• Journal of Plant Biotechnology
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• v.2 no.2
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• pp.61-71
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• 2000

By screening a cDNA library of auxin-treated mung bean (Vigna radiata L.) hypocotyls, we have isolated two full-length cDNA clones, pVR-ACS6 and pVR-ACS7, for 1-aminocyclopropane-1-carboxylate (ACC) synthase, the rate-limiting enzyme in the ethylene biosynthetic pathway. While PVR-ACS6 corresponds to the previously identified PCR fragment pMBA1, pVR-ACS7 is a new cDNA clone. A comparison of deduced amino acid sequences among auxin-induced ACC synthases reveal that these enzymes share a high degree of homology (65-75%) to VR-ACS6 and VR-ACS7 polypeptides, but only about 50% to VR-ACS1 polypeptide. ACS6 and ACS7 are specifically induced by auxin, while ACS1 is induced by cycloheximide, and to lesser extent by excision and auxin treatment. Results from nuclear run-on transcription assay and RNA gel blot studies revealed that all three genes were transcriptionally active displaying unique patterns of induction by IAA and various hormones in etiolated hypocotyls. Particularly, 24-epibrassinolide (BR), an active brassinosteroid, specifically enhanced the expression of VR-ACS7 by distinct temporal induction mechanism compared to that of IAA. In addition, BR synergistically increased the IAA-induced VR-ACS6 and VR-ACS7 transcript levels, while it effectively abolished both the IAA- and kinetin-induced accumulation of VR-ACS1 mRNA. In light-grown plants, VR-ACS1 was induced by IAA in roots, whereas W-ACS6 in epicotyls. IAA- and BR-treatments were not able to increase the VR-ACS7 transcript in the light-grown tissues. These results indicate that the expression of ACC synthase multigene family is regulated by complex hormonal and developmental networks in a gene- and tissue-specific manner in mung bean plants. The VR-ACS7 gene was isolated, and chimeric fusion between the 2.4 kb 5'-upstream region and the $\beta$-glucuronidase (GUS) reporter gene was constructed and introduced into Nicotiana tobacum. Analysis of transgenic tobacco plants revealed the VR-ACS7 promoter-driven GUS activity at a highly localized region of the hypocotyl-root junction of control seedlings, while a marked induction of GUS activity was detected only in the hypocotyl region of the IAA-treated transgenic seedlings where rapid cell elongation occurs. Although there was a modest synergistic effect of BR on the IAA-induced GUS activity, BR alone failed to increase the GUS activity, suggesting that induction of VR-ACS7 occurs via separate signaling pathways in response to IAA and BR.

• Journal of the Korean Society of Tobacco Science
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• v.1 no.1
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• pp.1-8
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• 1979

For the preliminary experiments of mass Production of tobacco cells in tank culture, the effects of nutritional conditions on the growth of suspended cells were investigated ; 1. The tobacco cell growth was affected by concentrations of sucrose or inorganic phosphate, type of nitrogen source, and plant hormone, especially 2, 4-D. 2. The optimum level of sucrose concentration was 3% and the level of inorganic phosphate was 0.3mg /ml, which was about twice as high as the level of Linsmaier - Skoog medium. 3. The best growth was observed when the ratio of nitrate nitrogen to ammonium nitrogen was 2 : 1, where the total nitrogen content was equal to that of nitrogen source. 4. To find out the mechanism of promotive effects of 214-D and inorganic phosphate on the tobacco cell growth, the respiration and metabolism of $^{14}\textrm{C}$-91ucose were investigated. Addition of 2, 4 -D in culture medium increased if 2, 4-D (0.2ppm )was added to medium or the level of inorganic Phosphate was raised 2.5 times as high as standard. In cultures with high inorganic phosphate and 2, 4-D, the absorbed 14C-glucose was converted to amino acids and organic acids rather than remained as sugars.

