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Agrobacterium Mediated Transformation of Rehmannia glutinosa L. with Glutathione S-Transferase Gene (Gh-5)
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  • Agrobacterium Mediated Transformation of Rehmannia glutinosa L. with Glutathione S-Transferase Gene (Gh-5)
저자명
Lim. Jung-Dae,Sung. Eun-Soo,Yang. Deok-Chun,Yun. Song-Joong,Chung. Ill-Min,Kim. Myong-Jo,Yu. Chang-Yeon
간행물명
韓國藥用作物學會誌
권/호정보
2003년|11권 4호|pp.289-297 (9 pages)
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한국약용작물학회
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

Using Agrobacterium-me야ated transformation method the auxin-regulated cotton GST (Gh-5) constructs were used to transform Rehmannia glutinosa L. The PCR analysis was conducted to verify transgenicity. Based on the PCR analysis, there was verified that the 988 bp DNA band had showed in transgenic plant genomes in PCR anaJysis using Gh5-1 and Gh5-2 primers. The effects of cocultivation with Agrobacterium tumefaciens, regeneration and selection conditions on the transformation efficiency of Chinese foxglove (Rehmannia glutinosa L.) were investigated. Factors such as cocultivation period, use of acetosyringone, postcultivation in darkness, and different kanamycin concentrations for selection were assessed. In vitro regeneration, the number of leaves, shoot lengths and numbers on MS medium were superior to on B5 and WPM medium, and the shoot formation rate was highest level of 95% in cultured base part containing leaf stalk. Addition of acetosyringone at concentration of $200{mu}M$ to cocultivation medium and 3-day of cocultivation improved transformation frequencies. Exposure of explants to darkness for 4 weeks on selection medium resulted in further increased the regeneration frequency of transgenic shoots. In PCR analysis, the amplified fragments of Gh5 gene were detected (988 bp), and GST-expressing transgenic R. glutinosa L. plants had approximately three-fold higher activity in leaf extracts compared with control plant.