• Title/Summary/Keyword: herbicide resistant plant

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Molecular and Cytogenetic Analysis of Transgenic Plants of Rice(Oryza sativa L.) Produced by Agrobacterium-mediated Transformation

  • Cho, Joon-Hyeong;Kim, Yong-Wook
    • Plant Resources
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    • v.7 no.1
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    • pp.39-46
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    • 2004
  • To demonstrate the importance of transformation efficiency in independent event, molecular and cytogenetic analysis were conducted with genomic DNA and chromosome of transgenic plants produced by Agrobacterium tumefeciens LBA4404 (pSBM-PPGN: gusA and bar). Selection ratios of putative transgenic calli were similar in independent experiments, however, transformation efficiencies were critically influenced by the type of regeneration media. MSRK5SS-Pr regeneration mediun, which contains 5 mgL$^{-1}$ kinetin, 2% (w/v) sucrose in combination with 3% (w/v) sorbitol, and 500 mgL$^{-1}$ proline, was efficient to produce transgenic plant of rice from putative transgenic callus in the presence of L-phosphinotricin (PPT). With MSRK5SS-Pr medium, transformation efficincies of Nagdongbyeo were significantly enhanced from 3.7% to 6.3% in independent callus lines arid from 7.3% to 19.7% in plants produced, respectively. Stable integration and expression of bar gene were confirmed by basta herbicide assay, PCR amplification and Southern blotting of bar gene, and fluorescence in situ hybridization (FISH) analysis using pSBM-PPGN as a probe. In Southern blot analysis, diverse band patterns were observed in total 44 transgenic plants regenerated from 20 independent PPT resistant calli showing from one to five copies of T-DNA segments, however, the transformants obtained from one callus line showed the same copy numbers with the same fractionized band patterns.

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Development of Bialaphos Resistant Transgenic Tabacco Plants by Pollination and Utilization of Fertilization Cycle (수분ㆍ수정 시기를 이용한 Bialaphos 저항성 형질전환 담배의 개발)

  • ;;;;;;Toshiaki KAMEYA
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.2
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    • pp.99-103
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    • 1994
  • The herbicide bialaphos is a potent inhibitor of glutamine synthetase in higher plants. A bialaphos resistance (bar) gene encoding for an acetyltransferase was isolated from genomic DNA of Pseudomonas syringae pv tabaci. The bar gene was ligated to the binary vector pBI121. Pistils of tobacco plane were heated with the bar gene containing plasmid DNA at various times after pollination. When the treatment was applied at 30 and 40 h after pollination, a number of transgenic plants were obtained. Premary transformation (T$_{0}$ generation) and their progenies (T$_1$T$_2$) were resistant to both bialaphos and kanamycin at a dosage lathal to untransformed control plants. Stable integration of bar gene into chromosomal DNA was proven by Southern blot analysis of genomic DNA isolated from T$_1$progenies. These results show that the bialaphos resistant plane could be obtained by treatment to pistils with the exgenous bar gene through the fertilization cycle of tobacco.o.

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Selection of Herbicide - Tolerant Rice(Oryza sativa L.) Callus by Tissue Culture (조직배양(組織培養)을 통한 수도내성(水稻耐性) 카루스 선발(選拔))

  • Shin, D.H.;Moody, K.;Zapata, F.J.;Kim, K.U.
    • Korean Journal of Weed Science
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    • v.10 no.4
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    • pp.285-293
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    • 1990
  • The response of callus growth of rice (Oryza sativa L.) cultivars which showed different responses to herbicides as seedlings was investigated to select resistant or tolerant calli. Callus growth of IR28 which was susceptible to thiobencarb (S-[(4-chlorophenyl)methyl]diethylcarbamothioate) during callus induction was not inhibited by $10^{-5}$ M and $10^{-6}$ M thiobencarb, indicating that there was a difference in tolerance among callus induction and growth, and the intact plant level. A similar result was obtained with IR31917-45-3-2-2 to butachlor [N - (buthoxymethyl) -2-chloro-N- (2, 6-diethylphenyl) acetamide]. The fresh weight of IR28 callus transferred into $10^{-5}$ M thiobencarb after treatment at $10^{-6}$ M for 30 days was not affected by the herbicide, indicating that transferring callus into gradually higher herbicide concentrations can be a useful method for selection of herbicide-tolerant cell lines.