• 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 CROP SCIENCE
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• v.50 no.3
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• pp.156-160
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• 2005

We studied the effect of SMP (solid matrix priming) treatment, seed pretreatment for germination enhancement, to tobacco seeds by measurement of germination rate and contents of fatty acid, energy source for tobacco germination. The results were as follows. In result of germination test, germinative ability was the highest in KF109 at nine day treatment and in KB108 at seven day treatment. The composition of fatty acid in tobacco seed confirmed by gas chromatography were palmitic acid, stearic acid, oleic acid+elaidic acid, linoleic acid+linole­laidic acid, and $\alpha-linolenic$ acid. Palmitic acid, stearic acid, oleic acid and elaidic acid, and linoleic acid and linole-laidic acid were highest in KF109 at eight or nine day treatment and in KB108 at six or seven day treatment. Especially, content of oleic acid + elaidic acid, and linoleic acid+linolelaidic acid were changed largely by treatment, so these might be used for index to examine treatment effect.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.41 no.4
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• pp.475-481
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• 1996

This study was carried out to acquire the basic information of root growth under different pot size, imposing different space limitation on rhizosphere. Different size of pots that had same surface area but different depth, 5cm(Iength)$\times$5cm(width)$\times$30, 15, 5cm(depth), were used during the seedling stage of tobacco plant. Space limitation on rhizosphere affected not only the aerial growth, stem height, leaf area and shoot dry weight, but also root growth and root architecture. Aerial growth was highly related to growth of underground part, so space limitation on rhizosphere decreased aerial growth. Limitation on pot volume by reducing pot depth induced new rooting on crown. Root number and relative multiplication rate were higher in small pot that had 5cm depth than large pot, but total root length and mean extension rate showed reverse patterns. Root numbers of 1st order and 2nd order were increased as pot depth was increased, but the root number of 3rd order was increased in small pot. Root system of seedling grown in large pot distributed more horizontally than that in small pot at 20 days after temporary planting (DAT), but the root architecture of seedling was reversed at 25 DAT.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.31 no.4
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• pp.454-464
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• 1986

To find main factors to affect tobacco culture and quality, NC 2326 (Nicotiana tabacum L.) was cultivated in Korea and in the United States under different plant density, fertilization, mulching and curing. Among the chemical characteristics of the both experimental soils, the organic matters were similar concentration in both locations but effective phosphorous contents were higher in Oxford in the United States. Plant height, length and width of the largest leaf, leaf thickness, and midrib ratios were larger in Oxford than in Suwon in Korea. Also they were larger in non mulching system than in mulching system. But the total numbers of the leaves were decreased in non mulching system. The content of nicotine was higher in the plant grown in Suwon than in Oxford. The concentrations of nicotine and sugar tend to increase in mulching system comparing of non mulching system. During the growing, the concentration of non-volatile organic acids was higher in Suwon, while it was lower in cured leaf produced in Suwon. Also the contents of total fatty acids were lower in the harvested leaf grown in Suwon, but not in cured tobacco. Forty three compounds identified among the volatile oils from these experimental samples were quantified. The concentrations of the major components related to the tobacco flavour such as damascone, damascenone, solanone, nor-solanadione, and megastigmatrienones were higher in the cured tobacco produced in Oxford rather than in Suwon.

• Korean Journal of Environmental Agriculture
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• v.17 no.3
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• pp.215-219
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• 1998

This experiment was conducted to investigate effects of NaCl concentration on photosynthetic rate, free proline content and ion content in tobacco. As NaCl concentration was increased growth was retarded. The decrease growth characteristics(shoot/root ratio was 2.0) at 90mM NaCl indicated that this concentration could be a limiting level. As NaCl concentration was increased photosynthetic rate, transpiration rate, and water use efficiency were decreased. Photosynthetic rate was highly decreased at 60mM NaCl. There was no significant difference between transpiration rate and water use efficiency. Leaf water potential was decreased as NaCl concentration was increased, in that twice lower at 30mM than that of control and drop largely at 120mM NaCl. Free proline content was increased as NaCl increased until 120mM NaCl and drop at 150mM NaCl. The $Ca^{2+}$, $Mg^{2+}$, and $K^+$ contents were increased until NaCl concentration was 120mM. The $Na^{2+}$ content was slowly increased as NaCl concentration increased until 120mM NaCl, and largely increased at 150mM NaCl. There was no significant difference between $Cl^-$ and NaCl treatments except 30 mM NaCl in which $Cl^-$ content was higher than that of control. As NaCl concentration was increased $K^+/Na^+$ ratio was decreased. The negative correlation between $K^+$ and $Na^+$, and positive correlation between $K^+/Na^+$ and protein content were found.

