• Title/Summary/Keyword: nicotiana tabacum

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Development of Black Shank Resistant Burley Tobacco Germplasm, KB 104 and KB 106, and Their Characteristics (버어리종 역병저항성 계통 KB 104와 KB 106의 육성경과 및 특성)

  • 조천준;김대송;정석훈;최상주;조명조
    • Journal of the Korean Society of Tobacco Science
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    • v.17 no.1
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    • pp.57-61
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    • 1995
  • Black shank(Phytophthora parasitira roar. nicotianae) resistant burley tobacco (Nicotiana tabacum L.) germplasms, KB 104 and KB 106, were developed by Korea Ginseng & Tobacco Research Institute. KB 104 was developed from the single cross of Burley 21$\times$Newton 77, using a modified pedigree method. KB 104 was highly resistant to black shank, and its agronomic characteristics and chemical contents were comparable to those of Burley 21, and value per 10a was slightly higher than Burley 21, KB 106 is a maternally derived doubled haploid made by N. africana method from the single cross of Burley 21$\times$ Va 509. KB 106 was also highly resistant to black shank, had two more harvestable leaves per plant and flowered three days later than Burley 21 did. Total alkaloid and nicotine contents of KB 106 were significantly lower than those of Burley 21. But its nornicotine content was higher than Burley 21 5. Key wads : Burley tobacco germplasm, Black shank resistance.

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Expression of Glutathione Reductase Gene in Transgenic Tobacco Plant (형질전환 담배 식물체에서 Glutathione Reductase 유전자의 발현)

  • 이효신;조진기
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.2
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    • pp.87-90
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    • 2001
  • BcGRl gene encoding cytosolic glutathione reductase of Chinese cabbage (Brassica campestris var. Pekinensis cv. Seoul) was placed under the control of the CaMV 35S promoter and introduced into tobacco (Nicotiana tabacum L. cv. Samsun) via Agrobacterium-mediated transformation. T$_{0}$ 32 independent plants transformed with BcGRl gene were selected with kanamycin and they were confirmed by polymerase chain reaction (PCR) and Southern blot analysis. Northern blot analysis revealed that the constitutive expression of BcGRl gene and there was no relationship between the copy number of introduced gene and the levels of BcGRl transcripts.

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Generation and Retention of Defective RNA3 from Cucumber Mosaic Virus and Relevance of the 2b Protein to Generation of the Subviral RNA

  • Emi Ota;Chikara Masuta;Minoru Takeshita
    • The Plant Pathology Journal
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    • v.39 no.6
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    • pp.592-599
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    • 2023
  • A defective RNA3 (D3Yα) of strain Y of cucumber mosaic virus (CMV-Y) was examined on host-specific maintenance, experimental conditions, and a viral factor required for its generation in plants. D3Yα was stably maintained in cucumber but not in tomato plants for 28 days post inoculation (dpi). D3Yα was generated in Nicotiana tabacum or N. benthamiana after prolonged infection in the second and the third passages, but not in plants of N. benthamiana grown at low temperature at 28 dpi or infected with CMV-Y mutant that had the 2b gene deleted. Collectively, we suggest that generation and retention of D3Yα depends on potential host plants and experimental conditions, and that the 2b protein has a role for facilitation of generation of D3Yα.

Plant Regeneration and Protein Analysis from Cadmium Resistant Callus of Tobacco (Nicotiana tabacum cv. BY4) (담배 (Nicotiana tabacum cv. BY4)카드뮴 저항성 캘러스로부터 식물체 재생과 단백질 분석)

  • 오승철;소웅영;조덕이;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.7-13
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    • 2001
  • Calli were induced from diploid and haploid tobacco after 4 weeks and maintained on MS medium with combination of 2.0 mg/L 2,4-D,0.1 mg/L BAP and 2.0 mg/L kinetin. Suspension cells were screened through 65 $\mu$m-nylon mesh and 100 $\mu$m-mesh, then they were smeared on selection medium combined with cadmium and PFP by using the low melting agarose of 0.8%. After 30days smeared cultures of the medium the cell was treated with 500 $\mu$M and 1000 $\mu$M to select the resistant cell line were selected. Plant regeneration was induced from the selected cell lines on medium with 0.5, 1.5, 2.0 mg/L BAP and on media with combination of auxin and BAP under 500 $\mu$M and 1000 $\mu$M cadmium. At this time, plant regeneration was achived on cadmium free medium. In case of haploid, occurred from the cell line which is selected in medium with cadmium and PFP. In case of diploid regeneration occurred is in the medium with cadmium alone. The plantlet regenerated from cadmium resistant calli grew well in cadmium 500 $\mu$M. Protein pattern of leaf, root, stem of regenerated plants was analyzed. The quantum was 6.5188 ug/mg.fr.wt in the leaf of plant, 5.3611 ug/mg.fr.wt in the stem, 3.0213 ug/mg.fr.wt in the root. On the other hand, 5.9652 ug/mg.fr.wt. in the leaf of control, 3.5974 ug/mg.fr.wt in the stem of the control, 4.3766 ug/mg.fr.wt. in the root of the control. The one dimension bends regenerated from cadmium resistant calli resistant to cadmium in leaf were 49 involving 198.7KD etc. Disappeared were 4 involving 160.5KD etc, The protein bends were combinized were 3 involving 83.4KD etc. The bends resistant to cadmium stress in stem were 41 involving 4.3KD etc. Disappeared were 5 involving 114.8KD etc. The protein bends combinized were 6 involving 128.7KD etc. The bends which had the resistance to cadmium stress in root is 27 in volving 166,9KD etc. The bends which disappeared were 198.7KD etc. There were 5 involving 83.4KD etc.

