• Title/Summary/Keyword: tobacco plants

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Overexpression of a Rice Diacylglycerol Kinase Gene OsBIDK1 Enhances Disease Resistance in Transgenic Tobacco

  • Zhang, Weidong;Chen, Jie;Zhang, Huijuan;Song, Fengming
    • Molecules and Cells
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    • v.26 no.3
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    • pp.258-264
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    • 2008
  • A rice diacylglycerol kinase (DGK) gene, OsBIDK1, which encodes a 499-amino acid protein, was cloned and characterized. OsBIDK1 contains a conserved DGK domain, consisting of a diacylglycerol kinase catalytic subdomain and a diacylglycerol kinase accessory subdomain. Expression of OsBIDK1 in rice seedlings was induced by treatment with benzothiadiazole (BTH), a chemical activator of the plant defense response, and by infection with Magnaporthe grisea, causal agent of blast disease. In BTH-treated rice seedlings, expression of OsBIDK1 was induced earlier and at a higher level than in water-treated control seedlings after inoculation with M. grisea. Transgenic tobacco plants that constitutively express the OsBIDK1 gene were generated and disease resistance assays showed that overexpression of OsBIDK1 in transgenic tobacco plants resulted in enhanced resistance against infection by tobacco mosaic virus and Phytophthora parasitica var. nicotianae. These results suggest that OsBIDK1 may play a role in disease resistance responses.

Food Attractancy of the Oriental Tobacco Budworm, Heliothis assulta, Larvae (담배나방(Heliothis assulta)유충의 먹이 유인성)

  • 최광식;부경생
    • Korean journal of applied entomology
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    • v.28 no.2
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    • pp.88-92
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    • 1989
  • The Oriental tobacco budworm, Heliothis assulta (Guenee), larvae usually feed on the hot-pepper fruits and young tobacco leaves or buds in Korea. This experiment was conducted t(} compare their attraction to their host plants including fruits of the sweet-pepper and solanum uk kwang, besides the other two mentioned above. First instar larvae were most attracted to the hot-pepper fruits. Flowers or leaves of the hot pepper plants were not attractive either. When the hot-pepper fruits were compared for attractancy, based on developmental stages, relatively mature fruits, just before ripening, were the most attractive. Their extract with or without 10% ethanol also gave similar result to 1st instar larvae or 2~4 instar larvae, suggesting a possibility that there is an attractant(s) in the hot-pepper fruits.

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Study of a Tobacco MADS-Box Gene Triggering Flower Formation

  • Chung, Yong-Yoon;N, Gynheung-A
    • Proceedings of the Botanical Society of Korea Conference
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    • 1996.07a
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    • pp.10-18
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    • 1996
  • Recently, we have reported a rice MADS-box gene, OsMADS1, as a molecular factor triggering flower formation; this has been well studied in a heterologous system (Chung et al., 1994). In order to study whether the OsMADS1 homolog exists in other plant species, the OsMADS1 cDNA was used as a probe to screen a tobacco cDNA library, and a potential homolog, NtMADS3, was isolated. Sequence analysis revealed that the gene shares 56.1% identity in whole amino acids with OsMADS1. Like OsMADS1, the NtMADS3 gene starts to express at a very early stage of flower development, and the expression continues up to flower maturation. In the tobacco flower, the gene is expressed in whorl 2,3 and 4, corresponding to the petal, stamen, and carpel, respectively. Upon ectopic expression in the homologous system, NtMADS3 caused a trasition from inflorescence shoot meristem into floral meristem, reducing flowering time dramatically. These phenotypes strongly suggest the NtMADS3 gene is the OsMADS1 homolog of tobacco. Hybrids between the OsMADS1 and the NtMADS3 plants were also generated. The hybrids flowered even earlier than these two transgenic plants. The detailed studies are discussed here.

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Generation of Transgenic Plant (Nicotiana tabacum var. Petit Havana SR1) harboring Bacillus thuringiensis Insecticidal Crystal Protein Gene, cry II A (Bacillus thuringiensis 살충성 결정단백질 유전자(cry II A)의 형질전환 식물 제작)

