论文总字数：19904字

摘 要

离子液体（ILs）是不对称催化、有机化学、电化学等领域应用较多的一类化合物，并且具有非常好的应用潜力^[1]。但由于某些离子液体的水溶性和缓慢降解的特性使其对水环境造成一定影响。近年来，离子液体对微藻的毒性反应的研究已多有报道^[2]，本课题以处于一定浓度下的微藻为研究对象，分析了离子液体对微藻不同方面的影响^[3]。

微藻是淡水及海水中均存在的微生物，其生长状况能直接反应水质情况，可以用来检测环境污染。离子液体全部由阴离子和阳离子组成。离子液体是绿色的化工溶剂，由于离子液体没有明显的蒸汽压，可以减少对大气的污染以及对操作人员的危害，因而又有“绿色溶剂”之称。如今，离子液体由于具有许多传统溶剂所不具备的优点而正在逐渐取代传统的有机溶剂，并在多个领域普及。但随着研究的深入，人们发现离子液体对动物、植物、酶、藻类等具有一定的毒性。本课题将探讨微藻在不同浓度离子液体下培养时其生长速率、活性氧水平、叶绿素含量和细胞超微结构的变化等情况。

本课题选择离子液体(EMIM)采用摇瓶来培养鱼腥藻和小球藻，以4天为一个周期，每24h取样测定。采用UV来进行测定，并分别对时间作图，得到了它们的生长曲线和叶绿素含量变化曲线。

关键词：微生物培养，HPLC 分析，紫外光度计分析，数理统计。

Physiological response of microalgae to ionic liquids

Abstract

Ionic liquids (ILs) are a class of compounds that are widely used in asymmetric catalysis, organic chemistry, electrochemistry and other fields, and have very good application potential. However, due to the water solubility and slow degradation characteristics of some ionic liquids, it has a certain impact on the water environment. In recent years, there have been many reports on the toxic reactions of ionic liquids on microalgae. This subject takes microalgae at a certain concentration as the research object, and analyzes the effects of ionic liquids on different aspects of microalgae, such as growth rate, chlorophyll content, etc.

Microalgae are microorganisms present in both freshwater and seawater. Their growth status can directly reflect the water quality and can be used to detect environmental pollution. Ionic liquids are composed entirely of anions and cations.Ionic liquid is a green chemical solvent. Because ionic liquid has no obvious vapor pressure, it can reduce the pollution to the atmosphere and the harm to the operator, so it is also called "green solvent". Nowadays, ionic liquids are gradually replacing traditional organic solvents due to many advantages that traditional solvents do not have, and are popular in many fields. But with the deepening of research, it was found that ionic liquids have certain toxicity to animals, plants, enzymes, algae, etc. This topic will discuss the growth rate, active oxygen level, chlorophyll content and changes in cell ultrastructure of microalgae when cultured in ionic liquids with different concentrations.

In this project, ionic liquids (EMIM) were selected to cultivate Anabaena and Chlorella using shake flasks, taking 4 days as a cycle and sampling every 24h. UV was used for the measurement, and the time was plotted separately to obtain their growth curve and chlorophyll content change curve.

Keywords:Microbial culture, HPLC analysis, UV photometer analysis, mathematical statistics

目 录

摘 要 I

Abstract II

第一章 文献综述 1

1.1微藻 1

1.2离子液体 1

1.2.1离子液体的发展 1

1.2.2离子液体的性质 2

1.2.3离子液体的分类 2

1.3离子液体对微藻的毒性 3

1.3.1生长速率 3

1.3.2光合作用与叶绿素含量 4

1.3.3细胞膜通透性与细胞形态 4

1.3.4 ROS水平及抗氧化酶 5

1.4影响离子液体对微藻毒性的因素 5

1.4.1阳离子烷基链长度 5

1.4.2阴离子 6

1.4.3手性 7

1.5总结与展望 7

第二章 实验内容 9

2.1实验仪器及试剂 9

2.1.1实验所用主要试剂 9

2.1.2实验所用主要仪器 9

2.2实验方法 10

2.2.1 BG11培养基的配制与灭菌 10

2.2.2接种藻种及其细胞密度和叶绿素含量的测定 10

2.2.3藻种的培养及其受离子液体的影响的测定 10

2.2.4鱼腥藻叶绿素含量的计算 11

2.2.5小球藻叶绿素含量的计算 11

2.3实验结果 11

2.3.1鱼腥藻 11

2.3.2小球藻 14

2.4实验结论 18

2.4.1离子液体对微藻生长情况的影响 18

2.4.2离子液体对微藻叶绿素含量的影响 18

2.5讨论 19

参考文献 24

致 谢 26

第一章 文献综述

1.1微藻

微藻并不是分类学上的一个专属名称，而是对一类细胞粒径微小、只能用显微镜才能看清其形态特征的藻类的统称。该类藻细胞可以通过光合作用来满足自身生长所需的各种能量，它们非常微小，有应用价值，可以应用生物技术进行大量生产或培养，本次实验用到了两种微藻，分别是鱼腥藻（蓝藻）和小球藻（绿藻）。相对而言，小球藻对外界压力的耐受性要高于鱼腥藻，可能能够承受更高浓度的离子液体。

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