• Current Research on Agriculture and Life Sciences
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• v.6
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• pp.7-12
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• 1988

This study was conducted to establish the most appropriate transplanting time which can attribute to increase of aromatic substances in tobacco variety, Hyangcho, maintaining its original aroma. The highest content of total alkaloid, total nicotine and total nitrogenous compounds were observed in the late transplanting time such as July 5, showing 4.16%, 3.83% and 3.68%, respectively. However, the petroleum ether extract was highest in the early transplanted one such as April 5, showing 5.77% and total sugar content, 12% in April 20. The content of petroleum ether extract decreased as the transplanting time delayed. The early transplanting of Hyangcho on April 20 under vinyl mulching showed the increase of fatty acid content, but the decrease of non-volatile organic acids such as malic and citric acids than those of the conventional transplanting time on March 5. The earlier transplanting time also increased the content of volatile acids such as 2,3-methylbutanoic acid and 3-methyl-pentanoic acid, which seem to be related to tobacco aroma.

• Korean Journal Plant Pathology
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• v.3 no.4
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• pp.291-298
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• 1987

Samples showing mosaic symptom of cowpea (Vigna sesquipedalis) with vein banding, chlorotic spot, vein yellow were collected from Chinju areas in Korea, Two viruses were distinguishable by stability in sap, host range, and relations with cells and tissues were examined under an electron microscope, Blackeye cowpea mosaic(BICMV) was sap-transmissible to 7 plant species in 2 families, Of the plants, only leguminous species were systemically infected. This virus was inactivated by heating at $50-65^{\circ}C$ for 10 min, by diluting at $10^{-4}-10^{-5}$, and aging at room temperature for 1-6 days. Preparations examined under the electron microscope by direct negative staining method(DN -method) always showed particles of flexuous filament bout 750nm in length and cytopasmic inclusions. Cytoplasmic inclusions and virus particles were also confirmed to present in the cytoplasm of a mesophyll cell by ultrathin sections of BICMV infected cowpea leaves. Cucumber mosaic virus (CMV) was transmitted by sap- inoculation on inoculated leaves of Chenopodium amaranticolor, C. quinoa producing local lesions, but non-inoculated upper leaves of Nicotiana glutinosa, Cucurbita pepo and Vigna sesquipedalis producting systemic mosaic symptoms. Electron microscopic examination of virus preparation by direct negative staining showed spherical particles of about 30nm in diameter. In ultrathin sections of CMV infected tissues, virus particles of crystalline array were found in the vacuole and a large number of virus particles were found in the cytoplasm and the plasmodesmata of mesophyll cells.

• The Plant Pathology Journal
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• v.36 no.4
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• pp.323-334
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• 2020

Lysin motif (LysM) proteins are reported to be necessary for the virulence and immune response suppression in many herbaceous plant pathogens, while far less is documented in woody plant pathogens. In this study, we preliminarily characterized the molecular function of a LysM protein LtLysM1 in woody plant pathogen Lasiodiplodia theobromae. Transcriptional profiles revealed that LtLysM1 is highly expressed at infectious stages, especially at 36 and 48 hours post inoculation. Amino acid sequence analyses revealed that LtLysM1 was a putative glycoprotein with 10 predicted N-glycosylation sites and one LysM domain. Pathogenicity tests showed that overexpressed transformants of LtLysM1 displayed increased virulence on grapevine shoots in comparison with that of wild type CSS-01s, and RNAi transformants of LtLysM1 exhibited significantly decreased lesion length when compared with that of wild type CSS-01s. Moreover, LtLysM1 was confirmed to be a secreted protein by a yeast signal peptide trap assay. Transient expression in Nicotiana benthamiana together with protein immunoblotting confirmed that LtLysM1 was an N-glycosylated protein. In contrast to previously reported LysM protein Slp1 and OsCEBiP, LtLysM1 molecule did not interact with itself based on yeast two hybrid and co-immunoprecipitation assays. These results indicate that LtLysM1 is a secreted protein and functions as a critical virulence factor during the disease symptom development in woody plants.