• Title/Summary/Keyword: Transgenic Plant

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Overproduction of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) confers resistance to the herbicide glyphosate in transgenic rice

  • Lee, Soo-In;Kim, Hyun-Uk;Shin, Dong-Jin;Kim, Jin-A;Hong, Joon-Ki;Kim, Young-Mi;Lee, Yeon-Hee;Koo, Bon-Sung;Kwon, Sun-Jong;Suh, Seok-Chul
    • Journal of Plant Biotechnology
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    • v.38 no.4
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    • pp.272-277
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    • 2011
  • Plants expressing Agrobacterium sp. strain CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) are known to be resistant to glyphosate, a potent herbicide that inhibits the activity of the endogenous plant EPSPS. In order to develop herbicide-resistant rice, we prepared transgenic rice plants with CP4 EPSPS gene under the control of CaMV 35S promoter for over-expression. A recombinant plasmid was transformed into rice via Agrobacterium-mediated transformation. A large number of transgenic rice plants were obtained with glyphosate and most of the transformants showed fertile. The integration and expression of CP4 EPSPS gene from regenerated plants was analyzed by Southern and northern blot analysis. The transgenic rice plants had CP4 EPSPS enzyme activity levels more than 15-fold higher than the wild-type plants. EPSPS enzyme activity of transgenic rice plants was also identified by strip-test method. Field trial of transgenic rice plants further confirmed that they can be selectively survived at 100% by spay of glyphosate (Roundup$^{(R)}$) at a regular dose used for conventional rice weed control.

Selection of Transgenic Potato Plants Expressing Both CuZnSOD and APX in Chloroplasts with Enhanced Tolerance to Oxidative Stress (CuZnSOD와 APX를 엽록체에 발현시킨 산화스트레스 내성 형질전환 감자의 선발)

  • Tang, Li;Kwon, Suk-Yoon;Sung, Chang-K;Kwak, Sang-Soo;Lee, Haeng-Seoon
    • Journal of Plant Biotechnology
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    • v.31 no.2
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    • pp.109-113
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    • 2004
  • In order to develop transgenic potato plants with enhanced tolerance to multiple stress, we constructed the transformation vector expressing both superoxide dismutase and ascorbate peroxidase genes in chloroplasts under the control of a stress-inducible SWPA2 promoter. Transgenic potato plants (cv. Superior and Atlantic) were generated using an Agrobacterium-mediated transformation system. Transgenic potato plants were regenerated on MS medium containing 100mg/L kanamycin. Genomic Southern blot analysis confirmed the incorporation of foreign genes into the potato genome. When potato leaf discs were subjected to methyl viologen (MV) at 10 $\mu$M, transgenic plants showed higher tolerance than non-transgenic or vector-transformed plants. To further study we selected the transgenic plant lines with enhanced tolerance against MV. These plants will be used for further analysis of stress-tolerance to multiple environmental stresses.

Overexpression of a Chromatin Architecture-Controlling ATPG7 has Positive Effect on Yield Components in Transgenic Soybean

  • Kim, Hye Jeong;Cho, Hyun Suk;Pak, Jun Hun;Kim, Kook Jin;Lee, Dong Hee;Chung, Young-Soo
    • Plant Breeding and Biotechnology
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    • v.5 no.3
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    • pp.237-242
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    • 2017
  • AT-hook proteins of plant have shown to be involved in growth and development through the modification of chromatin architecture to co-regulate transcription of genes. Recently, many genes encoding AT-hook protein have been identified and their involvement in senescence delay is investigated. In this study, soybean transgenic plants overexpressing chromatin architecture-controlling ATPG7 gene was produced by Agrobacterium-mediated transformation and investigated for the positive effect on the important agronomic traits mainly focusing on yield-related components. A total of 27 transgenic soybean plants were produced from about 400 explants. $T_1$ seeds were harvested from all transgenic plants. In the analysis of genomic DNAs from soybean transformants, ATPG7 and Bar fragments were amplified as expected, 975 bp and 408 bp in size, respectively. And also exact gene expression was confirmed by reverse transcriptase-PCR (RT-PCR) from transgenic line #6, #7 and #8. In a field evaluation of yield components of ATPG7 transgenic plants ($T_3$), higher plant height, more of pod number and greater average total seed weight were observed with statistical significance. The results of this study indicate that the introduction of ATPG7 gene in soybean may have the positive effect on yield components.

