Isolation and Regeneration of Protoplasts of the Moss Physcomitrella patens

David J. Cove; Pierre-François Perroud; Audra J. Charron; Stuart F. McDaniel; Abha Khandelwal; Ralph S. Quatrano

doi:10.1101/pdb.prot5140

Protocol

Isolation and Regeneration of Protoplasts of the Moss Physcomitrella patens

Department of Biology, Washington University, St. Louis, MO 63130, USA

↵¹Corresponding author (rsq{at}wustl.edu)

This article is also available in Emerging Model Organisms: A Laboratory Manual, Vol. 1. CSHL Press, Cold Spring Harbor, NY, USA, 2009.

INTRODUCTION

This protocol describes how to isolate individual protoplasts from young gametophyte tissue of the moss Physcomitrella patens and how to regenerate them into plants if desired. Protonemal tissue is grown on cellophane-overlay plates, harvested, and treated with 0.5% Driselase to produce protoplasts. After filtration and washing, the protoplasts are regenerated on solid medium.