• Journal of Plant Biotechnology
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• v.31 no.4
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• pp.285-293
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• 2004

FMDV is a viral pathogen that caused foot-and-mouth disease in animals. VP1 is a major capsid protein of FMDV. It is known as one of best materials for the FMDV diagnosis and for the development of protein vaccine. In this study, 633 bp of VP1 gene was modified for the expression of VP1 in plant, based on the VP1 DNA sequence from FMDV taiwan O type and from FMDV isolated vietnam. The. deduced DNA fragment was artificially synthesized using the multiple fragment extension with long-nucleotides. A new plant transgenic vector system, pCAMBIA139011 was constructed on the basis of pBI12l and pCAMBIA1390. Using this vector system and GFP gene or modified VP1 gene, each target gene was introduced into Nicotiana tabacum. The insertion of whole target gene was successfully confirmed in each transgenic plant named GFP-A7 and VP1-4, respectively. The expression level of each gene was estimated by RT-PCR and Real-Time PCR using VP1, GFP specific primers.

• Journal of the Korean Society of Tobacco Science
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• v.1 no.1
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• pp.56-62
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• 1979

Boring the process of curing of the tobacco ( Nicotiana tabacum L. clicks) leaf, the and other fundamental components of the tobacco leaf was observed The amount of total chlorophyll was reduced from 2902$\mu\textrm{g}$/gr to 1, 131$\mu\textrm{g}$/gr as a function of curing time. In the case of total carotene, the concentration was increased from 373-712$\mu\textrm{g}$/gr. to 3, 003~2, 215$\mu\textrm{g}$/gr. up to 70 hours of curing period. However, after 70 hours of curing, the concentration of total carotene stared to be decreased up to 1, 050~1, 039$\mu\textrm{g}$/gr. Simultaneously, the greenish yellow color becomes to be close an intermediate color between yellow and orange. The amount of total nicotine and nitrogen (2~2%) in the green harvested leaf was essentially identical after the curing process. However, the amount of total sugar was enhanced from 17%, in the green harvested leaf, to 35% at the time of 70 hours curing. At the end of the curing, the amount of total sugar was again reduced up to 16%. The correlation coefficient (r) between the axis of the color coordinate and the ratio of chlorophyll/carotene was estimated to be r=-0.96. This result indicates a linear relationship between those two parameters and the following linear equation is obtained as y=28.87-68.38 x. From the above equation, x=0.442 could be estimated when chlorophyll was completely decomposed (ylongrightarrowO). As a consequence of this result, we can suggest the tobacco leaf of Hicks belongs to the high quality level.

• Korean journal of applied entomology
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• v.56 no.1
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• pp.19-28
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• 2017

Polydnaviruses (PDVs) are a group of double-stranded DNA viruses symbiotic to some endoparasitoid wasps. Cotesia plutellae bracovirus (CpBV) is a PDV symbiotic to an endoparasitoid wasp, C. plutellae, parasitizing young larvae of Plutella xylostella. An early expressed gene, CpBV-ELP1, plays an important role in the parasitism by suppressing host cellular immunity by its cytotoxic activity against hemocytes. This study aimed to test its oral toxicity against insect pest by expressing it in a recombinant tobacco plant. A recombinant CpBV-ELP1 protein was produced using a baculovirus expression system and secreted to cell culture medium. The cell cultured media were used to purify CpBV-ELP1 by a sequential array of purification steps: ammonium sulfate fractionation, size exclusion chromatography, and ion exchange chromatography. Purified rCpBV-ELP1 exhibited a significant cytotoxicity against Spodoptera exigua hemocytes. CpBV-ELP1 was highly toxic to the fifth instar larvae of S. exigua by injection to hemocoel. It also showed a significant oral toxicity to fifth instar larvae of S. exigua by a leaf-dipping assay. CpBV-ELP1 was cloned into pBI121 vector under CaMV 35S promoter with opaline synthase terminator. Resulting recombinant vector (pBI121-ELP1) was used to transform Agrobacterium tumefaciens LBA4404. The recombinant bacteria were then used to induce callus of a tobacco (Nicotiana tabacum Xanthi) leaves and subsequent generation (T1) plants were selected. T1 generation tobacco plants expressing CpBV-ELP1 gave significant insecticidal activities against S. exigua larvae. These results suggest that CpBV-ELP1 gene can be used to control insect pests by constructing transgenic crops.

