• Title/Summary/Keyword: Transgenic Plant

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The Function of ArgE Gene in Transgenic Rice Plants

  • Guo, Jia;Seong, Eun-Soo;Cho, Joon-Hyeong;Wang, Myeong-Hyeon
    • Korean Journal of Plant Resources
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    • v.20 no.6
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    • pp.524-529
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    • 2007
  • We carried out to study the function of ArgE in transgenic rice plants, which were confirmed by PCR analysis and hygromycin selection. Transgenic rice plants were with selectable marker gene(HPT) inserted in genome of the rice. Southern analysis with hpt probe confirmed by two restriction enzymes that copy numbers of the selectable gene was introduced into the plant genome. We displayed that the relationship between drought stress and ArgE gene with the overexpressing rice plants. From this result, we observed that the degree of leaves damage has no difference in control and transgenic lines. The total RNAs were extracted from 6 weeks-seedling in normal condition in order to examine their expression levels with ArgE-overexpressed transgenic rice. In particular, expression patterns of genes encoding enzymes involved in abiotic stress, including drought and salt stresses. OsGF14a and OsSalt were investigated by reverse transcription-PCR(RT-PCR). Expression levels of the OsSalt gene decreased significantly in transgenic rice plants compared to control plant. However, ion leakage measurement did not demonstrate any leaves damage change between control and ArgE transgenic plants exposure to mannitol treatment. These results suggest that expression of the ArgE is not involved in tolerance for drought stress in rice but may playa role of signaling networks for salt-induced genes.

형질전환 벼에서 Cyanobacterial Sucrose-Phosphate Synthase 유전자의 발현

  • Sang-Kyu Lee;Soo-Jung Lee;Na-Yeon Ryoo;Jang-Wook Lee;Seok-Yoon Yoon;Woon-Chul Shin;Se-Ho Ko;Deok Chun Yang;Youn-Hyung Lee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.126-126
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    • 2003
  • Sucrose-phosphate synthase (SPS) is a key regulatory enzyme in sucrose synthesis. To investigate the role of SPS in carbon partitioning, we produced transgenic rice plants overexpressing a cyanobacterial SPS from Synechocystis sp. PCC 6803. The gene was expressed under the control of the maize Ubil promoter in transgenic plants. Southern and Northern blot analyses confirmed the integration and the expression of the transgene in four transgenic rice lines. All of the four transgenic! lines analyzed showed abnormal vegetative and reproductive developments. Analysis of SPS activities and primary metabolites in the transgenic rice plants will be presented.

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Responses of Transgenic Tobacco Plants Overexpressing Superoxide Dismutase and Ascorbate Peroxidase in Chloroplasts to Water Stress (Superoxide Dismutase와 Ascorbate Peroxidase를 엽록체에 과발현하는 형질전환 담배의 수분스트레스에 대한 반응)

  • 최선미;권석윤;곽상수;박용목
    • Journal of Environmental Science International
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    • v.10 no.1
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    • pp.79-84
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    • 2001
  • To assess resistance of transgenic tobacco plants which overexpress superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts to water stress, changes in leaf water potential, turgor potential, stomatal conductance and transpiration rate were measured. Leaf water potential in all plants remained high up to day 4 after withholding water but thereafter decreased markedly. In spite of a remarkable decrease in leaf water potential, some of transgenic plants maintained higher turgor potential compared with control plant on day 12. In particular, the transgenic plant expressing MnSOD showed an outstanding maintenance in turgor pressure by osmotic adjustment throughout the experiment, resulting in high stomatal conductance and transpiration rate. However, among transgenic plants, osmotic potential was reduced more effectively in multiple transformants such as the double transformant expressing both MnSOD and APX, and the triple transformant expressing CuznSOD, MnSOD and APX than single transformants. Consequently, further research is needed to get general agreement on the tolerance of transgenic plants to water stress at different growth stages for each transgenic plant.

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Cloning of Superoxide Dismutase (SOD) Gene of Lily 'Marcopolo' and Expression in Transgenic Potatoes

  • Park, Ji-Young;Kim, Hyun-Soon;Youm, Jung-Won;Kim, Mi-Sun;Kim, Ki-Sun;Joung, Hyouk;Jeon, Jae-Heung
    • Journal of Applied Biological Chemistry
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    • v.49 no.1
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    • pp.1-7
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    • 2006
  • Differential display reverse transcription PCR (DDRT-PCR) analysis was performed on lily 'Marcopolo' bulb scale for isolation of expressed genes during bulblet formation. Cu/Zn lily-superoxide dismutase (LSOD) of 872 bp gene, with ability to scavenge reactive oxygen in stress environment, was isolated. Northern blot analysis showed expression levels of LSOD maximized 12 days after bulblet formation. Ti plasmid vectors were constructed with sense and antisense expressions of LSOD gene and transformed into potato. Southern blot analysis of transgenic potatoes revealed different copies of T-DNA were incorporated into potato genome. In transgenic potatoes, lily SOD gene was overexpressed in sense lines and not in antisense lines. In native polyacrylamide gel electrophoresis analysis, additional engineered LSOD was detected in sense overexpressed transgenic line only. Transgenic potatoes were subjected to oxidative stress, such as herbicide methyl viologen (MV). Transgenic potato lines with sense orientation exhibited increased tolerance to MV, whereas in antisense lines exhibited decreased tolerance. In vitro tuberization of transgenic potato with sense orientation was promoted, but was inhibited in transgenic potato with antisense orientation.

