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http://dx.doi.org/10.5352/JLS.2006.16.2.289

Optimization of Genetic Transformation Conditions for Korean Soybean Cultivars  

Lee Ki-Jung (Dept. of Genetic Engineering, Dong-A University)
Seo Jen-Kyung (Dept. of Genetic Engineering, Dong-A University)
Lee Hye-Young (Dept. of Genetic Engineering, Dong-A University)
Jeon Eun-Hee (Dept. of Genetic Engineering, Dong-A University)
Shin Sang-Hyun (Dept. of Genetic Engineering, Dong-A University)
Lee Jai-Heon (Dept. of Genetic Engineering, Dong-A University)
Kim Doh-Hoon (Dept. of Genetic Engineering, Dong-A University)
Ko Jong-Min (Yeongnam Agricultural Research Institute, NICS, RDA)
Hahn Won Young (Yeongnam Agricultural Research Institute, NICS, RDA)
Baek In-Youl (Yeongnam Agricultural Research Institute, NICS, RDA)
Oh Boung-Jun (Jeonnam Biotechnology Research Center)
Chung Young-Soo (Dept. of Genetic Engineering, Dong-A University)
Publication Information
Journal of Life Science / v.16, no.2, 2006 , pp. 289-296 More about this Journal
Abstract
In order to establish highly efficient gene transfer condition at early stage of soybean transformation, various experiments were performed and compared their efficiencies by transient GUS analysis; those conditions are genotype determination of Korean soybean cultivars for amenability to Agro-infection, appropriate agar and selective agent concentration, orientation of explant placement, hormone pre-culture, and liquid selection condition. In the genotype screen of Korean soybean varieties, 14 amenable genotypes were selected. For efficient Agrobacterium washing, cefotaxime was chosen and hygromycin at the concentration of 10 and 15 ppm was used as selection agent in the media. Agar concentration was slightly better in 0.6% and 0.8% for both shoot and callus formation, and explant placement with adaxial side down showed high frequency of GUS expression. For wounding treatment, oriental needle was efficient than scalpel for shoot formation and gene transfer. To increase the frequency of gene transfer, hormone pre-treatment was applied. BA at the concentration of 5 and 10 ppm resulted in better survival at the late stage of selection in shoot elongation media. Selection in liquid media after hormone pre-treatment seemed to be effective to remove the escaped non-transformants at early stage of procedure. Considering the results obtained, Eunhakong could be the first choice as a material for soybean transformation among Korean soybean genotypes.
Keywords
Gene transformation; soybean; GUS; Agrobacterium;
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