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Vol 36(2020) N 3 p. 16-24; DOI 10.21519/0234-2758-2020-36-3-16-24
L.N. Konovalova1, S.R. Strelnikova1, N.E. Zlobin1, P.N. Kharchenko1, R.A. Komakhin1

Efficiency of Transient Expression in Protoplasts of Various Potato Varieties

1All-Russia Research Institute of Agricultural Biotechnology, Moscow, 127550 Russia

*recombination@iab.ac.ru
Received - 20.02.2020; Accepted - 15.05.2020

References

Production of mesophilic protoplasts in potato (Solanum tuberosum L.) for transient gene expression is a mandatory technological stage in testing efficiency of new genetic constructs and CRISPR/Cas genome editing. In this study, isolation of protoplasts was carried out from leaves of potato plants of nine Russian cultivars grown in vitro for 6-7 weeks in vessels with foil caps, which prevented gas exchange with the environment. We first ever demonstrated the possibility of obtaining 2.4∙106 to 4.6∙106 viable protoplasts from one gram of aseptic plant leaves depending on the used potato cultivar. We established that the level of transfection of protoplasts with the genetic construct pHBT-sGFP-NosT amounts from 10 to 49% depending on the cultivar, which is sufficient for subsequent successful analysis of the efficiency of genetic constructs and potato genome editing.

potato, Solanum tuberosum, protoplasts, plasmid DNA, transfection, transient expression, gfp

The work was financially supported by the Comprehensive Research Program «Development of Potato Breeding and Seed Production».



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