Method for rapid identification of plant pathogenic bacteria

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To establish the rapid, precise, and yet simple method for the identification of plant pathogenic bacteria using hybridization methods, the proper probes were searched. These probes can be divided into two groups. First group is the ones for the test of microbiological phenotypes. Here, we have chosen gram reaction, utilization of lactose, urease reaction, utilization of galactose as the model systems. The common regions in the genes responsible for those phenotypes were first searched from genetic data base. After confirming the applicability of these region as the unique region only among the specific genes among several sources, the oligomers were synthesized, used as the probes. As a general conclusion from the experiment using these probes, it became clear that those strains known to show positive reaction in the microbiological characters can be judged properly as positive by hybridization. On the other hand, some strains which have been reported to be negative were judged as positive. To increase accuracy of the identification, second group of probes were chosen with the special emphasis on the involvement in the pathogenicity. Here, we have tested the genes for the synthesis of cyclic glucan, avr/pth group, hrp group and pectate lyase. As a result, it was found that large grouping of plant pathogenic bacteria and finite grouping was possible in dot-blot hybridization and in RFLP analysis, respectively. In conclusion, it is possible for us to identify plant pathogenic bacteria rapidly by combining these two types of probes.