Objective: To establish genetic transformation system of active fibroblast growth factor (aFGF) in Carthamus tinctorius.
Method: The culture condition was optimized by orthogonal experiment design with cotyledon of C. tinctorius as the explant. The aFGF was transferred into safflower through Agrobacterium-mediated transformation and screened under different concentrations of antibiotics, and then PCR was identified.
Result: It confirmed the optimal differentiation medium: MS + BA 1.0 mg x L(-1) + NAA 0.2 mg x L(-1), the optimal root medium: 1/4 MS + NAA 2.0 mg x L(-1) + IAA0.5 mg x L(-1). The bacteriostatic effect of the three antibiotics showed slight difference. From them Tim was selected with the concentration of 400 mg x L(-1). It showed the bacteriostatic effect and promoted also differentiation. The selective concentration of hyg was confirmed to be 6 mg x L(-1). The eight transformed plants were identified, the positive rate was 25%.
Conclusion: It was determined the best hormones and the ratios for the differentiation and rooting of the safflower by organogenesis. It was identified the optimal concentration of inhibitory antibiotics and selection antibiotics. The aFGF gene was cloned in a part of plant by PCR analysis. It is shown that the aFGF gene has been integrated into safflower genome.