Thermotolerant Transgenic Ginseng (Panax ginseng C.A. Meyer) by Introducing Isoprene Synthase Gene through Agrobacterium tumefaciens-mediated Transformation

  • Kim, Ok-Tae;Hyun, Dong-Yun;Bang, Kyong-Hwan;Jung, Su-Jin;Kim, Young-Chang;Shin, Yu-Su;Kim, Dong-Hwi;Kim, Swon-Won;Seong, Nak-Sul;Cha, Seon-Woo;Park, Hee-Woon
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.2
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    • pp.95-99
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    • 2007
  • The cost of conventional cultivation of ginseng (Panax ginseng C.A. Meyer) is very expensive, because shadow condition should be maintained during cultivation periods owing to inherently weak plant for high-temperature. Therefore, application of plant biotechnology may be possible to overcome these difficulties caused by conventional breeding of ginseng. Transgenic plants were produced via Agrobacterium tumefaciens Gv3101, both carrying the binary plasmid pBI121 mLPISO with nptII and Iso (isoprene synthase) gene. Integration of the transgenes into the P. ginseng nuclear genome was confirmed by PCR analysis using nptII primers and Iso primers. RT-PCR result also demonstrated the foreign isoprene synthase gene in three transgenic plant lines (T1, T3, and T5) which was expressed at the transcriptional level. When whole plants of transgenic ginseng were exposed to high temperature at $46^{\circ}C$ for 1 h, a non-transformed plant was wilted from heat shock, whereas a transgenic plant appeared to remain healthy. We suggest that the introduction of exogenous isoprene synthase is considered as alternative methods far generating thermotolerance ginseng.

Resistance to the Fungal Pathogen Phytophthora infestans of Transgenic Potato Plants Harboring of Chitinase Gene (Chitinase 유전자 도입 형질전환 감자식물체의 역병저항성)

  • Choi, Kyung-Hwa;Yang, Duk-Chun;Kim, Hyun-Soon;Choi, Kyung-Ja;Cho, Kwang-Yeon;Jung, Hyuk
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.3
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    • pp.177-182
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    • 1999
  • A fungal infection assay between normal and transgenic potato harboring chitinase gene in cultivar Belchip was investigated. In the first stage of experiment, seven transgenic lines having 12cm tall were tested for their resistance against potato late blight pathogen Phytophthora infestans by infection with the zoospores, artificially, Susceptibility to potato late blight infection could be classified into three types based on the rate. In terms of resistance to the disease, two lines were higher, two lines were more suppressive, and three lines were similar as compared with the control. In the following experiment, only 2 risistant lines and 1 suppressed line were used to confirm the resistance again. The results of both experiments were similar. Furthermore, two highly resistant transgenic lines grown in field exhibited a higher resistance than control under the conditions of natural ocurrence of the fungal disease.

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Cloning And Characterization of Pathogen-Inducible EREBP-Like Transcription Factor(CaNR19) From Hot Pepper (Capsicum annuum L.)

  • Yi, So-Young;Kim, Jee-Hyub;Yu, Seung-Hun;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.77.2-78
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    • 2003
  • An EREBP/AP2-type transcription factor (CaPFl) was isolated by DDRT-PCR following inoculation of soybean pustule pathogen Xanthomonas axonopodis pv. glycines Bra which induces HR on pepper leaves. Genomic Southern blot analysis revealed that the CaPFl gene is present as a single copy within the hot pepper genome. The deduced amino acid sequence of CaPFl has two potential nuclear localization signals, a possible acidic activation domain, and an EREBP/AP2 motif that could bind to a conserved cis- element present in promoter region of many stress-induced genes. The mRNA level of CaPFl was induced by both biotic and abiotic stresses. We observed higher-level transcripts in resistance-induced pepper tissues than diseased tissues. Expression of CaPFl is also induced upon various abiotic stresses including ethephon, MeJA, cold stress, drought stress and salt stress treatments. To study the role of CPFI in plant, transgenic Arabidopsis and tobacco plants which express higher level of pepper CaPFl were generated. Global gene expression analysis of transgenic Arabidopsis by cDNA microarray indicated that expression of CaPFl in transgenic plants affect the expression of quite a few GCC box and DRE/CRT box-containing genes. Furthermore, the transgenic Arabidopsis and tobacco plant, expressing CaPFl showed tolerance against freezing temperature and enhanced resistance to Pseudomonas syrnigae pv. tabaci. Taken together, these results indicated that CaPFl is a novel EREBP/AP2 transcription factor in hot pepper plant and it may has a significant role(s) in regulation of biotic and abiotic stresses in plant.

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Altered Expression of Pyrophosphate: Fructose-6-Phosphate 1-Phosphotransferase Affects the Growth of Transgenic Arabidopsis Plants

