Clarification of microscopic soil ecosystem of methane-producing bacteria and sulfate-reducing bacteria

众所周知，自然环境下的甲烷产气量在很大程度上受产甲烷菌和硫酸盐还原菌的竞争控制。因此，在本研究中，我们开发了一种新的图像检测系统，通过荧光法和显微镜来澄清土壤中的这种竞争。1995年4月至1996年3月，通过测定模型系统的pH、Eh、产甲烷量、S^<2->浓度和有机质浓度，明确了水稻土产甲烷菌的生态系统。2)利用荧光探针5-（和6-）磺基荧光素双乙酸酯和6-羧基荧光素双乙酸乙酰氧基甲基酯建立了土壤中所有活菌的可视化方法。3)利用荧光显微镜对东京湾海边人工土壤中的产甲烷活菌（Mathanosarcina sp.）进行了拍摄。4)产甲烷菌的荧光光谱峰为420nm和460nm。

It is well known that the methane gas production in natural environments is largely controlled by the competition between methane-producing bacteria and sulfate-reducing bacteria. Therefore, in this study we developed a new image-detection system by fluorometry and microscopy for clarifying this competition in soil.From April in 1995 through March in 1996 we obtained the following results : 1) Ecosystem of the methane producing-bacteria in paddy soil was clarified by measuring pH, Eh, methanegas production, and concentrations of S^<2-> and organic materials in model systems. 2) Two visualization methods for all living bacteria in soil were developed by using fluorescence probes, 5- (and 6-) sulfofluorescein diacetate and 6-carboxyfluoresceindiacetate acetoxymethy1 ester.3) Photographs of living methane-producing bacteria (Mathanosarcina sp.) in an artificial soil of the seaside of Tokyo Bay were taken by fluorescence microscopy. 4)Peaks of fluorescence spectra of the methane-producing bacteria were found to be 420nm and 460nm.