Bio-Environment Control
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• v.18 no.1
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• pp.23-28
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• 2009

This study was conducted to evaluate the effect of optimized environment conditions with growth stage in photoautotrophic micropropagation on the growth of potato (Solanum tuberosum L. cv. Dejima) plantlets and energy efficiency. Optimum environment conditions at each stage were decided in our previous study. For the evaluation of optimized environment control, potato plantlets were cultured under four different conditions: photoautotrophic optimum conditions of photosynthetic photon flux density (PPFD) and $CO_2$ levels with growth stage (POG), photoautotrophic constant condition with average PPFD and $CO_2$ levels (PCA), photoauototrophic constant condition with maximum PPFD and $CO_2$ levels (PCM), and photomixotrophic conventional condition with 3% sucrose (PMC) as control. As a result, environment control with growth stage (POG) significantly promoted all the growth characteristics such as the number of nodes and unfolded leaves, shoot height, shoot diameter, and fresh and dry weights of potato grown in vitro. In addition, based on dry weight consumed electricity and $CO_2$ were the lowest in POG suggesting the highest energy efficiency among the treatments. After transferring potato plantlets to greenhouse, the plantlets under POG showed vigorous growth, which was pretty similar with those under PMC. The accumulations of dry matter in POG were 4.7 times in vitro and 3.8 times in greenhouse as much as those in the conventional control (PCM). Thus, we concluded that in vitro environment control with growth stage induced vigorous growth of potato plantlets both in vitro and in greenhouse with less energy consumption.

• Korean Journal of Plant Tissue Culture
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• v.26 no.3
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• pp.183-187
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• 1999

To establish an efficient screening system for new herbicides using plant cultured cells, responses of tobacco photomixotrophic cultured (PH) cells to various herbicides with different modes of action were surveyed by measuring the cell growth and ion conductivity in medium. The cells were cultured in Murashige and Skoog (MS) medium containing 0.7mg/L 2,4-D, 0.3mg/L kinetin and 30 g/L sucrose at $25^{\circ}C$ in the light (100 rpm). Chemicals were treated to suspension cultures of tobacco PH cells at the time of subculture. The cell growth and ion conductivity in the medium were investigated on 12 days after chemical treatment. The ion conductivity assay gave well correlated results to the cell growth inhibition data. The responses of tobacco PM cells were dependent on the modes of action of chemicals tested. Atrazine, an inhibitor of photosynthetic electron transport (PET), strongly inhibited both the cell membrane and cell growth ($IC_{50}$/, about 1 $\mu$M). Butachlor (an inhibitor of cell division), glufosinate (an inhibitor of amino acid biosynthesis), and fluridone (an inhibitor of carotenoid biosynthesis) showed a dose-dependent inhibition. However, Quinclorac, a herbicide with an auxin activity, did not affect the cell growth and ion leakage. These results suggested that tobacco PM cells is suitable materials for the simple screening of new herbicides such as PET, amino acid biosynthesis, ceil division inhibitors by measuring the cell growth and ion conductivity.

• Journal of Bio-Environment Control
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• v.13 no.1
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• pp.51-55
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• 2004

