๊ฒฝ์ ์ ์ผ๋ก ์ค์ํ ์์ฉ์๋ฌผ์ธ ์ฝฉ(Glycine max)์ ์ด๊ธฐํ ์ง์ ํ ํจ์จ์ฆ๋๋ฅผ ์ํ์ฌ ์ฐ๊ตฌ๋ฅผ ์ํํ์๋ค. ๋์ ์ด๊ธฐํ ์ง์ ํ ์กฐ๊ฑด ํ๋ฆฝ์ ์ํ์ฌ ๊ธฐ์ด์คํ์ผ๋ก Agrobacterium ๊ฐ ์ผ์ ์์์ ์ธ ๊ตญ๋ด ์ฝฉ ํ์ข
์ ์คํฌ๋ฆฐ ํ์์ผ๋ฉฐ, ์ ์ ํ agar ๋๋์ ์ ๋ฐํญ์์ hygromycin ๋๋ ๊ท๋ช
, ์์ฒ๋ฐฉ๋ฒ, explant์ ์น์๋ฐฉํฅ, ํธ๋ฅด๋ชฌ ์ ์ฒ๋ฆฌ, ์ก์ฒด๋ฐฐ์ง ์ ๋ฐ ๋ฑ ํ์ง ์ ํ ์ด๊ธฐํจ์จ์ ๋ฏธ์น๋ ์ํฅ์ transient GUS ๋ถ์์ ํตํ์ฌ ์์๋ณด์๋ค. Agrobacterium ๊ฐ์ผ์ ์์์ ์ธ ํ์ข
์ ์ ๋ฐํ๊ธฐ ์ํ์ฌ 32๊ฐ์ ๊ตญ๋ด ์ฝฉ ์ฅ๋ คํ์ข
์ ์คํฌ๋ฆฐํ ๊ฒฐ๊ณผ, ์ผํ๊ฒ์ ์ฝฉ, ์ํ์ฝฉ, ๋ง๋ฆฌ์ฝฉ, ๋์์ฝฉ ๋ฑ 14๊ฐ์ ์์ํ ํ์ข
์ ์ ๋ฐํ์์ผ๋ฉฐ, ํจ์จ์ ์ธ ์ ๊ท ์ ๋ก๋ cefotaxime์ด, ํจ์จ์ ์ธ ์ ๋ฐํญ์์ ๋ก์๋ hygromycin์ด ์ ํ๋์๋ค. Hygromycin ๋๋๋ 10-15 ppm์ด ํจ์จ์ ์ด์๊ณ , agar ๋๋ 0.6-0.8%, explant์ ์น์๋ฐฉํฅ์ ํฅ๋ฐฐ์ถ(adaxial side)์ downํ์ฌ ์น์ํ์์ ๋๊ฐ ๋์ GUS (<TEX>$\beta$</TEX>-glucuronidase)๋ฐํ๋น๋๋ฅผ ๋ํ๋๊ณ ์์ฒ๋ฐฉ๋ฒ์ scalpel๋ณด๋ค. ๋์์นจ์ ์ฌ์ฉํ์์ ๋ ๋๊ฒ ๋ํ๋ฌ๋ค. ํ์ง์ ํ ์ฒด๊ณํ๋ฆฝ์ ์ํ์ฌ ํธ๋ฅด๋ชฌ ์ ์ฒ๋ฆฌ๋ฅผ ํ ๊ฒฐ๊ณผ, BA 5 ppm ๊ณผ 10 ppm์ ์ฒ๋ฆฌํ์์ ๋ ํ๊ธฐ ์ ๋ฐ์์ ๋์ ์์กด์จ์ ๋ณด์๊ณ , ์ ์ฒ๋ฆฌ ํ์ ์ก์ฒด๋ฐฐ์ง์์ ์ ๋ฐ์ ํ์์ ๋ ๋นํ์ง ์ ํ์ฒด(escape)์ ๋น๋๋ฅผ ํฌ๊ฒ ๋ฎ์ถ ์ ์์๋ค. ๋ํ ์ฌ๋ฌ ์กฐ๊ฑด์ ๊ฒฐ๊ณผ๋ฅผ ๋น๊ตํ์ฌ ๋ณผ ๋, ๊ตญ๋ด ์ฝฉ ํ์ข
์ค์์ ํ์ง์ ํ์ ์ํ ์ฌ๋ฃ๋ก์จ๋ ์ํ์ฝฉ์ด ๊ฐ์ฅ ์ ํฉํ ๊ฒ์ผ๋ก ์๊ฐ๋๋ค. 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.