• 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 Biology
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• v.32 no.4
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• pp.275-283
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• 1989

$\beta$-1, 4-endoglucanase (EC 3, 2, 1, 4) was isolated and purified from Nicotiana tabaccum L. Var. Virginia 115 suspensin cultured cells. The molecular weight as estimated by Sephadex G-100 was about 14, 000. The optimum pH for activity was 5.4. The Km value for carboxymethyl cellulose was 0.18 mg/unit and the enzyme was quite resistant to heating. Polyamine did not affect the activity of $\beta$-1, 4-endoglucanase in vitro but the activity increased drastically when polyamines were added in the suspension culture medium. It is suggested that increase in $\beta$-1, 4-endoglucenase activity due to polyamine might be related to growth of the plant.

• KSBB Journal
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• v.13 no.4
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• pp.399-403
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• 1998

This study was carried out to optimize the concentration of Ca2+ and pH of fusion medium which affected electrofusion frequency of protoplasts isolated from Nicotiana tabacum L. (cv. BY4) mesophyll cells and callus. The protoplasts were electrofused in the fusion media containing two different Ca2+ concentrations and three different pH regions. Fusion frequency was lower in the fusion medium containing only 13% mannitol as osmotic stabilizer. However, higher degree of fusion frequency (47.3%) was observed in the fusion medium containing 50mM CaCl2 at pH 10.5 than any other conditions. Cell viability was decreased by Ca2+ and high pH treatment in the fusion media, while fusion frequency was increased. It is concluded that Ca2+ is involved in electrofusion of protoplasts.

• Journal of Bio-Environment Control
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• v.11 no.3
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• pp.139-143
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• 2002

Cotton Glutathione S-Transferase (GST: EC 2.5.1.18) was cloned and overexpressed in tobacco (Nicotiana tabacum) plants. Northern blot analysis confirmed the successful transformation of cotton gst gene in tobacco plant. Type I and Type ll transcript patterns were identified in transgenic tobacco plants and only Type I transcripts were discussed in this paper, The activity of GST in the type II transgenic plants was about 1.5-fold higher than those of the wild type and non-expresser by using 1-chloro-2,4-dinitrobenzene (CDNB) and reduced glutathione as the substrate. The expression of cotton GST in tobacco plants proved that Gh-5 could be translated into functional protein. Type II transgenic plants produced functional GST in the cells. The effects of cotton GST in the seedlings was evaluated by growing the control and transgenic seedlings at $15^{\circ}C$ in the growth chamber in the light. Overexpressors were grown well compared to the control plants (non-expressors). lo test far tolerance to salinity, seeds of Gh-5 overexpressors and the wild type Xanthi seedlings were grown at 0, 50, 100, 150, and 200 mM NaCl solution. Gh-5 transgenic seedlings showed higher growth rate over control seedlings on 50 and 100 mM NaCl solution. There was no difference in growth rate at 150 and 200mM NaCl concentration.

• Korean Journal of Plant Resources
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• v.21 no.2
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• pp.139-143
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• 2008

Enzyme activity and expression of cytochrome P450 gene involved in the pathway of capsidiol biosynthesis were compared in five different solanaceae plants such as red pepper, green pepper, tobacco, potato and egg plant. Base on genomic DNA and/or RT-PCR results, four solanaceae plants such as red pepper, green pepper, tobacco and egg plant possess P450 gene in the genome and specifically expressed by elicitor treatment. However, potato was appeared to have neither P450 nor cyclase gene in the genome. P450 genes did not show any expression in the plants under normal condition, but showed highly specific expression under elicitation condition in various organs and tissue such as leaf, root, stem and culture cells.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.3
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• pp.1519-1529
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• 2016

