Influence of bacterial strain genotype on transient expression of plasmid DNA in plant protoplasts

Abstract

It is demonstrated that transient expression of plasmid DNA in plant protoplasts can be strongly influenced by the bacterial strain used for plasmid propagation. Four different promoter constructs containing two light-responsive and two fungal elicitor-responsive parsley promoters translationally fused to the reporter gene, β-glucuronidase (GUS), were amplified in bacterial strains MC1061, DH5α or GM2163 and tested individually. Marked differences in basal expression levels and in fold inducibilities were observed upon transfection of parsley protoplasts. Low levels of basal expression and strong light or elicitor inducibilities were observed only with plasmid DNA derived from the methylation-deficient GM2163 strain. In vitro methylation of DNA prior to transformation also drastically increased basal expression levels. The results suggest that DNA methylation may be partly responsible for deregulating promoter activity in the transient expression system.