Optimization of Agrobacterium tumefaciens-Mediated Immature Embryo Transformation System and Transformation of Glyphosate-Resistant Gene 2mG2-EPSPS in Maize (Zea mays L.)

作者：Guirong Yu, Yan Liu, Yan Liu, Wen-Ping Du, Jun Song, Min Lin, Xu LiYuan, Fangming Xiao, Yongsheng Liu, Yongsheng Liu · 发表于：Journal of Integrative Agriculture · 年份：2013 · DOI：10.1016/s2095-3119(13)60567-5 · 被引用次数：15 · 研究领域：Plant tissue culture and regeneration、CRISPR and Genetic Engineering、Transgenic Plants and Applications

Since maize is one of the most important cereal crops in the world, establishment of an efficient genetic transformation system is critical for its improvement. In the current study, several elite corn lines were tested for suitability of Agrobacterium tumefaciens-mediated transformation by using immature embryos as explants. Infection ability and efficiency of transformation of A. tumefaciens sp. strains EHA105 and LBA4404, different heat treatment times of immature embryos before infection, influence of L-cysteine addition in co-cultivation medium after transformation, and how different ways of selection and cultivation influence the efficiency of transformation were compared. Glyphosate-resistant gene 2mG2-EPSPS was transformed into several typical maize genotypes including 78599, Zong 31 and BA, under the optimum conditions. Results showed that the hypervirulent Agrobacterium tumefaciens sp. strain EHA105 was more infectious than LBA4404. Inclusion of L-cysteine (100 mg L−1) in co-cultivation medium, and heating of the immature embryos for 3 min prior to infection led to a significant increase in the transformation efficiency. Growth in resting medium for 4-10 d and delaying selection was beneficial to the survival of resistant calli. During induction of germination, adding a high concentration of 6-BA (5 mg L−1) and a low concentration of 2,4-D (0.2 mg L−1) to regeneration medium significantly enhanced germination percentage. Using the optimized transformation procedure,...