• Title/Summary/Keyword: Transgenic Arabidopsis

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Enhanced Resistance to Botrytis cinerea Mediated by Transgenic Expression of the Spider Chitinase Gene AvChit in Arabidopsis

  • Hur, Yeon-Jae;Kim, Doh-Hoon
    • International Journal of Industrial Entomology and Biomaterials
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    • v.19 no.2
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    • pp.259-264
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    • 2009
  • The AvChit gene encodes for a chitinase from the spider, Araneus ventricosus. This spider, A. ventricosus, is an abundant species in Korea. Arabidopsis thaliana plants were transformed with the AvChit gene using Agrobacterium tumefaciens. Thirteen transgenic lines expressing the AvChit gene were obtained. Functional expression of the AvChit gene in transgenic Arabidopsis was confirmed by Southern, northern and western blot analysis. The AvChit cDNA was expressed as a 61 kDa polypeptide in baculovirus-infected insect Sf9 cells. AvChit protein extracted from transgenic Arabidopsis exhibited high levels of chitinase activity. Phytopathological tests showed that two transgenic Arabidopsis lines expressing the AvChit gene displayed high levels of resistance to gray mold disease (Botrytis cinerea).

Overexpression of the Metal Transport Protein1 gene (MTP1) in Arabidopsis Increased tolerance by expression site (금속전달 유전자(MTP1)의 과발현 애기장대에서 발현 위치에 따른 내성 증가 연구)

  • Kim, Donggiun
    • The Journal of the Convergence on Culture Technology
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    • v.5 no.3
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    • pp.327-332
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    • 2019
  • Today's scientists try to remove heavy metals with many new technologies such as phytoremediation. One of the best cutting edge technologies is developing transgenic plants to remove certain heavy metal in soil. I constructed the transformation vector expressing T. goesingense Metal Transport Protein1 gene and TgMTP1: GFP genes. The transgenic plants were selected and confirmed the transformed genes into Arabidopsis thaliana genome. Expression was confirmed in several parts in Arabidopsis cells, tissues and organs. When TgMTP1 overexpressing Arabidopsis thaliana were subjected, transgenic plants showed higher heavy metal tolerance than non-transgenic. For further study I selected the transgenic plant lines with enhanced tolerance against four different heavy metals; Zn, Ni, Co, Cd. The accumulation of these metals in these plants was further analyzed. The TgMTP1 overexpressing Arabidopsis thaliana plant of selected lines are resistant against heavy metals. This plant is characterized by the expression of the MTP1 gene accumulating heavy metal in the vacuole and being simultaneously expressed on the plasma membrane. In conclusion, these plants may be used in plant purification applications, and as a plant with increased tolerance.

Regulation of methionine biosynthesis in plants; transgenic study

  • Kim, Jungsup;Thomas Leustek
    • Proceedings of the Botanical Society of Korea Conference
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    • 2002.04a
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    • pp.73-82
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    • 2002
  • The committing step in Met and S-adenosyl-L-methionine (SAM) synthesis is catalyzed by cystathionine ${\gamma}$ -synthase (CGS). Transgenic Arabidopsis thaliana overexpressing CGS under control of 35S promoter show increased soluble Met and its metabolite S-methylmethionine, but only at specific stages of development. CGS-overexpressing seedlings are resistant to ethionine. Similar results were obtained with transgenic potato plants overexpressing Arabidopsis CGS. Several of the transgenic lines show silencing of CGS resulting in deformed p]ants with a reduced capacity for reproductive growth similar as transgenic plants by antisense RNA (CGS[-]). Exogenous feeding of Met to the CGS[-] and CGS[+] silenced plants partially restores their growth. Similar morphological deformities are observed in plants cosuppressed for SAM synthetase, even though such plants accumulate 250 fold more soluble Met than wild type and they overexpress CGS. The results suggest that the abnormalities associated with CGS and SAM synthetase silencing are due in part to a reduced ability to produce SAM, and that SAM may be a regulator of CGS expression.

