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KMID : 0613820060160020289
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Journal of Life Science
2006 Volume.16 No. 2 p.289 ~ p.296
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Optimization of Genetic Transformation Conditions for Korean Soybean Cultivars
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Lee Ki-Jung
Seo Jen-Kyung
Lee Hye-Young
Jeon Eun-Hee
Shin Sang-Hyun
Lee Jai-Heon
Kim Do-Hoon
Ko Jong-Min
Han Won-Young
Baek In-Youl
Oh Boung-Jun
Chung Young-Soo
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Abstract
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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.
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KEYWORD
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Gene transformation, soybean, GUS, Agrobacterium
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