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Efficiency of Transient Expression in Protoplasts of Various Potato Cultivars

Efficiency of Transient Expression in Protoplasts of Various Potato Cultivars The production of mesophilic protoplasts of potato (Solanum tuberosum L.) for transient gene expression is a required technological stage in the testing of the efficiency of the new genetic constructs, including CRISPR/Cas genome editing. In this study, the leaves of potato plants of nine Russian cultivars were used to isolate protoplasts. These plants were cultivated in vitro for 6–7 weeks in vessels with foil caps to prevent gas exchange with the environment. We demonstrated for the first time that 2.4 × 106 to 4.6 × 106 viable protoplasts can be obtained from 1 g of aseptic plant leaves depending on the used potato cultivar. We found that the level of protoplast transfection with the pHBT-sGFP-NosT genetic construct ranged from 10 to 49% depending on the cultivar, which is sufficient for a successful subsequent analysis of the efficiency of genetic constructs and editing of the potato genome. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Applied Biochemistry and Microbiology Springer Journals

Efficiency of Transient Expression in Protoplasts of Various Potato Cultivars

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Publisher
Springer Journals
Copyright
Copyright © Pleiades Publishing, Inc. 2021. ISSN 0003-6838, Applied Biochemistry and Microbiology, 2021, Vol. 57, No. 7, pp. 800–807. © Pleiades Publishing, Inc., 2021. Russian Text © The Author(s), 2020, published in Biotekhnologiya, 2020, Vol. 36, No. 3, pp. 16–24.
ISSN
0003-6838
eISSN
1608-3024
DOI
10.1134/s0003683821070048
Publisher site
See Article on Publisher Site

Abstract

The production of mesophilic protoplasts of potato (Solanum tuberosum L.) for transient gene expression is a required technological stage in the testing of the efficiency of the new genetic constructs, including CRISPR/Cas genome editing. In this study, the leaves of potato plants of nine Russian cultivars were used to isolate protoplasts. These plants were cultivated in vitro for 6–7 weeks in vessels with foil caps to prevent gas exchange with the environment. We demonstrated for the first time that 2.4 × 106 to 4.6 × 106 viable protoplasts can be obtained from 1 g of aseptic plant leaves depending on the used potato cultivar. We found that the level of protoplast transfection with the pHBT-sGFP-NosT genetic construct ranged from 10 to 49% depending on the cultivar, which is sufficient for a successful subsequent analysis of the efficiency of genetic constructs and editing of the potato genome.

Journal

Applied Biochemistry and MicrobiologySpringer Journals

Published: Dec 1, 2021

Keywords: potato; Solanum tuberosum; protoplasts; plasmid DNA; transfection; transient expression; gfp

References