  • Lim, Hyemin;Cho, Man-Ho;Jeon, Jong-Seong;Bhoo, Seong Hee;Kwon, Yong-Kook;Hahn, Tae-Ryong
    • Molecules and Cells
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    • v.27 no.6
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    • pp.641-649
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    • 2009
  • Pyrophosphate:fructose-6-phosphate 1-phosphotransferase (PFP) catalyzes the reversible interconversion of fructose-6-phosphate and fructose-1,6-bisphosphate, a key step in the regulation of the metabolic flux toward glycolysis or gluconeogenesis. To examine the role of PFP in plant growth, we have generated transgenic Arabidopsis plants that either overexpress or repress Arabidopsis PFP subunit genes. The overexpressing lines displayed increased PFP activity and slightly faster growth relative to wild type plants, although their photosynthetic activities and the levels of metabolites appeared not to have significantly changed. In contrast, the RNAi lines showed significantly retarded growth in parallel with the reduced PFP activity. Analysis of photosynthetic activity revealed that the growth retardation phenotype of the RNAi lines was accompanied by the reduced rates of $CO_2$ assimilation. Microarray analysis of our transgenic plants further revealed that the altered expression of $AtPFP{\beta}$ affects the expression of several genes involved in diverse physiological processes. Our current data thus suggest that PFP is important in carbohydrate metabolism and other cellular processes.

Localization of Single Chain Fv Antibodies (scFv) in Transgenic Tobacco Ptants Showing Resistance against Tomato Bushy Stunt Virus

  • Jeun, Y.C.;Boonrod, K.;Nagy, P.;Conrad, U.;Krczal, G.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.75.2-75
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    • 2003
  • To develop an effective protection strategy against tomato bushy stunt virus (TBSV), tobacco plants expressing single-chain Fv antibodies (scFv), were established. A previous had shown that the replication activity of viral replicase was inhibited by the selected scFvs. Moreover, no systemic symptom was found after virus inoculation on leaves of wt N. benthamiana infiltrated with an Agrobacterium suspension resulting i3l expression of the scFvs. However, control plants showed systemic symptoms. In this study the localization of the scFvs within two transgenic plant lines, (CP28H3, CP-P55) was demonstrated using immunogold labelling. The gold particles, indicating the presence of scFv, were mostly found In the cytoplasm of the plant cells including chloroplasts and in the cell walls. However, they were hardly found in the vacuole, nucleoplasm and intercellular spaces. Gold particles often accumulated in either the cytosol or chloroplasts showing a specific labeling, There was no difference in type of gold labeling between both transgenic lines. The localization of the scFv in the cytoplasm further conforms the inhibition of the RNA-dependent RNA polymerase (RdRp) by the selected scFv because it is known that the RdRp is localized to membraneous cytosolic structures.

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Transcriptome analysis of a transgenic Arabidopsis plant overexpressing CsBCAT7 reveals the relationship between CsBCAT7 and branched-chain amino acid catabolism

  • Kim, Young-Cheon;Lee, Dong Sook;Jung, Youjin;Choi, Eun Bin;An, Jungeun;Lee, Sanghyeob;Lee, Jeong Hwan
    • Journal of Plant Biotechnology
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    • v.48 no.4
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    • pp.228-235
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    • 2021
  • The amino acids found in plants play important roles in protein biosynthesis, signaling processes, and stress responses, and as components in other biosynthesis pathways. Amino acid degradation helps maintain plant cells' energy states under certain carbon starvation conditions. Branched-chain amino acid transferases (BCATs) play an essential role in the metabolism of branched-chain amino acids (BCAAs) such as isoleucine, leucine and valine. In this paper, we performed genome-wide RNA-seq analysis using CsBCAT7-overexpressing Arabidopsis plants. We observed significant changes in genes related to flowering time and genes that are germination-responsive in transgenic plants. RNA-seq and RT-qPCR analyses revealed that the expression levels of some BCAA catabolic genes were upregulated in these same transgenic plants, and that this correlated with a delay in their senescence phenotype when the plants were placed in extended darkness conditions. These results suggest a connection between BCAT and the genes implicated in BCAA catabolism.

Expression of Rice Chitinase Gene in Genetically Engineered Tomato Confers Enhanced Resistance to Fusarium Wilt and Early Blight

  • Jabeen, Nyla;Chaudhary, Zubeda;Gulfraz, Muhammad;Rashid, Hamid;Mirza, Bushra
    • The Plant Pathology Journal
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    • v.31 no.3
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    • pp.252-258
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    • 2015
  • This is the first study reporting the evaluation of transgenic lines of tomato harboring rice chitinase (RCG3) gene for resistance to two important fungal pathogens Fusarium oxysporum f. sp. lycopersici (Fol) causing fusarium wilt and Alternaria solani causing early blight (EB). In this study, three transgenic lines TL1, TL2 and TL3 of tomato Solanum lycopersicum Mill. cv. Riogrande genetically engineered with rice chitinase (RCG 3) gene and their R1 progeny was tested for resistance to Fol by root dip method and A. solani by detached leaf assay. All the R0 transgenic lines were highly resistant to these fungal pathogens compared to nontransgenic control plants. The pattern of segregation of three independent transformant for Fol and A. solani was also studied. Mendelian segregation was observed in transgenic lines 2 and 3 while it was not observed in transgenic line 1. It was concluded that introduction of chitinase gene in susceptible cultivar of tomato not only enhanced the resistance but was stably inherited in transgenic lines 2 and 3.