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노경희 ( Kyung Hee Roh ),곽보경 ( Bo Kyoung Kwak ),김현욱 ( Hyun Uk Kim ),이경렬 ( Kyeong Ryeol Lee ),김순희 ( Sun Hee Kim ),서미정 ( Mi Chung Suh ),김효진 ( Hyo Jin Kim ),김종범 ( Jong Bum Kim ) 한국응용생명화학회 2011 Journal of Applied Biological Chemistry (J. Appl. Vol.54 No.1
To improve genetic transformation of Brassica napus winter cultivar ``Youngsan``, factors influencing shoot regeneration and transformation from cotyledonary petioles were investigated. Shoot induction was enhanced in the combination of 0.5 mg/L NAA and 2~4 mg/L kinetin. Silver nitrate was essential for successful shoot regeneration, ranging from 5 to 9 mg/L. The addition of GA3 promoted plant regeneration. Among the tested Agrobacterium strains, co-cultivation times, and antibiotic selection regimes, choice of appropriate Agrobacterium strain was the most critical factor for efficient transformation of B. napus cv. ``Youngsan``. The EHA105 succinamopine strain was the most efficient and the maximum transformation efficiency was 26.8%. Transgenic shoots were selected on 10 mg/L phosphinothricin (PPT) containing media. The transgenic plants expressing bar and gus genes were resistant for commercial herbicide Basta and stained with X-Gluc. Southern blot hybridization indicated that the presence of one to three gus gene copies per genome and inheritance of the gus gene into the T1 generation.