  • 이정민;류종석;권무식
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.5
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    • pp.305-311
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    • 1997
  • Bacillus thuringiensis, a gram-positive soil bacterium, is characterized by its ability to produce crystalline inclusions during sporulation. The crystal proteins exhibit a highly specific insecticidal activity. An insecticidal crystal protein (ICP), Cry II A, is specifically toxic to both lepidopteran and dipteran insects. In this study, tobacco plants transformed by the cry II A gene have been generated. The Cry II A crystal protein was purified from E. coli JM103 harboring cry II A gene by differential solubility. The activated Cry II A was prepared by tryptic digestion. The purified protoxin (70 kDa) and the activated toxin (50 kDa) were analyzed by SDS-PAGE. To generate the transgenic tobacco having cry II A gene, the cry II A gene was subcloned to a plant expression vector, pSRL2, having two CaMV 35S promoters. The recombinant plasmid was transformed into tobacco (N. tabacum var. Petit Havana SR1) by Agrobacterium-mediated leaf disc transformation. Through the regeneration, six putative transgenic tobacco plants were obtained and three transformants were confirmed by Southern blot analysis. It has been found that one plant had single copy of cry II A gene, another had two copies of the gene, and the third had a truncated gene. After the immunochemical confirmation of cry II A expression in plants, the transgenic tobacco plants will be used to study the genetics of future generation with the insecticidal crystal protein gene cry II A.

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Expression of de novo Designed High Nutritional Peptide (HEAAE) in Tobacco

  • Kim, Jae-Ho;Lee, Chang-Kook;Hong, Bun-Shik
    • Journal of Microbiology and Biotechnology
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    • v.7 no.2
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    • pp.138-143
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    • 1997
  • We have designed and constructed a gene encoding novel high essential amino acid encoding protein(HEAAE). The resultant DNA fragment was tested for in vitro and in vivo expression and then cloned into plant expression vector pBI121, under the control of the cauliflower mosaic virus 35S promoter. Agrobacterium tumefaciens, strain LBA4404, was subsequently transformed with this new construct and Nicotiana tabacum var. Xanthi transgenic plants were obtained. DNA analysis by Southern procedure confirmed the presence of the multi-copy number of genes in the transformed plants. Analysis of RNA and protein synthesized in these transgenic plants demonstrated the stable expression of this gene.

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Functional Analysis of a Grapevine UDP-Glucose Flavonoid Glucosyl Transferase (UFGT) Gene in Transgenic Tobacco Plants (담배 형질전환체를 이용한 포도 UDP-glucose flavonoid glucosyl transferase (UFGT) 유전자의 기능 분석)

  • Park, Ji-Yeon;Park, Sung-Chool;Pyee, Jae-Ho
    • Journal of Life Science
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    • v.20 no.2
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    • pp.292-297
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    • 2010
  • Anthocyanin, a phenolic compound, is a pigment that shows blue or red color in the fruit, petal and other tissues. It is an important factor in grape berry skin pigment and accumulates only in the skin. This skin-specific accumulation of anthocyanin has been reported to be regulated by the ufgt gene which encodes UDP-glucose: flavonoid 3-O-glucosyltransferase that participates in the biosynthesis of anthocyanin. The ufgt gene is expressed only in berry skin, while the other genes involved in the biosynthetic pathway are expressed in both skin and flesh tissues. In order to determine whether anthocyanin accumulation is primarily regulated by compartment of UFGT, a ufgt cDNA clone was isolated from grape berry, its open reading frame was ligated in pBI121 vector in either a sense or an antisense orientation under the control of the CaMV35S promoter and the recombinant constructs were incorporated into tobacco plants. Several transgenic lines were selected and characterized to determine the level of expression of the grapevine ufgt transcript and endogenous homologs of tobacco. Compared to the wild-type, the amount of anthocyanins in sense transgenic plants increased by 44%, while the amount of anthocyanins in antisense transgenic plants decreased by 88%. In addition, the color of flowers became intense in the sense transgenic plants. These results suggest that over-expression or repression of the ufgt gene affected the accumulation of anthocyanin in flowers of tobacco.

Expression of Human Interleukin-ll and Granulocyte-Macrophage Colony-Stimulating Factor in Transgenic Plants

  • LEE BO-YE;LEE JEONG-HYUN;YOON HOON-SEOK;KANG KYUNG HO;KIM KYUNG-NAM;KIM JAE-HONG;KIM JU-KON;KIM JEONG-KOOK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1304-1309
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    • 2005
  • The production of therapeutic proteins for human diseases in plants results in many economic benefits, including reduced risk of animal virus contamination, high yields, and reduced production and storage costs. Human cytokines, interleukin-11 (hlL-11) and granulocyte-macrophage colony-stimulating factor (hGM-CSF), cDNAs were introduced into rice or tobacco, using either the maize ubiquitin promoter or the 35S promoter. The primary hIL-11 transgenic rice plants exhibited stunted growth and a sterile phenotype, whereas the hIL-11 transgenic tobacco plants did not. This suggests that hIL-11 expression in rice disrupts the normal growth and development of the plant. The regeneration efficiency of rice calli transformed with hGM-CSF was found to be approximately a quarter of that seen with the hIL-11, suggesting that hGM-CSF expression is more deleterious to the regeneration of rice calli than is hIL-11. However, the surviving hGM-CSF transgenic rice plants exhibited a normal phenotype of growth. Therefore, it appears that only those transgenic rice lines that expressed the human cytokines in small quantities were able to survive the selection process.