The most commonly used inorganic nutrient compositions such as Murashige & Skoog medium have been optimized for heterotrophic growth. Therefore, they may not be optimal for photomixotrophic and photoautotrophic growth of plantlets. In photomixotrophic micropropagation, emdium sugar level is often lowered, while light and $CO_2$ levels in vessel are raised, and chlorophyllous explants are used to facilitate photosynthetic carbon acquisition. In a factorial experiment effect of addition (+) and omission(_) of organic materials (OM, 0.5 g ${\cdot}$ $m^{-3}$ each of thiamine, nicotinic acid and pyridoxine and 100 ${\cdot}$ $m^{-3}$ myo-inositiol) combined with three sucrose levels (0, 15, and 30 kg ${\cdot}$ $m^{-3}$) was tested on the growth of potato plantlets. Each of nodal cuttings with a leaf was cultured on 0.1${\times}$$10^{-4}m^{-3}$) MS agar ( 8 kg ${\cdot}$ $m^{-3}$) medium (pH 5.80 before autoclave) in glass test tubes (100 mm${\times}$25mm) capped with a sheet of transparent film with a 6 mm diameter gas permeable filter (5.1 air exchanges ${\cdot}$$h^{-1}$). Cultures were maintained in a room for 27 days at $23^{\circ}C$, 50% RH, 350-450${\mu}mol\;{\codt}\;mol^{-1}CO_2$, 16 h ${\cdot}$ $d^{-1}$ photoperiod at 13${\mu}mol\;{\codt}\;m\;{\codt}\;s^{-1}$ PPFD provided by white cool fluorescent lamps. Growth of potato plantlet in the +OM and -OM treatments were similar, while medium pH was 0.2 scale lower in the latter. Dry weight, % dry matter, and stem diameter enhanced, while shoot to root dry weight ratio, leaf area, chlorophyll concentration per gram dry weight, and medium pH decreased with increasing initial sucrose level. Interaction between OM and sucrose levels was observed in shoot length and medium pH. Results indicate that OM can be omitted from the medium without detrimental effect while addition of sucrose was beneficial for the photomixotrophic growth of potato plantlets under raised light and $CO_2$.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2002.11a
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• pp.29-30
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• 2002

The goal of this research was to develop the effectiveness of in vitro culture method for A. formosanus and study the environment in vitro conditions affecting on growth. The first series of experiments were examined to investigate the response of three different basal media, MS (Murashige and Skoog, 1962), Knudson (KC; Knudson, 1946) and modified hyponex on growth and multiplication during in vitro culture. Multiple shoot proliferation was induced in shoot tip explants on Hyponex (H3) media supplemented with BA (1 mg1$\^$-1/) or TDZ (1-2 mg1$\^$-1/). Addition of activated charcoal (1%) to the TDZ containing medium promoted rapid shoot tip proliferation (11.1 shoots per explant) but the same medium had an opposite effect resulting in poor proliferation in the nodal explants. However, the regenerated shoots had slow growth rate and failed to elongate. This problem was overcome by transferring the shoot clumps to a hormone free H3 media supplemented with 2% sucrose and 0.5% activated charcoal. Using bioreactor culture for scaling up was also shown the best way for multiple shoot induction and growth of this plant. The second series of experiments was studied to investigate the effect of physical environment factors on growth of in vitro plantlets. The Anoectochilus formosanus plantlets were cultured under different air exchange rate (0.1, 0.9, 1.2h$\^$-1/), without sucrose or supplement 20g.1$\^$-1/ (photoautotrophic or photomixotrophic, respectively), and different photosynthesis photon flux (40, 80, 120 ,${\mu}$mol.m$^2$.s$\^$-1/- PPF). Under non-enrichment CO$_2$ treatment, slow growth was observed in photoautotrophical condition as compared with photomixotrophical condition on shoot height, fresh weigh and dry weight parameters; High air exchange (1.2.h-l) was found to be inadequate for plant growth in photomixotrophical condition. On the contrary, under CO$_2$, enrichment treatment, the plant growth parameters were sharply (visibly) improved on photoautotrophic treatments, especially on the treatment with air exchange rate of 0.9.h-1. The growth of plant in photoautotrophic condition was not inferior compared with photomixotrophic, and the best growth of plantlet was observed in treatment with low air exchange rate (0.9.h-1). Raising the PPF level from 80 to 120${\mu}$mol.m$\^$-2/.s$\^$-1/ decreased the plant height, particularly at 120${\mu}$mol.m$\^$-2/.s$\^$-1/ in photoautotrophic condition, fresh weight and dry weight declined noticeably. At the PPF of 120${\mu}$mol.m$\^$-2/,s$\^$-1/, chlorophyll contents lowed compared to those grown under low PPF but time courses of net photosynthesis rate was decreased noticeably. Light quality mainly affected morphological variables, changes of light quality also positively affected biomass production via changes in leaf area, stem elongation, chlorophyll content. Plant biomass was reduced when A. formosanus were grown under red LEDs in the absence of blue wavelengths compare to plants grown under supplemental blue light or under fluorescent light. Stem elongation was observed under red and blue light in the present experiment. Smaller leaf area has found under blue light than with other lighting treatments. Chlorophyll degradation was more pronounced in red and blue light compared with white light or red plus blue light which consequent affected the photosynthetic capacity of the plant. The third series of experiment were studied to investigate the effect of physical environment factors on growth of ex vitro plants including photosynthesis photon flux (PPF), light quality, growing substrates, electrical conductivity (EC) and humidity conditions. In the present experiments, response of plant on PPF and light quality was similar in vitro plants under photosynthesis photon flux 40${\mu}$mol.m,$\^$-2/.s$\^$-1/ and white light or blue plus red lights were the best growth. Substrates testing results were indicated cocopeat or peat moss were good substrates for A. formosanus growth under the greenhouse conditions. In case of A. formosanus plants, EC is generally maintained in the range 0.7 to 1.5 dS.m-1 was shown best results in growth of this plant. Keeping high humidity over 70% under low radiation enhanced growth rate and mass production.

