Transformation of the Moss Physcomitrella patens Using T-DNA Mutagenesis

Abstract

INTRODUCTIONIn 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).