• Journal of the Korean Society of Tobacco Science
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• v.21 no.1
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• pp.64-69
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• 1999

The effect of phytohormones, light and phosphate on in vitro production of ubiquinone 10 from the suspension cultures of Nicotiana tabacum cv. Xanthi callus was investigated. The inoculum size and cultured time in the suspension culture had to be at least over 2 % of medium volume at 15 days for the excellent growth of Xanthi callus. The growth of Xanthi callus in the suspension culture was improved by addition of NAA and 2,4-D, especially NAA 1.0mg/1 alone, at the light condition. The optimal concentration of phytohormone was 0.1 mg/l 2.4-D and 1.0 mg/l NAA for productivity of ubiquinone 10 in the suspenseion of Xanthi callus. Addition of 3mM KH$_2$PO$_4$ to the medium was more effective in promoting ubiquinone-l0 formation than other concentration in the light condition. Content and production of ubiquinone-l0 in the suspension cultures of Xanthi callus were the highest at the MS media containing 0.5mg/L kinetin, 0.5mg/L 2,4-D, 1.0mg/1 NAA, 3mM phosphate and 2 % inoculum in the light.

• Korean Journal of Plant Tissue Culture
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• v.21 no.6
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• pp.341-345
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• 1994

The effect of phytohormones on in vitro production of ubiquinone 10 from the callus cultures of Nicotiana tabacum cv Xanthi was investigated. The growth of callus cultures of Xanthi was in proved by addition of NAA and 2,4-D, especially NAA 0.5 mg/L alone, at the light condition. Ubiquinone 10 was detected by HPLC, and confirmed from Xanthi callus cultured on the all of uppermedia. The ubiquinone 10 content in Xanthi tobacco callus cultured on the medium with NAA 0.5 mg/L only was higher than that of other mixed medium with NAA and 2,4-D. However addition of IBA 1 mg/L and NAA 0.5 mg/L to the medium was more effective in promoting ubiquinone 10 formation than that of NAA 0.5 mg/L only As the callus growth of Xanthi was considerabley restrained at concentration of kinetin, Content and production of ubiquinone low as the highest at kinetin 0.5mg/L and 2,4-D 0.5mg/L in the light.

• Korean Journal of Weed Science
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• v.8 no.1
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• pp.53-58
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• 1988

This study was carried out to determine effect of butachlor [N.-(buthoxymethyl)-2-chloro-N-(2,6-diethylphenyl) acetamide] and bialaphos [2-amino-4(hydroxy)(methyl) phosphionyl] butyryl-alanylalanine sodium salt on the germination of tobacco seed, induction and growth of callus from tobacco. Further, fatty acids and ammonia content of tobacco calli were determined. Bialaphos had no effect on tobacco seed germination, but the growth of seedling was markedly affected by an application of 10 ppm bialaphos. However, regardless of varieties tested, tobacco seed germination was completely inhibited by $5{\times}10^{-5}M$ of butachlor. At an application of $5{\times}10^{-5}M$ butachlor, tobacco seeds were to some extent germinated and showed further growth. Hyangcho among varieties tested, showed the most tolerant response to butachlor. In induction of callus from various tobacco varieties and their growth, aromatic type of tobacco varieties exhibited the most tolerance against bialaphos. However, no distinct varietal differences were determined in the treatment of butachlor. The major fatty acids identified in tobacco calli were palmitic, oleic and linoleic acid. No marked difference in terms of fatty acids was observed among tobacco varieties used, but it was observed that there was the higher ratio of quantity in unsaturated fatty acids over saturated one, bialaphos treatment accumulated about 9 times higher ammonia content than that of the untreated control, giving an evidence that bialaphos might inhibit glutamine synthetase activity.

• Korean journal of applied entomology
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• v.56 no.1
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• pp.19-28
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• 2017

Polydnaviruses (PDVs) are a group of double-stranded DNA viruses symbiotic to some endoparasitoid wasps. Cotesia plutellae bracovirus (CpBV) is a PDV symbiotic to an endoparasitoid wasp, C. plutellae, parasitizing young larvae of Plutella xylostella. An early expressed gene, CpBV-ELP1, plays an important role in the parasitism by suppressing host cellular immunity by its cytotoxic activity against hemocytes. This study aimed to test its oral toxicity against insect pest by expressing it in a recombinant tobacco plant. A recombinant CpBV-ELP1 protein was produced using a baculovirus expression system and secreted to cell culture medium. The cell cultured media were used to purify CpBV-ELP1 by a sequential array of purification steps: ammonium sulfate fractionation, size exclusion chromatography, and ion exchange chromatography. Purified rCpBV-ELP1 exhibited a significant cytotoxicity against Spodoptera exigua hemocytes. CpBV-ELP1 was highly toxic to the fifth instar larvae of S. exigua by injection to hemocoel. It also showed a significant oral toxicity to fifth instar larvae of S. exigua by a leaf-dipping assay. CpBV-ELP1 was cloned into pBI121 vector under CaMV 35S promoter with opaline synthase terminator. Resulting recombinant vector (pBI121-ELP1) was used to transform Agrobacterium tumefaciens LBA4404. The recombinant bacteria were then used to induce callus of a tobacco (Nicotiana tabacum Xanthi) leaves and subsequent generation (T1) plants were selected. T1 generation tobacco plants expressing CpBV-ELP1 gave significant insecticidal activities against S. exigua larvae. These results suggest that CpBV-ELP1 gene can be used to control insect pests by constructing transgenic crops.