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.
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