Expression of diligent protein and Pinoresinol/Lariciresinol reductase genes of forsythia in transgenic potatoes

  • Chuong, Tran-Van;Kim, Hyun-Soon;Park, Ji-Young;Joung, Jae-Youl;Youm, Jung-Won;Jeon, Jae-Heung
    • Plant Resources
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    • v.4 no.3
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    • pp.181-188
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    • 2001
  • We tried to introduce two forsythia genes related in lignan biosynthesis, dirigent protein and pinoresinol/lariciresinol (Ph) reductase, into potatoes for accumulation of lignans in transgenic potatoes. We made binary vectors overexpressing dirigent protein gene and P/L reductase gene driven by a CaMV35S promoter and transformed into potatoes via Agrobacterium mediated transformation. And in order to control the metabolic flux of lignan biosynthesis pathway, we tried to inhibit chalcone synthase genes of potatoes by antisense inhibition technique also. We tried to use PCR screening method for selection of transgenic plants of different vectors. We tried to determine and compare lignan contents from different transgenic potato lines.

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Expression of Indica rice OsBADH1 gene under salinity stress in transgenic tobacco

  • Hasthanasombut, Supaporn;Ntui, Valentine;Supaibulwatana, Kanyaratt;Mii, Masahiro;Nakamura, Ikuo
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.75-83
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    • 2010
  • Glycine betaine has been reported as an osmoprotectant compound conferring tolerance to salinity and osmotic stresses in plants. We previously found that the expression of betaine aldehyde dehydrogenase 1 gene (OsBADH1), encoding a key enzyme for glycine betaine biosynthesis pathway, showed close correlation with salt tolerance of rice. In this study, the expression of the OsBADH1 gene in transgenic tobacco was investigated in response to salt stress using a transgenic approach. Transgenic tobacco plants expressing the OsBADH1 gene were generated under the control of a promoter from the maize ubiquitin gene. Three homozygous lines of $T_2$ progenies with single transgene insert were chosen for gene expression analysis. RT-PCR and western blot analysis results indicated that the OsBADH1 gene was effectively expressed in transgenic tobacco leading to the accumulation of glycine betaine. Transgenic lines demonstrated normal seed germination and morphology, and normal growth rates of seedlings under salt stress conditions. These results suggest that the OsBADH1 gene could be an excellent candidate for producing plants with osmotic stress tolerance.

Overexpression of Cotton Glutathione S-Transferase (GST) cDNA and Increase of low Temperature and Salt Tolerance in Plants

  • Kang, Won-Hee;Jong Hwa kim;Lim, Jung-Dae;Yu, Chang-Yeon
    • Journal of Plant Biotechnology
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    • v.4 no.3
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    • pp.117-122
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    • 2002
  • Cotton Glutathione S-Transferase(GST: EC 2.5.1.18) was cloned and Gh-5 cDNA was overexpressed in tobacco (Nicotiana tabacum) plants. The transformation of cotton GST in tobacco plant was confirmed by northern blot analysis. Type I and Type II transcript patterns were identified in Gh-5 transgenic tobacco plants. Type I transcripts was only discussed in this paper. Glutathione and 1-chloro-2,4-dinitrobenzene (CDNB) were used as the substrates, and the activity of GST in the type I transgenic plants was about 2.5-fold higher than the non-expressers and wild type tobacco plants. The expression of cotton GST in tobacco plants proved that Gh-5 could be translated into functional protein. Type I transgenic plants produced functional GST in the cells. Type I showed higher GST specific activity than Type II in the transgenic plants. Control and transgenic seedlings were grown in the growth chamber and under the light at 15$^{\circ}C$, and the effects of cotton GST in the seedlings was evaluated. The growth rate of Gh-5 overexpressors was better than the control and non-transgenic tobacco plants. Salinity tolerance was also analyzed on the seeds of transgenic plants. Seeds of Gh-5 overexpressors and the wild type tobacco seedlings were germinated and grown at 0, 50, 100, 150, and 200 mM NaCl solution. Gh-5 transgenic seedlings showed higher growth rate over control seedlings at both 50 and 100 mM NaCl solution. But at 0, 150, and 200 mM NaCl concentration, the difference in growth rate was not detected.