Mutation Induction and Selection of Salt-tolerant Plants by Heavy-ion Beam Irradiation in Tobacco Proembryo (중이온 빔 조사에 의한 담배의 돌연변이 유도와 내염성 식물의 선발)

  • ;Abe Tomoko
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.2
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    • pp.103-107
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    • 1998
  • Tobacco proembryos were irradiated with 100 Gy of heavy-ion beams($^{14}\textrm{N}$, $^{20}\textrm{Ne}$: 135 Mev/u) after 24 to 96 hours of pollination as a mutagen and screened $\textrm{M}_{1}$ generation for morphological mutants and salt-tolerant plants. Morphological and physiological characteristics of the salt-tolerant plants derived from the irradiated proembryo are discussed in this report. Mutants irradiated proembryos with the beams after pollination produced various kinds of morphological variation. A total of 17 salt-tolerant plants were selected from tobacco cultivar (BY-4) by treatment with $^{14}\textrm{N}$ beam. Shapes of filament and pollen grain of most salt-tolerant plants were abnormal compared with non-irradiated wild type, and seeds weight and fertility obviously decreased. The germination rates of the several $\textrm{M}_{2}$ lines on the saline and the mannitol condition were higher than that of wild type.

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EFFECT OF TWO PVY STRAINS ON YIELD AND CHEMICAL CONSTITUENTS OF TOBACCO PLANT (두가지 계통의 PVY가 잎담배 수량 및 내용성분에 미치는 영향)

  • 박은경;김정화;이영근
    • Journal of the Korean Society of Tobacco Science
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    • v.7 no.1
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    • pp.7-13
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    • 1985
  • Effect of the two PVY strains(PVY-VB and PVY-VN) on yield and chemical constituents of tobacco plants was determined in roar. NC2326 and Burley 21. The virus was inoculated 6 and 8 weeks after transplanting, respectively. The strains and time of inoculation were most critical factors for yield reduction. The greatest reduction was caused by the necrotic strain (PVY-VN) inoculated 6 weeks after transplanting, accounting for the loss of 40% in var. NC2326 and of 45% in var.. Burley21. When inoculated 8 weeks after transplanting with the necrotic strain, only 17% reduction was recorded in roar. Burley21, but no reduction occurred in roar. NC2326. Generally less reduction was caused by the PVY-VB strain, ranging 12-16% depending on the time of inoculation and/or variety. Compared to healthy tobacco, PVY infected tobacco contained higher concentrations of total nitrogen, protein nitrogen, nitrate nitrogen, and lower total sugar in cured leaf of roar. NC2326. Total alkaloid, P, Mg, Ca, and K levels were not altered. In var. Burley21, protein nitrogen and nitrate nitrogen increased, but other chemical components were not changed. Necrotic strain-infected tobacco with a severe symptom had higher nitrogen than did mild strain-infected tobacco.

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Expression of Canavalia Iineata Leghemoglobin cDNA in Transgenic Nicotiana tabacum (형질전환된 담배에서 해녀콩 Leghemoglobin cDNA의 발현)

  • 이선영
    • Journal of Plant Biology
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    • v.38 no.2
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    • pp.203-209
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    • 1995
  • Tobacco (Nicotiana tahacum L. cv. Wisconsin 38) leaf discs were cocultivated with Agrohacterium carrying a leghemoglobin (Lb) cDNA from Canavalia lineata. Seven plants were regenerated from the transformed leaf discs on MS media supplemented with 0.5 mg/L BAP, 0.1 mg/L ${\alpha}-NAA$, 200 mg/L kanamycin and 500 mg/L carbenicillin. Southern hybridization and PCR of genomic DNA from transgenic plants showed that the Lb cDNA was stably integrated into the genome of the tobacco. Total RNA from the transgenic tobacco showed northern hybridization signal at 1,000 nt and PCR of the first strand cDNA synthesized from the total RNA amplified 0.5 kb Lb cDNA. Furthermore, western hybridization using a polyclonal antibody against soybean Lb showed a 15.8 kD LB-like band on SDS-PAGE of proteins from the transformed tobacco. These results demonstrated that the Lb cDNA of C. lineata was not only incorporated into the genome of tobacco, but also transcribed into mRNA and translated into Lb protein in the transformed tabacco.

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