• Analytical Science and Technology
• /
• v.28 no.2
• /
• pp.98-105
• /
• 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
• /
• v.2 no.2
• /
• pp.79-82
• /
• 2000

We have isolated a cDNA encoding a calcium-dependent protein kinase (CDPK) in Nicotiana tabacum, which was designated NtCDPK1. Accumulation of the NtCDPK1 mRNA was stimulated by various stimuli, including phytohormones, CaCl$_2$ wounding, fungal elicitors, chitin and methyl jasmonate. The NtCDPK1 gene encodes a functional Ser/Thr protein kinase of which phosphorylation activity is strongly induced by calcium. By analyzing expression of the NtCDPK1-GFP fusion protein and by immunoblotting with antibody which reacts with NtCDPK1, we found that NtCDPK1 is localized in membrane and nucleus in plant cells. Silencing expression of the NtCDPK1 transgene resulted in marked decrease of lateral root development in the transgenic tobacco plants. Yeast two hybrid screening using NtCDPK1 as a bait identified a tobacco homologue of proteasome regulatory subunit 21D7, designated Nt21D7. The 21D7 mRNA has been shown to be predominantly expressed in proliferating tissues in the cell cycledependent manner in carrot. The recombinant NtCDPK1 protein associated with Nt21D7 in vitro, and could phosphorylate the Nt21D7 protein in vitro in the presence of calcium, suggesting that Nt21D7 protein is a natural substrate of NtCDPK1 in tobacco. These results suggest that NtCDPK1 may regulate tell proliferation processes, such as lateral root formation, by regulating specificity and/or activity of proteasome-mediated protein degradation pathway.

• Korean Journal of Environmental Biology
• /
• v.19 no.3
• /
• pp.211-217
• /
• 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
• /
• v.22 no.3
• /
• pp.215-220
• /
• 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 Plant Tissue Culture
• /
• v.22 no.3
• /
• pp.149-155
• /
• 1995

The coat protein (CP) gene was cloned from RNA genome of the Cucumber Mosaic Virus strain ABI (CMV-ABI) isolated in Korea. The comparisons of the nucleotide sequence of the cloned CP gene and its deduced amino acid sequences with other CP genes revealed that the CMV-ABI belongs to subgroup I (type I), CMV-ABI developed the typical mosaic symptom in infected plants. Tobacco plants (Samsun and NC82) were transformed by leaf-disc transformation via Agrobacterium, temefaciens LB4404 harboring pVCP, witch CMV-ABI CP gene was inserted into the pBI121, and a number of mature transgenic tobacco plants were developed. Southern and PCR analysis of genomic DNA from the transgenic plants showed that the CP gene was integrated into the genomes of the most of the transgenic plant. Result of the segregation patterns of resistance in T1 seedlings of the plants to kanamycin showed that the transgenic plants containing l,2 and 3 copies of CP gene were50%, 39% and 11% of the total transgenic plants, respectively.

• Korean Journal of Plant Tissue Culture
• /
• v.27 no.6
• /
• pp.441-445
• /
• 2000

In order to improve phosphate use efficiency of rice using phosphate transporter (PT), transgenic rice plants containing a tobacco PT gene were developed. Calli from Dongjinbyeo (Oryza sativa L.) were cocultured with A. tumefaciens LBA 4404 harboring PT gene. Multiplied calli were transferred to MS medium supplemented with 50 mg/L hygromycin, 500 mg/L carbenicillin, 2 mg/L kinetin, 0.1 mg/L NAA. After 2 weeks, hygromycin resistant shoots were obtained from the calli on the selection medium. The putative transgenic shoots were transferred to rooting MS medium supplemented with 250 mg/L cabenicillin. Plant regeneration rate from the calli was about 52%. Stable incorporation of the tobacco PT gene into rice genomic DNA was confirmed by PCR and Southern blot analysis.

