IDENTIFICATION OF TWIN-ARGININE TRANSLOCATION SYSTEM IN PSEUDOMONAS SYRINGAE AND ITS CONTRIBUTION TO PATHOGENICITY AND FITNESS

Authors: Bronstein, Philip; MARRICHI, MATTHEW - CORNELL UNIVERSITY; Cartinhour, Samuel; Schneider, David; DELISA, MATTHEW - CORNELL UNIVERSITY

Submitted to: Journal of Bacteriology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/30/2005
Publication Date: 12/1/2005
Citation: Bronstein, P., Marrichi, M., Cartinhour, S.W., Schneider, D.J., Delisa, M.P. 2005. Identification of twin-arginine translocation system in pseudomonas syringae and its contribution to pathogenicity and fitness. Journal of Bacteriology. 187(24):8450-8461.

Interpretive Summary: The bacterial plant pathogen Pseudomonas syringae causes disease in the model plant Arabidopsis thaliana and various crops. The ability to cause disease depends upon a special protein secretion system and proteins that are injected into plant cells. Additional factors are also known or suspected. Therefore, we explored the contribution of another secretion system known as the twin-arginine translocation (Tat) pathway. We found that a Tat mutant displayed a number of altered characteristics including loss of motility, deficiency in iron acquisition, increased sensitivity to copper, significant loss of extracellular phospholipase activity and decreased ability to cause disease. We showed that the decreased ability to cause disease likely arises from a combination of a compromised fitness of bacteria in planta and a decreased efficiency of protein secretion. Finally, we designed a general detection system based on a fluorescent protein for the identification of Tat-secreted proteins. Our evidence supports the notion that the ability of Pseudomonas syringae to cause disease is a multifactorial process and that the Tat system is important component of this.

Technical Abstract: The bacterial plant pathogen Pseudomonas syringae pv. tomato DC3000 (DC3000) causes disease in Arabidopsis thaliana and tomato, and it elicits the hypersensitive response (HR) in non-host plants such as Nicotiana tabacum and Nicotiana benthamiana. While these events chiefly depend upon the type III protein secretion system (TTSS) and the effector proteins that this system translocates into plant cells, additional factors have been shown to contribute to DC3000 virulence and still many others are likely to exist. Therefore, we explored the contribution of the twin-arginine translocation (Tat) system to the physiology of DC3000. We found that a tatC mutant strain of DC3000 displayed a number of phenotypes including loss of motility on soft agar plates, deficiency in siderophore synthesis and iron acquisition, sensitivity to copper, loss of extracellular phospholipase activity and attenuated virulence in host plant leaves. In the latter case, we provide evidence that decreased virulence of tatC mutants likely arises from a synergistic combination of: (i) compromised fitness of bacteria in planta; (ii) decreased efficiency of type III translocation; and (iii) cytoplasmically retained virulence factors. Finally, we demonstrate a novel broad-host-range genetic reporter based on the green fluorescent protein (GFP) for the identification of Tat-targeted secreted virulence factors that should be generally applicable to any Gram-negative bacterium. Collectively, our evidence supports the notion that virulence of DC3000 is a multifactorial process and that the Tat system is an important virulence determinant of this phytopathogenic bacterium.