Cite as: Cold Spring Harb. Protoc.; 2009; doi:10.1101/pdb.prot5144

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Transformation of the Moss Physcomitrella patens Using T-DNA Mutagenesis

David J. Cove, Pierre-François Perroud, Audra J. Charron, Stuart F. McDaniel, Abha Khandelwal, and Ralph S. Quatrano1

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

In this protocol, the transformation of moss (Physcomitrella patens) protoplasts is performed via Agrobacterium-mediated transfer of T-DNA. Protoplasts are incubated with Agrobacterium and acetoseringone in regeneration medium. They are then washed and plated on antibiotic-containing medium to select for T-DNA insertion in stable transformants. The transformation rate for this protocol is typically 10–4 (expressed as the frequency of stable transformants among regenerants surviving the transformation procedure).


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The Moss Physcomitrella patens: A Novel Model System for Plant Development and Genomic Studies
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Cold Spring Harb Protoc 2009: 115. [Abstract] [Full Text]

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Home page
 Cold Spring Harb ProtocHome page
D. J. Cove, P.-F. Perroud, A. J. Charron, S. F. McDaniel, A. Khandelwal, and R. S. Quatrano
The Moss Physcomitrella patens: A Novel Model System for Plant Development and Genomic Studies
Cold Spring Harb Protoc, February 1, 2009; 2009(2): pdb.emo115 - pdb.emo115.
[Abstract] [Full Text]

Home page
 Cold Spring Harb ProtocHome page
D. J. Cove, P.-F. Perroud, A. J. Charron, S. F. McDaniel, A. Khandelwal, and R. S. Quatrano
Isolation and Regeneration of Protoplasts of the Moss Physcomitrella patens
Cold Spring Harb Protoc, February 1, 2009; 2009(2): pdb.prot5140 - pdb.prot5140.
[Abstract] [Full Text]