• Title/Summary/Keyword: GUS histochemical assay

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Agrobacterium-mediated Transformation of PAT into Platycodon grandiflorum A. De. candolle (Agrobacterium을 이용한 Phosphinothricin Acetyl Transferase의 도라지로의 형질전환)

  • Park, Jae-Seong;Kim, Ik-Hwan;Hong, Eui-Yon;Yun, Tae;Lee, Cheol-Hee;Jeong, Jae-Hun;Yang, Deok-Chun;Yun, Jong-Sun
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.4
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    • pp.285-290
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    • 2007
  • This study was conducted to introduce phosphinothricin acetyl transferase (PAT) gene, resistant to basta which was non-selective herbicide, into balloon flower (Platycodon grandiflorum A. De. candolle). Seeds were germinated on MS medium, and 10-day-old immature cotyledon explants and 30-day-old leaf explants were cocultured with Agrobacterium tumefaciens strain MP 90 (pBinSyn) on 1/10 MS medium for 48 hours in the dark at $25^{\circ}C$. The cultures were transferred for selection of kanamycin-resistant shoots to the MS medium supplemented with 0.2 $mg/{\ell}$ NAA, 1.0 $mg/{\ell}$ BA, 3% sucrose, 100 $mg/{\ell}$ kanamycin, 500 $mg/{\ell}$ carbenicillin. Shoots were obtained from 10-day-old immature cotyledon explants after 4 weeks of culture. The shoots were subcultured twice every 4 weeks on the same medium for growth of transgenic shoots. Successful transformation was confirmed by histochemical GUS assay, PCR analysis, RT-PCR analysis, 10 $mg/{\ell}$ phosphinothricin treatment and 0.3% basta spray. The basta-resistant transgenic plants flowered normally.