Background: Matrix metalloproteinase -2 (gelatinase-A, Mr 72,000 type IV collagenase, MMP-2) and -9 (gelatinase-B, Mr 92,000 type IV collagenase, MMP-9) are key molecules that play roles in tumor growth, invasion, tissue remodeling, metastasis and stem-cell regulation by digesting extracellular matrix barriers. MMP-2 and -9 are well known to impact on solid cancer susceptibility, whereas, in hematological malignancies, a paucity of data is available to resolve the function of these regulatory molecules in bone marrow mononuclear cells (BM-MNCs) and stromal cells of myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). Objectives: The present study aimed to investigate mRNA expression and gelatinase A and B secretion from BM-MNCs in vitro and genotypic associations of MMP-2 (-1306 C/T; rs243865), MMP-9 (-1562 C/T; rs3918242), tissue inhibitor of metalloproteinase -1 (TIMP-1) (372T/C; rs4898, Exon 5) and TIMP-2 (-418G/C; rs8179090) in MDS and AML. Results: The study covered cases of confirmed MDS (n=50), AML (n=32) and healthy controls (n=110). MMP-9 mRNA expression revealed 2 fold increased expression in MDS-RAEB II and 2.5 fold in AML M-4 (60-70% blasts). Secretion of gelatinase-B also revealed the MMP-9 mRNA expression and ELISA data also supported these data. We noted that those patients having more blast crises presented with more secretion of MMP-9 and its mRNA expression. In contrast MMP-9 (-1562 C/T) showed significant polymorphic associations in MDS (p<0.02) and AML (p<0.02). MMP-9 mRNA expression of C/T and T/T genotypes were 1.5 and 2.5 fold increased in MDS and AML respectively. In AML, MMP-2 C/T and T/T genotypes showed 2.0 fold mRNA expression. Only MMP-9 (-1306 C/T) showed significant 4 fold (p<0.001) increased risk with chemical and x-ray exposed MDS, while tobacco and cigarette smokers have 3 fold (p<0.04) risk in AML. Conclusions: In view of our results, MMP-9 revealed synergistic secretion and expression in blast crises of MDS and AML with 'gene' polymorphic effects and is significantly associated with increased risk with tobacco, cigarette and environmental exposure. Release and secretion of these enzymes may influence hematopoietic cell behavior and may be important in the clinical point of view. It may offer valuable tools for diagnosis and prognosis, as well as possible targets for the treatments.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.1
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• pp.33-41
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• 1997

This study was carried out to establish simple and easy methods to judge the survival, senescence and death of the protoplasts in culture system by identifying the marker substance related to metabolic status of the cells. When rice and tobacco protoplasts were cultured in MS and KM-8P media containing 2,4-D or coconut milk ABA decreased especially in the media containing coconut milk, but GA$_3$, IAA and zeatin increased as the cultures progressed. The decrease of ABA and increase of zeatin was especially remarkable. When the supraoptimal amount of osmoticum (mannitol) was added to the culture media ABA decreased after a momentary increase, but other growth hormones slowly increased as the concentration of the osmoticum increased. Contents of individual hormones were contrasted when protoplasts rice and tobacco were cultured on the same medium containing 10mM super mine or NaCl. Tobacco protoplasts were more sensitive to NaCl stress and stopped protoplast division at the late stage of culture. Protoplast viability decreased greatly in 48 hours when the protoplast were at 32$^{\circ}C$ on a medium lacking several components. ABA content increased up to 10 days from incubation in negative proportion to the protoplast viability. On the other hand contents of other growth hormones, especially zeatin, decreased. The present results clearly showed that the contents of individual growth hormones in the plant protoplasts in culture varied sensitively in response to environmental factors that they are faced with. This indicates that the physiological states of the protoplast, such as survival, senescence or death can be simply judged based on the quantitative analysis of those hormones by ELISA.

