Distribution analysis and the application of the nitrifying bacteria which inhabits in

硝化细菌的分布分析及应用

• 批准号: 17580073
• 负责人: TAKAHASHI Reiji
• 金额: $ 1.86万
• 依托单位: Nihon University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17580073/
• 关键词: nitrifying bacteria; rhizoplane; phylogenetic analysis; distribution; community structure; norB; cbbL; pyk; gap; pgk

In this research we focused on plant rhizoplane of plants which utilize different formed nitrogen compounds ("ammonia plant": rice, blueberry and so on, "nitrate plant"; barley, eggplant and so on), to designate the distribution of nitrifying bacteria and to definite dominant strain in these environment as final accomplishment. Recently for non-cultured analysis of ammonia-oxidizing bacteria (AOB) from several environmental sources such as DGGE, the CTO primer and the specific region originated from ammonia mono oxygenase (AMO) gene which is specific to AOB as another available primer. We tried to develop the other primers of more effective and high resolution for nitrifying bacteria analysis and to establish more efficient analysis.<Research result>(1) Development of unique probe for nitrifying bacteria screening : We examined that norB (nitrite oxidoreductase gene) showed the possibility of minute classification internal genus Nitrobacter of nitrite ozidizing bacteria (NOB). We showed that three of housekeeping gene ( gap, pgk, pyk), especially pyk phylogenetic tree of the AOB showed high resolution compared with the tree based on 16SrRNA gene sequence. It made that 3 subclasses of Nitrosospira, Nitrosolobus and Nitrosovibrio based on their morphological difference which it cannot identify with classification of 16SrRNA gene sequence.(2) Analysis of distribution on the plant rhizoplane : On the basis of the previous section, for DGGE analysis, we attempt to generate the primers.(3) Pure isolation, its breeding and functional analysis of new isolate : Newly isolated AOB and NOB were obtained from the rhizoplane of rice, blueberry, barley and eggplant. Especially the strain isolated from rhizoplae of leucaena grown in fields of an acid sulfate soil in Thailand which showed high urease activity and copper tolerance, so is very useful strain for functional analysis.

在这项研究中，我们重点研究利用不同形成氮化合物的植物（“氨植物”：水稻、蓝莓等，“硝酸盐植物”；大麦、茄子等），以指定硝化细菌的分布和在这些环境中确定主导应变作为最终成就。最近，对于来自多种环境来源（例如 DGGE）的氨氧化细菌（AOB）的非培养分析，CTO 引物和源自氨单加氧酶（AMO）基因的特定区域作为另一种可用的引物，该基因对 AOB 具有特异性。我们试图开发其他更有效和高分辨率的硝化细菌分析引物，并建立更高效的分析。<研究结果>（1）开发用于硝化细菌筛选的独特探针：我们检查了norB（亚硝酸氧化还原酶基因）显示亚硝酸盐氧化细菌（NOB）内部硝化细菌属的微小分类的可能性。我们发现，与基于 16SrRNA 基因序列的树相比，AOB 的三个管家基因（gap、pgk、pyk），特别是 pyk 系统发育树显示出高分辨率。根据16SrRNA基因序列分类无法识别的形态差异，将其分为亚硝螺菌属、亚硝化叶菌属和亚硝化弧菌属3个亚类。 (2)植物根面分布分析：在上节的基础上，进行DGGE分析， (3)新分离物的纯分离、选育及功能分析：新分离的AOB和NOB是从水稻、蓝莓、大麦和茄子的根面。特别是从生长在泰国酸性硫酸盐土壤田地的银合欢根际中分离得到的菌株，显示出高脲酶活性和铜耐受性，因此是用于功能分析的非常有用的菌株。

项目成果

微生物の世界

微生物世界

• 发表时间: 2006
• 作者: Karatani;H.;Heguri;A.;Hirayama;S.;Matsumura-Inoue;T;横田 明
• 通讯作者: 横田 明

Phylogenetic relationshipa smong ammonia-oxidizing bacteria as revealed by gene sequences of glyceraldehyde 3-phosphate dehydrogenease and phosphoglycerate kinase

3-磷酸​​甘油醛脱氢酶和磷酸甘油酸激酶基因序列揭示的氨氧化细菌的系统发育关系

• 发表时间: 2005
• 期刊: Journal of Bioscience and Bioengineering 99(6)
• 作者: Takeshi Ida;Michimoto Kigimiya;Mina Kogure;Reiji Takahashi;Tatsuaki Tokuyama
• 通讯作者: Tatsuaki Tokuyama

Classification of chemoautotrophic ammonia-oxidizing bacteria using pyruvate kinase genes as high resolution phylogenetic markers

使用丙酮酸激酶基因作为高分辨率系统发育标记对化学自养氨氧化细菌进行分类

• 发表时间: 2006
• 期刊: Soil Science and Plant Nutrition 52
• 作者: Takeshi Ida;Reiji Takahashi;Kazuo Satoh;Mina Kogure;Tatsuaki Tokuyama
• 通讯作者: Tatsuaki Tokuyama

Phylogenetic relationships among ammonia-oxidizing bacteria as revealed by gene sequences of glyceraldehyde 3-phosphate dehydrogenase and phosphoglycerate kinase

3-磷酸​​甘油醛脱氢酶和磷酸甘油酸激酶基因序列揭示氨氧化细菌之间的系统发育关系

• 发表时间: 2005
• 期刊: Journal of Bioscience and Bioengineering 99
• 作者: Takeshi Ida;Michitomo Kugimiya;Mina Kogure;Reiji Takahashi;Tatsuaki Tokuyama
• 通讯作者: Tatsuaki Tokuyama