Streptomyces turgidiscabies secretes a novel virulence protein, Nec1, which facilitates infection

Emergence of new, economically important plant-pathogenic species in the mostly saprophytic genus Streptomyces involves acquisition of a large, mobile pathogenicity island (PAT). Biosynthetic genes for a phytotoxin, thaxtomin A, are contained on this PAT. The Nec1 protein has necrogenic activity on excised potato tuber tissue, and the encoding gene is highly conserved in plant-pathogenic Streptomyces spp. The G+C content of nec1 indicates lateral transfer from an unrelated taxon; however, the nucleic acid and protein databases have not yielded homologs. Data presented in this article demonstrate that the Nec1 protein is necrogenic when expressed in Escherichia coli and that an active 16-kDa form of Nec1 is secreted from the plant pathogen Streptomyces turgidiscabies. Deletion analysis of nec1 demonstrated that the 151-amino-acid C-terminal region of the Nec1 protein is sufficient to confer necrogenic activity. Analysis of nec] transcriptional start sites indicates that two mRNA species are produced and that the site of transcription initiation is influenced by glucose. S. turgidiscabies containing a nec1 deletion was greatly compromised in virulence on Arabidopsis thaliana, Nicotiana tabacum, and Raphanus sativus seedlings. The wild-type strain, S. turgidiscabies Car8, aggressively colonized and infected the root meristem of radish, whereas the Delta nec1 mutant Car811 did not. Taken together, these data suggest that Nec1 is a secreted virulence protein with a conserved plant cell target that acts early in plant infection.