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

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 protoco

SUBMITTER: Neubauer A 

PROVIDER: S-EPMC9039799 | biostudies-literature | 2022 Mar-Apr

REPOSITORIES: biostudies-literature