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Selection of Herbicide Resistant Potatoes Transformed with Phosphinothricin Acetyltransferase Gene (Phosphinothricin Acetyltransferase 유전자 도입에 의한 제초제 저항성 감자의 선발)

  • Han, S.S.;Jeong, J.H.;Bang, K.S.;Yang, D.C.
    • Korean Journal of Weed Science
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    • v.17 no.4
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    • pp.390-399
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    • 1997
  • This experiment was conducted to introduce PAT (phosphinothricin acetyltransferase, non-selective herbicide bialaphos resistant gene) gene into potato (Solanum tuberosum. cv. Desiree). Optimal shoot regeneration from leaf discs and stem segments was obtained in MS medium supplemented with 0.1 mg/L IBA and 0.5 mg/L BA, and the frequency of shoot regeneration was 54% in left discs and 46% in stem segments. In this condition, leaf discs and stem segments of potato were co-cultivated with A. tumefaciens MP90 which contained binary vector with GUS: :NPTII gene and PAT gene. Transgenic shoots were regenerated from leaf and stem-derived calli on selection medium with 100mg/L kanamycin. The 100${\mu}M$ acetosyringone treatment during the co-cultivation highly enhanced(4 times than the control) the shoot regeneration on selection medium. When the putative transgenic plants were transferred to medium with 10mg/L basta, all of them were survived. After PCR. GUS test, and Southern blot analysis of the survived plant, we confirmed that the gene was stably integrated into the potato genome and expressed. After the transgenic plants were transplanted in soil, and the transgenic plants were sprayed with the herbicide basta (300ml/10a), the transgenic plants remained green but control plants were died.

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Current Research Status on the Development of Genetically Modified Plants in Korea (유전자변형식물의 국내 연구 현황)

  • Lee, Shin-Woo
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.1-6
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    • 2003
  • In an attempt to evaluate the current research status of genetically modified (GM) plants, the scientific research publications in Korea as well as in international SCI journals were screened. About 190 research articles related to the development of GM plants were searched from 10 different domestic journals in the last 12 years (Jan. 1990 to Sept. 2002), The researches in 65 articles were carried out with tobacco plant, 20 with rice, 19 with potatoes, and less then 9 articles from each other plant species, respectively, In total, 38 different plant species were being subjected for the development of GM plants. In particular, there was only one article for each major staple grains such as wheat, barley, soybean, and maize. In more than 47% of total published articles, scientists mainly focused on the basic research such as developing transformation system (46 articles), gene expression study in transgenic plants (34), and vector constructions (10). In addition, 28 articles which main authors are Korean scientists were searched from 11 different international SCI journals. Again, major plants for GM research were tobacco (10) and rice (7). More than 50% of published articles were focused on the basic research, gene expression study with transgenic plants (16). The publications on the research of disease-resistant plants were 7 articles, 3 for the development of stress-resistant and 2 for the herbicide-resistant plants, respectively. It is believed that the last 10 year's investment through government organizations has just strengthen the capacity for the next big stride on agricultural biotechnology in Korea.

