• 한국연초학회지
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• 제4권2호
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• pp.23-30
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• 1982

Six flue- cured cultivars of Nicotiana tabacum L., 15 possible $F_1$ hybrids and $F_2$ Populations among them, and 15 haploid Populations from Fl hybrids and haploids from Parents, were evaluated. Comparisons of the $F_1$, hybrids and $F_2$ Populations with the Parents indicated that heterosis values were small but significant for yield, plant height, days to flower, leaf length and width and total alkaloids from-6.0% to 5.4% in $F_1$ hybrids, and from -3.4% to 3.6% for Plant height days to flower leaves per plant in $F_2$ populations, respectively. There were positive correlations for yield, plant height, days to flower, leaves per plant and total alkaloids between diploid and haploid populations. Increase or decrease ratio of haploids to diploids of total alkaloids was appeared to 30.3%. Those for yield, leaf length and width, value arid reducing sugar were ranged from -4.1% to -27.6%.

• KSBB Journal
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• 제6권3호
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• pp.289-297
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• 1991

The insecticidal protein gene in the pKL-20-1 clone derived from Bacillus thuringiensis serovar. kurstaki plasmid was subcloned in the plant shuttle vector, pGA643. The 7.3 kb fragment was cloned in the BglII and Hpal sites of pGA643 vector and expressed in E. coli S17-1, which produced insecticidal proteins killing Bombyx mori larvae. The clone was named pHL-20. The protoplast formation, calli induction and plantlet regeneration of Nicotiana tabacum was carried out. A tremendous number of mesophyll protoplasts of N. tabacum were formed, up to 7$\times$105 protoplast per ml, for 20 hours in darkness in the enzyme solution of 0.5% cellulase and 0.1% macerosin, pH 5.8. The viabilities of the protoplasts were maintained above 80% for 6 days in the media containing 2mg/1 of NAA and 1mg/1 of kinetin. Calli were induced from the protoplasts and leaves of the N. tabacum on MS medium containing 0.5mg/1 BAP. Under the culture conditions the protoplasts underwent repeated cell division into calli. Plantlets were regenerated from callus cultures derived from protoplast and leaves. Shoots were induced in a medium containing 1mg/1 of BAP.

• KSBB Journal
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• 제19권5호
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• pp.374-380
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• 2004

Production of therapeutic proteins by transgenic plant cell suspension cultures is an attractive system alternative to the other expression system. However, plant cell cultures have shown low expression level of foreign proteins and decreased cell viability by the changes of culture conditions. Therefore, it is necessary to enhance cell viability during the culture period. In this study, a quantitative analysis technique was designed to measure relative cell viability for plant suspension cells which have cell wall and aggregates. It was found that the programmed cell death of plant cells by apoptosis was essentially linked with the apoptotic pathway of animal cells. Therefore, effects of nicotinamide, 3-aminobenzamide and antioxidants on cell viability and apoptosis were examined in transgenic Nicotiana tabacum cells producing hGM-CSF. With those additives, cell viability could be maintained and apoptosis could be redued. In the result, the extracellular production of hGM-CSF could be enhanced 2.5 fold. It was also found that the supplementation of glutathione and ascorbic acid suppressed both the cold stress-induced decrease in cell viability and the increase of total genomic DNA fragmentation.

• 한국연초학회지
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• 제30권2호
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• pp.85-93
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• 2008

This report describes a set of seven informative single-nucleotide polymorphisms (SNPs) and one insertion-deletion (INDEL) distributed over 24 cultivars that can be used for tobacco (Nicotiana tabacum L.) cultivar identification. We analyzed 163,000 genomic DNA sequences downloaded from Tobacco Genome Initiative database and assembled 31,370 contigs and 60,000 singletons. Using relatively long contigs, we designed primer sets for PCR amplification. We amplified 61 loci from 24 cultivars and sequenced the PCR products. We found seven significant SNPs and one INDEL among the sequences and we classified the 24 cultivars into 10 groups. SNP frequency of tobacco, 1/8,380 bp, was very low in comparison with those of other plant species, between 1/46 bp and 1/336 bp. For exact identification of tobacco cultivars, many more SNP markers should be developed. This study is the first attempt to identify tobacco cultivars using SNP markers.

• 한국자원식물학회지
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• 제23권6호
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• pp.535-540
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• 2010

Phenylalanine ammonia-lyase (PAL), a key enzyme of the phenylpropanoid biosynthesis pathway, is activated by a number of developmental and environmental cues. The coding region of the NtPAL4 gene was 2,154 bp in length, and its deduced protein was composed of 717 amino acids. Sequence analysis of NtPAL4 cDNA from tobacco (Nicotiana tabacum L.) revealed high structural similarity to PAL genes of other plant species. The NtPAL4 gene exists as a single copy in the tobacco plant, and its transcripts were strongly expressed in flowers and leaves. NtPAL4 expression was significantly induced in response to NaCl, mannitol, and cold treatments, but it was not induced by abscisic acid (ABA). NtPAL4 expression decreased gradually after treatment with ABA and $H_2O_2$; however, NtPAL4 transcripts accumulated after treatment with methyl viologen (MV). Our results suggest that the NtPAL4 gene may function in response to abiotic stresses.

