Step-by-step protocol for the isolation and transient transformation of hornwort protoplasts.
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Authors
Nötzold, Svenja I
Publication Date
2022-02-11Journal Title
Applications in plant sciences
ISSN
2168-0450
Volume
10
Issue
2
Language
eng
Type
Article
This Version
VoR
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Neubauer, A., Ruaud, S., Waller, M., Frangedakis, E., Li, F., Nötzold, S. I., Wicke, S., et al. (2022). Step-by-step protocol for the isolation and transient transformation of hornwort protoplasts.. Applications in plant sciences, 10 (2) https://doi.org/10.1002/aps3.11456
Abstract
<h4>Premise</h4>A detailed protocol for the protoplast transformation of hornwort tissue is not yet available, limiting molecular biological investigations of these plants and comparative analyses with other bryophytes, which display a gametophyte-dominant life cycle and are critical to understanding the evolution of key land plant traits.<h4>Methods and results</h4>We describe a detailed protocol to isolate and transiently transform protoplasts of the model hornwort <i>Anthoceros agrestis</i>. The digestion of liquid cultures with Driselase yields a high number of viable protoplasts suitable for polyethylene glycol (PEG)-mediated transformation. We also report early signs of protoplast regeneration, such as chloroplast division and cell wall reconstitution.<h4>Conclusions</h4>This protocol represents a straightforward method for isolating and transforming <i>A. agrestis</i> protoplasts that is less laborious than previously described approaches. In combination with the recently developed stable genome transformation technique, this work further expands the prospects of functional studies in this model hornwort.
Keywords
Protoplasts, Model organism, Hornworts, Transient Transformation, Anthoceros
Sponsorship
Swiss National Science Foundation (160004, 184826)
Identifiers
35495192, PMC9039799
External DOI: https://doi.org/10.1002/aps3.11456
This record's URL: https://www.repository.cam.ac.uk/handle/1810/337814
Rights
Attribution-NonCommercial-NoDerivatives 4.0 International
Licence URL: https://creativecommons.org/licenses/by-nc-nd/4.0/
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