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


Type
Article
Change log
Abstract

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.

Description
Keywords
Anthoceros, hornworts, model organism, protoplasts, transient transformation
Journal Title
Appl Plant Sci
Conference Name
Journal ISSN
2168-0450
2168-0450
Volume Title
10
Publisher
Wiley
Sponsorship
Swiss National Science Foundation (160004, 184826)