• Title/Summary/Keyword: PCR-Southern blot

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Transformation of Bottle Gourd Rootstock (Lagenaria siceraria Standl.) using GFP gene (GFP유전자를 이용한 대목용 박 형질전환)

  • Lim, Mi-Young;Park, Sang-Mi;Kwon, Jung-Hee;Han, Sang-Lyul;Shin, Yoon-Sup;Han, Jeung-Sul;Harn, Chee-Hark
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.33-37
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    • 2006
  • Bottle gourd (Lagenaria siceraria Standl.) has been used as a rootstock for the watermelon cultivation because of better growth ability at low temperature and avoidance from contamination of the soil disease. Since the genetic source for the elite rootstock is limited in nature, the genetic engineering method is inevitable to develop new lines especially to obtain the functionally important or multi-disease resistant bottle gourd. Recently, our lab has set up a successful system to transform the bottle gourd. in order to monitor the transformation process, GFP gene is used. Cotyledons of the inbred line 9005, 9006 and G5 were used to induce the shoot under the selection media with MS + 30 g/L sucrose + 3.0 mg/L BAP + 100 mg/L kanamycin + 500 mg/L cefotaxime + 0.5 mg/L $AgNO_3$, pH 5.8. The shoot was developed from the cut side of the explants after 3 weeks on the selection media. The shoot was incubated in the rooting media with 1/2 MS + 30 g/L sucrose + 0.1 mg/L IAA + 50 mg/L kanamycin + 500 mg/L cefotaxime, pH 5.8 and moved to pot for acclimation. Although the shoot development rate was depended on the genotype, the G5 was the best line to be transformed. Monitoring GFP expression from the young shoot under microscope could make the selection much easier to distinguish the transformed shoot from the non-transformed shoots.