Step‐by‐step protocol for the isolation and transient transformation of hornwort protoplasts

Premise 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 ev...

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Published inApplications in plant sciences Vol. 10; no. 2; pp. e11456 - n/a
Main Authors Neubauer, Anna, Ruaud, Stéphanie, Waller, Manuel, Frangedakis, Eftychios, Li, Fay‐Wei, Nötzold, Svenja I., Wicke, Susann, Bailly, Aurélien, Szövényi, Péter
Format Journal Article
LanguageEnglish
Published United States John Wiley & Sons, Inc 01.03.2022
John Wiley and Sons Inc
Wiley
Subjects
Anthoceros
Anthoceros agrestis
Cell division
Cell walls
Chloroplasts
Comparative analysis
Flowers & plants
Genetic transformation
genome
Genomes
hornworts
Life cycles
liquids
Localization
model organism
Plant sciences
Polyethylene glycol
Proteins
Protocol
Protocol Note
Protoplasts
transient transformation
hornworts
transient transformation
model organism
Anthoceros
protoplasts
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Summary:Premise 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. Methods and Results We describe a detailed protocol to isolate and transiently transform protoplasts of the model hornwort Anthoceros agrestis. 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. Conclusions This protocol represents a straightforward method for isolating and transforming A. agrestis 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.
Bibliography:This article is part of the special issue “Methodologies in Gametophyte Biology.”
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ISSN:2168-0450
2168-0450
DOI:10.1002/aps3.11456