• Title/Summary/Keyword: BASTA

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Bioloistic-mediated Transformation of Cotton (Gossypium hirsutum L.): Embryogenic Calli as Explant

  • Haq Ikram-ul;Asad Shaheen;Zafar Yusuf
    • Journal of Plant Biotechnology
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    • v.7 no.4
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    • pp.211-218
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    • 2005
  • Genetic transformation was carried out by using biolistic gun method. The hypocotyl derived embryogenic calli (explants) of cotton (Gossypium hirsutum L.) cv. Cocker-312 were transformed with a recombinant pGreen II plasmid, in which both, bar (selection marker) and GUS (${\beta}$-glucuronidase) reporter genes were incorporated. Explants were arranged on osmoticum-containing medium (0.5M mannitol) 4 hours prior to and 16 hours after bombardment that was resulted into an increase about >80% for GUS stable expression. 3 days after bombardment, GUS assay was performed, which exhibited, $18.36{\pm}1.00$ calli showed blue spots. The transformed embryogenic calli were cultured on selection medium (@ 6 mg/L basta) for 3 months. The putative transgenic plants were developed via selective somatic embryogenesis (@1.50 mg/L basta); maximum $27.58{\pm}1.25$ somatic embryos were obtained while $17.47{\pm}1.00$ embryos developed into plantlets (@ 0.75mg/L basta). In five independent experiments, up to 7.24% transformation efficiency was recorded. The presence of the transgenes was analyzed by using PCR and southern hybridization analysis. The transgenic plants were developed with in 6-7 months, but mostly transformants were abnormal in morphology.

Expression and Inheritance of bar Gene in Petunia hybrida Transformed with Agrobacterium (Petunia hybrida에 Agrobacterium으로 도입된 bar Gene의 발현과 후대검정)

  • Ha, Young-Min;Kim, Jong-Chul;Lee, Sang-Woo;Lee, Shin-Woo;Kim, Zhoo-Hyeon
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.143-149
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    • 2003
  • This experiment was carried out to confirm the stability of bar gene introduced into petunia plant through Agrobacerium-mediated transformation. Twenty-five transgenic plants T$_{0}$ plants, back cross (BC$_1$) populations to wild type and F$_1$plants between different T$_{0}$ plants were prepared, and polymerase chain reaction(PCR), PCR-Southern blot analysis, and field test with 0.1% Basta treatment were done. The results of PCR, PCR-Southern blot hybridization, and field test indicated that NPTII and bar gene introduced into the genome of petuina plants were stably transmitted to their progenies, and conferred the plants resistance to herbicide, Basta.sta.

A Case of Acute Glufosinate Ammonium ($BASTA^{(R)}$) Intoxication Associated with Various Neurological Abnormalities (다양한 신경학적 이상을 나타낸 glufosinate ammonium(바스타$^{(R)}$) 급성 경구 중독 1례)

  • Paik Jin Hui;Kim Jun Sig;Yi Hyeon Gyu;Park Hyun Joo;Ha Choong-Kun;Roh Hyung-Keun
    • Journal of The Korean Society of Clinical Toxicology
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    • v.3 no.2
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    • pp.103-106
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    • 2005
  • BASTA is an herbicide which contains glufosinate ammonium as a main component with an anionic surfactant, polyoxyethylene alkylether sulfate, and nonselectively inhibits glutamine synthetase. It became a wildly used herbicide in Korea and its intoxication is now increasing. A 42-year old woman ingested about 300ml of BASTA in a suicide attempt. She showed unconsciousness and respiratory distress in the beginning, and later developed multiple generalized convulsions, low blood pressure, fever and diabetes insipidus. Although she became alert 12 days after the ingestion, she showed retrograde amnesia for a period of about recent 10 years. A neuropsychological test on day 22 revealed frontal lobe dysfunction, visual memory disturbance and slight decrease in visuospatial function. All these neurological abnormalities that might occur due to glufosinate ammonium were almost improved in the follow-up test performed a month later.

