• Applied Biological Chemistry
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• v.38 no.5
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• pp.387-392
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• 1995

To study the regulatory expression mechanism of soybean glycinin gone, Gy2, the 5' upstream region of the gene was searched for the presence of putative regulatory elements by nucleotide sequencing. It revealed various kinds of regulatory sequence elements commonly found in plant storage protein genes. There were canonical promoter sequences, TATA box (TATAAT) and AGGA box (GAAT) which are common in the 5' upstream region of the plant genes. The embryo factor binding sequence, RY repeat, CACA sequences, ${\alpha}$-conglycinin enhancer-like sequences were also found. To delineate the function of these sequences, 5' upstream deletion mutants of Gy2 were prepared and fused to the ${\alpha}$-glucuronidase (GUS) gene. Each chimeric construct was transferred into soybean protoplasts for transient assay, which led to the identification of the sequences between -281 and -223, -170 and -122, of Gy2 promoter as negative regulatory elements, and the sequences between -223 and -170, -122 and -16 as positive regulatory elements. These results are consistent in transformed tobacco plants as well. The serially deleted promoter fragments fused to the GUS were transformed into Nicotiana tabacum by Agrobacterium tumefaciens using the binary vector system. GUS activity of Gy2 promoter deletion constructs was detected only in seeds but not in leaves with different levels of expression as in transient assay. These results suggest that the glycinin Gy2 promoter drives a tissue-specific expression in transgenic tobacco plants.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.34 no.2
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• pp.198-203
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• 1989

Field experiment was conducted to find out the effect of lime materials application on yield, and chemical composition of Burley 21 in 1986. Lime materials and application rate were CaSO$_4$ : Ca 35kg/l0a. CaCO$_3$ : Ca 35kg/l0a and Liming: pH6.5. Contents of Ca and Mg in fresh or cured leaves were increased, but AI, Fe and Mn were decreased by applying lime materials. Yield were increased by applying lime materials. Among lime materials treatment, yield of limed and CaCO$_3$ plot were higher than that of CaSO$_4$ plot. Value per kg of cured leaves was not affected by applying lime materials. Cured leaves of CaSO$_4$ plot contained higher NH$_3$-nitrogen and alkalinity number of water insoluble ash than those of unlimed plot. Cured leaves of CaCO$_3$, plot contained higher alkalinity number of water soluble ash, insoluble ash and volatile neutral constituent, but lower protein-nitrogen and petroleum ether extracts than those of unlimed plot. Cured leaves of limed plot contained higher alkalinity number of water soluble and insoluble ash, NO$_3$-nitrogen and volatile neutral constituents, but lower protein -nitrogen, nicotine and petroleum ether extracts than those unlimed plot. Yield was increased, however leaf Quality in respect to chemical and organoleptic characteristics were not affected considerably by applying lime materials. Therefore, it suggests that controlling the soil pH about 6.5 by liming might be necessary for tobacco cultivation.

• Journal of Life Science
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• v.13 no.1
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• pp.83-89
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• 2003

To investigate the function of chloroplast small heat shock protein (HSP), transgenic tobacco plants (Nicotiana tabacum L, cv. SR-1) that constitutively overexpress the rice chloroplast small HSP (Oshsp26) were generated. Effects of constitutive expression of the Oshsp26 on thermotolerance were investigated with the chlorophyll fluorescence. After 5-min incubation of leaf discs at high temperatures, an increase in the Fo level, indication of separation of LHCII from PSII, was mitigated by constitutive expression of the chloroplast small HSP When tobacco plantlets grown in Petri dishes were incubated at $20^{\circ}C$/TEX> for 45 min and subsequently incubated at $20^{\circ}C$/TEX> leaf color of wild-type plant became gradually white and all plantlets were finally died. Under the conditions in which all the wild-type plants died, more than 80% of the transformants remained green and survived. It was also found that the levels of Oshsp26 protein accumulated in transgenic plants were correlated with the degree of thermotolerance. These results suggest that the chloroplast small HSP plays an important role in protecting photosynthetic machinery, as a results, increases thermotolerance of whole plant during heat stress.

