The virulence of soft-rot Erwinia species is dependent mainly upon secreted enzymes such as pectinases, pectin lyases, and proteases that cause maceration of plant tissue. Some soft-rot Erwinia spp. also harbor genes homologous to the hypersensitive reaction and pathogenesis (hrp) gene cluster, encoding components of the type III secretion system. The hrp genes are essential virulence determinants for numerous nonmacerating gram-negative plant pathogens but their role in the virulence of soft-rot Erwinia spp. is not clear. We isolated and characterized 11 hrp genes of Erwinia carotovora subsp. carotovora. Three putative sigmaL-dependent Hrp box promoter sequences were found. The genes were expressed when the bacteria were grown in Hrp-inducing medium. The operon structure of the hrp genes was determined by mRNA hybridization, and the results were in accordance with the location of the Hrp boxes. An E. carotovora strain with mutated hrcC, an essential hrp gene, was constructed. The hrcC- strain was able to multiply and cause disease in Arabidopsis, but the population kinetics were altered so that growth was delayed during the early stages of infection.
The occurrence of bacteria in different tissues was studied using field-grown strawberries, in vitro-grown strawberries, wild strawberries, and aseptically germinated strawberry seedlings. Strawberry has a number of endophytic bacteria in its the internal tissue, most of which appear to be nonpathogenic. In the in vitro-grown strawberries, all identified isolates were in the genus Pantoea. In field-grown garden and wild strawberries the most common genera were Pantoea and Pseudomonas. Location of eubacterial inhabitants within strawberry tissue sections was studied by in situ hybridization. Bacteria were detected in flower stalks, leaf stalks, leaves, stolons, berries and aseptically germinated seedlings. The existence of bacteria in seeds and seedlings suggests that bacteria are able to move up to the generative tissue and, ultimately, to the next generation, forming a symbiosis-like chain of plant-bacteria coexistence.
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