• Korean Journal of Plant Tissue Culture
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• v.26 no.3
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• pp.197-203
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• 1999

The albino plants of tobacco (Nicotiana tobacum L. cv. BY-4) were isolated from seed populations that were induced by heavy-ion ($^{14}N$) beam irradiation to proembryo and the in vitro growth and differentiation have been characterized. The in vitro cultured albino plants showed significant reduction of chlorophyll content and possessed larger number of stomata on both upper and lower epidermis than that of wild-type plants. Stem growth of the mutants remained dwarfed, however, the internode recovered its normal length after GA$_3$ treatment (10.0mg/L) on the MS medium containing sucrose under continuous light. When explants of leaf blades of albino plants were cultured, multiple shoots formed directly on MS medium containing 1.0mg/L of BAP or kinetin and a large number of calli were induced on the MS medium containing 1.0mg/L NAA or 1.0 mg/L 2,4-D. The albino calli regenerated multiple albino plantlets in the MS medium containing 0.1mg/L NAA + 1.0 mg/L BAP. No significant differences between the wild-type and albino plants were detected in the multiple shoot induction, callus formation from the explants and the plantlets regeneration from calli. In addition, albino plants have a similar organogenesis Pattern to that of the wild-type in the media with different combinations of NAA (0 to 5.0mg/L) and BAP (0 to 5.0mg/L) treatment. These results indicate that the albino mutant has the same normal regeneration ability as that of wild-type, although the mutant has lost functions in photosynthesis, such as pigmentation.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.5
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• pp.632-639
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• 1997

The experiment which imposed the water stress to tobacco(Nicotiana tabacum L.) plant was carried at the late of maximum growth period. In order to know the influence of drought stress on the growth and developmemt of tobacco leaves of different position and to elucidate the physiological response of plant to various soil water content, stomatal conductance, and leaf water potential were measured. The drought stress at the maximum growth period negatively affected to the overall growth characteristics of shoot. The response of the growth was small at the middle and the lower leaves, and great at the upper leaves. The relative water content of upper, middle, and lower leaves at the fifth day after treatment were 74, 64, and 59%, respectively, as soil water content was reduced by 4.3%. This suggested that the wilting point of tobacco leaf was about 75%. The leaf water potential was -0.58 MPa in control and dropped to -1.20 MPa at the fifth day after treatment. This indicated that wilting of leaf may occur at the condition in which the difference of water potential between treatment and control, well watered, was greater than about 20%. Stomatal conductance at the fifth day after treatment dropped from 12 mol /$\textrm{m}^2 sec^{-1}$ to 0.8 mol /$\textrm{m}^2 sec^{-1}$ in the middle and the upper leaves. Stomatal conductance of lower leaves already matured were not affected highly by drought stress at the maximum growth period, but maturing leaves, middle and upper leaves, were highly affected by limitation of soil water.

• Korean Journal of Weed Science
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• v.18 no.1
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• pp.69-75
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• 1998

Responses of tobacco(Nicotiana tabacum cv. Xanthi) leaves of different age to diphenyl ether herbicide oxyfluorfen were evaluated with respect to cellular leakage, chlorophyll loss, and membrane lipid peroxidation. When tobacco leaves of different age were incubated under light condition at $25^{\circ}C$ following 12hr dark incubation. Significant electrolyte leakage from the treated tissues into the bathing medium occurred. The change of electrolyte leakage was proportional to the oxytluorfen concentration and the duration of light exposure to the tissues. Electrolyte leakage from the tissues treated with oxyfluorfen was highly dependent on the leaf age. From the tissues of younger age, more electrolyte leakage occurred and lag period was greatly reduced. Chlorophyll loss and membrane lipid peroxidation, as measured by malondialdehyde production, caused by oxyfluorfen treatment were also dependent an the age of treated leaf tissues. In conclusion, physiological responses of tobacco leaves to oxyfluorfen greatly varied with the age of treated tissues, and thus tobacco plants could be used as appropriate materials for studying the mechanisms of tolerance to diphenyl ether herbicides.