• 한국응용곤충학회지
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• 제18권1호
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• pp.11-14
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• 1979

짙은 연색의 모자익병징을 나타내는 이병 시금치를 채집하여 Broad bean wilt virus(BBWV)를 분리동정 하였다. 분리된 Broad bean wilt virus (BBWV)를 지표식물에 즙액접종한 결과, 명아주(Chenopodium amaranticolor), 명아주(Chenopodium quinoa) ,잠두(Vicia faba)에서는 접종엽에 국부병반이 나타났고 접종상엽에서는 모자익(전신감염)병징이 나타났으며 꽈리(Physalis floridana), 시금치, 담배 (W.B), 담배 (Nicotiana glutinosa)에는 모자익 병징이 나타났다. 이병시금치로부터 분리한 병원바이러스와 BBWV의 항혈청을 한천내확산법(Agar gel-diffusion test)으로 반응시킨 결과 양성 반응이 나타났다. 이병엽을 Dip법으로 시료를 제작하여 전자현미경에서 검경한 결과 구형의 입자가 관찰되었으며 직경은 25nm이었다. 시금치 재배포장에서 BBWV의 발생분포는 수원, 안양, 대구 진주 등 거의 전국적으로 발생하였다.

• 한국연초학회지
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• 제14권1호
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• pp.3-11
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• 1992

The present study examined various environmental and cultural media conditions for in vitro "rescue" of cross-fertilized ovules formed through sexual crosses between Nicotiana rustica and N. tabacum cv. BY4. The response ovules to two cultural procedures was compared; ovules were cultured either separately or left attached to the placenta. Total yield of seedlings and percent of normal seedlings were increased by culturing individual ovules separately, rather than on excised placenta. Total yield of seedlings and number of normal seedlings were produced following in vitro culture of individual fertilized ovules of N. rustica X M tabacum cv. BY4 at four days post-pollination on NN medium containing 2% sucrose. In the in vitro culture of fertilized ovules, high sucrose concentration increased the frequency of seedlings of abnormal appearance. Therefore, sucrose should be supplied to developing ovules at gradually decreased concentrations. Culture of fertilized ovules from three to eight days after pollination gave increased number of seedlings, but with delayed cultral time the number of morphologically normal seedling were decreased. Hybrids were uniform in appearance and showed vegetative heterosis but flower characteristics were generally intermediate between those of the parents. All hvbrids evaluated were self-sterile.f-sterile.

• BMB Reports
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• 제34권5호
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• pp.408-414
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• 2001

Ornithine decarboxylase (EC 4.1.1.17) is an essential enzyme for polyamine synthesis and growth in mammalian cells and plants. We compared the biochemical and immunological properties of rat and Nicotiana glutinosa ODC by cloning and expressing the recombinant proteins. The primary amino acid sequence between rat and N. glutinosa ODC had a 40% homology The molecular weight of the overexpressed rat ODC was 53 kDa, and that of N. glutinosa was 46.5 kDa. Adding 1 mM of putrescine to the enzyme reaction mixture inhibited both rat and N. glutinosa ODC activity to 30%. Agmatine had an inhibitory effect only on N. glutinosa ODC. Cysteine and lysine modifying reagents reduced both ODC activities, verifying the key roles of cysteine and lysine residues in the catalytic mechanism of ODC. ELISA was performed to characterize the immunological difference between the rat and plant ODC. Both the rat and N. glutinosa ODC were recognized by the polyclonal antibody that was raised against purified N. glutinosa ODC, but the rat ODC was 50-fold less sensitive to the antibody binding. These results indicate that even though both ODCs have the same evolutionary origin, there seems to be a structural distinction between the species.

• BMB Reports
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• 제29권5호
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• pp.462-467
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• 1996

Acetolactate synthase (ALS, EC 4.1.3.18) is the first common enzyme in the biosynthesis of leucine, isoleucine, and valine. It is the target enzyme for several classes of herbicides, including the sulfonylureas, the imidazolinones, the mazolopyrimidines, the pyrimidyl-oxy-benzoates, the pyrimidyl-thio-benzens, and the 4,6-dimethoxypyrimidines. An amino-terminal fragment of the sulfonylurea-resistant ALS gene (SurB) from Nicotiana tabaccum was cloned into the bacterial expression vector pGEX-2T. The resulting recombinant plasmid pGEX-ALS1 was used to transform Escherichia coli strain BL21, and the tobacco ALS was expressed in the bacteria as a protein fused with glutathione S-transferase (GST). Polyclonal antibodies against the fusion product (GST-ALS) were produced, and the sensitivity of GST-ALS with the rabbit anti-GST-ALS IgG was up to 50 ng. This antibody was used for Western blot analysis of the partially purified ALS from barley shoots. The results suggest that the polyclonal antibody produced in this study can be used to detect plant ALS.

• KSBB Journal
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• 제8권2호
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• pp.97-103
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• 1993

세포질 유전형질의 도입을 성공적으로 수행코자 세질 웅성 불임(eMS; cytoplasmic male sterili­t ty) 계통의 담배 MS(male sterility) Burley 21의 callus tissue에서 cytoplast를 분리시키고 이와 Ni­c cotiana tabacum Burley 64의 mesophyll protoplast 를 PEG방법에 의한 세포융합으로 cybrid cell올 유도할 수 있었다. cytoplast의 분라는 density gradient solution의 osmolarity를 일정하게 하는 것보다 gradient간의 차이가 더 효과적이였으며 분리된 cytoplast의 양은 재료로 이용된 protoplast의 20-30% 정도를 나타 내었다. 또한 cytoplast와 protoplast의 융합은 P PEG농도 50%에셔 효과적이었다.