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1、降萘菌株 DQ-1 的种子培养组分优化摘要随着人类活动对土壤环境的干扰日益加剧,工农业生产常可导致土壤污染也越来越受到人们的高度关注。多环芳烃就是其中的一种环境污染物,具有致癌性和致突变性。其中萘是被美国环保署列为优先控制污染物的 16 种多环芳烃之一，具有长效性，不可挥发的污染物。目前萘污染相当严重，工业排放和大量燃烧使得土壤中含有大量萘。生物修复(Bioremediation)是新兴的污染治理方法，是利用生物特别是微生物将环境中的污染物降解或转化为其他无害物质的过程。世界上几乎每一种化学物质，都有相应的细菌能将其分解，利用细菌来消除污染即生物修复是一种高效、经济且生态可承受的清洁技术。萘作
2、为研究对象，从污染土壤中筛选到一株萘降解活性较高的细菌菌株，经菌种鉴定，定名为 Pseudomonas stutzeri DQ-1。研究表明，利用乙醇无机盐培养基培养液体种子是适宜的, 菌体浓度可达 108CFU/mL。本论文以 DQ-1 为实验材料，乙醇为种子液培养底物，先制作 DQ-1 的生长曲线，再对它的培养基各组分进行单因素优化实验，筛选出种子生长的最适条件。研究了不同乙醇浓度、不同氮源、氮源浓度、不同磷源、磷源总量及不同酵母粉浓度对 DQ-1种子液生长的影响,结果表明：乙醇浓度 1%,氮源为(NH 4)2SO4 ,氮源浓度0.06mol/L，磷源比例(加上磷源种类)1:1，磷总量 0
3、.03mol/L 时为最佳培养条件。关键词：萘降解，乙醇，单因素，生物修复Drop naphthalene strains DQ-1 A seed culture of component optimizationABSTRACTWith the interference of human activities on soil environment is becoming more and more serious, the industrial and agricultural production can often lead to soil pollution has become i
4、ncreasingly subject to peoples attention. A kind of environmental pollutants is one of the polycyclic aromatic hydrocarbons, with carcinogenicity and mutagenicity. The United States Environmental Protection Agency which napHthalene was listed as one of 16 priority pollutant polycyclic aromatic hydro
5、carbons, and long, not volatile pollutants. At present, the napHthalene pollution is very serious, industrial emissions and combustion makes the soil contains large amounts of napHthalene.Bioremediation (Bioremediation) is a new method of pollution control, is to use biology especially microbial pol
6、lutants in the environment degradation or converted to other harmless substances. Almost every kind of chemical substances on the world, have corresponding bacteria can decompose them, to eliminate the pollution that bioremediation is a kind of high efficient, economic and ecological sustainable cle
7、an technology using bacteria.NapHthalene as the research object, screened from the polluted soil to a bacterial strain with high activity of napHthalene degradation, the identification of bacteria, called Pseudomonas stutzeri DQ-1. Research shows that, by using ethanol inorganic salt culture medium
8、liquid seed is appropriate, the cell concentration reached 108CFU/mL. In this paper, DQ-1 as experimental material, ethanol as seed liquid culture medium, the growth curve to make DQ-1, and medium groups on the technology was optimized by single factor experiment, selected the optimum conditions of
9、seed growth. Effects of different ethanol concentration, different nitrogen sources, nitrogen source concentration, different pHospHorus source, pHospHorus source volume and different concentration of yeast powder, DQ-1 seed liquid growth results show that: ethanol concentration 1%, (NH4) 2SO4 as ni
10、trogen source, nitrogen source concentration ratio (0.06mol/L, P Sources and types of pHospHorus source) 1:1, as the optimum culture conditions of total pHospHorus 0.03mol/L.Key words: Soil NapHthalene Pollution DQ-1, Ethanol, Single Factor, Degradation, Bacteria Concentration目 录前 言 .1第一章 文献综述 .21.
11、土壤多环芳烃的污染 .21.1.1 概述 .21.1.2 土壤多环烃污染的来源 .21.1.3 土壤多环芳烃污染状况 .31.1.4 土壤多环芳烃的危害 .31.2 多环芳烃降解细菌的种类 .41.3 土壤多环芳烃污染修复技术.41.3.1 物理方法修复 .51.3.2 化学方法修复 .61.3.3 生物方法修复 .61.3.4 植物方法修复.71.4 多环芳烃的细菌对萘降解 .71.5 微生物修复的优点和它的局限性.81.6 多环芳烃污染的生物修复和它的应用前景 .81.6.1 生物修复.91.6.2 生物修复的应用前景91.7 本实验研究的内容 .9第二章 实验部分 .132.1 前言 .132.2 材料与方法 .132.2.1 泥样与菌株 .132.2.2 材料与培养基 .132.2.3 试剂与仪器 .142.3 菌种的分离筛 .172.3.1 降萘菌株的筛选 .172.4 16SrDNA 的扩增培养基组分的研究.182.4.1 小量提细取菌 DNA.182.4.2 种子液碳源的确定182.5 种子培养组分的优化
