• 한국연초학회지
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• 제33권1호
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• pp.1-7
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• 2011

Major diseases of tobacco plants were surveyed throughout the Korea in 2011. Mosaic, bacterial wilt and hallow stalk were most severe during the harvest season on not only flue-cured tobacco plants but also burley tobacco plants. On flue-cured tobacco plants, mosaic caused by potato virus Y were more severe than those by tobacco mosaic virus or cucumber mosaic virus. The mosaic caused by potato virus Y was severe at Yeongwol and Chungju. On burley tobacco plants, mosaic were more severe at Jeongeup and Gochang than those at Chungnam and Jeonnam. A negative correlation between the mosaic incidence and the precipitation was recognized. On the other hand, there was a positive correlation between the incidence of hallow stalk incidence of flue-cured tobacco plants in harvesting stage and the precipitation during June was recognized significantly.

• BMB Reports
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• 제32권1호
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• pp.1-5
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• 1999

In order to understand the biological role of calmodulin in plants, transgenic tobacco plants expressing a calmodulin mutant (VU-4 calmodulin, lys to ile-115) gene have been analyzed. SDS-PAGE and Western-blot analyses showed that the foreign calmodulin mutant is stably and highly expressed in the transgenic tobacco plants. The levels of $H_2O_2$were elevated approximately 2-fold in the transgenic plants. Furthermore, the transgenic tobacco plants have more than 6-fold higher levels of NADPH compared to control tobacco plants. The present findings, combined with previous data showing differences in the susceptibility of the transgenic tobacco seeds and normal tobacco seeds to fungal contamination (Oh and Yang, 1996), suggest that the expression of the calmodulin derivative gene in tobacco plants could increase resistance to infection by fungal pathogens.

• 한국식물병리학회지
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• 제11권4호
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• pp.374-379
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• 1995

The cDNA of tobacco mosaic virus-pepper strain (TMV-P) coat protein (CP) genes were introduced into tobacco plants (Nicotiana tabacum cv. Samsun nn) using a binary Ti plasmid vector of Agrobacterium tumefaciens. these cDNAs introduced into tobacco plants were detected by polymerase chain reaction. Symptom development was distinctly suppressed in the transgenic plant introduced buy sense CP cDNA when the plant was inoculated with TMV-P, while in transgenic tobacco plants of antisense CP gene, symptom development was not suppressed as in non-transgenic plants. TMV-P concentration in the sense CP transgenic tobacco plant was decreased to 1/14 of the concentration in non-transgenic plants. Expression of the kanamycin resistance gene of these transgenic plants could be detected in the progeny.

• 한국식물병리학회지
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• 제11권1호
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• pp.80-86
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• 1995

The cDNA of CMV-As satellite RNA was introduced into tobacco plants (Nicotiana tabacum cv. Samsun NN) using a binary Ti plasmid vector system of Agrobacterium tumefaciens. The cDNA of satellite RNA introduced into tobacco plants was detected by polymerase chain reaction (PCR) and molecular hybridization analyses. Symptom development was distinctly suppressed in the transgenic tobacco plants when inoculated with CMV-Co. CMV concentration in the transgenic tobacco plants was decreased to 1/40 of non-transgenic tobacco plants. The kanamycin resistance gene of the transgenic tobacco plants was also detected in the progeny.

• Journal of Plant Biotechnology
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• 제34권1호
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• pp.31-36
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• 2007

Though higher plants car not metabolize D-amino acid, many prokaryotes and eukaryotes have the D-amino acid metabolism. Therefore, we transformed tobacco plants with D-amino acid oxidase (DAO), which can metabolize D-amino acid, and confirmed that transgenic tobacco plants might metabolize D-amino acid. Transgenic tobacco plants were survived a high concentration of D-serine, however non-transgenic plants were not grown on D-serine medium. From Southern and Northern blot analysis, transgenic tobacco plants selected on D-serine medium were confirmed by insert and expression of transgene. $T_{1}$ tobacco seeds derived $T_{0}$ tobacco plants selfing were grown on D-serine medium and showed normal phenotype compared to wild tobacco plants. Transgenic tobacco plants displayed the metabolic capability of D-serine. Therefore, we suggested that DAO is useful selectable marker gene for plant transformation.

• 한국연초학회지
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• 제18권1호
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• pp.21-29
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• 1996

Tobacco plants resistant (cvs. Xanthi-nc and Samsun-NN) and susceptible (cv. NC 82) to tobacco mosaic virus (TMV) were inoculated with TW to obtain basic information about the characteristics of resistance expression in tobacco plants by examining the viral populations, symptom development and gene expression of pathogenesis-related proteins (PR-proteins) such as PR-1 and $\beta$-1, 3-glucanase in different temperature conditions. TMV populations in resistant plants increased more at 37$^{\circ}C$ than at 27$^{\circ}C$, while the viral populations increased continuously and were not significantly influenced by the temperature conditions in the susceptible tobacco plants. Infection sites of resistant tobacco leaves were remarkably expanded in proportion with increased time at the high temperature.