Genetic transformation of Sedum erythrostichum via Agrobacterium-mediated transformation by introducing herbicide-resistant gene (아그로박테리움을 통한 제초제 저항 꿩의비름(Sedum erythrostichum) 형질전환체 개발)

  • 윤의수;정재훈;최용의
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2002.11b
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    • pp.30-30
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    • 2002
  • 꿩의비름 (Sedum erythrostichum)은 매우 우수한 지피식물이며 건조에 강한 대표적 식물로 바위정원 (rock garden)을 가꾸는데 있어서 중요한 수종으로 이용되며, 유럽등지에서는 지붕에 식재하기도 하며 최근에는 빌딩옥상녹화의 대표적 수종으로 식재되고 있다. 또한 한방에서는 경천이라 불리우기도 하는데 피부상처 치유 및 미백효과가 탁월하다고 알려져 있다. 본 연구에서는 Agrobacterium을 매개로한 꿩의비름의 형질전환 시스템을 개발하고 아울러 phosphinothricin-N-acetyltransferase (PAT) 유전자를 도입하여 제초제 저항 식물을 개발하고자 수행되었다. 꿩의비름 잎을 Agrobacterium에 담근후 0.5 mg/l NAA와 2 mg/1 BA가 첨가된 MS 배지에 3일간 공동배앙 하였다. 그 후 300 mg/1 cefotaxime이 첨가된 같은 배지에 옮겨 계대하면서 Agrobacterium을 제거하였다. 약 3주후에 잎 절편으로 부터 직접적으로 부정아가 형성되기 시작 하였는데 이 시기부터 잎 절편을 25 mg/1 kanamycin이 첨가된 선발배지에 옮겨 주었다. 이 결과 배양된 잎 절편 절편 중 3.75%에서 kanamycin에 저항하는 부정아를 얻을 수 있었다. 형질전환체는 X-gluc 반응, PCR, Southern, Nothern analysis를 통하여 확인하였다. 약 94%의 형질전환 식물체는 성공적으로 토양에 옮길 수 있었으며 약 3개월후에 꽃을 피웠다. 형질전환체는 제초제인 Basta ($^{(R)}$ phosphinothricine at 200 mg/1)를 살포하여 주었을 경우 생존함을 확인 하였다.

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Progress and Prospect of Rice Biotechnology in Korea

  • Tae Young, Chung
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 1997.06a
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    • pp.23-49
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    • 1997
  • This is a progress report of rice biotechnology including development of gene transformation system, gene cloning and molecular mapping in rice. The scope of the research was focused on the connection between conventional breeding and biotech-researches. Plant transformation via Agrobacterium or particle bombardment was developed to introduce one or several genes to recommended rice cultivars. Two chimeric genes containing a maize ribosome inactivating protein gene (RIP) and a gerbicide resistant gene (bar) were introduced to Nipponbare, a Japonica cultivar, and transmitted to Korean cultivars. The homozygous progenies of herbicide resistant transgenic plant showed good fertility and agronomic characters. To explore the genetic resourses in rice, over 8,000 cDNA clones from immature rice seed have been isolated and sequenced. About 13% of clones were identified as enzymes related to metabolic pathway. Among them, twenty clones have high homology with genes encoding enzymes in the photorespiratory carbon cycle reaction. Up to now about 100 clones were fully sequenced and registered at EMBL and GenBank. For the mapping of quantitative tarits loci (QTL) and eternal recombinant inbred population with 164 F13 lines (MGRI) was developed from a cross between Milyang 23 and Gihobyeo, Korean rice cultivars. After construction of fully saturated RFLP and AFLP map, quantitative traits using MGRI population were analyzed and integrated into the molecular map. Eighty seven loci were determined with 27 QTL characters including yield and yield components on rice chromosomes. Map based cloning was also tried to isolate semi-dwarf (sd-1) gene in rice. A DNA probe, RG 109, the most tightly linked to sd-1 gene was used to screen from bacterial artifical chromosome (BAC) libraries and five over lapping clones presumably containing sd-1 gene were isolated. Rice genetic database including results of biotech reasearch and classical genetics is provided at Korea Rice Genome Server which is accessible with world wide web (www) browser. The server provides rice cDNA sequences and map informations linked with phenotypic images.

