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