• Journal of Biosystems Engineering
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• v.32 no.2 s.121
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• pp.109-114
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• 2007

In mass production of seed-potato plantlets, the processes for in vitro propagation and ex vitro acclimatization with a high cost should be improved by a culture system with environmental control using scaled-up culture vessels. The experiment was conducted to design a hydroponic culture system for enhancement of growth and development of seed-potato (Solanum tuberosum) plantlets cultured under photoautotrophic (without sugar in culture medium) conditions with controlled light intensity and ventilation rate. The culture system was consisted of scaled-up culture vessels, ventilation pipes, a multi-cell tray and an environmental control system (ECS) for optimum controlling in temperature, light intensity, ventilation rate, and culture-medium supply. Growth and development of the plantlets was significantly increased under the ECS compared with a conventional culture system (CCS) of photomixotrophic culture (with sugar in culture medium) using small scale vessels. For 21 days, leaf area of the plantlets was expanded more than 2 times, and number of internodes also approximately 4 times greate. under the ECS. In addition, the photoautotrophic growth in sweetpotato (Ipomoea batatas) and chrysanthemum (Chrysanthemum morifolium) plantlets was greater more than 2 times compared with the CCS.

• Molecules and Cells
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• v.19 no.2
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• pp.256-261
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• 2005

In silico analysis of genome of the cyanobacterium Synechocystis sp. PCC 6803 identified two genes, slr0329 and sll0593, that might participate in glucose (Glc) phosphorylation (www.kazusa.or.jp/cyano). In order to determine the functions of these two genes, we generated deletion mutants, and analyzed their phenotypes and enzymatic activities. In the presence of 10 mM Glc, wild-type (WT) and slr0329 defective strain (M1) grew fast with increased respiratory activity and NADPH production, whereas the sll0593 deletion mutant (M2) failed to show any of the Glc responses. WT and M1 were not significantly different in their glucokinase activity, but M2 had 90% less activity. Therefore, we propose that Sll0593 plays a major role in the phosphorylation of glucose in Synechocystis cells.

• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.115-119
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• 1999

