• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.469-474
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• 2000

Effect of the antibiotic carbenicillin on callus and shoot formation from the leaf disc culture of tobacco (Nicotiana tabacum L. cv. BY-4) was examined. The number of shoot induced from the leaf explants was decreased as the amount of carbenicillin increased from 250 mg/L to 2,000 mg/L on MS medium containing 0.5 mg/L of BAP or kinetin. In addition, calli formation from the leaf explants was increased by the treatment of 250 mg/L ∼ 500 mg/L carbenicillin with or without 0.5 mg/L of 2,4-D or NAA. However, the fresh weight of 4-week-cultured explants was decreased with increasing carbenicillin from 250 mg/L to 2,000 mg/L on MS medium containing 0.5 mg/L of 2,4-D or NAA. These results indicate that carbenicillin has an auxin-like effect, such as promoting callus formation and inhibiting shoot induction. It leads to the conclusion that the auxin-like property should be taken into account for the production of transgenic plants via Agrobacterium-mediated transformation.

• Korean Journal of Plant Tissue Culture
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• v.28 no.4
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• pp.179-184
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• 2001

We evaluated a possibility of the use of chloroplast number per guard cell pairs as a measure for ploidy level in the different ploidy levels of tobacco plant (Nicotiana tabacum L. cv. BY-4) . The guard-cell chloroplast numbers of leaves of haploid plant were a half of wild-type plant. Furthermore, the number of chloroplast per guard cell pairs of the leaves of doubled-haploid plant increased in two times compared with that of haploid plant. In addition, the chloroplast number was not changed in the F$_1$ progenies. The change of the chloroplast number by leaf stage was not observed. The results indicate that there is a strong relationship between ploidy level (2x and 4x) and chloroplast number per guard cell pairs. This relationship was also, observed in both in vitro and pot cultured plants. It was determined that the measurement of chloroplast number in guard cells of leaf epidermis is simple to use and less labour intensive, and hence can be considered a practical alternative to the chromosome counting methods or flow cytometry in the tobacco plant.

• Journal of Plant Biology
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• v.34 no.1
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• pp.37-43
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• 1991

Mesophyll protoplasts of Nicoliana labacum ($NR^{-}/SR^{+}$) and N glulinosa were electrofused with AC field of 0.5 MHz and 1 kV DC pulse for 2 ms. Fused protoplasts were selected and cultured to the green cell clusters in $MSNO_3$ medium containing 1.2 mg!ml streptomycin sulfate. Four plant lines regenerated from selected colonies showed both parental morphological characteristics of leaf and flower and these plant lines were confirmed as somatic hybrids based on electrophoretic patterns of leaf peroxidase. In XhoI restriction patterns of chloroplast DNA, these hybrid plant lines expressed both parent common restriction sites and parent specific sites. One of these hybrid lines exhibited interspecific pattern of both parental chloroplast genomes. indicating nine both parent common sites, one N labacum specific site and two N glutinosa specific sites. sites.

• Journal of the Korean Society of Tobacco Science
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• v.4 no.2
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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%.

• Journal of the Korean Society of Tobacco Science
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• v.30 no.2
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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.

• Korean Journal of Plant Resources
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• v.23 no.6
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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.

• Journal of the Korean Society of Tobacco Science
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• v.14 no.1
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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.

• Journal of the Korean Society of Tobacco Science
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• v.22 no.1
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• pp.25-30
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• 2000

Bacterial wilt(Ralstonia solanacearum) is one of the major diseases of flue-cured tobacco (Nicotiana tabacum L.) in the world. This study was conducted to investigate degree of dominance, selection, and correlation between leaf shape and degree of bacterial wilt resistance in flue-cured tobacco. The degree of disease caused by bacterial wilt was evaluated in parents, F$_1$, F$_2$ and F$_3$ populations of two crosses, BY 4 x NC 95 and BY 4 x Coker 86, in the infected field. The leaf shape index was also measured in parents and F$_2$ population of BY 4 x NC 95. The incidence of bacterial wilt was observed in the middle of June and peaked in late July, when the highest value of pathogen density reached 1.0 x 10$^{6}$ colony forming unit per gram. It was concluded that the inheritance mode of risestance to bacterial wilt in the above two crosses of susceptible and resistant varieties was recessive and polygenic. The resistance to bacterial wilt was significantly correlated with leaf shape in F2 generation of BY 4 x NC 95. But certain plants having narrower leaves were also resistant to bacterial wilt. It is considered that the bacterial wilt resistant lines having narrower leaves could be selected. The selection for bacterial wilt resistance in the F$_2$ population might be effective.

• Journal of the Korean Society of Tobacco Science
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• v.22 no.1
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• pp.31-38
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• 2000

Seven flue-cured tobacco(Nicotiana tabacum) varieties and their 21 progenies in Fl and F2 generation, half diallel crosses(no reciprocal), were tested for the degree of heterosis and combining abilities in sucker producing character according to topping and suckering treatment. Significant difference in number and weight of sucker was observed among seven cultivars. The values of general combining ability(GCA) were greater than those of specific combining ability(SCA) in weight and number of sucker in F$_1$ generation. In topping and suckering plot, Cocker 86, McNair 944, and Va 115 revealed the significant negative GCA effects for number and weight of sucker, but By 104 and Speight G-28 did positive GCA. Without topping and suckering, GCA effects for sucker weight in Cocker 139 and Cocker 86 were negative and those for sucker number in Va. 115 were also negative. Negative SCA values in sucker weight could be observed in F$_1$ generation of Cocker 86 X BY 104 and Speiht G-28 X Cocker 86, in F$_2$ of Hicks $\times$ Speight G-28 and McNair 944 $\times$ BY 104 in topping and suckering plot. SCA in F$_1$ of McNair 944 X Cocker 86 and Speight G-28 X Cocker 139 was negative without topping and suckering. The value of heterosis in sucker producing character of F$_1$ hybrids was positive

• Journal of the Korean Society of Tobacco Science
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• v.18 no.2
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• pp.126-131
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• 1996

In this study effects of isolation distance, transplanting time of maternal plants, and bagging of flower head with the gauze-cloth bag on the outcrossing of burley tobacco (Nicotiana tabacum L.) were investigated. Also the effect of fertilizer level and control of the number of capsules per plant on seed production and quality were examined. A male sterile line. produced 0.3 to 3.8 capsules Per plant when it was Planted with normally flowering tobacco with the average outcrossing of 7.2 plants, ranging from 2 to 18 out of 20 plants. With the farther the isolation distance between maternal plants and pollen donor plant, the lower the outcrossing occurred. Outcrossing occurred even at the isolation distance of 312 m. When the maternal plants were transplanted 35 days after transplanting the pollen donor ones, the outcrossed plants were not decreased significantly. The bagging of the flower head with the gauze-cloth bas (#0.9∼ 1.0 mm) decreased the outcrossed plants significantly, but couldn't prevent the outcrossing completely. The seed amount per plant was higher in the highly fertilized cultivation. The number of seed capsules per plant affected significantly on seed yield and quality. When the seed capsules was controlled by 30 or 50 capsules per plant, the weight of 1,000 seeds and germination rate were higher than those with 70 or 90 capsules per plant. Key words : Nicotiana tabacum, outcrossing, bagging.