Studies on genetic transformation of Rehmannia glutinosa mediated by Agrobacterium tumefaciens / 中草药

ABSTRACT

Objective: To determine the optimal concentration of hormone and hygromycin for seedling regeneration of Rehmannia glutinosa. Methods: Using the young leaf of sterile plantlet from R. glutinosa as explants, we conducted the transformation mediated by Agrobacterium tumefaciens, analyzed the efficiency of hygromycin and acetosyringone (AS) on resistant callus induction and plant regeneration. Results: The concentration of hygromycin had greatly affected the production of resistant callus and seedlings. The critical concentration of hygromycin on the resistant callus induction and shoot regeneration were 9 and 6 mg/L, respectively. The optimal concentration of hygromycin for the genetic transformation of Wen 85-5 was 12 mg/L. Adding 100 μmol/L AS could greatly improve the transformation efficiency of R. glutinosa. It was confirmed by PCR detection of hpt gene and GUS staining that the foreign gene was integrated into the genome of R. glutinosa. Conclusion: The stable genetic transformation system of R. glutinosa is established, which lays the foundations for the research on molecular pharmacognosy and genetic improvement.