• Analytical Science and Technology
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• v.28 no.2
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• pp.98-105
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• 2015

The development of an enzymatic biosensor for dopamine determination based on multiwall carbon nanotubes (MWCNTs) and peroxidase obtained from the crude extract of burley tobacco (Nicotiana tabacum L.) was proposed. Peroxidase catalyzes the oxidation of dopamine to dopamine quinone. The influence on the response of analytical parameters of biosensors such as enzyme concentration, dopamine concentration, pH, and phosphate buffer solution concentration were investigated. The analytical parameters obtained, including sensitivity, linearity, and stability, were investigated. The proposed method for dopamine determination presented good selectivity even in the presence of uric acid and ascorbic acid. The sensor presented a higher response for dopamine in 0.010 M phosphate buffer at pH 6.50, with an applied potential of -0.15 V. The detection limit of the electrode was 2.7×10⁻⁶ M (S/N = 3) and the relative standard deviation of the measurements, which were repeated 10 times using 5.0×10⁻² M dopamine, was 1.3%.

• Journal of Plant Biotechnology
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• v.4 no.4
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• pp.143-150
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• 2002

A strong oxidative stress-inducible peroxidase promoter (referred to as SWPA2 promoter) was cloned from tell cultures of sweetpotato (Ipomoea batatas) and characterized in transgenic tobacco cultured cells in terms of biotechnological applications. Employing a transient expression assay in tobacco protoplasts, with five different 5'-deletion mutants of the SWPA2 promoter fused to the $\beta$-glucuronidase (GUS) reporter gene, the 1314 bp deletion mutant showed approximately 30 times higher GUS expression than the CaMV 35S promoter. The expression of GUS activity in suspension cultures of transgenic cells derived from transgenic tobacco leaves containing the -1314 bp SWPA2 promoter-GUS fusion was strongly expressed following 15 days of subculture compared to other deletion mutants, suggesting that the 1314 bp SWPA2 promoter will be biotechnologically useful for the development of transgenic cell lines engineered to produce key pharmaceutical proteins. In this respect, we developed transgenic cell lines such as tobacco (Nicotiana tabacum L. BY-2), ginseng (Panax ginseng) and Siberian ginseng (Acanthopanax senticosus) using a SWPA2 promoter to produce a human lactoferrin (hLf) and characterized the hLf production in cultured cells. The hLf production monitored by ELISA analysis in transgenic BY-2 cells was directly increased proportional to cell growth and reached a maximal level (up to 4.3% of total soluble protein) at the stationary phase in suspension cultures. The SWPA2 promoter should result in higher productivity and increased applications of plant cultured cells for the production of high-value recombinant proteins.

• Journal of the Korean Society of Tobacco Science
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• v.9 no.2
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• pp.3-9
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• 1987

This study was been conducted to determine chemical characteristics of soils in the major districts cultivating flue-cured tobacco plant. Also native soil productivities were measured by means of bioassay planting tobacco plant without fertilizer at 87 selected lolls through pot and field experiments. Inorganic nutrient in soils affecting the dry weight of tobacco leaves cultivated in the field were investigated. The results obtained are summarized as follows; 1. Among soil chemical characteristics, pH, $NO_3$-N, $NH_4$-N , $P_2O_5$, and Mg Influenced significantly the dry weight of tobacco loaves In pot experiment, whole In the field experiments, pH, $NO_3$-N, $NO_3$-N+$NH_4$-N, and Ca had influence. 2. Correlation coefficients between soil chemical characteristics and dry weight of tobacco leaves were higher in pot experiment than field. The results revealed that soil morphological characteristics might more close influence on dry weight of tobacco leaves than chemical characteristics. 3. For prediction of dry weight (Productivity) of tobacco leaves without fertilizer multiple regression analysis were introduced using troll chemical characteristics. A combination of pH, $NO_3$-N, and Ca was very reliable for prediction of productivity as equation. y=5.02+18.07$x_1$ +2.61$x_2$ +5.36$x_3$ R=0.444** Where $x_1$ : pH, $x_2$ : $NO_3$-N, $x_3$:Ca

• Korean Journal of Environmental Biology
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• v.19 no.3
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• pp.211-217
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• 2001

