Establishment of transient expression system of eggplant cotyledon by agrobacterium-mediated injection

doi:10.12307/2023.747

Abstract

Abstract: BACKGROUND: The production of oral vaccines by transgenic plants has great advantages and broad application prospects. At present, there are few reports on the study of the tomato transient expression system. It is necessary to optimize tomato transient expression conditions, which can greatly improve the genetic transformation efficiency of plants.
OBJECTIVE: The transient expression system of eggplant cotyledon was established by Agrobacterium tumefaciens-mediated injection to detect the transcription level of exogenous genes in plant expression vector pCAMBIA-E8-APB-DOCK8.
METHODS: Through the cultivation of tomato bacterial-free seedlings, significant differences in the number of sprouts, seedlings and agronomic traits of different tomato varieties were observed, and the tomato varieties suitable for this experiment were selected. The plant expression vector pCAMBIA-E8-APB-DOCK8 constructed earlier was transformed into Agrobacterium tumefaciens EHA105, and then the tomato cotyledons, the concentration of Agrobacterium tumefaciens injected, the amount of Agrobacterium tumefaciens injected, the age of seedlings and the days of coculture were screened. The transient expression system of eggplant cotyledon mediated by Agrobacterium tumefaciens was established to observe the transcriptional level of the reporter gene.RESULTS AND CONCLUSION: The transient expression system of eggplant cotyledon was preliminarily established. The suitable tomato variety was selected as European Dahong (apple type), and the selected material was tomato cotyledon. The concentration of Agrobacterium for injection was A600:0.2-0.7, and the amount of Agrobacterium for injection was 50 μL. The seedling age was 7-10 days, and the total culture time was 24-48 hours. It provides a convenient, fast and effective method for the analysis of exogenous gene protein expression in the later stage.

Key words: tomato, Agrobacterium mediated, instantaneous expression system construction, E8-APB-DOCK8, plant vaccine, transcription analysis

CLC Number:

Lang Yaoling, Wang Qian, Chen Bin, Bai Guohui, Guan Xiaoyan, Liu Jianguo. Establishment of transient expression system of eggplant cotyledon by agrobacterium-mediated injection[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(34): 5462-5468.

Figures/Tables 13

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