• 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.

• The Plant Pathology Journal
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• v.22 no.2
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• pp.131-138
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• 2006

Two strains of Cucumber mosaic virus (CMV) isolated in Kuwait were confirmed their infectivity based on symptomatology and host range on different cultivars of tomato (Lycopersicon esculentum), tobacco(Nicotiana tabacum L.) and squash (Cucurbita pepo). The pattern of symptoms differed for the two CMV strains in tomato and tobacco, showing severe stunting and mosaic symptoms with one strain designated KU2, and almost symptomless with the other strain designated KU1. A satellite RNA 5 (sat-RNA) was found to be associated with the KU1 strain and was characterized as a benign viral satellite RNA. Using reverse transcription and polymerase chain reaction (RT-PCR) with sat-RNA specific primers, an amplified PCR product of about 160bp was determined and analyzed by gel electrophoresis. This naturally occurring benign viral satellite RNA was successfully used as a biological control agent to protect tomato plants against the severe KU2 strain. Tomato plants grown in plant-growth chambers, were preinoculated with KU1 containing the benign viral satellite and then challenge inoculated with the severe KU2 strain at different time intervals. All plants challenged three weeks after preinoculation showed nearly complete protection from subsequent infection by the severe strain. This biological control technology using plant viruses was found protective and could be successfully established sooner after the preinoculation.

• KSBB Journal
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• v.7 no.3
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• pp.155-160
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• 1992

To improve the nutritional quality of plant proteins, a synthetic gene, called HEAAE (high essential amino acid encoding)-DNA, was introduced and expressed in tobacco plants. The synthetic gene, which is 292 basepair-long, codes for a protein composed of about 80% essential amino acids. To improve its expression level in plants, Cauliflower Mosaic Virus (CaMV) 355 and CaMV duplicate 35S promoters which are known as strong promoters were used with Nopaline Synthase promoter as a control. Transformed and regenerated tobacco plants were subject to analysis for introduction and expression of this gene. Integration of the gene into the plant genome and its expression into mRNAs and its proteins have been demonstrated using Southern, northern blot analysis and amino acid analysis. The differences of expression levels among CaMV duplicate 35S, CaMV 35S and Nopaline Synthase promoters are significant in term of mRNAs, but not in terms of proteins.

• Horticultural Science & Technology
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• v.35 no.3
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• pp.323-332
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• 2017

Homology-specific transcriptional and post-transcriptional silencing, an intrinsic mechanism of gene regulation in most eukaryotes, can be induced by anti-sense, co-suppression, or hairpin-based double-stranded RNA. Hairpin-based RNA interference (RNAi) has been applied to analyze gene function and genetically modify crops. However, RNAi vector construction usually requires high-cost cloning steps and large amounts of time, or involves methods that are protected by intellectual property rights. We describe a more effective method for generating intron-spliced RNAi constructs. To produce intron-spliced hairpin RNA, an RNAi cassette was ligated with the first intron and splicing sequences of the Brassica rapa ssp. pekinensis histone deacetylase 1 gene. This method requires a single ligation of the PCR-amplified target gene to SpeI-NcoI and SacI-BglII enzyme sites to create a gene-specific silencing construct. We named the resulting binary vector system pKHi and verified its functionality by constructing a vector to silence DIHYDROFLAVONOL 4-REDUCTASE (DFR), transforming it into tobacco plants, and confirming DFR gene-silencing via PCR, RT-qPCR, and analysis of the accumulation of small interfering RNAs. Reduction of anthocyanin biosynthesis was also confirmed by analyzing flower color of the transgenic tobacco plants. This study demonstrates that small interfering RNAs generated through the pKHi vector system can efficiently silence target genes and could be used in developing genetically modified crops.

• The Journal of Natural Sciences
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• v.10 no.1
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• pp.33-37
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• 1998

Cytokinin is one of major growth regulators in plants. In this study, the gene isopentenyl transferase (jpt) which encodes a key enzyme involved in the biosynthesis of the growth regulator cytokinin isolated from Agrobacterium tumefaciens was introduced ito tobacco plant via Agrobacterium-mediated transformation. The jpt gene was modulated using the proteinase inhibitor II (PI-IIK) promotor. In general, this promoterlipt gene fusion resulted in overproduction of cytokinins throughout the transgenic plants. The overproduction of cytokinin caused dramatic changes in morphology of the plant, including stem thickness and reduced root development. The studies reported in this paper were initiated to examine the consequences of overproduction of cytokinin in plant.

