Specific Maceration and Induction of PR-3 Gene in Potato Tuber Tissue by Pectobacterium carotovorum subsp. Atrosepticum Type III Secretion System Mutants

Abstract

The exact function of type III secretion system in some phytopathogenes including Pectobacterium carotovorum subsp. atrosepticum (Pca) is not understood and is a matter of debate. The aim of this study were to determine specific effect of type III secretion system on potato tubers and to reveal the connection of this system with potato resistant genes such as PR-3. A Pca hrpW fragment was subcloned into a low-copy-number cloning vector (pZH448). The resulting plasmid (pAS19) was then conjugated into the wild-type and mutant strains of Pca by type III secretion system. The virulence property of different Pca strains was studied and the influence of over expression of hrpW on maceration activity was also investigated. Furthermore, the effect of mentioned mutation on the maceration of carrot-root was evaluated. Finally, using real-time PCR, the copy-number of PR-3 gene in potato tuber tissue was assessed. In conclusion, for type III secretion system mutant strains, in contrast with the wild-type, the maceration amount of potato tuber tissue decreased after over expression of hrpW while inoculation of tubers by mutants, increased this amount. In the case of potato, HrpN and DspE proteins appeared to be avirulent factors. Compared with the wild-type strains, Pca nominated mutants significantly reduced potato PR-3 expression thus, PR-3 expression level in potato tuber tissue in answer to infiltration by Pca, depends on functional type III secretion system in the bacterium.