• 한국연초학회지
• /
• 제14권2호
• /
• pp.159-167
• /
• 1992

It would be clear that the constituents of the leaf surface lipid is ye비 important as an evaluation index of tobacco leaf quality since the quality of tobacco specific aroma with leaf species depends on the contents of the lipid and the strength of the aroma is determined by the amounts of the lipid secreted. For the reason, a rapid and peproducible method to quantify DVT, which is a kind of lipid, has been studied. The biosynthesis procedure of DVT in leaf growing processes, and the volatile or decompositional characters of DVT in leaf drying processes were also discussed. In consequence, it might be possible to get the data available to the cultivation of better tobacco leaf and the manufacture of cigarettes with better aroma and taste. The results obtained from this study are as follows. 1. Chloroform/dichloromethane solvent was better than chloroform alone for DVT extraction. The extraction yields of the leaf surface lipid were about 5% 2. The extractives with dichloromethane were treated by silylation with BSTPa and the quantitative analysis of DVT was carried out using SE -54 fused silica capillary column. It was found that rapid and reproducible data could be obtained from these methods. 3. In flue - cured tobacco species, DVT contents were $30.3\mu\textrm{g}/cm^2$ in the beginning stage of leaf drying processes and $12.1\mu\textrm{g}/cm^2$ corresponded to 30% levels of the beginning stage, in the end stage. 4. DVT contents in Burley mere 2 times as large as those in fluecured tobacco. DVT in the upper stalk position of leaf was 3 times larger than that in the lower stalk position. 5. DVT of tobacco leaves was decomposed by $SO_2$ gas or the sun light. The decomposition rate was largest in the sample used methanol as a extraction solvent.

• 한국연초학회지
• /
• 제14권2호
• /
• pp.116-125
• /
• 1992

This experiment was conducted to examine characteristics of agronomic characters and estimate of gene effect for several mutant characters. The genetic populations were derived from cross between 83H-5 and Hicks. There were significant difference for plant height, stlk height, leaf shape and bacterial wilt disease index except leaf number, leaf length, and what is more, F3 variance is more than Bl and B2 generation from cross 83H-5 X Hicks. Gene actions for stalk height and bacterial wilt disease were estimated by 3-parameter, and by 6- parameter model for all characters except above two characters but stalk height and bacterial wilt disease index are not significant in the additive and dominance effects. Dominant$\times$dominant epitasis for plant height, dominant and dominant$\times$dominant epistasis for leaf length, additive and additive$\times$additive and dominant$\times$dominant epistasis for leaf width, and additive and additive$\times$dominant epistasis for days to flower were appeared significant in gene action.

• 한국연초학회지
• /
• 제20권1호
• /
• pp.57-65
• /
• 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.

• 한국연초학회지
• /
• 제14권2호
• /
• pp.126-132
• /
• 1992

This experiment was conducted to examine estimates of heterosis, degree of dominance, heritability and phenotypic and genotypic correlation for several mutant characters in flue-cured tobacco variety. Significant heterosis was noted, recording 2.73 and 6.16% for leaf width, -6.86 and -4.72 for leaf shape index, 19.8 and 23.4% for bacterial wilt disease index in Fl an4 F2 generation, respectively, and -1.44 for days to flower in F2 generation. And also leaf width was appeared partial dominance in Fl, and leaf shape index in Fl and F2, Leaf width in F2 was appeared overdominance. Estimated heritabilities in the broad sense ranged from 0.77 to 0.87 for plant and stalk height and days to flower, while those of leaf number, leaf length and width, leaf shape index and bacterial wilt disease index ranged from 0.50 to 0.65. And the heritabilities in the narrow sense were appeared 0.64 and 0.72 for stalk and plant height, respectively, and the others were ranged from 0.32 to 0.47 Positive genotpic correlations appeared among plant height, stalk height, leaf length and leaf width, and between leaf shape index ands day to flower. And negative genotypic correlations appeared between leaf shape index and plant height and stalk height and leaf width, and between bacterial wilt disease index and leaf length.

