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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.

为建立快速、准确、简便的植物病原细菌杂交鉴定方法，寻找合适的探针。这些探针可以分为两组。第一组是微生物表型检测组。本文选择了革兰氏反应、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相关的基因的共同区域。在确认这些区域仅在几个来源中的特定基因中作为唯一区域的适用性后，合成寡聚体，用作探针。作为使用这些探针的实验的一般结论，很明显，那些已知在微生物学特征中显示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，一些已报告为阴性的菌株被判定为阳性。为了提高鉴定的准确性，选择第二组探针，特别强调参与致病性。在此，我们检测了环葡聚糖合成基因、avr/pth基因组、hrp基因组和果胶酸裂解酶基因。结果表明，在斑点杂交和RFLP分析中，植物病原菌的大类群和有限类群分别是可能的。因此，将这两种探针结合起来进行植物病原细菌的快速鉴定是可行的。