Development of analytical technique of change in microbial population for remediation in the polluted soil

污染土壤修复微生物种群变化分析技术的开发

• 批准号: 11558075
• 负责人: FUJIE Koichi
• 金额: $ 8万
• 依托单位: Toyohashi University of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11558075/
• 关键词: Microbial community structure; Quinone profile; FISH; CTC reduction; Soil microorganism; Tidal flat ecosystem; Remediation; Bacteria extraction; DTAF; CTC; 土壤汚染; FISH; キノン量; PCR; 土壌汚染; バイオマス; 微生物多様性

Microorganisms perform an important role for remediation in various polluted environment. To optimize the activity of microorganisms on contaminated sites, the ability of microorganisms must be clarified on the optimal condition for decomposing bacteria. The behavior of specific bacteria and microbial population should be clarified. The objective of this study is to develop the analytical technique for clarification of the behavior of specific bacteria and microbial community structure in natural waters and soils.The direct application of molecular genetic methods on soil bacteria is often prevented by the large amounts of soil colloids. Hence, an extraction of bacteria cells from soil is usually required before analysis of their genetic material. For this reason, the selection of a suitable extractant was investigated. The extraction efficiency of bacteria and the representation of the microbial population extracted from soil by using quinone-profiling method were evaluated in the exp … More eriments. The fraction of the respiratory active cell extracted from soil was also evaluated by using CTC (5-cyano-2, 3-ditolyl tetorazolium chloride) reduction and plate count techniques. 1/15M Na-phosphate, 0.03M Na-oxalate and 0.03M Na-pyrophosphate were used as the extractants. The extraction efficiency of bacteria was defined as the ratio of quinone contents in the extract to that in soil. Compared with other two extractants, Na-pyrophosphate demonstrated the highest extraction efficiency of bacteria and respiratory active cells in the extract from soil. In addition, cell numbers of Proteobacteria, and γ-subclass in extraction with Na- pyrophosphate was measured by FISH. Almost all bacteria of Proteobacteria α, β and γ-subclass contain UQ-10, UQ-8, UQ-9, respectively. The experimental results showed that there was very similar relationship between ratio of Proteobacteria β, γ to α-subclasses and ratio of UQ-8, UQ-9 to UQ-10, respectively. It indicates that specific bacteria in soil sample can be quantified by FISH technique.In addition, the quinone profile and CTC reduction can be used to clarify the effect of particle size of sediment on microbial-ecosystem and water purification using an experimental facility of the tidal flat, and to analyze microbial community structure in sediment. Less

微生物在各种污染环境的修复中发挥着重要作用。为了优化污染场地中微生物的活性，必须明确微生物在最佳分解条件下的分解能力。应明确特定细菌和微生物种群的行为。本研究的目的是发展一种分析技术，用于阐明自然沃茨和土壤中特定细菌的行为和微生物群落结构。因此，在分析其遗传物质之前，通常需要从土壤中提取细菌细胞。为此，研究了合适的萃取剂的选择。本实验对土壤中细菌的提取效率和醌型微生物种群的代表性进行了评价， ...更多信息 实验。从土壤中提取的呼吸活性细胞的分数也通过使用CTC（5-氰基-2，3-二甲苯基氯化四唑）还原和平板计数技术来评估。使用1/15 M磷酸钠、0.03 M草酸钠和0.03 M焦磷酸钠作为萃取剂。细菌提取效率定义为提取液中醌含量与土壤中醌含量的比值。结果表明，焦磷酸钠对土壤细菌和呼吸活性细胞的提取效率最高。此外，用荧光原位杂交技术（FISH）测定了焦磷酸钠提取液中变形菌门和γ亚门的细胞数.变形菌门α、β、γ亚类细菌几乎都含有UQ-10、UQ-8、UQ-9。实验结果表明，变形菌β、γ和α亚类的比值分别与UQ-8、UQ-9和UQ-10的比值之间存在非常相似的关系。结果表明，荧光原位杂交技术可以定量分析土壤中的特定细菌，醌含量和CTC还原率可以用于研究沉积物粒径对微生物生态系统和水质净化的影响，并分析沉积物中微生物的群落结构。少

项目成果

Katsuaki Saitou, Koichi Fujie and Arata Katayama: "Detection of microbial groups metabolizing a substrate in soil based on the [^<14>C] quinone profile"Soil Science and Plant Nutrition. 45(3). 669-679 (1999)

Katsuaki Saitou、Koichi Fujie 和 Arata Katayama：“基于 [^14C] 醌谱检测土壤中代谢底物的微生物群”土壤科学与植物营养。

A.Katayama, K.Funasaka, K.Fujie: "Changes in the respiratory quinone profile of a soil treated with pesticides"Biol.Fertil.Soils. 33. 454-459 (2001)

A.Katayama、K.Funasaka、K.Fujie：“用农药处理的土壤呼吸醌谱的变化”Biol.Fertil.Soil。

Hong-Ying Hu: "Development of a novel solid phase extraction method for the analysis of bacterial quinones in activated sludge with a higher reliability"J.Bioscience and Bioeng.. 87(3). 378-382 (1999)

胡红英：“开发一种新型固相萃取方法，用于分析活性污泥中细菌醌，具有更高的可靠性”J.Bioscience and Bioeng.. 87(3)。

Katsuaki Saitou: "Detection of microbial groups metabolizing a substrate in soil based on the[^<14>C]quinone profile"Soil Science and Plant Nutrition. 45(3). 669-679 (1999)

Katsuaki Saitou：“基于[^ 14 C]醌谱检测土壤中代谢底物的微生物群”土壤科学与植物营养。

Katsuaki Saitou: "Linear relation between the amount of respiratory quinones and the microbial biomass in soil"Soil Science and Plant Nutrition. 45(3). 775-778 (1999)

Katsuaki Saitou：“呼吸醌量与土壤中微生物量之间的线性关系”土壤科学与植物营养。