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An attempt to establish an Agrobacterium-mediated transient expression system in medicinal plants

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Abstract

Genetic transformation has always been an important method for studying medical plant secondary metabolic regulation, among which stable transformation has a good reproducibility. However, it was time-consuming to obtain a stable transformed hairy root or transgenic plants, which was difficult to satisfy the great demand of researches on medical plant secondary metabolism–related genes. Moreover, Agrobacterium tumefaciens–mediated transient transformation has been extensively applied in studies of functional genes because of its simpleness, low cost, and short period. However, presently, researches on medical plant functional genes commonly used stable genetic transformation and some high-cost and high-difficulty transient transformation methods, such as gene gun and protoplast transformation. Thus, in this study, we selected the seedlings of Nicotiana benthamiana, Salvia miltiorrhiza, and Prunella vulgaris as the experimental material, with the methods of Agrobacterium tumefaciens injection, fast Agrobacterium-mediated seedling transformation (FAST), and FAST and mechanical damage. The results demonstrated that the injection transient transformation system of pCAMBIA1301 vector mediated by A. tumefaciens and the transient transformation of seedling system were not established in S. miltiorrhiza. In addition, the instantaneous transformation system of N. benthamiana and P. vulgaris seedlings was basically set up by FAST method. Besides, using the method of FAST and mechanical damage, the transient genetic transformation system of P. vulgaris seedlings was established for the first time. A. tumefaciens–mediated transient transformation of seedlings with pEAQ vectors provided an effective way and reference for the further study of functional genes of the medicinal plants N. benthamiana and P. vulgaris.

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Abbreviations

FAST:

Fast Agrobacterium-mediated seedling transformation

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Funding

This work was supported by National Natural Science Foundation of China (81703641, 31871694), Zhejiang Province Public Welfare Technology Application Research Project (CN) (LGN19H280004), China Postdoctoral Science Foundation (2019 M652146) and Fundamental Research Funds of Zhejiang Sci-Tech University (2019Q046).

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Authors and Affiliations

  1. Laboratory of Plant Secondary Metabolism and Regulation of Zhejiang Province, College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, 310018, China

    Pengguo Xia, Wanying Hu, Tongyao Liang, Dongfeng Yang & Zongsuo Liang

  2. State Key Laboratory of Membrane Biology, Innovation Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing, 100084, China

    Pengguo Xia

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  1. Pengguo Xia
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  2. Wanying Hu
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  3. Tongyao Liang
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  4. Dongfeng Yang
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  5. Zongsuo Liang
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Contributions

P.X. and Z.L. conceived and designed the research. W.H., T.L., and P.X. analyzed the results. D.Y. collected the plant samples. T.L. and P.X. preformed the experiments. W.H. and P.X. wrote the manuscript.

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Correspondence to Pengguo Xia or Zongsuo Liang.

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Supplementary figure 1

The pattern of pEAQ-GFP-HT (PNG 161 kb)

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Xia, P., Hu, W., Liang, T. et al. An attempt to establish an Agrobacterium-mediated transient expression system in medicinal plants. Protoplasma 257, 1497–1505 (2020). https://doi.org/10.1007/s00709-020-01524-x

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  • DOI: https://doi.org/10.1007/s00709-020-01524-x

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