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Optimization of Biolistic Transformation Parameters for Nicotiana tabacum

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Abstract

Biolistics is one of the widely used methods to deliver nucleic acids into plant cells. In this work, we optimized the protocol for biolistic transformation of Nicotiana tabacum with the gene gun PDS-1000/He Biolistic Particle Delivery System. The Green Fluorescent Protein (GFP) gene was used as a marker. The optimal parameters for the transformation of N. tabacum leaf cells were determined as: pressure, 1350 psi; tungsten particle size, 1.3 μm; plasmid DNA purification method, ethanol re-precipitation. The results should be useful for the development of biolistic transformation protocols for plant cells, including application for plant genome editing of agricultural species.

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ACKNOWLEDGMENTS

Authors are grateful to the Laboratory of Plant Stress Tolerance of the All-Russia Research Institute of Agricultural Biotechnology for providing the pHBT95::sGFP(S65T)-NOS plasmid. This work was performed using the experimental climate control facility in the Institute of Bioengineering, Research Center of Biotechnology, Russian Academy of Sciences.

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  1. Skryabin Institute of Bioengineering, Federal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences, 117312, Moscow, Russia

    A. A. Davlekamova, A. V. Zubritsky, T. A. Timofeeva, I. V. Yakovleva & A. M. Kamionskaya

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  1. A. A. Davlekamova
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  2. A. V. Zubritsky
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  3. T. A. Timofeeva
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  4. I. V. Yakovleva
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  5. A. M. Kamionskaya
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Correspondence to T. A. Timofeeva.

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The authors declare that they have no conflicts of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.

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Davlekamova, A.A., Zubritsky, A.V., Timofeeva, T.A. et al. Optimization of Biolistic Transformation Parameters for Nicotiana tabacum. Appl Biochem Microbiol 59, 368–372 (2023). https://doi.org/10.1134/S0003683823030055

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