<i>Agrobacterium tumefaciens</i>-mediated transformation of hybrid sweetgum embryogenic callus

Jiang Shuaifei; Cui Ying; Zhao Ruirui; Qi Shuaizheng; Kong Lisheng; Zhao Jian; Li Shanshan; Zhang Jinfeng

doi:10.12171/j.1000-1522.20210032

Journal of Beijing Forestry University > 2021 > 43(8): 9-17. > DOI: 10.12171/j.1000-1522.20210032

Jiang Shuaifei, Cui Ying, Zhao Ruirui, Qi Shuaizheng, Kong Lisheng, Zhao Jian, Li Shanshan, Zhang Jinfeng. Agrobacterium tumefaciens-mediated transformation of hybrid sweetgum embryogenic callus[J]. Journal of Beijing Forestry University, 2021, 43(8): 9-17. DOI: 10.12171/j.1000-1522.20210032

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Agrobacterium tumefaciens-mediated transformation of hybrid sweetgum embryogenic callus

1.
National Engineering Laboratory for Tree Breeding, Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education, Key Laboratory of Forest Trees and Ornamental Plants Biological Engineering of State Forestry Administration, School of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China
2.
Department of Biology, University of Victoria, Victoria BC V8P5C2, Canada
3.
Shenzhou Lüpeng Agricultural Technology, Beijing 101105, China

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Received Date: January 28, 2021
Revised Date: February 21, 2021
Available Online: July 14, 2021
Published Date: August 30, 2021

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Abstract

Abstract

Objective Hybrid sweetgum is an important timber and ornamental tree resources in China, but its genetic transformation system has not been established yet. Establishing genetic transformation system of hybrid sweetgum provides a useful approach for trait improvement and allows us to conduct a functional identification of gene in hybrid sweetgum.
Method Based on the efficient somatic embryogenesis of hybrid sweetgum, the embryogenic calluses were transformed by Agrobacterium tumefaciens-mediated genetic transformation. Factors influencing transformation were studied by orthogonal experiment, including hygromycin selective pressure, concentration of Agrobacterium, infection time, co-culture time and co-culture temperature.
Result Results showed that the minimum lethal concentration of hygromycin to embryogenic callus was 10 mg/L. The number of Gus positive spots was highest when bacterium solution (OD₆₀₀) was 0.8, co-culture time was 3 d, infection time was 10 min, co-culture temperature was 25 ℃. The most transgenic positive resistant calluses were obtained when bacterium solution (OD₆₀₀) was 0.2, co-culture time was 2 d, infection time was 10 min, co-culture temperature was 23 ℃. And the optimal treatment combination was 0.2 bacterium solution (OD₆₀₀), 2 d co-culture time, 20 min infection time, 23 ℃ co-culture temperature.
Conclusion A total of 210 positive transgenic callus were obtained by molecular identification. The genetic transformation system was established, which provides more feasible foundation for the transformation of broadleaf tree callus.
- hybrid sweetgum,
- embryogenic callus,
- Agrobacterium tumefaciens,
- genetic transformation

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References

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Agrobacterium tumefaciens-mediated transformation of hybrid sweetgum embryogenic callus