牛粪堆肥中氨氧化细菌群落结构及其与环境因子相关性研究  

Research of ammonia oxidizing bacterial community structure and its correlation with environmental factors in cow manure composting

作　　者：孙雪微 许修宏[1] 孟庆欣[1] 成利军[1] 张文浩[1] 门梦琪 许本姝 孙瑜[1] 

机构地区：[1]东北农业大学资源与环境学院,哈尔滨150030

出　　处：《农业环境科学学报》2017年第1期189-197,共9页Journal of Agro-Environment Science

基　　金：国家自然科学基金项目(31372351;31672469)~~

摘　　要：采用PCR-DGGE(聚合酶链反应-变性梯度凝胶电泳)和Real-time PCR技术,分析牛粪堆肥期间氨氧化细菌群落结构和数量的变化及其与理化因子的相关性。结果表明,在牛粪堆肥过程中氨氧化细菌数量变化范围为1.62×106~3.80×107copies·g-1,在第14d氨氧化细菌数量达到最高值;DGGE图谱显示,堆肥各个时期氨氧化细菌群落组成存在明显的差异;Shannon指数随着温度的变化而变化,在第7 d达到最大值2.671 8;系统发育分析表明,氨氧化细菌均隶属于β-变形菌纲不可培养的亚硝化螺菌属Uncultured Nitrosospira sp.和不可培养的亚硝化单胞菌属Uncultured Nitrosomonas sp.,其中与Uncultured Nitrosospira sp.相似性高的氨氧化细菌为优势菌,占60%左右;冗余分析(Redundancy analysis,RDA)显示,温度和铵态氮等对氨氧化细菌群落结构的影响较大。研究表明在牛粪堆肥期间氨氧化细菌群落结构发生了明显的变化,同时证实了氨氧化细菌群落结构的变化受堆肥理化因子的影响。The dynamic succession of ammonia oxidizing bacterial community structure and quantity were investigated using PCR-DGGE（polymerase chain reaction and denaturing gradient gel electrophoresis）and real-time PCR, the correlations between ammonia oxidizing bacterial community structure and physic-chemical factors were evaluated using redundancy analysis（RDA） in cow manure composting.The results showed that during composting the range of ammonia oxidizing bacteria were 1.62×106~3.8×107copies·g-1, the amount of ammonia oxidizing bacteria reached the peak on 14 day; DGGE profile analysis showed that the community composition of ammonia oxidizing bacteria obviously varied in different periods of composting. Shannon index of ammonia oxidizing bacteria changed with composting temperature, reaching the maximum（2.671 8）on the 7th day. Phylogenetic analysis showed that ammonia oxidizing bacteria were clustered into two groups, Uncultured Nitrosomonas sp. and Uncultured Nitrosospira sp., which belonged to the β-Proteobacteria. the Uncultured Nitrosospira sp. which accounted for about 60% of ammonia oxidizing bacteria were predominant. RDA showed that ammonia and temperature（P〈0.05）effected significantly on the community structure of ammonia oxidizing bacteria. Significant changes of ammonia oxidizing bacterial community structure occurred in cow manure composting, and it is confirmed that the changes of ammonia oxidizing bacterial community structure were affected by environmental factors.

关 键 词：牛粪堆肥 PCR-DGGE REAL-TIME PCR 氨氧化细菌 冗余分析 

分 类 号：X713[环境科学与工程—环境工程]