Establishment of Genetic Transformation System for Suspension Cells of Taraxacum koksaghyz Rodin

Key words: 
• Taraxacum koksaghyz Rodin /
• suspension cells /
• genetic transformation /
• Agrobacterium tumefaciens

Abstract: Taraxacum koksaghyz Rodin, belonging to the genus Taraxacum F. H. Wigg. and the family Composite,is a perennial herb. It is considered to be a potential alternative source of natural rubber and an ideal model plant for the study of rubber biosynthesis. In order to establish a genetic transformation system for suspension cells of T.koksaghyz Rodin, the sensitivity of suspension cells of T. koksaghyz Rodin to Ticarcillin（a fungicide for Agrobacterium tumefaciens） and Hygromycin B were determined. The suspension cells were infected with Agrobacterium tumefaciens GV3101 which carried the vector p CAMBIA1302, and then the gene expression of GFP and Hyg in the suspension cells was detected to observe the success of the genetic transformation. The result showed that Ticarcillin at a concentration range of 0-300 mg·L^-1 had no effect on the suspension cells, and that the suitable concentration of Hygromycin for selective calli was 10 mg·L^-1. GFP fluorescent detection and PCR detection of the Hyg and GFP gene showed that transgenic positive calli were efficiently produced when A.tumefaciens at OD₆₀₀=0.6, carrying the vector p CAMBIA1302, was mixed with the suspension cells at a ratio of 1∶1.2 for 30 mins, transferred into a co-culture medium for culture for 10 days, and then to a selective culture medium for culture. This study lays the foundation for further optimization of the genetic transformation system for the suspension cells of T. koksaghyz Rodin.