• Korean Journal of Plant Tissue Culture
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• v.25 no.4
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• pp.245-250
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• 1998

Tobacco mosaic virus(TMV) coat protein cDNA was transformed to Nicotiana tabacum cv. NC82 and the transgenic tobacco plants resistant to TMV infection were isolated in the next generation. The expression of TMV coat protein cDNA and genetic stability of the fifth generation of TMV resistant transgenic tobacco plants at the higher temperature were investigated. The TMV coat protein cDNA was amplified by genomic PCR in all the TMV resistant transgenic tobacco plants. The TMV coat protein expressed in the transgenic tobacco plants was detected at very low level by immunoblot hybridization. Even in tansgenic plants that showed the viral symptom only on very late sucker growth (delay type plants), the coat protein expression in the suckers was much less than that of susceptible tobacco infected with TMV. The TMV coat protein expressed in the transgenic tobacco plants was below 0.01% of total protein. Transcription and expression of the coat protein cDNA in delay type plants were observbed at high temperature (38$^{\circ}C$), and TMV replication was suppressed at both 28$^{\circ}C$ and 38$^{\circ}C$. This indicates that unlike the resistance conferred by 'N' gene. TMV resistance of transgenic tobacco plant won't break down at high temperature.

• Korean Journal of Plant Tissue Culture
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• v.25 no.2
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• pp.135-139
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• 1998

Reciprocal interspecific hybrids between N. glauca(2n=24) and N. langsdorffii(2n=18) were obtained by intercrossing. One hundred percent of F$_1$ seeds was produced from intercrossing of N. glauca $\times$ N. langsdorffii, whereas the frequency of F$_1$ hybrid seed formation from N. langsdorffii $\times$ N. glauca was very low. However, all the hybrid seeds were germinated well and then grown to normal plantlets. All the plants of F$_1$ hybrids have chromosome number of interspecific hybrids (2n=21). From observation of morphological characteristic, the structure of petrol, leaf, flower, and the morphology of pollen have characteristics of F1 hybrid. Spontaneous tumors (genetic tumor) were formed from each F$_1$ hybrid; the genetic tumor arose at the reproductive phase when the maternal type of F$_1$ hybrid came from N. glauca, while the genetic tumor arose only after reproductive phase when the maternal type of F$_1$ hybrid came from N. langsdorffii. The genetic tumor actively proliferated on hormone-free medium and produced numerous teratoma shoots. In addition, normal leaf or stem explants of F$_1$ hybrid produced calli on hormone-free medium after 15 days of culture, the calli produced new numerous teratoma shoots after 30 days. The frequency of teratoma shoot formation from rnterspecific hybrid was higher in the N. glauca $\times$ N. langsdorffii than in the N. langsdorffii $\times$ N. glauca. Root development from the teratoma shoots was hardly obtained. Teratoma shoots without roots in vitro can form genetic tumor at the vegetative growth phase after tissue culture.

• Journal of Plant Biotechnology
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• v.6 no.1
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• pp.25-37
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• 2004

Successful genetic transformation of plants requires non-chimeric selection of transformed tissues and their subsequent regeneration. With rare exceptions, most transformation protocols still rely heavily on antibiotics for selecting transgenic cells that contain an antibiotic-degrading selectable marker gene. Here, the morphogenic capacity of in-vitro explants of chrysanthemnum and tobacco stems and leaves (control and transgenic) changed with the addition of aminoglycoside antibiotics (AAs), In a test of 6 AAs, phytotoxicity occurred at concentrations of 10 to 25 and 50 to 100$\mu\textrm{g}$ $mL^{-1}$ in chrysanthemum and tobacco explants, respectively. Light conditions as well as explant source and size also had significant effects. The use of transverse thin cell layers (tTCLs), in conjunction with high initial AA selection levels, supported the greatest regeneration of transgenic material (adventitious shoots or callus) and the lowest number of escapes. Flow-cytometric analyses revealed no endodu-plication in chrysanthemum, even at high AA levels. However, this phenomenon was observed in tobacco calli(8C or more), even at low AA concentrations (i.e., 5 to 10 $\mu\textrm{g}$ mL$^{-1}$ ).

