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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 Microbiology – Springer Journals

Published: Dec 1, 2021

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

References

  • Grand challenges in plant physiology: the underpinning of translational research
    Huber, S.C.
  • Novel perspectives for the engineering of abiotic stress tolerance in plants
    Cabello, J.V.; Lodeyro, A.F.; Zurbriggen, M.D.
  • Improving crop disease resistance: lessons from research on Arabidopsis and tomato
    Piquerez, S.J.; Harvey, S.E.; Beynon, J.L.; Ntoukakis, V.
  • Efficient targeted multiallelic mutagenesis in tetraploid potato (Solanum tuberosum) by transient CRISPR-Cas9 expression in protoplasts
    Andersson, M.; Turesson, H.; Nicolia, A.
  • Synthetic biology approaches for improving photosynthesis
    Kubis, A.; Bar-Even, A.
  • An innovative platform for quick and flexible joining of assorted DNA fragments
    De Paoli, H.C.; Tuskan, G.A.; Yang, X.
  • Enzymatic assembly of DNA molecules up to several hundred kilobases
    Gibson, D.G.; Young, L.; Chuang, R.Y.
  • Standards for plant synthetic biology: a common syntax for exchange of DNA parts
    Patron, N.J.; Orzaez, D.; Marillonnet, S.
  • Loop assembly: a simple and open system for recursive fabrication of DNA circuits
    Pollak, B.; Cerda, A.; Delmans, M.
  • Expression of plant antimicrobial peptide pro-SmAMP2 gene increases resistance of transgenic potato plants to Alternaria and Fusarium pathogens
    Vetchinkina, E.M.; Komakhina, V.V.; Vysotskii, D.A.
  • Advancing crop transformation in the era of genome editing
    Altpeter, F.; Springer, N.M.; Bartley, L.E.
  • A high-throughput transient expression system for rice
    Page, M.T.; Parry, M.A.J.; Carmo-Silva, E.
  • Transient expressions of synthetic biology in plants
    Sainsbury, F.; Lomonossoff, G.P.
  • An in planta, Agrobacterium-mediated transient gene expression method for inducing gene silencing in rice (Oryza sativa L.) leaves
    Andrieu, A.; Breitler, J.C.; Sire, C.
  • Endogenous silencing of Puccinia triticina pathogenicity genes through in planta-expressed sequences leads to the suppression of rust diseases on wheat
    Panwar, V.; McCallum, B.; Bakkeren, G.
  • Promoters pro-SmAMP1 and pro-SmAMP2 from wild plant Stellaria media for the biotechnology of dicotyledons
    Madzharova, N.V.; Kazakova, K.A.; Strel’nikova, S.R.
  • Reliable transient transformation of intact maize leaf cells for functional genomics and experimental study
    Kirienko, D.R.; Luo, A.; Sylvester, A.W.
  • Transient gene expression in epidermal cells of plant leaves by biolistic DNA delivery
    Ueki, S.; Magori, S.; Lacroix, B.; Citovsky, V.
  • Applications of new breeding technologies for potato improvement
    Hameed, A.; Zaidi, S.S.; Shakir, S.; Mansoor, S.
  • Evaluation of methods to assess in vivo activity of engineered genome-editing nucleases in protoplasts
    Nadakuduti, S.S.; Starker, C.G.; Ko, D.K.
  • Targeted gene mutation in tetraploid potato through transient TALEN expression in protoplasts
    Nicolia, A.; Proux-Wera, E.; Ahman, I.
  • A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity
    Jinek, M.; Chylinski, K.; Fonfara, I.
  • The CRISPR/Cas9 system for plant genome editing and beyond
    Bortesi, L.; Fische, R.
  • Improving cold storage and processing traits in potato through targeted gene knockout
    Clasen, B.M.; Stoddard, T.J.; Luo, S.
  • A revised medium for rapid growth and bioassays with tobacco tissue cultures
    Murashige, T.; Skoog, F.
  • Purification and viability determinations of plant protoplasts
    Larkin, P.J.
  • Engineered GFP as a vital reporter in plants
    Chiu, W.; Niwa, Y.; Zeng, W.
  • Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis
    Yoo, S.D.; Cho, Y.H.; Sheen, J.
  • Signal transduction in maize and Arabidopsis mesophyll protoplasts
    Sheen, J.
  • Tobacco protoplast transformation and use for functional analysis of newly isolated genes and gene constructs
    Fischer, R.; Hain, R.
  • Replication stress checkpoint signaling controls tRNA gene transcription
    Nguyen, V.C.; Clelland, B.W.; Hockman, D.J.
  • A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes
    Zhang, Y.; Su, J.; Duan, S.
  • Transient gene expression in electroporated Solanum protoplasts
    Jones, H.; Ooms, G.; Jones, M.G.
  • Effect of protoplast source and media on growth and regenerability of protoplast-derived calluses of Solanum tuberosum L
    Anjum, M.A.
  • Stoloniferous shoot protoplast, an efficient cell system in potato for somatic cell genetic manipulations
    Sharma, S.; Sarkar, D.; Pandey, S.K.
  • Direct gene transfer in potato: a comparison of particle bombardment of leaf explants and PEG-mediated transformation of protoplasts
    Craig, W.; Gargano, D.; Scotti, N.
  • Secretion marker proteins and cell-wall polysaccharides move through different secretory pathways
    Leucci, M.R.; Di Sansebastiano, G.; Gigante, M.