一株锰氧化细菌的分离、鉴定及其锰氧化特性  

Isolation and characterization of a manganese-oxidizing bacterium from soils

作　　者：郑洁[1] 孟佑婷[1] 方瑶瑶[2] 杨素玲[1] 王平[1] 

机构地区：[1]北京市辐射中心,北京100875 [2]北京市环境保护科学研究院,北京100037

出　　处：《微生物学报》2016年第11期1699-1708,共10页Acta Microbiologica Sinica

基　　金：国家自然科学基金(41203059);北京市科学技术研究院青年骨干计划(2013-014);北京市科学技术研究院萌芽项目(201501)~~

摘　　要：【目的】获得锰氧化细菌,对锰矿周边土壤中生物所参与的锰氧化过程进行初探。【方法】依据细菌是否能氧化Mn(Ⅱ),形成棕褐色锰氧化物进行筛选。利用染料LBB对生成的锰氧化物进行检测。通过考察分离菌株的形态、生理特征和16S r RNA基因、gyr B基因、gyr A基因序列的同源性对分离菌株进行鉴定。分析筛选菌与所在属已知锰氧化菌的亲缘关系。利用LBB显色法检测氧化锰的动态生成,通过扫描电镜-能谱分析和X射线衍射技术分析生物氧化锰的表征。【结果】获得1株锰氧化细菌菌株,命名为CP133,综合形态、生理及分子分析结果,鉴定为蜡样芽孢杆菌(Bacillus cereus),分离菌株与多株分离自海洋及土壤的芽孢类锰氧化菌在进化上具有一定的差异。与其他菌株比较菌株CP133具有较强的锰氧化能力,进入稳定期后可生成紧密结合在菌体周围的无定形态生物氧化锰。【结论】从锰矿周边土壤分离出1株具有较强锰氧化功能的蜡样芽孢杆菌,丰富了土壤芽孢类锰氧化菌的资源,同时也为锰矿周围土壤与锰氧化菌间的生物地球化学循环提供了线索及材料。[Objective] The aim of this study was to isolate and characterize manganese（Ⅱ）-oxidizing bacteria from surrounding area of manganese mine. [Methods] Mn（Ⅱ）-oxidizing strains were isolated based on the ability to produce brown Mn oxides on agar plates. The presence of Mn oxides was confirmed by using the leucoberbelin blue（LBB） assay. The isolate was identified by morphological and physiological characterization and sequence analyses of16 S rRNA gene, gyr B gene and gyr A gene. The phylogenetic relationship between the isolated strain and reported Mn（Ⅱ） oxidizers was also analyzed. LBB assay was used to indicate the kinetics of Mn（Ⅱ） oxides formation. The Mn oxides morphology and chemical contents were determined by scanning electron microscope with energy dispersive spectrometer and X-ray diffraction. [Results] An isolate, named strain CP133, with high manganese oxidizing activity was obtained and identified as Bacillus cereus. There were some phylogenetic differences between strain CP133 and other Mn（Ⅱ）-oxidizing Bacillus species isolated from deep sea and soils. Strain CP133 produced amorphous manganese oxides that adhered to spore surface after the stationary phase of the cell culture. About 0.3 mmol/L manganese oxides was obtained after 12 days. [Conclusion] A Mn（Ⅱ）-oxidizing Bacillus cereus was successfully isolated. Our results suggest the diversity of Mn（Ⅱ）-oxidizing Bacillus species, and help understanding biogeochemical cycles in manganese mine and surrounding soils.

关 键 词：锰氧化菌 生物氧化锰 蜡样芽孢杆菌 土壤 LBB显色法 

分 类 号：Q93[生物学—微生物学]