• Journal of Korean Society for Atmospheric Environment
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• v.22 no.E2
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• pp.89-98
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• 2006

Two tobacco cultivars (Nicotiana tabacum L.), Bel-B and Bel-W3, tolerant and sensitive to ozone, respectively, were grown in a greenhouse supplied with charcoal filtered air and exposed to 200 ppb ozone for 4 hr. Effects on chlorophyll fluorescence, net photosynthesis, and stomatal conductance are described. Quantum yield was calculated from chlorophyll fluorescence and the initial slope of the assimilation-light curve measured by the gas exchange method. Only the sensitive cultivar, Bel-W3, developed visual injury symptoms on up to 50% of the $5^{th}$ leaf. The maximum net photosynthetic rate of ozone-treated plants was reduced 40% compared to control plants immediately after ozone fumigation in the tolerant cultivar; however, photosynthesis recovered by 24 hr post fumigation and remained at the same level as control plants. On the other hand, ozone exposure reduced maximum net photosynthesis up to 50%, with no recovery, in the sensitive cultivar apparently causing permanent damage to the photosystem. Reductions in apparent quantum efficiency, calculated from the assimilation-light curve, differed between cultivars. Bel-B showed an immediate depression of 14% compared to controls, whereas, Bel-W3 showed a 27% decline. Electron transport rate (ETR), at saturating light intensity, decreased 58% and 80% immediately after ozone treatment in Bel-B and Bel-W3, respectively. Quantum yield decreased 28% and 36% in Bel-B and Bel-W3, respectively. It can be concluded that ozone caused a greater relative decrease in linear electron transport than maximum net photosynthesis, suggesting greater damage to PSII than the carbon reduction cycle.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.140.1-140
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• 2003

An Isometric virus was isolated from Paprika (Capsicum annuum var. glossum) showing necrosis spot and malformation on the fruit and the leaves, respectively, at yecheon in Korea. The virus could infect locally on Chenopodium amaranticolr, C. quinoa, Petunia x hybrida and Nicotiana glutinosa, but could not infect on Gomphrena globosa and Physalis floridana. The virus could infect systemically on red pepper and Lycopersicon esculentum. Datura stramonium, N. cleuarandii, N. rustim and N. tabacum cvs. were produced necrosis or necrotic ring spot lesions on the inoculated leaves and mosaic, vein necrosis or lethal death on the upper leaves. The virus was not related serologically to cucumber mosaic virus (CMV). In RT-PCR assay, it could not detected with specific primers of CMV and BBWV-II. The virions contain one molecule of genomic RNA, Which was approximately 3.8Kb and the coat protein (CP) of the purified virion migrated as a single band with molecular wight of about 29KDa in SDS-PAGE.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.139.2-140
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• 2003

Ornithogalum showing mosaic symptoms were collected from the isolated field of National Plant Quarantine Service in Sengrimmyon of Kyungnam province. Electron microscopic examination of negatively strained preparation was filamentous particle of 740nm in lenght. Indirect-ELISA determined that the virus was serologically related to potyvirus. A single major protein band of Mr 30,000 was observed after sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Indicator plant test showed masaic, necrotic local lesion and sunken areas in leaves of Nicotiana clevelandii and Tetragonia expansa, while the others of indicator plants did not infect. An enzyme-aided purification protocal was used, which eliminated a highly viscous mucilage from extracts of the Omithogalum. Total RNA extracted from infected Omithogalum leaves were amplified of 411b.p fragment in reverse transcription (RT)-PCR when primers specilic for the coat protein gene. An isolate of Omithogalum mosaic virus (OrMV) of the genus Potyvirus was identified as the casual agent of the disease on the basis of electron microscopic, biological and serological reaction.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.74.1-74
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• 2003

Differential display (DD) of mRNA is a technique in which mRNA species expressed by a cell population are reverse transcribed and then amplified by many separate polymerase chain reactions (PCR). Using DD-RT-PCR we obtained many genes that expressed differentially in healthy and PVX-infected Nicotiana benthamima, using total RNAs extracted from healthy and PVX-infected N. benthamiana plants. Three hundred and twenty-five DNA fragments isolated from DD-RT-PCR were cloned and sequenced for further characterization. Several host genes including SKPI-like protein, heat shock transcription factor and Avr9/Cf-9 rapidly elicited protein were selected to obtain full-length open reading frame and to characterize their potential involvement in virus disease development and/or host's defense against virus infection employing PVX-based expression vector. Transcrips from wild-type and clones containing each selected gene were inoculated onto N. benthamiana Levels of virus replication were confirmedby RT-PCR and RNA blot analysis, Expression profiles and potential role(s) of selected genes upon PVX infection will be discussed.