• Korean Journal of Environmental Agriculture
• /
• v.26 no.3
• /
• pp.239-245
• /
• 2007

The effect of ultraviolet-B (UV-B) radiation on photosynthesis was studied by the simultaneous measurements of $O_2$ evolution and chlorophyll (Chl) fluorescence in tobacco leaves. When the tobacco leaves were teated with UV-B (1 $W{\cdot}m^{-2}$), the maximal photosynthetic $O_2$, evolution (Pmax; 4.60 ${\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) at 200 ${\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) was decreased with increasing time of UV-B treatment showing 80% decline after 4 h treatment. Chl fluorescence parameters were also affected by ultraviolet-B. Fo was increased while both Fm and Fv were decreased, resulted in the decreased of photochemical efficiency of PSII (Fv/Fm). Non-radiative dissipation of absorbed light as heat as estimated as NPQ (Fm/Fm' - 1) was also decreased with increasing time of UV-B treatment while the extent of photochemical quenching (qP) was not changed. Thus, the ratio of (1-qP)/NPQ parameter was also increased with increasing time of UV-B treatment indicating PSII is under the threat of photoinhibition. The result indicate that UV-B primarily decreases the capacity to dissipate excitation energy by trans-thylakoid pH, which in turn inhibits PSII activity.

• Proceedings of the Botanical Society of Korea Conference
• /
• 1999.07a
• /
• pp.51-56
• /
• 1999

Plastids, which are organelles unique to plant cells, bear their own genome that is organized into DNA-protein complexes (nucleoids). Regulation of gene expression in the plastid has been extensively investigated because this organelle plays an important role in photosynthesis. Few attempts, however, have been made to characterize the regulation of plastid gene expression at the chromosomal structure, using plastid nucleoids. In this report, we summarize the recent progress in the characterization of DNA-binding proteins in plastids, with special emphasis on CND41, a DNA binding protein, which we recently identified in the choloroplast nucleoids from photomixotrophically cultured tobacco cells. CND41 is a protein of 502 amino acids which consisted of a transit peptide of 120 amino acids and a mature protein of 382 amino acids. The N-terminal of the 'mature' protein has lysine-rich region which is essential for DNA-binding. CNA41 also showed significant identities to some aspartyl proteases. Protease activity of purified CND41 has been recently confirmed and characterized. On the other hand, characterization of accumulation of CND41 both in wild type and transgenic tobacco with reduced amount of CND41 suggests that CND41 is a negative regulator in chloroplast gene expression. Further investigation indicated that gene expression of CND41 is cell-specifically and developmentally regulated as well as sugar-induced expression. The reduction of CND41 expression in transgenic tobacco also brought the stunted plant growth due to the reduced cell length in stem. GA3 treatment on apical meristem reversed the dwarf phenotype in the transformants. Effects of CND41 expression on GA biosynthesis will be discussed.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.31 no.2
• /
• pp.143-149
• /
• 1986

Anatomical changes in the forming and germinating processes of tobacco seeds were investigated to obtain basic information on the ecological characteristics of tobacco seeds. Seed development studied through the longitudinal section of the fertilized ovule clarified that the cell division of the zygote was initiated after 7 days of flowering. After 12 days of flowering, perfect seed constituents such as cotyledon, epicotyle and radicle were formed and those were expressed to recognizable level of germinability. After 15 days of flowering germination rate reached higher than 30% and 17 and 21 days after flowering a perfect seed which have 70% or higher germinability were produced. Seed size was ranged between 0.3-0.6 mm and varietal differences were noted in the given seed size range. Under the light treatment, the morphological changes were observed by elongation of radicle after 2 days of imbibition and apparent germination after 3 days of imbibition. But no responses of the seeds imbibed 6 days under the dark condition were observed.

• Korean Journal of Plant Tissue Culture
• /
• v.25 no.2
• /
• pp.103-107
• /
• 1998

Tobacco proembryos were irradiated with 100 Gy of heavy-ion beams($^{14}\textrm{N}$, $^{20}\textrm{Ne}$: 135 Mev/u) after 24 to 96 hours of pollination as a mutagen and screened $\textrm{M}_{1}$ generation for morphological mutants and salt-tolerant plants. Morphological and physiological characteristics of the salt-tolerant plants derived from the irradiated proembryo are discussed in this report. Mutants irradiated proembryos with the beams after pollination produced various kinds of morphological variation. A total of 17 salt-tolerant plants were selected from tobacco cultivar (BY-4) by treatment with $^{14}\textrm{N}$ beam. Shapes of filament and pollen grain of most salt-tolerant plants were abnormal compared with non-irradiated wild type, and seeds weight and fertility obviously decreased. The germination rates of the several $\textrm{M}_{2}$ lines on the saline and the mannitol condition were higher than that of wild type.