• Title/Summary/Keyword: Agrobacterium-mediated Transformation

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Efficient shoot regeneration using cotyledon explants and Agrobacterium tumefaciens - mediated transformation of bottle gourd (Lagenaria sicraria Standl.) (박 자엽조직을 이용한 효율적인 식물체 분화와 Agrobacterium에 의한 형질전환)

  • Kim, Soo-Yun;Ahn, Yul-Kyun;Huh, Yun-Chan;Lee, Hye-Eun;Kim, Do-Sun
    • Journal of Plant Biotechnology
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    • v.41 no.3
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    • pp.140-145
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    • 2014
  • This study were carried out for selection of proper transformation variety and development of efficient regeneration and transformation methods. The number of shoot in commercial varieties of gourd plant were 0 ~ 7.3. and fusarium wilt resistant pure lines were 2.0 ~ 6.5 per dish containing on MS medium supplemented with 3 mg/L BA. The shoot regeneration frequency of fusarium wilt resistant pure lines were wide variation on the deviation. The expression of GFP was high 67% and 100% at the co-cultivation with Agrobacterium. The effective shoot regeneration plant hormone were combination BA and 2,4-D. The number and elongation condition of shoot was good after 4 weeks change with MS medium supplemented with 1 mg/L BA. Effective callus production plant hormone were combination of 3 mg/L BA and 0.1 mg/L 2.4-D.

Production of Transgenic Birdsfoot Trefoil (Lotus corniculatus L.) Plants by Introduction of E35S Promoter + AtNDPK2 Gene (E35S 프로모터 + AtNDPK2 유전자 도입에 의한 버즈풋 트레포일 (Lotus corniculatus L.) 형질전환체 생산)

  • Kim Ki-Yong;Jang Yo-Soon;Choi Gi-Jun;Sung Byung-Ryeol;Kim Won-Ho;Seo Sung;Lee Byung-Hyun;Kwak Sang-Soo
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.26 no.2
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    • pp.83-90
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    • 2006
  • To develop transgenic birdsfoot trefoil (Lotus corniculatus L.) plants tolerant to environmental stress, Arabidopsis NDPK gene (AtNDPK) was introduced into birdsfoot trefoil plants using Agrobacterium-mediated transformation and expressed powerfully under the control of the E35S promoter. The expression vector, pEN-K was used for introduction of AtNDPK gene into birdsfoot trefoil plaits. The transformed calli were selected on kanamycin containing medium and then regenerated. The transformed birdsfoot trefoil plants were cultivated for 4 months on BOi2Y medium. Genomic DNA PCR and Southern blot analysis confirmed the incorporation of AtNDPK into the birdsfoot trefoil genome.

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.

Introduction of rolC gene into Petunia hybrida (Petunia hybrida 세포내로의 rolC 유전자의 도입)

  • 정재동;김경민;남윤연;김창길;정원일
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.1
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    • pp.21-26
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    • 1999
  • These experiments were attempted to introduce rolC gene in the Petunia hybrida cv. Titan white by Agrobacterium mediated. The maximum frequency of shoot regeneration was obtained by 60% on MS medium containing 1.0 mg/L BA, 0.1 mg/L NAA, 200 mg/L kanamycin, 500 mg/L carbenicillin, 30 g/L sucrose, and 8 g/L agar. Kanamycin-resistant calli were selected from petunia leaf discs by cocultivation with Agrobacterium suspension cultures on MS medium. The addition of AgNO$_3$ and KMnO$_4$ in the medium increased the shoot regeneration by 31.3% from leaf disc as compared with non-treated leaf disc. Among clones exhibiting kanamycin resistance, only 3 clones were confirmed by southern hybridization analysis.

