Production of Transgenic Plants in Brassica napus Winter Cultivar 'Youngsan'

Abstract

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 GA 3 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 T 1 generation.

Effects of different types and ages of explants and cytokinins on shoot regeneration in Brassica juncea L.

“…특 히 petiole explant를 이용한 direct shoot induction에 TDZ가 매우 효과적이라는 연구결과가 최근 꾸준히 발표되고 있 다 (Kumar and Reddy 2010;Nada et al 2011;Zhao et al 2012). 이제까지 Brassica 속 작물의 재분화에 BAP가 가장 많이 사용되어 왔으며 (Moloney et al 1989;Tang et al 2003;Bhuiyan et al 2009), 최근 들어 B. napus (Roh et al 2011)와 B. juncea (Guo et al 2005 …”