• Journal of Applied Biological Chemistry
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• 제42권1호
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• pp.19-24
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• 1999

Plants have been shown to contains $Ca^{2+}$/calmodulin-stimulated GAD and NAD kinase. To test how calmodulin and calmodulin methylation affect the activation of GAD and NAD kinase, GAD and NAD kinase were partially purified from tobacco plants. GAD was also partially purified from E. coli transformed with a plasmid carrying a cloned tobacco GAD gene. We find that GAD from the transformed E. coli showed 60-fold $Ca^{2+}$/calmodulin-dependent activation. However, GAD from tobacco plants was stimulated only about 3.8-fold by the addition of calmodulin in the presence of calcium, suggesting high background activity of the enzyme was possibly due to bound endogenous tobacco calmodulin. There were no significant differences in the tobacco GAD activator properties between calmodulins. A monoclonal antibody against petunia GAD interacted strongly with both GAD from tobacco plants and GAD from cloned gene. NAD kinase from tobacco plants showed a complete $Ca^{2+}$/calmodulin dependency for activity. Unmethylated calmodulins activated GAD in a manner similar to methylated calmodulin. However, the maximum level of NAD kinase activation obtained with unmethylated calmodulins is approximately 4-fold higher than methylated calmodutins. These data suggested that endogenous tobacco calmodulin may interact more tightly with GAD than NAD kinase and that calmodulin methylation affects the activator properties of calmodulins for tobacco NAD kinase but not for GAD.

• 한국연초학회지
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• 제20권1호
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• pp.57-65
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• 1998

This study was conducted to develop a resistant tobarro against Potato virus Y (PVY) by transformation of the plants with genetically engineered viral genes. The complimentary DNAs (cDNAS) of potato virus Y-necrosis strain (PVY-Vn) replicase mutant genes (3'-deleted, 5'-deleted and ADD-mutant Nlbs) were synthesized through RT-PCR by using purified PVY-VN RNA and synthesized primers, and cloned in the sense orientation into a plant expression vector (pMBPI), The cDNAS of the genes were transferred into Agrobacterium tumefaciens LBA 4404, and then transformed into tobacco (Nicotiana tabacum cv. Burley 21) plants. Regenerated plants were tested for PVY resistance by inoculation test; 13 transgenic plants including 7 for 3'-deleted Nlb, 3 for 5'-deleted Nlb, and 3 for ADD-mutant Nlb appeared to be resistant at 4 weeks after inoculation with PVY-VN. Among the 13 transgenic tobacco plants, 8 plants had no symptom up to 14 weeks after inoculation. The progenies ($T_1$) from self-fertilization of the transgenic lines varied 0.0% to 81.2% in their resistance (% of resistant plants). The analysis of Nlb-31deleted, -5'deleted and -ADD mutant in the $T_1$ plants by polymerase chain reaction (PCR) showed that Nlb-3'deleted, -5'deleted and -ADD mutants were detected in all of the resistant plants. These results suggest that the PVY resistance was inherited in the $T_1$ generation.

• The Plant Pathology Journal
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• 제16권5호
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• pp.264-268
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• 2000

A fluorescent material was accumulated in inoculated leaves showing necrotic local lesions of tobacco plants with N gene, Nicotiana tabacum cvs. Xanthi-nc NN, Samsun NN, Burley 21 and KF 114, and N. glutinosa, and Datura stramonium at the early growth stages by the inoculation of Tobacco mosaic virus (TMV). It was identified as a coumarin phytoalexin, scopoletin. Although the material was most prominently produced in TMV-inoculated tobacco leaves with local necrotic lesions, its accumulation was also noted in uninoculated leaves of TMV-inoculated plants. Its accumulation was somewhat greater in high resistance-induced leaves than low resistance-induced and intact leaves. Scopoletin treatment induced the expression of a pathogenesis-related protein, PR-1, prominently at the concentration of 500 or 1000 ${\mu}$g/ml. This suggests that scopoletin is a phytoalexin abundantly accumulating in N gene-containing resistant plants in response to TMV infection, and may be related to hypersensitive responses (HR) and systemic acquired resistance (SAR) in the resistant tobacco plants.

• 한국연초학회지
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• 제19권1호
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• pp.11-17
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• 1997

A flue-cured tobacco variety (Nicotiana tabacum cv. Wisconsin) was used for Plant transformation with the complementary DNA (cDNA) of potato virus Y-necrosis strain (PVY-VN) replicase gone (Nb) which was synthesized through reverse-transcription Primed with oligo(dT) and Polymerization using RNase H-digested template. The cDNA was cloned into Plant expression vector Plasmid (PMBP2), and introduced into tobacco plants by co-culturing tobacco leaf disks with Agrobacterium tumefaciens LBA4404 containing the plasmid before Plant regeneration. Eight Plants, in which the inserted cDNA fragment was detected by Polymerase chain reaction (PCR), out of 70 putative transformants inserted with sense-oriented Mb cDNA showed no symptom at 3 weeks after inoculation, while the other 62 plants, and all plants with vector gone only and antisense-oriented NIb cDNA had susceptible vein-necrosis symptoms. However, only 2 of the 8 resistant plants were highly resistant, which remained symptomless up to 10 weeks after inoculation. Among the first progenies (T1) from self-fertilized seeds of the two resistant transgenic plants, less than 10 % of 71 plants appeared highly resistant (with no symptom), 70% moderately resistant (with mild symptoms on 1 - 2 leaves), and about 20% susceptible (with susceptible symptoms on 3 or more leaves) at 3 weeks after inoculation. These results suggest that the PVY resistance was inherited in the 71 generation. Key words : potato virus Y. viral replicase gene, transgenic tobacco Plants, resistance.