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Aims: To develop a strain-specific TaqMan PCR method for detecting and quantifying the biocontrol strain Lysobacter enzymogenes 3.1T8., Methods and Results: A primer-probe combination was designed on the basis of a strain-specific sequence selected using REP-PCR (repetitive extragenic palindromic-polymerase chain reaction). The specificity of this combination was demonstrated by 14 other Lysobacter strains that did not react with the selected primer-probe combination. To quantify strain 3.1T8 in cucumber root samples, a calibration curve was prepared by spiking roots with a 10-fold dilution series of the strain. Detection of the biocontrol strain 3.1T8 with this method showed that the strain survived well for 22 days on root tips as well as on older cucumber roots. Survival was higher when the strain was inoculated to younger plants. In a cucumber production system with large volumes of substrate, strain 3.1T8 was detected in high numbers on cucumber roots 3 weeks after inoculation., Conclusions: The primer-probe combination developed was strain specific, because it did not react with other strains of the same species and genus. The TaqMan PCR method successfully quantified the inoculated biocontrol strain on cucumber roots grown in different cropping systems., Significance and Impact of the Study: The developed TaqMan PCR method is a strain-specific real-time detection method that can be used to assess the population dynamics of L. enzymogenes strain 3.1T8 for further optimization of its biocontrol efficacy.

The exploration of new source materials and the use of alternative isolation and identification methods have led to rapid expansion in the knowledge of diversity; in Lysobacter, 11 new species having been described since 2005, and in Stenotrophomonas with six new species since 2000. The new species of Lysobacter, isolated by dilution and direct plating on standard media, differ in several key phenotypic properties from those obtained by enrichment on complex polysaccharides in the original description of the genus. Revision of the definition of the genus will be required. Both culture-dependent and culture-independent methods to assess community structure, in a variety of host and nonhost environments, have established that some species of Lysobacter are a dominant component of the microflora, where previously their presence had not been suspected. Culture-independent studies have generally not added new information on the occurrence and distribution of Stenotrophomonas maltophilia and other members of the genus, which are readily isolated on standard media from source materials. Lysobacter enzymogenes and Sten. maltophilia produce similar antibiotics and share some enzyme activities which, subject to safety considerations, may make them attractive candidates for use in biological control of plant diseases and of nematodes.