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Environ. Microbiol. 7: 1809-1817 Abstract Our group studies tomato foot and root rot, a plant disease caused by the fungus Forl (Fusarium oxysporum f. sp. radicis-lycopersici). Several bacteria have been described to be able to control the disease, using different mechanisms. Here we describe a method that enables us to select, after application of a crude rhizobacterial mixture on a sterile seedling, those strains that reach the root tip faster than our best tomato root colonizer tested so far, the Pseudomonas fluorescens biocontrol strain WCS365. Of the five tested new isolates, four appeared to be able to reduce the number of diseased plants. Analysis of one of these strains, P. fluorescens PCL1751, suggests that it controls the disease through the mechanism ‘competition for nutrients and niches’, a mechanism novel for biocontrol bacteria. Moreover, this is the first report describing a method to enrich for biocontrol strains from a crude mixture of rhizobacteria. Another advantage of the method is that four out of five strains do not produce antifungal metabolites, which is preferential for registration as a commercial product. 26 Introduction Many plant diseases are caused by phytopathogenic fungi. In order to decrease the input of agrochemicals in agriculture, biocontrol microbes are used as possible alternatives (Xxxxxxxxx et al., 1987; Xxxxxxxxxx and Xxxxxxxxx, 2004). We study tomato foot and root rot (TFRR), which is a disease caused by the fungus Fusarium oxysporum f. sp. radicis-lycopersici (Forl). This disease cannot efficiently be prevented by chemicals. The isolation of biocontrol bacteria involves a labor intensive screening process that, in the case of antibiosis, can be enhanced by introducing a screening step for strains that produce antifungal metabolites (AFMs) in vitro. To our knowledge, no procedures have been described, which facilitate the selection of biocontrol microbes that act through other mechanisms. Xxxxxx and colleagues (2001) described a method to select enhanced grass root tip colonizing bacteria. In this method a mixture of rhizosphere bacteria is applied on a sterile seedling. After plant growth in a gnotobiotic system (Simons et al., 1996), those bacteria that have reached the root tip are isolated. These are subsequently used to inoculate a fresh sterile seedling, which again is allowed to grow. After three of these enrichment cycles, excellent competitive root tip colonizers were obtained (Xxxxxx et al., 2001). In the present ...