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Expression of yeast Hem1 gene controlled by Arabidopsis HemA1 promoter improves salt tolerance in Arabidopsis plants

  • Zhang, Zhi-Ping;Yao, Quan-Hong;Wang, Liang-Ju
    • BMB Reports
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    • v.43 no.5
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    • pp.330-336
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    • 2010
  • 5-Aminolevulinate (ALA) is well-known as an essential biosynthetic precursor of all tetrapyrrole compounds, which has been suggested to improve plant salt tolerance by exogenous application. In this work, the gene encoding aminolevulinate synthase (ALA-S) in yeast (Saccharomyces cerevisiae Hem1) was introduced into the genome of Arabidopsis controlled by the Arabidopsis thaliana HemA1 gene promoter. All transgenic lines were able to transcribe the YHem1 gene, especially under light condition. The chimeric protein (YHem1-EGFP) was found co-localizing with the mitochondria in onion epidermal cells. The transgenic Arabidopsis plants could synthesize more endogenous ALA with higher levels of metabolites including chlorophyll and heme. When the $T_2$ homozygous seeds were cultured under NaCl stress, their germination and seedling growth were much better than the wild type. Therefore, introduction of ALA-S gene led to higher level of ALA metabolism with more salt tolerance in higher plants.

Expression of Bacillus subtilis proBA Genes and Reduction of Feedback Inhibition of Proline Synthesis Increases Proline Production and Confers Osmotolerance in Transgenic Arabidopsis

  • Chen, Mingqing;Wei, Hongbo;Cao, JunWei;Liu, Ruijie;Wang, Youliang;Zheng, Congyi
    • BMB Reports
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    • v.40 no.3
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    • pp.396-403
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    • 2007
  • Proline accumulation has been shown to correlate with tolerance to drought and salt stresses in plants. We attempt to introduce the wild-type, mutant, and fusion proBA genes derived from Bacillus subtilis into Arabidopsis thaliana under the control of a strong promoter cauliflower mosaic virus 35S (CaMV35S). The transgenic plants produced higher level of free proline than control and the overproduction of proline resulted in the increased tolerance to osmotic stress in transgenic plants. Besides, the mutation in proBA genes, which were proved to lead $\alpha$-glutamyl kinase ($\alpha$-GK) reduces sensitivity to the end-product inhibition and the fusion of proB and proA also result in increasing proline production and confer osmotolerance in transgenic lines.

An Arabidopsis Homologue of Human Seven-in-Absentia-interacting Protein Is Involved in Pathogen Resistance

  • Kim, Youn-Sung;Ham, Byung-Kook;Paek, Kyung-Hee;Park, Chung-Mo;Chua, Nam-Hai
    • Molecules and Cells
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    • v.21 no.3
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    • pp.389-394
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    • 2006
  • Human seven-in-absentia (SIAH)-interacting protein (SIP) is a component of the E3 ligase complex targeting beta-catenin for destruction. Arabidopsis has one SIP protein (AtSIP) with 32% amino acid sequence identity to SIP. To investigate the functions of AtSIP, we isolated an atsip knockout mutant, and generated transgenic plants overexpressing AtSIP. The growth rates and morphologies of the atsip and transgenic plants were indistinguishable from those of wild type. However, atsip plants were more susceptible to Pseudomonas syringae infection, and the transgenic plants overexpressing AtSIP were more resistant. Consistent with this, RNA blot analysis showed that the AtSIP gene is strongly induced by wounding and hydrogen peroxide treatment. In addition, when plants were infected with P. syringae, AtSIP was transiently induced prior to PR-1 induction. These observations show that Arabidopsis AtSIP plays a role in resistance to pathogenic infection.

Identification of a Promoter Motif Involved in Curtovirus Sense-Gene Expression in Transgenic Arabidopsis

  • Hur, Jingyung;Choi, Eunseok;Buckley, Kenneth J.;Lee, Sukchan;Davis, Keith R.
    • Molecules and Cells
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    • v.26 no.2
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    • pp.131-139
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    • 2008
  • Expression of the seven open reading frames (ORFs) of single-stranded DNA Curtoviruses such as Beet curly top virus (BCTV) and Beet severe curly top virus (BSCTV) is driven by a bi-directional promoter. To investigate this bidirectional promoter activity with respect to viral late gene expression, transgenic Arabidopsis plants expressing a GUS reporter gene under the control of either the BCTV or BSCTV bi-directional promoter were constructed. Transgenic plants harboring constructs showed higher expression levels when the promoter of the less virulent BCTV was used than when the promoter of the more virulent BSCTV was used. In transgenic seedlings, the reporter gene constructs were expressed primarily in actively dividing tissues such as root tips and apical meristems. As the transgenic plants matured, reporter gene expression diminished but viral infection of mature transgenic plants restored reporter gene expression, particularly in transgenic plants containing BCTV virion-sense gene promoter constructs. A 30 base pair conserved late element (CLE) motif was identified that was present three times in tandem in the BCTV promoter and once in that of BSCTV. Progressive deletion of these repeats from the BCTV promoter resulted in decreased reporter gene expression, but BSCTV promoters in which one or two extra copies of this motif were inserted did not exhibit increased late gene promoter activity. These results demonstrate that Curtovirus late gene expression by virion-sense promoters depends on the developmental stage of the host plant as well as on the number of CLE motifs present in the promoter.

