Method for rapid identification of plant pathogenic bacteria

为建立一种快速、准确、简便的杂交法鉴定植物病原菌的方法，寻找合适的探针。这些探头可以分为两组。第一组为微生物表型检测组。在这里，我们选择了革兰氏反应、乳糖利用、尿素酶反应、半乳糖利用作为模型体系。首先从遗传数据库中搜索与这些表型有关的基因的共同区域。在确认这些区域作为几个来源的特定基因中唯一的区域的适用性后，合成寡聚体作为探针。从使用这些探针的实验中得出的总体结论是，那些已知的微生物学特性呈阳性反应的菌株可以通过杂交正确地判断为阳性。另一方面，一些被报告为阴性的菌株被判断为阳性。为了提高鉴定的准确性，选择了第二组探针，特别强调了与致病性的参与。在这里，我们测试了环状葡聚糖、AVR/PTH基团、HRP基团和果胶裂解酶的合成基因。结果发现，在斑点杂交和RFLP分析中，植物病原菌的大群和有限群是可能的。综上所述，将这两种探针结合起来，可以快速鉴定植物病原菌。

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.