Infection of Embryonic Callus with Agrobacterium Enables High-Speed Transformation of Maize
National Key Laboratory of Crop Genetic Improvement and College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China
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Int. J. Mol. Sci. 2019, 20(2), 279; https://doi.org/10.3390/ijms20020279
Received: 14 December 2018
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Revised: 6 January 2019
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Accepted: 6 January 2019
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Published: 11 January 2019
(This article belongs to the Section Molecular Plant Sciences)
Several approaches have recently been adopted to improve Agrobacterium-mediated transformation of maize; however, about eight months of in vitro culture are still required to isolate transgenic plants. Furthermore, genetic transformation of maize depends on immature embryos, which greatly increases costs. Here, we report a method that ensures the competency of an embryogenic callus secondary culture under laboratory conditions for Agrobacterium-mediated transformation. Moreover, pretreatment of the cell wall with a mixed lytic enzyme solution prior to Agrobacterium infection, significantly improved transformation efficiency and stability. Average stable transformation efficiency was approximately 30.39%, with peaks of 94.46%. Expression and phenotypic analysis of the Rsc reporter gene were tested in the T0 generation of transgenic plants. Using this system, we successfully regenerated transgenic maize plantlets within three months of the emergence of the embryogenic callus. Additionally, we reduced somaclonal variation accompanying prolonged culture of maize cells in the dedifferentiated state, thus facilitating the molecular breeding of maize.
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Keywords:
Agrobacterium; callus; maize; mixed enzyme solution; transformation
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MDPI and ACS Style
Du, D.; Jin, R.; Guo, J.; Zhang, F. Infection of Embryonic Callus with Agrobacterium Enables High-Speed Transformation of Maize. Int. J. Mol. Sci. 2019, 20, 279. https://doi.org/10.3390/ijms20020279
AMA Style
Du D, Jin R, Guo J, Zhang F. Infection of Embryonic Callus with Agrobacterium Enables High-Speed Transformation of Maize. International Journal of Molecular Sciences. 2019; 20(2):279. https://doi.org/10.3390/ijms20020279
Chicago/Turabian StyleDu, Dengxiang, Ruchang Jin, Jinjie Guo, and Fangdong Zhang. 2019. "Infection of Embryonic Callus with Agrobacterium Enables High-Speed Transformation of Maize" International Journal of Molecular Sciences 20, no. 2: 279. https://doi.org/10.3390/ijms20020279
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