Construction of genetic transformation system of ‘Jingzao 39’ callus

Feng Xuejing; Ma Ling; Yang Shuang; Bo Wenhao; Chen Xuexun; Pang Xiaoming

doi:10.12171/j.1000-1522.20240055

Journal of Beijing Forestry University > 2024 > 46(10): 74-80. > DOI: 10.12171/j.1000-1522.20240055

Feng Xuejing, Ma Ling, Yang Shuang, Bo Wenhao, Chen Xuexun, Pang Xiaoming. Construction of genetic transformation system of ‘Jingzao 39’ callus[J]. Journal of Beijing Forestry University, 2024, 46(10): 74-80. DOI: 10.12171/j.1000-1522.20240055

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Construction of genetic transformation system of ‘Jingzao 39’ callus

1.
State Key Laboratory of Tree Genetics and Breeding, National Engineering Laboratory of Tree Breeding, School of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China
2.
Shuangjing Forest Farm of Aohan Banner, Chifeng 024308, Inner Mongolia, China

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Received Date: March 06, 2024
Revised Date: May 04, 2024
Accepted Date: October 09, 2024
Available Online: October 20, 2024

Graphical Abstract

Abstract

Abstract

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.
- Ziziphus jujuba ‘Jingzao 39’,
- callus,
- genetic transformation,
- eYGFPuv

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References (23)

References

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Construction of genetic transformation system of ‘Jingzao 39’ callus