Construction of genetic transformation system of ‘Jingzao 39’ callus

Objective This paper aims to establish a genetic transformation system of jujube callus and optimize the identification method for selecting positive plants using the callus of ‘Jingzao 39’ as explants.

Method Using ‘Jingzao 39’ callus as plant material, the modified vector containing the reporter gene eYGFPuv was transformed by agrobacterium tumefaciens mediated method, genetic transformation was performed by agrobacterium tumefaciens mediated method, the effects of pre-culture time, agrobacterium concentration, infection time, acetosyringone (AS) mass concentration and co-culture time on genetic transformation of ‘Jingzao 39’ were investigated to establish a genetic transformation system of jujube callus and optimize the identification method for selecting positive plants.

Result (1) The minimum lethal concentration of kanamycin on callus was 30 mg/L. (2)The optimal treatment conditions of genetic transformation included that pre-culture time was 4 d, bacterial solution concentration OD₆₀₀ was 0.6, infection time was 20 min, AS mass concentration was 100 μmol/L, co-culture time was 4 d. (3) The positive callus group showed bright fluorescent green under 365 nm ultraviolet light and the transformation rate was averaged 21.2%.

Conclusion Through comparative experiments, the ‘Jingzao 39’ callus genetic transformation system is successfully established, which provides a new method for accelerating the genetic transformation of jujube.