Pseudosymptom Analysis Induced by Geminivirus L4 Gene in Transgenic Arabidopsis (Geminivirus L4 유전자 도입에 따른 형질전환 애기장대의 감염유사증상 분석)

  • Lee, Suk-Chan;Lee, Kyu-Bae;Park, Jong-Bum
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.129-133
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    • 2003
  • The factors for symptom development caused by Beat curly top virus(BCTV) have been analyzed by using a molecular genetic approach based on expressing BCTV encoded proteins in transgenic plants. BCTV open reading frame (ORF) L4 expression in transgenic Arabidopsis resulted in abnormal plant development and the production of callus inflorescence stems and bumpy trichomes, confiming that this gene alone is a primary symptom determinant. The L4 gene expression by northern hybridization in transgenic plants and a range of phenotypes were analyzed.

Heterologous expression of the Arabidopsis DREB1A/CBF3 gene enhances drought and freezing tolerance in transgenic Lolium perenne plants

  • Li, Xue;Cheng, Xiaoxia;Liu, Jun;Zeng, Huiming;Han, Liebao;Tang, Wei
    • Plant Biotechnology Reports
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    • v.5 no.1
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    • pp.61-69
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    • 2011
  • The dehydration-responsive element binding proteins (DREB1)/C-repeat (CRT) binding factors (CBF) function as transcription factors and play an important role in agricultural biotechnology and molecular biology studies of drought and freezing stress tolerance. We generated transgenic Lolium perenne plants containing the PCR-cloned Arabidopsis DREB1A/CBF3 gene (AtDREB1A/CBF3) to study the function of this gene construct in drought and freezing tolerance in a species of turfgrass. Compared to the control, AtDREB1A/CBF3 transgenic L. perenne plants showed enhanced drought and freezing stress tolerance. The activities of the enzymes superoxide dismutase (SOD) and peroxidase (POD) were higher in transgenic plants than in the non-transgenic plant control. These results demonstrate that the expression of the AtDREB1A/CBF3 gene in transgenic L. perenne plants enhanced drought and freezing tolerance and that the increased stress tolerance was associated with the increased activities of antioxidant enzymes. These results are relevant to stress biology and biotechnology studies of turfgrass.

Arabidopsis cyclin D2 expressed in rice forms a functional cyclin-dependent kinase complex that enhances seedling growth

  • Oh, Se-Jun;Kim, Su-Jung;Kim, Youn Shic;Park, Su-Hyun;Ha, Sun-Hwa;Kim, Ju-Kon
    • Plant Biotechnology Reports
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    • v.2 no.4
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    • pp.227-231
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    • 2008
  • D-class cyclins play important roles in controlling the cell cycle in development and in response to external signals by forming the regulatory subunit of cyclin-dependent kinase (CDK) complexes. To evaluate the effects of D-class cyclins in transgenic rice plants, Arabidopsis cyclin D2 gene (CycD2) was linked to the maize ubiquitin1 promoter (Ubi1) and introduced into rice by the Agrobacterium-mediated transformation method. Genomic deoxyribonucleic acid (DNA), ribonucleic acid (RNA), and Western blot hybridizations of the Ubi1:-CycD2 plants revealed copy number of transgene and its increased expression in leaf and callus cells at messenger RNA (mRNA) and/or protein levels. The H1 kinase assay using the immunoprecipitates of protein extracts from the Ubi1:CycD2 plants and nontransgenic controls demonstrated that the introduced Arabidopsis CycD2 forms a functional CycD2/CDK complex with an unidentified CDK of rice. Shoot and root growth was enhanced in the Ubi1:CycD2 seedlings compared with nontransgenic controls, together, suggesting that Arabidopsis cyclin D2 interacts with a rice cyclin-dependent kinase, consequently enhancing seedling growth.