Agrobacterium-mediated transformation of Lycopersicon esculentum (cv. MicroTom) with two pathogen-induced hot pepper transcription factors

  • Seong, Eun-Soo;Oh, Sang-Keun;Eunsook Chung;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.79.1-79
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    • 2003
  • Two pathogen-induced hot pepper transcription factors (CaNACl and CapIfl) were introduced into‘MicroTom’tomato by Agrobacterium tumefaciens-mediated transformation. We used to nptII containing kanamycin resistance gene as a selection marker. Both transformed and non-transformed plants were transferred to pot after rooting test in vitro. To approximate the levels of caNACl transcript in leaves of wild-type and transgenic plants, RNA blots were hybridized with double-stranded full-length CaNACl probe at moderate stringency, Although the relative signal strength for hybridization fluctuated among the samples on different blots, transgenic plant lines N-1, N-2 and N-3 consistently displayed increased levels of CaNACl transcript relative to other transgenic lines and wild-type plants. Of all the transgenic lines examined, line N-7 had the least amount of CaNACl transcript. Role of these transcription factors in pathogen defense will be examined by overexpression in tomato.

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Physiological Response and Cadmium Accumulation of MuS1 Transgenic Tobacco Exposed to High Concentration of Cd in Soil: Implication to Phytoremediation of Metal Contaminated Soil (토양 중 고농도 카드뮴에 노출된 MuS1 형질전환 담배 (Nicotiana tabacum cv. Xanthi)의 생리적 반응 및 카드뮴 축적: 식물학적 오염토양정화를 위한 형질전환 식물 탐색)

  • Jeoung, Yoon-Hwa;Kim, Young-Nam;Kim, Kwon-Rae;Kim, Kye-Hoon
    • Korean Journal of Soil Science and Fertilizer
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    • v.46 no.1
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    • pp.58-64
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    • 2013
  • The objective of this study was to understand the physiological response and cadmium accumulation of MuS1 transgenic tobacco exposed to high concentration of Cd in soil. For this, a pot experiment was carried out in a greenhouse for a month, with two lines of MuS1 transgenic tobaccos (S4 and S6) and non-transgenic tobacco cultivated in the soils spiked at three different Cd concentrations (0, 60 and 180 mg $kg^{-1}$). Both transgenic and non-transgenic tobacco showed visible toxic symptoms such as chlorosis and leaf roll as treated concentration increased. The net photosynthetic rates of MuS1 plants (S4 and S6) exposed at 180 mg $kg^{-1}$ Cd were 6.3 and $7.7{\mu}mol\;m^{-2}s^{-1}$, being higher than those of the non-transgenic plant ($4.8{\mu}mol\;m^{-2}s^{-1}$). Values of stomatal conductance of MuS1 transgenic plants (0.05 and 0.008 mmol $H_2O\;m^{-2}s^{-1}$) were also higher than those of non-transgenic plant (0.03 mmol $H_2O\;m^{-2}s^{-1}$). In addition, fresh and dry weights of MuS1 transgenic plants were heavier than those of non-transgenic plant. Likewise, MuS1 transgenic plants appeared to be better physiological performance than non-transgenic tobacco when exposed at high concentration of Cd in soil. With regard to metal accumulation, MuS1 transgenic tobaccos accumulated more Cd in their roots than non-transgenic tobacco implying that MuS1 transgenic tobacco is suggested to be used for phytostabilization of heavy metals.

Expression of Aβ-Fc Fusion Protein in Transgenic Potato

  • Kim, Hyun-Soon;Youm, Jung Won;Lee, Jeong-Hwan;Jeon, Jae-Heung;Ko, Kisung
    • Horticultural Science & Technology
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    • v.32 no.3
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    • pp.375-381
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    • 2014
  • Transgenic potato was generated to express recombinant 5 repeated ${\beta}$-amyloid ($A{\beta}$) peptides, potential antigens to be applied as a preventive accine for Alzheimer's disease using Agrobacterium mediated transformation. The $A{\beta}$ peptides were fused to the human IgG Fc fragment enhancing protein and KDEL, which is the endoplasmic reticulum (ER) retention signal ($5A{\beta}$-FcK). The $5A{\beta}$-FcK, was expressed under the control of the duplicated 35S promoter. PCR analysis confirmed the presence of the transgene in several transgenic potato lines. Southern blot analysis showed only a single gene copy number in transgenic line 22, whereas multiple gene copy numbers were shown for transgenic lines 31 and 44. Northern blot analysis showed that line 22 had stronger mRNA levels when compared to lines 31 and 44. Immunoblot analysis confirmed that the $5A{\beta}$-FcK protein was expressed in the transgenic potato plant. These results indicate that $5A{\beta}$ fused to Fc can be expressed in potato plants.