Development of Efficient Transformation Protocol for Soybean (Glycine max L.) and Characterization of Transgene Expression after Agrobacterium-mediated Gene Transfer

作者：Lee Kijong; Yi Bu Young; Kim Kyung Hwan; Kim Jung Bong; Suh Seok Cheol; Woo Hee Jong; Shin Kong Sik; Kweon Soon Jong^*

来源：Journal of the Korean Society for Applied Biological Chemistry, 2011, 54(1): 37-45.

DOI：10.3839/jksabc.2011.005

• soybean
• tocopherol
• transformation

Increasing vitamin E activity in economically important oil crops such as soybean will enhance the nutritional value of these crops. An improved soybean transformation system involving pre-culture of soybean seed on medium supplemented with 1 mg/L 6-benzylaminopurine under dark conditions was established. To improve the nutritional value of soybean by increasing the alpha-tocopherol content, soybeans were transformed with gamma-tocopherol methyltransferase (gamma-TMT) gene by Agrobacterium-mediated transformation. Frequency of soybean transformation was significantly increased from 0.5 to 4.3% by this modified system, and 13 lines of transgenic soybean plants containing gamma-TMT were obtained. The molecular characterization using polymerase chain reaction (PCR), reverse transcriptase PCR, and Southern blot analysis confirmed insertion and inheritance of the transgene in the transgenic plants and their progeny. Morphologically normal and fertile transgenic plants were analyzed, and the majority of transgenic soybean transmitted herbicide resistance at 3:1 or 15:1 ratios to their progeny. Alpha-tocopherol content in transgenic soybean seeds was determined by high-performance liquid chromatography; over-expression of gamma-TMT resulted in a 41-fold increase in alpha-tocopherol over wild-type soybean seeds.

• 出版日期2011-2