• 한국연초학회지
• /
• 제20권1호
• /
• pp.66-70
• /
• 1998

TMV resistant lines (TRLs) originated from the Blo plant of Nicotiana tabacum cv. NC82 transformed with TMV coat protein cDNA which initially showed delayed disease symptom were selected for increased resistance in each subsequent generation. The result of field experiment of the transgenic tobacco lines in the fifth generation for TMV resistance and their response to other tobacco diseases (black shank, bacterial wilt, and powdery mildew) is described in this report. When fifteen TRLs of the fifth generation were tested for TMV resistance by mechanically inoculating the individual plants, over 95 percent of the plants of 6 lines showed complete resistance even 8 weeks after the inoculation. Average frequency of the resistant plants in TRLs of the fifth generation 8 weeks after the inoculation was 87%. Stable insertion and expression of TMV coat protein cDNA in the fifth generation of the transgenic tobacco plant were confirmed by PCR and immunoblot hybridization, respectively. All TRLs were resistant to the black shank but were susceptible to the bacterial wilt disease and the powdery mildew to the same degree as non-transgenic NC82 was. Therefore, it was indicated that the phenotypes related at least to disease resistance were not changed in the transgenic tobacco. Key words : TMV CP cDNA, TMV resistant tobacco plant, transformation.

• BMB Reports
• /
• 제34권4호
• /
• pp.316-321
• /
• 2001

Dehydroascorbate (DHA) reductase (DHAR, EC 1.8.5.1) catalyzes the reduction of DHA to reduced ascorbate (AsA) using glutathione (GSH) as the electron donor in order to maintain an appropriate level of ascorbate in plant cells. To analyze the physiological role of DHAR in environmental stress adaptation, we developed transgenic tobacco (Nicotiana tabacum cv. Xanthi) plants that express a human DHAR gene isolated from the human fetal liver cDNA library in the chloroplasts. We also investigated the DHAR activity, levels of ascorbate, and GSH. Two transgenic plants were successfully developed by Agrobacterium-mediated transformation and were confirmed by PCR and Southern blot analysis. DHAR activity and AsA content in mature leaves of transgenic plants were approximately 1.41 and 1.95 times higher than in the non-transgenic (NT) plants, respectively In addition, the content of oxidized glutathione (GSSG) in transgenic plants was approximately 2.95 times higher than in the NT plants. The ratios of AsA to DHA and GSSG to GSH were changed by overexpression of DHAR, as expected, even though the total content of ascorbate and glutathione was not significantly changed. When tobacco leaf discs were subjected to methyl viologen at $5\;{\mu}M$, $T_0$ transgenic plants showed about a 50% reduction in membrane damage compared to the NT plants.

• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2002년도 생물공학의 동향 (X)
• /
• pp.325-328
• /
• 2002

Sucrose 의 경우 저농도에서는 후반부로 갈수록 세포생장 속도는 현저히 감소하는데 비해 hGM-CSF 생산은 후반부에 급격히 증가함을 확인하였다. 고농도 sucrose를 사용하는 경우에는 lag phase가 길어지는 동안에 hGM-CSF의 생산이 증가하였다. 따라서 배양 초기에는 고농도 sucrose가, 배양 후반에는 저농도 sucrose로 존재하는 경우에 hGM-CSF를 많이 얻을 수 있었다. Nitrogen source의 농도는 60.52 mM과 121.04 mM일 때가 세포의 생장이나 hGM-CSF의 생산을 증가시켰으며, phosphate의 경우에는 4.96 mM 일 때가 대조구인 2.48 mM 일 때보다 hGM-CSF의 생산을 3 배 증가시켰다.

