Method for rapid identification of plant pathogenic bacteria

一种植物病原菌的快速鉴定方法

基本信息

批准号：
06454060
负责人：
TSUYUMU Shinji
金额：
$ 3.9万
依托单位：
Shizuoka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06454060/
关键词：
probe hybridization genes for glucan hrp new pathogenicity gene pectate lyase identification of plant pethogenic bacteria Nisiyama's identification 植物病原細菌同定カンキツかいよう病菌 avr pth遺伝子 hrp遺伝子群植物体内増殖 firp ペクチンリアーゼ

项目摘要

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.

为了建立快速、准确、简便的杂交鉴定植物病原菌的方法，寻找合适的探针。这些探针可以分为两组。第一组是用于微生物表型测试的组。这里，我们选择克反应、乳糖利用、脲酶反应、半乳糖利用作为模型系统。首先从遗传数据库中搜索负责这些表型的基因中的共同区域。在确认这些区域仅适用于多个来源的特定基因中的唯一区域之后，合成寡聚物，用作探针。作为使用这些探针的实验的一般结论，很明显，那些已知在微生物特性中表现出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，一些被报道为阴性的菌株被判定为阳性。为了提高鉴定的准确性，选择了第二组探针，特别强调参与致病性。在这里，我们测试了环葡聚糖、avr/pth 基团、hrp 基团和果胶酸裂合酶合成的基因。结果发现，在斑点杂交和RFLP分析中，植物病原菌的大分组和有限分组分别是可能的。总之，结合这两类探针，我们可以快速识别植物病原菌。