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Expression of Recombinant Erythropoietin Gene in Transgenic Tobacco Plant (형질전환 담배 식물체에서 재조합 erythropoietin 유전자의 발현)

  • CHOI, Jang Won;PARK, Hee Sung
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.1
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    • pp.63-69
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    • 1997
  • Erythropoietin (EPO) is a glycoprotein that mediates the growth and differentiation of erythroid progenitors. In order to produce recombinant human erythropoietin in tobacco plant, the EPO genomic DNA (5.4 kb) was cloned into plant expression vectors, pBI$\Delta$GUS121, pBD$\Delta$GUS121 and pPEV-1, and introduced in Nicotiana tabacum (var. Xanthi) via Agrobacterium tumefaciens-mediated transformation. After selection on MS media containing kanamycin (Km), 10 Km-resistant plants were obtained per each construct. The correct integration of EPO genomic DNA in the genome of transgenic plant was confirmed by polymerase chain reaction (PCR). Northern blot showed that transcripts of 1.8 kb length were produced in leaves of the plants, but there was no difference of mRNA amount according to promoter number and 5'-untranslated sequence (UTS). The proteins obtained from leaves of transgenic plants were immunologically detected by Western blot using rabbit anti-human EPO polyclonal antibody. The expressed protein appeared as smaller band of apparent mass of 30 kDa as compared to the EPO protein from human urine (37 kDa), suggesting that the modification (glycosylation) system in tobacco plant might be different from that of mammalian cells.

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Construction of FMDV VP1 Gene Using Artificial DNA Synthesis and Transformation of Nicotiana tabacum Using Agrobacterium Vector System (유전자 인공합성을 이용한 구제역 유전자 VP1의 제작과 Agrobacterium Vector System을 이용한 담배 형질전환)

  • Lee, Eun-Jung;Lim, Hee-Young;Kim, Sung-Hoon;Kang, Kyung-Sun;Park, Young-Doo;Yun, Choong-Hyo;Yoon, Byoung-Su
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.285-293
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    • 2004
  • FMDV is a viral pathogen that caused foot-and-mouth disease in animals. VP1 is a major capsid protein of FMDV. It is known as one of best materials for the FMDV diagnosis and for the development of protein vaccine. In this study, 633 bp of VP1 gene was modified for the expression of VP1 in plant, based on the VP1 DNA sequence from FMDV taiwan O type and from FMDV isolated vietnam. The. deduced DNA fragment was artificially synthesized using the multiple fragment extension with long-nucleotides. A new plant transgenic vector system, pCAMBIA139011 was constructed on the basis of pBI12l and pCAMBIA1390. Using this vector system and GFP gene or modified VP1 gene, each target gene was introduced into Nicotiana tabacum. The insertion of whole target gene was successfully confirmed in each transgenic plant named GFP-A7 and VP1-4, respectively. The expression level of each gene was estimated by RT-PCR and Real-Time PCR using VP1, GFP specific primers.

A Study on the Variation of Color and its Components of the Flue-Cured Tobacco Leaf During the Process of Curing (Flue -cured 잎담배의 건조과정중 색상과 색소성분 변화에 관한 연구)

