• Korean Journal of Plant Tissue Culture
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• v.28 no.6
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• pp.311-315
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• 2001

The relationship among leaf size, leaf protoplast (cell) size, chloroplast size, and chloroplast number were investigated in tobacco and Arabidopsis thaliana at various developmental stages. In tobacco, protoplasts, less than 15.6 ${\mu}{\textrm}{m}$ in diameter had less than 20 chloroplasts, 0.93 ${\mu}{\textrm}{m}$ in thickness and 3.3 ${\mu}{\textrm}{m}$ in length on average. As protoplast size increased from 30 ${\mu}{\textrm}{m}$ to 45 ${\mu}{\textrm}{m}$ in diameter, chloroplast size remained the same (1.57 ${\mu}{\textrm}{m}$ in diameter and 5.55 ${\mu}{\textrm}{m}$ in length on average), but chloroplast number increase from 42 to 101 on average. A similar relationship was also observed in A. thaliana. The ratio of total chloroplast volume to protoplast volume was constant (0.105 in tobacco and 0.325 in A. thaliana) over various developmental stages.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.193-196
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• 2001

IL-18. formerly known as IGIF(interferon -gamma inducing factor), is structurally IL-l related but functionally IL-12 related pro-inflammatory cytokine. The human IL -18(hIL-lS), like IL-$1{\beta}$, is synthesized as a biologically inactive precursor of 24kDa lacking a signal peptide, and then cleaved into an active mature form by cystein protease IL-$1{\beta}$ converting enzyme (ICE: caspase- 1), We tested if the mature hIL -18 can be expressed and secreted into culture medium by transforming the forming gene construct consisting of a mature hIL-18 gene fused to signal peptide of rice amylase lA. Secondly, we were tested if the pro- IL-18 could be processed into a biologically active form by caspase-l like protease in plant. Cell suspension culture was established from the leaf-derived calli of transgenic tobacco plant. Southern and Northern blot analysis indicated the expression of both pro-hIL-18 and mature hIL-18 plant cells. Western blot analysis introduced the protein products of pro- hIL -18 and mhIL -18 were observed in transigenic cell lines. In addition, the molecular size of recombinant pro-hILl-18 and mhIL-18 were estimated to be 24kDa and 18kDa, respectively. ELISA revealed that the amount of pro- hIL -18 was 1.3ug per gram of fresh weight calli. Moreover, the presence of mhIL-18 was detected in the culture medium and it appeared to be 25ug/L.

• Mycobiology
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• v.47 no.1
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• pp.126-133
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• 2019

Isaria javanica pf185 is an important entomopathogenic fungus with potential for use as an agricultural biocontrol agent. However, the effect of I. javanica pf185 on plant growth is unknown. Enhanced tobacco growth was observed when tobacco roots were exposed to spores, cultures, and fungal cell-free culture supernatants of this fungus. Tobacco seedlings were also exposed to the volatiles of I. javanica pf185 in vitro using I-plates in which the plant and fungus were growing in separate compartments connected only by air space. The length and weight of seedlings, content of leaf chlorophyll, and number of root branches were significantly increased by the fungal volatiles. Heptane, 3-hexanone, 2,4-dimethylhexane, and 2-nonanone were detected, by solid-phase micro-extraction and gas chromatography-mass spectrophotometry, as the key volatile compounds produced by I. javanica pf185. These findings illustrate that I. javanica pf185 can be used to promote plant growth, and also as a biocontrol agent of insect and plant diseases. Further studies are necessary to elucidate the mechanisms by which I. javanica pf185 promotes plant growth.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.1
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• pp.12-16
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• 2004

Partitioning of human granulocyte-macrophage colony stimulating factor (hGM-CSF) was achieved in the aqueous two-phase systems (ATPSs) using a crude extract of transgenic tobacco cell suspension culture. This study examined the effects of polyethylene glycol (PEG) molecular weight and concentration and the effects of sodium phosphate concentration in different PEG/sodium phosphate systems on the partition coefficient, K. The best ATPS system was 5% PEG 8,000/1.6 M sodium phosphate after 2 h of incubation at room temperature. In this system, hGM-CSF was partitioned in the PEG-rich phase with a yield of 57.99% and K$\_$hGM-CSF/ of 8.12. In another system, 3% PEG 10,000/1.6 M sodium phosphate, hGM-CSF was also partitioned primarily in the top phase with a yield of 45.66% and K$\_$hGM-CSF/ of 7.64 after 2 h of incubation at room temperature.

• KSBB Journal
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• v.16 no.6
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• pp.568-572
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• 2001

Light is one of the most important environmental factors controlling plant physiology. The human granulocyte-macrophage colony-stimulating factor (hGM-CSF) was produced from cell suspension cultures of transgenic tobacco under different light conditions (24 hr light, 18 hr light/dark cycle, dark). Under 24 hr light condition, cell growth was best and dry cell weight reached 14.4 g/L. Light did not influenced the secretion of total proteins. However, in the dark condition, the ratio of secreted total protein/dry cell weight was 1.5 fold higher than those of ethel conditions. Production of hGM-CSF was highest with 18 hr light condition and reached 496.5 ug/L. In addition, the content of hGM-CSf in secreted total proteins was 1.8 fold higher than that of 24 hr light condition, which is beneficial for the purificationof the protein.

