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.

本研究以利用不同形态氮化合物的植物（“氨植物”：水稻、蓝莓等，“硝植物”：大麦、茄子等）根面为研究对象，明确硝化细菌的分布，确定硝化细菌在这些环境中的优势菌株。近年来，在非培养条件下，利用DGGE等技术对氨氧化细菌（AOB）进行分析时，CTO引物和来自AOB特异性的氨单加氧酶（AMO）基因的特异性区域作为另一个可用的引物。本研究试图开发出其他更有效、分辨率更高的硝化细菌分析引物，建立更高效的硝化细菌分析方法。<Research result>（1）硝化细菌特异性筛选探针的研制：我们检测了NORB（nitric oxidoreductase gene）基因作为亚硝酸盐氧化细菌（NOB）的微分类内属硝化杆菌（Nitrobacter）的可能性。结果表明，利用gap、pgk、pyk三个看家基因构建的AOB系统发育树，尤其是pyk系统发育树，与基于16SrRNA基因序列构建的系统发育树相比，分辨率更高。根据亚硝化螺旋菌属、亚硝化裂叶菌属和亚硝化弧菌属在形态上的差异，将亚硝化螺旋菌属划分为3个亚类，这些亚类与16SrRNA基因序列的分类不能完全一致。(2)在植物根面分布分析：在前一部分的基础上，我们尝试产生用于DGGE分析的引物。(3)从水稻、蓝莓、大麦和茄子根面分离到新分离的AOB和NOB。特别是从生长在泰国酸性硫酸盐土壤中的银合欢根际分离的菌株表现出较高的脲酶活性和铜耐受性，因此是非常有用的功能分析菌株。

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

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

Characteristics of newly isolated ammonia-oxidizing bacteria from acid sulfate soil and the rhizoplane of leucaena grown in that soil.

酸性硫酸盐土壤中新分离的氨氧化细菌及其生长的银合欢根面的特征。

• 发表时间: 2007
• 期刊: Soil Sci. Plant Nutr 53
• 作者: Satoh;K.;Itoh;C.;Kang;D.-J.;Sumida;H.;Takahashi;R.;Isobe;K.;Sasaki;S.;Tokuyama T
• 通讯作者: Tokuyama T

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