摘要本文以厌氧氨氧化（anaerobic ammonium oxidation，Anammox）混培物为研究对象，探究甜菜碱添加对高温Anammox活性的短期及长期影响。45℃条件下，批次实验中甜菜碱的添加量为0.1 mM和5 mM时，Anammox活性有明显升高现象；连续流实验中甜菜碱的添加（0.1、1、2 mM）并没有提高混培物的Anammox活性，同时，结合Anammox脱氮效果等生化指标及定量PCR（quantitive PCR，qPCR）对厌氧氨氧化菌（anaerobic ammonium oxidation bacteria，AnAOB）的定量结果分析表明甜菜碱的长期添加没有提高Anammox混培物的耐高温能力，反而给Anammox混培物造成较严重的伤害。51941

毕业论文关键词：甜菜碱；高温；厌氧氨氧化活性；厌氧氨氧化菌；定量PCR

Abstract With Anammox (anaerobic ammonium oxidation) mixed culture taken as the research objective, the purpose of this study was to explore the short- and long-term effects of glycine betaine on the activity of thermophilic Anammox. Under 45℃, each Anammox activity was obviously increased with the addtion of 0.1 mM and 5 mM glycine betaine in the batch experiment. However, the addtion of glycine betaine (0.1, 1, 2 mM) didn’t improve Anammox activity of the mixed culture; meanwhile, it indicated that the long-term addtion of glycine betaine  didn’t increase the heat-resisting ability of Anammox mixed culture combined the analysis of biochemical indexes (nitrogen removal effects, heme c) with the number of AnAOB investigated by qPCR (quantitive PCR).

Keywords: glycine betaine； high temperature； anammox activity; anammox bacteria；quantitive PCR

目  录

摘    要 2

引  言 4

1、材料和方法 5

1.1材料 5

1.1.1 实验装置 5

1.1.2 菌种 5

1.1.3 模拟废水 5

1.2方法 5

1.2.1   高温条件下甜菜碱的短期添加（批次实验） 5

1.2.2  高温条件下甜菜碱的长期添加（连续流实验） 6

1.2.2.1 反应器的运行 6

1.2.2.2  理化指标的测定 6

1.2.2.3 比厌氧氨氧化活性（SAA）分析 6

1.2.2.4  样品采集和DNA提取 6

1.2.2.5  PCR扩增 7

1.2.2.6  目的条带回收 7

1.2.2.7  目的片段连接 8

1.2.2.8  连接产物的热击转化 8

1.2.2.9  蓝白斑筛选及质粒回收 8

1.2.2.10  qPCR分析 8

2、结果与分析 9

2.1甜菜碱短期添加对高温Anammox活性的影响 9

2.2 甜菜碱长期添加对高温Anammox的影响 10

2.2.1甜菜碱添加下高温Anammox混培体系的脱氮效果 10

2.2.2  甜菜碱添加对高温Anammox活性的影响 12

2.2.3  甜菜碱添加对高温Anammox混培体系中AnAOB数量的影响 13

3、讨论 15

参考文献