• Journal of Plant Biotechnology
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• v.31 no.2
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• pp.163-167
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• 2004

The effects of sucrose concentration on the secretion of hGM-CSF, total protein and protease into the medium were investigated in transgenic tobacco cells. The dry cell weight (11.22 g/L), hGM-CSF (181.53 $\mu\textrm{g}$/L) and total protein (66.8 mg/L) were detected as highest at 30 g/L sucrose and protease activity (2660 U/L) was highest at 120 g/L sucrose after 5-day culture. However after 10-day culture, the maximum dry cell weight (28.36 g/L) was found at 60 g/L sucrose while the maximum hGM-CSF (95 $\mu\textrm{g}$/L) was appeared at 150 g/L sucrose. The total protein and protease activity was secreted as 52.28mg/L and 3430 U/L, respectively in the same culture.

• Journal of the Korean Society of Food Culture
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• v.18 no.2
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• pp.134-138
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• 2003

Quality stability of the herb pill coated with edible oils containing rosemary was investigated. Herb pills were made of herb powders such as Panax ginseng, Cinnamomum cassia, Lycium chinense, Zyzyphus jujuba and Zingiber officinale. Rapeseed oil and lubriol were used as edible coating oil. After herb pills coated with edible oils with or without rosemary were stored at $40^{\circ}C$ for 180 days, the microbial viable cell counts and peroxide values(POV) of the herb pill were investigated. After 180 day storage, POVs of herb pills with only rapeseed oil or lubriol were 0.51 and 0.49 meq/kg, respectively. However, when rosemary was added in herb pills the POVs were decreased to 0.30 and 0.39 meq/kg, respectively. The addition of rosemary to the rapeseed oil and lubriol tended to decrease the microbial viable cell counts of the herb pill. The microbial viable cell counts of rapeseed oil and lubriol were 940 and 820CFU/g, respectively after 180 days of storage. However, these levels were suppressed to 720 and 640CFU/g by the resemary addition. On the other hand, the ginseng saponin content of herb pills was not affected by the rosemary addition during storage.

• KSBB Journal
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• v.22 no.4
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• pp.185-190
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• 2007

Productivity of secreted recombinant protein depends largely on its stability in the extracellular environment with protease. Most hGM-CSF produced by transgenic tobacco cell cultures and secreted to the medium was confirmed to be rapidly degraded by protease in medium. To increase the productivity, therefore, various protein stabilizers such as gelling agents such as carrageenan and alginate, polymers, polyols, and amino acids have been tested. The stability of hGM-CSF in spent medium without cells was improved by the presence of gelling agents. However, the reason for the enhanced production by the addition of gelling agents may be due to the increased expression level and permeability rather than stability. The addition of DMSO inhibited the cell growth, but improved specific yield. The others were not effective for stability as well as hGM-CSF production.

• Korean Journal of Plant Tissue Culture
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• v.28 no.4
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• pp.179-184
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• 2001

We evaluated a possibility of the use of chloroplast number per guard cell pairs as a measure for ploidy level in the different ploidy levels of tobacco plant (Nicotiana tabacum L. cv. BY-4) . The guard-cell chloroplast numbers of leaves of haploid plant were a half of wild-type plant. Furthermore, the number of chloroplast per guard cell pairs of the leaves of doubled-haploid plant increased in two times compared with that of haploid plant. In addition, the chloroplast number was not changed in the F$_1$ progenies. The change of the chloroplast number by leaf stage was not observed. The results indicate that there is a strong relationship between ploidy level (2x and 4x) and chloroplast number per guard cell pairs. This relationship was also, observed in both in vitro and pot cultured plants. It was determined that the measurement of chloroplast number in guard cells of leaf epidermis is simple to use and less labour intensive, and hence can be considered a practical alternative to the chromosome counting methods or flow cytometry in the tobacco plant.

• Journal of Plant Biotechnology
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• v.48 no.4
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• pp.278-288
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• 2021

The development of bioenergy through biomass has gained importance due to the increasing rates of fossil fuel depletion. Biomass is important to increase the productivity of bioethanol, and production of biomass with high biomass productivity, low lignin content, and high cellulose content is also important in this regard. Inorganic salts are important in the cultivation of biomass crops for the production of biomass with desirable characteristics. In this study, the roles of various inorganic salts in biomass and bioethanol production were investigated using an in vitro tobacco culture system. The inorganic salts evaluated in this study showed dramatic effects on tobacco plant growth. For example, H₂PO₄ substantially improved plant growth and the root/shoot (R/S) ratio. The chemical compositions of tobacco plants grown in media after removal of various inorganic salts also showed significant differences; for example, lignin content was high after Mg²⁺ removal treatment and low after K⁺ treatment and H₂PO₄ removal treatment. On the other hand, NO₃^- and H₂PO₄ treatments yielded the highest cellulose content, while enzymatic hydrolysis yielded the highest glucose concentration ratio 24 h after NH₄⁺ removal treatment. The ethanol productivity after H₂PO₄ removal treatment was 3.95% (w/v) 24 h after fermentation and 3.75% (w/v) after 36 h. These results can be used as the basis for producing high-quality biomass for future bioethanol production.