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Auxin-like Effect of the Antibiotic Carbenicillin on Organogenesis of Leaf Discs of Tobacco (Nicotiana tabacum L. cv. BY-4) (담배 (Nicotiana tabacum L. cv. BY-4) 잎절편의 기관분화에 대한 항생제 carbenicillin의 auxin 유사효과)

  • 배창휴;양덕춘;이효연
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.6
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    • pp.469-474
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    • 2000
  • Effect of the antibiotic carbenicillin on callus and shoot formation from the leaf disc culture of tobacco (Nicotiana tabacum L. cv. BY-4) was examined. The number of shoot induced from the leaf explants was decreased as the amount of carbenicillin increased from 250 mg/L to 2,000 mg/L on MS medium containing 0.5 mg/L of BAP or kinetin. In addition, calli formation from the leaf explants was increased by the treatment of 250 mg/L ∼ 500 mg/L carbenicillin with or without 0.5 mg/L of 2,4-D or NAA. However, the fresh weight of 4-week-cultured explants was decreased with increasing carbenicillin from 250 mg/L to 2,000 mg/L on MS medium containing 0.5 mg/L of 2,4-D or NAA. These results indicate that carbenicillin has an auxin-like effect, such as promoting callus formation and inhibiting shoot induction. It leads to the conclusion that the auxin-like property should be taken into account for the production of transgenic plants via Agrobacterium-mediated transformation.

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Expression of Pea Superoxide Dismutase Gene in Transgenic Cucumber (Cucumis sativus L.) Plants (형질전환 오이(Cucumis sativus L.) 식물체에서 완두 Superoxide Dismutase 유전자의 발현)

  • 김재훈;오승용;이행순;조만현;이은모;우인식;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.201-206
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    • 1998
  • To develop the fruits of cucumber (Cucumis sativus L.) producing high yields of superoxide dismutase (SOD), the MnSOD cDNA from pea (Pisum sativum) under the control of the cauliflower mosaic virus 35S promoter was introduced into cucumber using Agrobacterium tumefaciens (strain LBA 4404)-mediated transformation. The kanamycin-resistant shoots were selected on the selection medium containing MS basal salt, 1.0 mg/L zeatin, 0.1 mg/L IAA, 300 mg/L claforan, and 100 mg/L kanamycin. After 6 weeks of culture on the selection medium, the shoots were transferred to MS medium containing 0.2 mg/L NAA to induce roots. PCR analysis using the primers for neomycin phosphotransferase (NPTII) gene revealed that three plantlets were transformed. The fruits of one transgenic plant had approximately 3.2-fold higher SOD activity than those of non-transgenic plants. MnSOD isoenzyme band was strongly detected on native gel in fruits of transgenic plants.

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Biotechnological Applications in Populus Species (포플러류(類)의 Biotechnology 응용(應用))

  • Chun, Young Woo;Klopfenstein, Ned B.;McNabb, Harold S. Jr.;Hall, Richard B.
    • Journal of Korean Society of Forest Science
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    • v.77 no.4
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    • pp.467-483
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    • 1988
  • The development of tissue culture systems for Populus species, and the utilization of tissue culture biotechnology will be reviewed and discussed. Special emphasis will be placed on prospects for genetic transformation by Agrobacterium-mediated gene transfer methods.

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Random Insertional Mutagenesis with Subtracted cDNA Fragments in Arabidopsis thaliana

  • Euna Cho;Kwon, Young-Myung;Lee, Ilha
    • Journal of Photoscience
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    • v.7 no.3
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    • pp.103-108
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    • 2000
  • We have evaluated a new mutagenesis strategy called random insertional mutagenesis with subtracted cDNA fragments. The cDNAs from long day Arabidopsis plants were subtracted by cDNAs from short day plants using PCR based cDNA subtraction. The subtracted cDNAs were inserted between 35S promoter and 3'-NOS terminator regardless of orientation. When the cDNA library was used for the random insertion into Arabidopsis genome by Agrobacterium-mediated transformation, approximately 15% of transformants showed abnormal development in leaf, floral organ, shoot apex. When 20 mutants were analyzed, 12 mutants showed single cDNA fragment insertion and 8 mutants showed more than 2 transgene insertions. Only two mutants among 12 mutants that have single cDNA insert showed consistent phenotype at T2 generation, suggesting the genetic instability of the mutants.

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