• Korean journal of applied entomology
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• v.23 no.4 s.61
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• pp.203-208
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• 1984

Four varieties of tobacco, Nicotiana tabacum var. NC 2326. NC 95, NC 744 and Havana. and two plant species, Physalis floridana Rydb., N. repanda Wild. were used for classification of ten isolates of potato virus Y(PVY) obtained Iron potato or tabacco plants showing various symptoms. Each of the 10 isolates could be distinguished by the host reactions showing necrosis, vein banding and/or mottling, or no symptom on these hosts. Five isolates of PVY, PVY-VN, PVY-N, PVY-NSNR, PVY-Chile, and PVY-Argentina, showed necrotic symptom on NC 2326, but others showed vein handing and/or mottling symptom. On NC95, each of the isolates tested showed similar symptons as on NC2326, except necrotic symptom by the isolate PVY-MSNR. Havana showed mottling reaction to the PVY-NSNR, PVY-MSNR, PVY-MSMR, PVY-VB, and necrosis to PVY-Chile, PVY-Argentina, but showed no symptom to the others. NC744 showed mottling symptom to the PVY-MSNR and PVY-MSMR, necrosis to the PVY-Chile and PVY-Argentina, and no symptom to the others. On N. repanda, necrotic reaction to PVY-Argentina. no symptom to PVY-VN and PVY-C, and nettling to others were observed.

• Korean Journal of Plant Tissue Culture
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• v.26 no.1
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• pp.31-35
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• 1999

Morphological characteristic during formation of tobacco crown gall tumor and genetic tumor, and their differential response to growth regulator were investigated in in vitro culture. Crown gall tumor was induced from tumor tissue transformed by infecting Agrobacterium tumefaciens C58. Genetic tumor was induced from tumor tissue which was induced spontaneously from reciprocal interspecific hybrids between Nicotiana glauca (2n=24) and Nicotiana langsdorffii (2n=18). Morphological characteristic of crown gall tumor, genetic tumor, and teratoma shoot was very similar, and they were actively proliferated on hormone-free medium. Typical tumor callus and teratoma shoot formed from crown gall tumor on the hormone-free medium. On the contrary, tumor callus derived from genetic tumor formed as a crown gall tumor callus on the medium supplemented with 0.5 mg/L of 2,4-D, and lots of teratoma shoots without any root formed on the hormone-free medium. Root development from the teratoma shoots was hardly obtained on the medium with IAA, GA and active carbon. However, teratoma shoots with roots, as normal shoots, were initiated occasionally on the hormone-free medium. These shoots also formed new genetic tumor on the stem, which leaves formed lots of teratoma shoot on the hormone-free medium in in vitro culture.

• Research in Plant Disease
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• v.17 no.2
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• pp.211-215
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• 2011

Cucumber mosaic virus (CMV)-like isolate was collected from Raphanus sativus (cv. Choon-hyang), which showed mosaic symptoms. The isolate was confirmed to a strain of CMV by host responses in Vigna unguiculata, Chenopodium amaranticolor and Gomphrena globosa, by viral genome composition with RT-PCR and PCR-RFLP, and by serological analysis. Symptom developed by the strain of CMV was severe in Nicotiana benthamiana, N. glutinosa, N. tabacum (cv. Samsun, cv. Xanthi), Cucumis melo (cv. Early hanover), Cucumis sativus (cv. White wonder), Capsicum annuum (cv. Chung-yang and cv. Geum-top), but mild symptom was developed in Raphanus sativus (cv. Choon-hyang), Brassica rapa ssp. pekinensis (cv. Bul-Am No. 3), and B. juncea (cv. Daenong Jukgot). Newly isolated strain of CMV could infect diverse crops including Solanaceae, Cucurbitaceae and Brassicaceae. We designated the new strain of CMV as Gn-CMV based on the novel infectivity of Brassicaceae. In double-stranded (ds) RNA analysis, Gn-CMV consisted of 3.3, 3.0, and 2.2 kb genomes likewise other strains of CMV. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) showed 28 kDa of the CMV coat protein. By restriction enzyme mapping using Cac8I, ClaI and MspI of RT-PCR products indicated that Gn-CMV belongs to CMV subgroup I.

