摘要本实验利用了异养氨氧化细菌培养基,从友谊河淤泥中分离、纯化出一支异养氨氧化细菌(未知菌X),在好氧时,其具有将铵离子氧化为亚硝酸盐的氨氧化能力,厌氧时具有反硝化能力。定量测定了5天内,铵离子及亚硝酸盐浓度随时间的变化情况。结果表明了该菌在5天内对铵离子的降解能力达到了58%。定性地测定了其降解有机氮源地能力,发现其在LB培养基内可以将有机氮转变为亚硝酸盐,其存在于自然环境中是氮元素循环不可缺少的一员。对该菌进行了基因组整DNA的提取,16SrDNA的扩增并测序,但是16SrDNA的鉴定的结果尚未得到。最终对其进行了零下80度的保藏63957
毕业论文关键词 异养氨氧化细菌 氨氮 亚硝酸盐 16SrDNA
毕业设计说明书(论文)外文摘要
Title The Isolation and Identification Of Heterotrophic Ammonia Oxidizing Bacteria .
Abstract In this research, we utilized the culture medium for heterotrophic ammonia oxidizing bacteria (AOB) to shuffle and isolate one strain of heterotrophic AOB, AOB-X, from the silts of Youyi River, which had ability to oxidizing ammonia to nitrite when exposed to air, while that could carry out anaerobic denitrification. We quantitatively measured the change of concentration of ammonia and nitrite of the culture for 5 days.In 5 days the ability of oxidizing of ammonia was about 58%. Also we measured qualitatively its ability of oxidizing of organic nitrogen and found its ability of generation of nitrite. Then we extracted the DNA of AOB-X,after that,16SrDNA was amplified by PCR and sequenced. However we did not got the result of sequencing. Then I preserved in the -80 centigrade refrigerator.
Keywords heterotrophic ammonia oxidizing bacteria ,ammonia, nitrite ,16SrDNA
目 次
1 绪论 1
1 . 1 研究背景 1
1 . 2 氨氧化细菌(Ammonia Oxidizing Bacteria ,氨氧化细菌) 3
1. 2. 1 氨氧化细菌的简介及其研究进展 3
1. 2. 2 各种因素对于氨氧化细菌在土壤中群落分布的影响 4
1. 2. 2. 1 氨浓度的影响 4
1. 2. 2. 2 pH的影响 4
1. 2. 2. 3 温度的影响 5
1 . 3 实验分析方法原理 5
1. 3. 1氨氮的测定——水杨酸-次氯酸法 5
1. 3. 2亚硝酸盐氮的测定——格里斯试剂分光光度法 5
2 实验内容及方法 5
2.1实验材料及仪器 5
2. 2 实验方法和步骤 7
2. 2. 1 培养基 7
2. 2. 2 实验样本采集 7
2. 2. 3 待分离菌种的筛选 7
2. 3 菌种的鉴定 8
2. 3. 1 未知菌X的单菌落的分离、验证及富集培养。 8
2. 3. 2 未知菌X的基因组整DNA的提取 8
2. 3. 3 未知菌X的基因组整DNA提取的验证 9
2 .3. 4 未知菌X的16SrDNA的PCR 9
2. 3. 5 16S rDNA 的复验与切胶回收