• Journal of Microbiology and Biotechnology
• /
• 제18권1호
• /
• pp.67-73
• /
• 2008

The potential of the exopolysaccharide (EPS) from a Serratia sp. strain Gsm01 as an antiviral agent against a yellow strain of Cucumber mosaic virus (CMV-Y) was evaluated in tobacco plants (Nicotiana tabacum cv. Xanthi-nc). The spray treatment of plants using an EPS preparation, 72h before CMV-Y inoculation, protected them against symptom appearance. Fifteen days after challenge inoculation with CMV-Y, 33.33% of plants showed mosaic symptoms in EPS-treated plants compared with 100% in the control plants. The EPS-treated plants, which showed mosaic symptoms, appeared three days later than the controls. The enzyme-linked immunosorbent assay (ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR) analyses of the leaves of the protected plants revealed that the EPS treatment affected virus accumulation in those plants. Analysis of phenylalanine ammonia lyase, peroxidase, and phenols in protected plants revealed enhanced accumulation of these substances. The pathogenesis-related (PR) genes expression represented by PR-lb was increased in EPS-treated plants. This is the first report of a systemic induction of protection triggered by EPS produced by Serratia sp. against CMV-Y.

• The Plant Pathology Journal
• /
• 제18권3호
• /
• pp.130-137
• /
• 2002

Potato virus Y (PVY) is one of the most important viruses in many field crops in Korea. In this study, 31 PVY isolates were isolated from infected potato (Solanum tuberosum), tobacco (Nicotiana tabacum), pea (Pisum sativum), and weeds (Veronica persica, Lamium amplexicause and Capsella bursa-pastoris) showing different mosaic symptoms in Jeonbuk, Chungnam, Gangwon, and Gyeongbuk areas in Korea. The 640 nucleotide region containing the C-terminal portion of coat protein (CP) gene and 3'non-translated region (NTR) was amplified by reverse transcription-polymerase chain reaction (RT-PCR) using PVY-specific oligonucleotide primers. Sequence analyses of the amplified DNA fragments showed that the C-terminal portion of CP gene was not significantly different from that of previously reported PVY strains from potato (PVY-OK and -T) and tobacco (PVY-VN) in Korea. Homologies of the deduced CP amino acid sequences were 93.3-99.0% to corresponding regions of the other PVY strains including PV $Y^{N}$, PV $Y^{o}$ , PV $Y^{OK}$ , PV $Y^{T}$ , and PV $Y^{VN}$ . In contrast the sequences located at the 3'-NTR showed more diverse sequence homologies (76.4-99.7%). These results indicate that the C-terminal portion of the CP gene was relatively conserved while sequences at the 3'NTR were more diverse and variable over the host species and the regions where they were isolated.e isolated.

• The Plant Pathology Journal
• /
• 제30권2호
• /
• pp.200-207
• /
• 2014

A viral disease causing severe mosaic, necrotic, and yellow symptoms on Vigna angularis var. nipponensis was prevalent around Suwon area in Korea. The causal virus was characterized as Cucumber mosaic virus (CMV) on the basis of biological and nucleotide sequence properties of RNAs 1, 2 and 3 and named as CMV-wVa. CMV-wVa isolate caused mosaic symptoms on indicator plants, Nicotiana tabacum cv. Xanthi-nc, Petunia hybrida, and Cucumis sativus. Strikingly, CMV-wVa induced severe mosaic and malformation on Cucurbita pepo, and Solanum lycopersicum. Moreover, it caused necrotic or mosaic symptoms on V. angularis and V. radiate of Fabaceae. Symptoms of necrotic local or pin point were observed on inoculated leaves of V. unguiculata, Vicia fava, Pisum sativum and Phaseolus vulgaris. However, CMV-wVa isolate failed to infect in Glycine max cvs. 'Sorok', 'Sodam' and 'Somyeong'. To assess genetic variation between CMV-wVa and the other known CMV isolates, phylogenetic analysis using 16 complete nucleotide sequences of CMV RNA1, RNA2, and RNA3 including CMV-wVa was performed. CMV-wVa was more closely related to CMV isolates belonging to CMV subgroup I showing about 85.1-100% nucleotide sequences identity to those of subgroup I isolates. This is the first report of CMV as the causal virus infecting wild Vigna angularis var. nipponensis in Korea.