The single node cuttings of sweet potato (cv. Mokpo #29) plantlets maintained in vitro were cultured with (MF+) or without membrane filter (MF-) under photomixotrophic (PM), hetrotrophic (HT) and autotrophic (AT) conditions. Shoot length was the greatest (11.9cm) in 3$0^{\circ}C$ (HT) treatment and it was the shortest (3.4 cm) in $25^{\circ}C$ (PM) treatment. Nodal explants cultured in 3$0^{\circ}C$ treatment looked more vigorous than those of $25^{\circ}C$ in appearance, and node number was the greatest (10.5 per plantlet) among the treatments. But plantlets grew in 3$0^{\circ}C$ (HT) treatment were observed all overgrown. The size in leaf area was about 2 times greater and shoot length was about 2 times shorter in PM than in HT condition. Percent dry matter of shoots was 5.9% (HT) and 7.4% (PM) in $25^{\circ}C$ treatment and 6.1% (HT) and 7.4% (PM) in 3$0^{\circ}C$ treatment. Plantlets cultured in the MF+ treatments were less succulent than those cultured in the MF- treatment. Vitrified plantlets were examinated 14.8% (both $25^{\circ}C$ and 3$0^{\circ}C$) in PM condition and 22.2% ($25^{\circ}C$) and 31.5% (3$0^{\circ}C$) in HT condition. Sucrose was necessary for the survival of in vitro plantlets. In the sucrose-free medium, explants cultured in the MF- had turned yellow and were dead after 30 days of culture. But explants cultured in the MF+ were alive and produced plantlets with shoot and root (AT). On the other hand, the survival of explants on the MS basal medium (sucrose-free and hormone-free) depended entirely upon the MF attachment.

• Journal of Plant Biotechnology
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• v.7 no.1
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• pp.67-74
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• 2005

To overcome various disadvantages of conventional cul-ture vessels for micropropagation, a novel disposable vessel, the 'Vitron', made of a multi-layered $OTP^{(R)}$ film and supported by a polypropylene frame, was developed. The film possesses superior properties such as: high light transmittance, low water vapor transmittance and thermal stability and in particular, high gas-permeability. Single nodal explants, which were excised from the multiple shoots derived from shoot-tip culture, were cultured in Vitron and polycarbonate vessels on $3\%$ sugar-containing agar on MS medium and placed at 3000 ppm $CO_2$-enrichment at a low photosynthetic photon flux density (PPFD) ($45{\mu}mol\;m^{-2}\;s^{-1}$). The in vitro and ex vitro growth, and the net photosynthetic rate of in vitro and ex vitro plantlets were significantly enhanced in the Vitron compared to those cultured in a polycarbonate vessel. Explants that were cultured on the same MS medium under low PPFD at various $CO_2$ concentrations were also cultured at 3000 ppm $CO_2$- enrichment at various PPFD: 30, 45, 60, 75 and $90{\mu}mol\;m^{-2}\;s^{-1}$. The best in vitro and ex vitro growth obtained for 3000 ppm $CO_2$-enrichment at $75{\mu}mol\;m^{-2}\;s^{-1}$ PPFD. The novel Vitron vessel, when placed under the two conditions, may replace conventional culture vessels for the successful micropropagation of sweetpotato.

• Korean Journal of Plant Tissue Culture
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• v.25 no.4
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• pp.257-261
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• 1998

The repeated subcultures of in vitro plant materials in wasabi became highly vitrified and the capacity for multiple shoot formation from the vitrified plant materials was very low. In order to improve the quality of in vitro propagated planting materials, the experiments were carried out using culture vessels capped with membrane filter(MF). When vitrified shoots were cultured on MS medium with 0.2mg/L BA in the vessels with MF or without MF for 60 days, the shoots in the vessels with MF did not vitrified. In contrast, the shoots grown in the vessels without MF vitrified at 65%. The stomates of vitrified leaves were circular and inflated, whereas those of normal leaves acclimatizated in the vessels with MF were ovate in shape. The hardened shoots were also cultured on MS media without sucrose containing 0.01mg/L IBA in vessels with(photoautotrophic culture) or without(control) MF. Sucrose was necessary for survival of the in vitro plantlets in the vessels without MF. After 20 days of culture, the shoots in the vessels without MF on the sucrose-free media turned yellow and died. But the shoots in the vessels with MF in the sucrose-free media produced a lot of roots. When shoots were cultured on MS medium with 2% sucrose containing 0.01mg/L IBA in the vessels with(photomixotrophic culture) or without(heterotrophic culture) MF, best growth occured in photomixotrophic culture.