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Herbicide Resistant Turfgrass(Zoysia japonica cv. 'Zenith') Plants by Particle bombardment-mediated Transformation

  • Lim Sun-Hyung;Kang Byung-Chorl;Shin Hong-Kyun
    • Asian Journal of Turfgrass Science
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    • v.18 no.4
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    • pp.211-219
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    • 2004
  • Transgenic zoysiagrass (Zoysia japonica cv. Zenith) plants have been obtained by particle bombardment of embryogenic callus with the plasmid pSMABuba, which contains hygromycin resistance (hpt) and bialaphos resistance (bar) genes. Parameters on DNA delivery efficiency of the particle bombardment were partially optimized using transient expression assay of a chimeric $\beta-glucuronidase$(gusA) gene driven by the CaMV 35S promoter. Stably transfarmed zoysiagrass plants were recovered with a selection scheme using hygromycin. Transgenic zoysiagrass plants were confirmed by PCR analysis with specific primer for bar gene. Expression of the transgene in transformed zoysiagrass plants was demonstrated by Reverse transcriptase (RT)-PCR analysis. All the tested transgenic plants showed herbicide BastaR resistance at the field application rate of $0.1\%-0.3\%$.

The effect of acute toxicity of marine toxicants on early life of coastal olive flounder (넙치의 초기 생활사에 미치는 해양 독성물질들의 급성독성 영향)

  • 탁건태;김중균
    • Journal of Life Science
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    • v.10 no.3
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    • pp.247-253
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    • 2000
  • The effects of acute of marine toxicants on early life of coastal olive flounder were investigated. An increasing order of acute toxicity on embryo- and larva-stages of Paralichtys olivaceus was CdCl2$\mu\textrm{g}$/L and 29 $\mu\textrm{g}$/L, respectively, and those values at larva-stage were estimated 3.5 ng/L, 16.0 nL/L, 10.5 $\mu\textrm{g}$/L and 15.0 $\mu\textrm{g}$/L, respectively.

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Transformation of Populus alba $\times$Populus glandulosa Using Phosphinothricin Acetyltransferase Gene (Phosphinothricin acetyltransferase 유전자를 이용한 현사시의 형질전환)

  • 오경은;양덕춘;문흥규;박재인
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.3
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    • pp.163-169
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    • 1999
  • This study was conducted to produce herbicide resistant plants by transferring phosphinothricin acetyltransferase (PAT) gene into Populus alba $\times$ Populus glandulosa No .3 using Agrobacterium tumefaciens MP 90/PAT. Leaf segments from in vitro grown shoots of hybrid poplar No. 3 were soaked in a AB medium containing Agrobacterium tumefaciens MP 90/PAT for 10 min and cocultivated for 2 days on MS medium containing 1.0 mg/L 2,4-D and 0.2mg/L kinetin (CIM). Putative transformed calli could be selected after cocultivation of leaf segments on CIM supplemented with 50mg/L kanamycin and 500mg/L cefotaxime for 3 weeks. The selected calli were cultured on CIM supplemented with 50 mg/L kanamycin and 500 mg/L cefotaxime for 5~8 weeks before transfer to WPM containing 1.0mg/L zeatin, 0.1mg/L BAP, 50 mg/L kanamycin and 500mg/L cefotaxime for shoot regeneration. Shoots were regenerated from the callus after 4 week cultivation, and the regenerants were grown on the same medium for 7~l0 weeks. The plants rooted on 1/2 WPM containing 0.2 mg/L IBA and 50 mg/L kanamycin. To confirm the gene insertion into plants, GUS activity was detected by histochemical assay in the transformed plants. Finally, the presence of both NPT II and PAT genes from the transgenic plants were confirmed by PCR amplification with the gene specific primers and subsequent PCR-Southern blot with DIG-labeled PAT gene probe. After acclimatization in pots for 4 weeks, the plants were sprayed by 3 mL/L of Basta to test resistance to the herbicide. The transgenic plants remained green, whereas all the control plants died after one week.

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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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Genetic Transformation of a Mushroom Forming Fungus Coprinellus congregatus to an Antibiotic Resistance Using Oidia Instead of Protoplast Generation (먹물버섯 Coprinellus congregatus에서 분열자를 사용한 형질전환)

  • Park, Nam-Mee;Kim, Dong-Sik;Choi, Hyoung-T.
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.59-61
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    • 2006
  • Genetic transformation of a mushroom-forming fungus Coprinellus congregatus to antibiotic resistance gene had been successfully carried out by electroporation to oidia instead of protoplasts. Since there was no protoplast generation step which required not only cell wall degrading enzymes but many skillful procedures, commercial herbicide (basta) could be used without any difficulty with simple procedure. The transformation yield was 10-20 $transformants/{\mu}g$ DNA, and the transformants were very stable even after 10 consecutive transfers through the non-selective medium.