• Korean Journal of Plant Tissue Culture
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• v.25 no.2
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• pp.89-93
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• 1998

Tomato phenylalanine ammonia-lyase 5 (tPAL5) was identified that alternate initiation sites were utilized differentially in response to environmental stimuli (Lee et al, 1992b). In this study, we tried to look into tissue -or cell- specific expression pattern of tPAL5 gene by fusing with ${\beta}-glucuronidase$ (GUS) gene in transgenic tobacco plants. In transgenic plants, root and stem extracts contained 8~12 fold higher levels of GUS activity than petiole or leaf tissue while the highest levels of induction was observed from leaf tissue by mechanical wounding (5~11 fold). In trans-sections of stems and petioles, GUS activity was restricted to phloem cells(outer region) of developing vascular bundle and mainly at apical tip region in the root tissues. The levels of GUS activity was drastically reduced (10~12 fold reduction) when the 5'-upstream region of tPAL5 gene (-1151bp from ATG codon) was deleted up to -665. The levels of GUS expression, however, raised up by 6~8 fold when deleted up to -455. Therefore, we conclude that there are positive cis-elements at the region -1151 to -1008 and at -455 to -195 while the negative cis-element is at -1008 to -455.

• Korean Journal of Plant Tissue Culture
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• v.27 no.3
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• pp.239-243
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• 2000

Helicobacter pylori, an etiologic agent of gastritis and peptic ulceration, produces urease which elicits a powerful immunoglobulin response in H. pylori-infected individuals. To establish a model plant vaccine agains H. pylori, 750 bp -ureA DNA amplified by polymerase chain reaction from pH 808 plasmid harboring urease gene cluster was cloned and manipulated to be expressed in tobacco plants. From the regenerated transgenic tobacco plants, ureA DNA integration,m its mRNA expression and protein synthesis were analyzed and confirmed by standard molecular techniques. The CaMV 35S promoter-driving ureA construct was expressed to produce a 30 kDa protein which was identical with bacterial UreA in size when detected on immunoblot of SDS polyacrylamide gel electrophoresis.

• Applied Biological Chemistry
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• v.37 no.4
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• pp.248-254
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• 1994

Bromoxynil is an antidicot herbicide widely used on cereal crops and has a short half life in the soil. A bxn gene, encoding a specific nitrilase that converts bromoxynil to its primary metabolite 3,5-dibromo-4-hydroxybenzoic acid, was inserted in plant binary vector pGA482, and then introduced into tobacco and lettuce plants via Agrobacterium mediated leaf-disc transformation method. Transgenic plants with the bxn gene were selected by kanamycin and regenerated to whole plants. The regenerated transgenic plants were determined level of expression of bxn gene by Northern blot analysis. Leaf-disc analysis and pot-assay confirmed that the transgenic tobacco and lettuce plants were resistant to high doses of bromoxynil.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.30 no.4
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• pp.347-351
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• 1985

The effect of leaf water content on apparent photosynthesis and respiration of tobacco plants(five varieties) was studied under the condition without the irrigation for 10 days after the plants were sufficiently watered on the first day. The wild race (N. longiflora) among varieties showed highest apparent photosynthesis (AP) and AP had a positive correlation with specific leaf weight. N. longiflora and Andongyeob were different in their AP from the other varieties significantly under the water stress condition. Respiration rate also decreased to be simillar to AP except slight increase at early stage of water deficit. The stomatal resistance and the mesophyll resistance increased in the stressed plants. The water stress resistant character seems to be mainly due to open stomata.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.41 no.6
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• pp.692-697
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• 1996

This experiment was conducted to determine the influences of growth stage, stalk position and growth environment on duvatrienediols(DVT-diols) and cis-abienol production in Nicotiana tabacum L. The leaves of plants at transplanting stage contained very small amount of duvatrienediols and cis-abienol. Comparing leaves on different stalk position at flowering stage, upper and middle leaves contained more DVT-diols and cis-abienol than lower leaves. Plants grown under controlled environmental room at 3$0^{\circ}C$ contained more DVT-diols and cis-abienols than room at 18$^{\circ}C$, $25^{\circ}C$ and glass room. Tobacco plant grown under field condition produced lower levels of duvatrienediols and cis-abienol contents than glass room-grown plants. The amounts of duvatrienediols and cis-abienol of tobacco leaves significantly decreased after rain under field conditions.