• Proceedings of the Plant Resources Society of Korea Conference
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• 1999.10a
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• pp.46-57
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• 1999

Ginseng(Panax ginseng C.A. Meyer) is important medicinal plant but requires 4-year cultivation for root harvest because of slow growth. In contrast, ginseng callus and hairy roots grow vigorously and may Produce the same or more biologically active compounds for human health than natural ginseng roots. Therefore, ginseng callus and hairy roots can be used for commercial purposes. Polyacetylene, one of anti-cancer compounds in ginseng, was not detected in the callus cultured on the medium containing 2, 4-B, but cells derived from the callus growth was excellent, The ginseng calli cultured on the medium containing 2mg11 CPA and 0.05mg/1 BA was grown vigorously and produced panaxydol, one of ginseng polyacetylene. The biosynthesis of polyacetylene in callus was not affected by addition of NAA and sucrose in media. The SH medium was better than the MS medium for ginseng callus growth and biosynthesis of panaxydol. Another ginseng anti-cancer compounds, ginsenoside-Rg$_3$, Rh$_1$and Rh$_2$ were detected in ginseng hairy roots by heat treatment. Those of Panax ginseng were obtained after root disks of three-year old roots were infected with Agrobacterium rhizogenes Rl000 $A_4$T in dark condition after one month of culture. The optimum growth of hairy roots was achieved in the culture of 1/2 MS liquid medium in dark(22$^{\circ}C$) under 60 rpm gyratory shaking. Hairy roots grew well in 5 ι Erlenmeyer flasks, 1ι roller drums, 10ι jar-fermenters, and especially in 20ι air-lift .culture vessels. All heat treatments had remarkably different ginsenoside contents. Eleven ginsenosides were determined in heat treatment, eight in freeze dried hairy roots. Contents of ginsenoside-Rbl , Rb2, Rc, Rd. Re, Rf, and Rg$_1$tested in all heat treatments were less than those of freeze dried hairy roots. Contents of glnsenoside-Rg$_2$ in heat treatment for 1 hour at 105$^{\circ}C$ was 4.92mg/g dry wt, 3.9 times higher than 1.27 mg/g dry wt of freeze dried hairy roots. The optimum condition of heat treatment for the production of ginsenoside-Rg$_3$and Rhl was 2 hours at 105$^{\circ}C$, and ginsenoside content was 2.58mg/g dry wt and 3.62mg/g dry wt, respectively. The production of ginsenoside-Rh2 was the highest in heat treatment for 2 hours at 105$^{\circ}C$ among treatments examined, and ginsenoside-Rh$_2$content was 1.08mg/g dry wt.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.38 no.3
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• pp.245-252
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• 1993

To investigate a simple and rapid screening method for photosynthetic inhibitory herbicides, responses of tobacco(Nicotiana tabacum L.) and liverwort(Marchantia polymorpha L.) PA(photoautotrophic) cells to various commercial herbicides with different modes of action and leaf extracts of four weed species were compared. PET (photosynthetic electron transport) inhibitory type of herbicides has greater inhibitory effect in liverwort photoautotrophic cells than the photomixotrophic and heterotrophic cultured cells. Similary, PET inhibitory type of herbicides inhibited the oxygen evolution more in liverwort PA cells than the other type of herbicides. Based on oxygen evolution, 60% inhibition was observed by the 10% aqueous extracts of Polygonum hydropiper, while there was 100% inhibition by the 10% methanol extracts of Polygonum hydropiper. This assay gave well correlated results to the Hill reaction data using isolated thylakoids. Thus liverwort photoautotrophic cells might be suitable materials for rapid screening method for photosynthetic inhibitory herbicides.

• Molecules and Cells
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• v.22 no.2
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• pp.233-238
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• 2006

Endogenous salicylic acid (SA) and its predominant conjugates, SA 2-O-${\beta}$-D-glucoside (SAG) and the glucose ester of SA (SGE), increase dramatically during plant defense responses. Here I report the isolation and characterization of an Arabidopsis thaliana UDP-glucose:SA glucosyltransferase1 (AtSGT1) gene using a tobacco SGT gene previously reported, whose product catalyzes the formation of both SAG and SGE. The recombinant AtSGT1 protein had significant activities with SA and benzoic acid, and synthesized SAG and SGE. Northern blot analysis showed that AtSGT1 was rapidly induced both by exogenous SA and infection with the bacterial pathogen Pseudomonas syringae, indicating that pathogen-inducible AtSGT1 expression is an early disease response and may be involved in the accumulation of glucosyl SA during pathogenesis.