• Korean journal of applied entomology
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• v.14 no.2 s.23
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• pp.59-63
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• 1975

A total of 40 virus infected tobacco plants (Nicotiana tabaccum L.) with various symptom types Were collected from Bucheon and Jeonju area by its symptoms were investigated on the incidence of tobacco mosaic virus (TMV), cucumber mosaic virus (CMV), alfalfa mosaic virus (AMV), potato virus X (PVX) and potato virus Y (PVY) by serological methods. van Slogteren's microprecipitin test was applied for the testing of PVX and PVY from infected plants and Ouchterlony agar double diffusion test was used for CMV, TMV and AMV. Results obtained are as follows: 1. TMV, CMV, AMV, PVX and PVY wcre found to occur on the tobacco plants growing in Korea. 2. The prevalence of each of these viruses among the 40 tobacco plants investigated was in the order of AMV: $(67.5\%)>CMV:(60.0\%)>TMY:(47.5\%)>PVY:(17.5\%)>PVX: (10.0\%).$ 3. In Burley variety, the percentage of infection by TMV was $15\%$, whereas it was as high as $80\%$ in Hicks variety. 4. Among the 40 tobacco plants investigated, $37.5\%$ showed infection with one kind of virus whereas the remaining $62.5\%$, revealed mixed infection with more than two different viruses.

• Research in Plant Disease
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• v.12 no.3
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• pp.221-225
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• 2006

Cucumber mosaic virus(CMV) and Broad bean wilt virus 2(BBWV2) were isolated from Gentiana scabra plants showing typical mosaic and yellowing symptoms. When the inoculum of mosaic symptom propagated in Nicotiana benthamiana was inoculated to primary leaves of Vigna sinensis, the local lesions of different types was developed. Type one produced a small necrotic spot(SNS) of pinpoint type, while the other one showed a large necrotic spot(LNS) of halo type. LNS on primary leaves of V. sinensis was also induced by inoculum from yellowing symptom on G scabra. Single lesion from SNS induced a typical mosaic symptom on N. Benthamiana. On the other hand, LNS produced a chlorotic ring symptom on inoculated leaves and mosaic plus necrotic ringspot on upper leaves of N. benthamiana. An isolate of CMV from SNS and BBWV2 from LNS were detected by using dsRNA analysis, RT-PCR and agar gel double-diffusion test. Thus, our results should provide a tool of a simple method for discrimination from mixed infected plants by CMV and BBWV2.

• Research in Plant Disease
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• v.15 no.2
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• pp.68-71
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• 2009

Cucumber green mottle mosaic virus (CGMMV) was not infected in figleaf gourd by sap inoculation. However CGMMV was detected by RT-PCR from the figleaf gourd collected from a field growing cucumber grafted onto figleaf gourd in Cheonan, Chungcheongnam Province in 2008. Which field showed 100% infection rate of the virus disease. In the experiment grafted with cucumber onto figleaf gourd, transportation of CGMMV through cucumber to figleaf gourd was confirmed by RT-PCR when the virus was mechanically inoculated on the leaves of the cucumber. The amplified DNA concentration of the virus on electrophoresis gel was much higher in the cucumber than in the figleaf gourd. However, the virus particles from the figleaf gourds were not observed under electron microscopy, also sap of the figleaf gourds was not transmittable to Nicotiana benthamiana. To identify the existence of CGMMV particle, the virus was purified from figleaf gourd and cucumber growing together in the graft system. CGMMV solution extracted from the cucumber represented a typical absorption spectrum of the virus but that from the figleaf gourd did not. Only a few CGMMV particles were observed in the purified preparation from the figleaf gourd. These results confirmed that CGMMV only passed through figleaf gourd in the grafting system. This study indicated that figleaf gourd is not a host of CGMMY.

• Korean Journal of Soil Science and Fertilizer
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• v.42 no.2
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• pp.70-78
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• 2009

Remote sensing technique in agriculture can be used to identify chlorophyll content, biomass, and yield caused from N stress level. This study was conducted to evaluate biomass, N stress levels, and yield of tobacco (Nicotiana tabacum L.) under sand culture in a plastic film house using ground-based remote sensors. Nitrogen rates applied were 40, 60, 80, 100, 120, and 140 percent of N concentration in the Hoagland's nutrient solution. Sensor readings for reflectance indices were taken at 30, 35, 40, 45, 50 and 60 days after transplanting(DAT). Reflectance indices measured at 40th DAT were highly correlated with dry weight(DW) of tobacco leaves and N uptake by leaves. Especially, green normalized difference vegetation index(gNDVI) from spectroradiometer and aNDVI from Crop Circle passive sensor were able to explain 85% and 84% of DW variability and 85% and 92% of N uptake variability, respectively. All the reflectance indices measured at each sampling date during the growing season were significantly correlated with tobacco yield. Especially the gNDVI derived from spectroradiometer readings at the 40th DAT explained 72% of yield variability. N rates of tobacco were distinguished by sufficiency index calculated using the ratio of reflectance indices of stress to optimum plot of N treatment. Consequently results indicate that the reflectance indices by ground-based remote sensor can be used to predict tobacco yield and recommend the optimum application rate of N fertilizer for top dressing of tobacco.