  • 김찬호;김정옥;석영선
    • Journal of the Korean Society of Tobacco Science
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    • v.1 no.1
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    • pp.56-62
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    • 1979
  • Boring the process of curing of the tobacco ( Nicotiana tabacum L. clicks) leaf, the and other fundamental components of the tobacco leaf was observed The amount of total chlorophyll was reduced from 2902$\mu\textrm{g}$/gr to 1, 131$\mu\textrm{g}$/gr as a function of curing time. In the case of total carotene, the concentration was increased from 373-712$\mu\textrm{g}$/gr. to 3, 003~2, 215$\mu\textrm{g}$/gr. up to 70 hours of curing period. However, after 70 hours of curing, the concentration of total carotene stared to be decreased up to 1, 050~1, 039$\mu\textrm{g}$/gr. Simultaneously, the greenish yellow color becomes to be close an intermediate color between yellow and orange. The amount of total nicotine and nitrogen (2~2%) in the green harvested leaf was essentially identical after the curing process. However, the amount of total sugar was enhanced from 17%, in the green harvested leaf, to 35% at the time of 70 hours curing. At the end of the curing, the amount of total sugar was again reduced up to 16%. The correlation coefficient (r) between the axis of the color coordinate and the ratio of chlorophyll/carotene was estimated to be r=-0.96. This result indicates a linear relationship between those two parameters and the following linear equation is obtained as y=28.87-68.38 x. From the above equation, x=0.442 could be estimated when chlorophyll was completely decomposed (ylongrightarrowO). As a consequence of this result, we can suggest the tobacco leaf of Hicks belongs to the high quality level.

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Insect Resistance of Tobacco Plant Expressing CpBV-ELP1 Derived from a Polydnavirus (폴리드나바이러스 유래 CpBV-ELP1 발현 담배의 내충성)

  • Kim, Eunseong;Kim, Yonggyun
    • 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.

Characterization of an Isolate of Cucumber mosaic virus from Raphanus sativus L. (열무에서 분리한 오이모자이크바이러스 분리주의 특성)

  • Rhee, Sun-Ju;Hong, Jin-Sung;Choi, Jang-Kyung;Kim, Eun-Ji;Lee, Gung-Pyo
    • Research in Plant Disease
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    • v.17 no.2
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    • pp.211-215
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    • 2011
  • Cucumber mosaic virus (CMV)-like isolate was collected from Raphanus sativus (cv. Choon-hyang), which showed mosaic symptoms. The isolate was confirmed to a strain of CMV by host responses in Vigna unguiculata, Chenopodium amaranticolor and Gomphrena globosa, by viral genome composition with RT-PCR and PCR-RFLP, and by serological analysis. Symptom developed by the strain of CMV was severe in Nicotiana benthamiana, N. glutinosa, N. tabacum (cv. Samsun, cv. Xanthi), Cucumis melo (cv. Early hanover), Cucumis sativus (cv. White wonder), Capsicum annuum (cv. Chung-yang and cv. Geum-top), but mild symptom was developed in Raphanus sativus (cv. Choon-hyang), Brassica rapa ssp. pekinensis (cv. Bul-Am No. 3), and B. juncea (cv. Daenong Jukgot). Newly isolated strain of CMV could infect diverse crops including Solanaceae, Cucurbitaceae and Brassicaceae. We designated the new strain of CMV as Gn-CMV based on the novel infectivity of Brassicaceae. In double-stranded (ds) RNA analysis, Gn-CMV consisted of 3.3, 3.0, and 2.2 kb genomes likewise other strains of CMV. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) showed 28 kDa of the CMV coat protein. By restriction enzyme mapping using Cac8I, ClaI and MspI of RT-PCR products indicated that Gn-CMV belongs to CMV subgroup I.

Evaluation of Biomass and Nitrogen Nutrition of Tobacco under Sand Culture by Reflectance Indices of Ground-based Remote Sensors (지상원격측정 센서의 반사율 지표를 활용한 사경재배 연초의 생체량 및 질소영양 평가)

  • Kang, Seong-Soo;Jeong, Hyun-Cheol;Jeon, Sang-Ho;Hong, Soon-Dal
    • Korean Journal of Soil Science and Fertilizer
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    • v.42 no.2
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    • pp.70-78
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    • 2009
  • Remote sensing technique in agriculture can be used to identify chlorophyll content, biomass, and yield caused from N stress level. This study was conducted to evaluate biomass, N stress levels, and yield of tobacco (Nicotiana tabacum L.) under sand culture in a plastic film house using ground-based remote sensors. Nitrogen rates applied were 40, 60, 80, 100, 120, and 140 percent of N concentration in the Hoagland's nutrient solution. Sensor readings for reflectance indices were taken at 30, 35, 40, 45, 50 and 60 days after transplanting(DAT). Reflectance indices measured at 40th DAT were highly correlated with dry weight(DW) of tobacco leaves and N uptake by leaves. Especially, green normalized difference vegetation index(gNDVI) from spectroradiometer and aNDVI from Crop Circle passive sensor were able to explain 85% and 84% of DW variability and 85% and 92% of N uptake variability, respectively. All the reflectance indices measured at each sampling date during the growing season were significantly correlated with tobacco yield. Especially the gNDVI derived from spectroradiometer readings at the 40th DAT explained 72% of yield variability. N rates of tobacco were distinguished by sufficiency index calculated using the ratio of reflectance indices of stress to optimum plot of N treatment. Consequently results indicate that the reflectance indices by ground-based remote sensor can be used to predict tobacco yield and recommend the optimum application rate of N fertilizer for top dressing of tobacco.