The Investigation of Gene Flows in Artificial Pollination between GM Rice and its Wild Relatives by RAPD Analysis (RAPD PCR에 의한 GM벼의 야생 근연종 벼로의 유전자 전이 분석법)

  • Kim, Yoon-Sik;Kim, Hyun-Soon;Joung, Hyouk;Jeon, Jae-Heung
    • Korean Journal of Plant Resources
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    • v.19 no.5
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    • pp.612-616
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    • 2006
  • In recent years, there has been increasing concerns in gene flow from GM crops to wild or weedy relatives as a potential risk in the commercialization of GM crops. To access the possibility of the environmental impacts by GM rice, small-scale experiments of gene transfer were carried out. Herbicide and drought stress resistant GM rice and non-GM rice Nakdongbyeo, wild rice Oryza nivara, and weedy rice Sharebyeo were used for artificial pollination experiments and bar gene was used as a tractable marker after pollination. The harvested putative hybrid seeds after artificial pollination were germinated and true hybrid plants were selected by basta treatment. The hybrid plants were verified again by PCR amplification of bar and trehalose-6-phosphate phosphatase (TPP) genes and RAPD PCR analysis.

Development of herbicide tolerant soybean using Agrobacterium tumefaciens (아그로박테리움을 이용만 제초제 저항성 콩 개발)

  • Lee, Ki-Jong;Park, Hong-Jae;Yi, Bu-Young;Lee, Kyeong-Ryeol;Kim, Myung-Sik;Woo, Hee-Jong;Jin, Yong-Moon;Kweon, Soon-Jong
    • Journal of Plant Biotechnology
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    • v.35 no.1
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    • pp.69-74
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    • 2008
  • This study aims to establish the efficient soybean transformation system and develop soybean [Glycine max (L.) Merill] transformants using cotyledonary node explants. The cotyledonary node of soybean were co-cultivated with Agrobacterium tumefaciens strains (KYRT1, EHA105). These strains contain the binary vector pCAMBIA3301 which carries a herbicide-resistant far gene. Korean cultivars (Danbaekkong, Eunhakong) and foreign cultivars (Jack, Peking) were the most efficient in regenerating cotyledonary node. Therefore, they were chosen for the transformation. Results showed that the T-DNA transfer reached up to 60% and transformation efficiency reached up to 3% in the cotyledonary node explants from Jack cultivar, co-cultivated with EHA105 strain. Histochemical GUS evaluation showed that 12 individual lines, transformed with the 현 gene, have positive response. The transformed soybeans have been confirmed in the $T_0$ generation through phenotypic assay using herbicide $Basta^{(R)}$ and Southern blot analysis.

Current status on the development and commercialization of GM plants (국내·외 GM식물의 개발 및 산업화 현황)

  • Lee, Shin-Woo
    • Journal of Plant Biotechnology
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    • v.37 no.3
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    • pp.305-312
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    • 2010
  • During a last decade, the introduced traits in commercialized GM crops have been diversified from a simple trait such as herbicide resistance gene or insectresistance gene which are related to the crop production into more complicated traits such as modification of fatty acid or essential amino acid composition, modified coloring pattern of flower. In addition, it was investigated that several other GM crops bearing more refined traits expected to lead next generation are also awaiting for risk assessment (RA) or under field test for the preparation of RA in the near future. These GM crops include abiotic stress resistance including drought or cold, increased biomass, production of bioethanol or diesel, production of pharmaceuticals or functional materials for industrial. In particular, in 2008 and 2009, it was reported that the highest number of GM crops for molecular farming are under developed in laboratory or green house level in all the world. Likewise, in Korea, 171 events from 49 plant species are under developed to introduce several important traits. At present, about 10 events are under field test to select elite lines for RA application. For the first time, herbicide resistance turfgrass developed by Korean research team has been submitted for RA and currently under requested for additional data. Moreover, GM rice resistant to leaf roll (folder) disease is expected as a next event to be submitted for RA application.