Salicylate is involved in the induction of pathogen-related proteins and plant defense response. The effects of salicylate on the activity isoperoxidase $A_3$ from tobacco callus (Nicotiana tabacum L.) and the protection against the enzyme inactivation by salicylate in the presence of $Fe^{2+}$ were examined. About 20% and 85% activity losses of peroxidase occurred at 0.48 mM and 0.6 mM salicylate, respectively, showing that isoperoxidase $A_3$ was inactivated by salicylate. The inactivation occurred depending on pH and showed noncompetitive inhibition mode. Moreover, inactivation of the enzyme by salicylate was completely protected in the presence of $Fe^{2+}$. Apoperoxidase without heme moiety was constructed and the effects of various metal ions on the recovery of enzyme activities were investigated. More than 80% of the activity was reconstituted by the addition of $Fe^{2+}$ or hemin. However, the enzyme activity was not recovered by $Cu^{2+},\;Zn^{2+},\;Co^{2+},\;or\;Mn^{2+}$.

• Korean Journal of Soil Science and Fertilizer
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• v.22 no.3
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• pp.215-220
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• 1989

For development of slow releaseing K-fertilizer using natural zeolite, two kinds of K adsorbed zeolite were made by ion exchange reaction in $K^+$ saturated aqueous solution. K-Zeolite was prepared by removing the excess salt after K adsorption on zeolite, however, NW. K-Zeolite was prepared only by K adsorption reaction without removing the excess salt. The adsorbed K on zeolite was not easily released, but continuously and slowly released from the surface of zeolite. By the results of application for tobacco cultivation, the K contents in soil treated with K-Zeolite and NW. K-Zeolite were lower than that of control (20kg/10a) at the early growth stage, but were higher at the late growth stage. Therefore, the results suggest that K adsorbed zeolite could be applied as a slow-releasing K fertilizer.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.44 no.3
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• pp.263-266
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• 1999

Tobacco (Nicotiana tabacum L. ‘KF109’) seeds were primed in polyethylene glycol 6000 (PEG 6000) solutions to determine a) what osmotic potential of the solution would be optimal for priming, i.e., critical potential level for preventing germination, and b) what temperature and duration would be the most effective in priming. The germination was completely prevented below -0.8 MPa of PEG 6000, that indicates a optimum water potential for seed priming. Seeds were primed for 0, 1, 2, 3, 5, 10 and 15 days at 15, 20 and $25^{\circ}C$, respectively, under the-0.8 MPa PEG 6000 solution to find out the most effective temperature and duration for priming. The effectiveness of priming, particularly in germination speed, was observed more distinctly when the primed seeds were germinated at 15$^{\circ}C$ than 2 5$^{\circ}C$. The greatest reduction of the time to 50% germination (T/sob 50/) was when the seeds were primed at $25^{\circ}C$. The reduction rate of the $T_{50}$ was rapid when primed from 1 day to 8 days and then slowed down in the seeds primed for longer than 8 days. The time from 10 to 90% germination ( $T_{10-90}$ increased in the primed seeds for longer than 8 days which showed the reversed effect of synchronous germination. However, $T_{50}$ was reduced continuously in the seeds even primed over 8 days. Thus, the optimum condition for tobacco seeds priming with PEG 6000 solution was -0.8 MPa in osmotic potential of the solution at $25^{\circ}C$ for 8 days.ays.

• Current Research on Agriculture and Life Sciences
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• v.17
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• pp.15-20
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• 1999

To investigate the function of the chloroplast small heat shock protein (small HSP), transgenic tobacco plants (Nicotiana tabacum L., cv. SRI) that show constitutive expression of the chloroplast small HSP were generated. Effects of constitutive expression of the introduced gene on thermotolerance were first probed with the chlorophyll fluorescence. After a 5-min incubation of leaf discs at high temperatures, an increase in the Fo level and a decrease in the Fv level, indications of separation of LHCII from PSII and inactivation of electron transport reactions in PSII, were mitigated by constitutive expression of the small HSP. When tobacco plantlets grown in Petri dishes were incubated at $52^{\circ}C$ for 45 min and subsequently incubated at $25^{\circ}C$, leaf color of nontransformants was gradually became white and all plantlets finally were died. Under conditions in which all nontransformants were dying, more than 80% of the transformants remained green and survived. These results suggest that the chloroplast small HSP plays an important role in protecting photosynthetic machinery during heat stress.