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1、 硕士硕士学位论文学位论文 大萍及其碱类提取物对铜绿微囊藻生长的抑制作用研究 THE STUDY ON INHIBITION EFFECTS OF MICROCYSTIS AERUGINOSA GROWTH BY PISTIA STRATIOTES AND ITS ALKALI EXTRACTS 辛文克辛文克 哈尔滨工业大学哈尔滨工业大学 2013 年年 6 月月 国内图书分类号：TU992.3 学校代码：10213 国际图书分类号：628.3 密级：公开 工工学学硕士硕士学位论文学位论文 大萍及其碱类提取物对铜绿微囊藻生长的抑制作用研究 硕 士 研 究 生 ： 辛文克 导 师 ： 张光明教授
2、 申 请学位 ： 工学硕士 学科 ： 城市水资源 所 在 单 位 ： 市政环境工程学院 答 辩 日 期 ： 2013 年 6 月 授予学位单位 ： 哈尔滨工业大学 Classified Index: TU992.3 U.D.C:628.3 Dissertation for the Master Degree in Engineering THE STUDY ON INHIBITION EFFECTS OF Microcystis Aeruginosa GROWTH BY Pistia Stratiotes AND ITS ALKALI EXTRACTS Candidate： Wenke Xin
3、 Supervisor： Prof.Guangming Zhang Academic Degree Applied for： Master of Engineering Speciality： Urban Water Resource Affiliation： School of Muni & Evn.Eng Date of Defence： June, 2013 Degree-Conferring-Institution： Harbin Institute of Technology 哈尔滨工业大学工学硕士学位论文 - I - 摘 要 水体富营养化引起的藻类过度生长问题，被称为水环境中的“生
4、态癌症”。研究一种低成本的、高效的、环保的控藻技术，一直是研究人员追求的目标。近年来，人们逐渐发现了大型水生植物在富营养化水体修复中的优势。随着研究的深入，水生植物对藻类的化感抑制作用浮出水面。研究水生植物对水中藻类的化感抑制作用，并且开发出一种新型的绿色控藻药剂，已经成为藻类控制技术开发的一个热点。 本文通过研究大萍的生长对水中的营养盐和有机污染物的浓度影响，分析了大萍对营养盐的吸收能力和有机污染物的去除能力，得出了大萍可以有效去除水中的营养盐和有机污染物。通过大萍与铜绿微囊藻的共培养实验，分析了大萍对铜绿微囊藻的藻细胞密度、叶绿素 a 的浓度、胞外蛋白质和多糖的浓度的影响，发现了大萍对铜绿
5、微囊藻的细胞增殖具有显著的抑制作用，对藻细胞的光合作用系统产生伤害，并且对细胞膜的完整性产生破坏。 此外，本文还研究了大萍对铜绿微囊藻的化感作用。利用大萍种植水培养铜绿微囊藻，得出了大萍种植水对铜绿微囊藻的生长呈现“低促高抑”的效应。通过藻萍隔膜培养实验，发现了大萍在生长过程中，会释放化感物质。抑制铜绿微囊藻的生长。并且当有铜绿微囊藻刺激时，抑制作用会加强。同时本文还研究了大萍初始生物量对其抑藻能力的影响，利用曲线拟合的方法计算出了大萍的达到半抑制率时所需的初始生物量。 通过分析大萍与铜绿微囊藻共培养时，培养液中 TN 和 TP 的变化情况， 得出了大萍对藻类的抑制作用不是通过营养性竞争导致的
6、，而是大萍化感作用引起的。 最后，利用直接法和间接法相结合的方法，从大萍植物体内提取了对藻类生长具有明显抑制作用的碱类提取物，并且利用 GC-MS 分析得出大萍对铜绿微囊藻生长产生抑制作用的有效成份为反-（2，3-二苯甲基环丙基）甲基苯亚砜和 9-(2,2-二甲基丙苯腙)- 3,6-二氯-2,7-双-2-（二乙胺）-乙氧基芴。除了大萍之外，又选取了黄菖蒲、水蓼、菱角和芦苇 4 种水生植物，对它们的碱性提取物进行抑藻实验。利用 GC-MS 对它们的碱类提取物进行分析，发现抑制藻类生长的有效成份不是单一的，而是多种化学物质混合作用的结果。 关键词：大萍；铜绿微囊藻；碱性提取物；化感作用；水生植物
7、哈尔滨工业大学工学硕士学位论文 - II - Abstract The excessive growth of algae caused by the eutrophication of natural water, is known as “ecological cancer“. Researching a new green methods of low cost as well as high efficiency, has always been the goal of researchers to pursue. In recent years, people gradually f
8、ound the advantages of large aquatic plants aboat the restoration to the eutrophication water. With the deepening of the research, the allelopathic effects of aquatic plants on algae inhibition has surfaced. Research on aquatic plants allelopathy inhibition of algae in water, and to develop a new ty
9、pe of green reagents to remove algae, has become a hot spot of algae controlling technology. This paper studied the effect on the concentration of the nutrient and the organic matter pollution in the water by the growth of Pistia stratiotes, analyzed the nutrient absorption capacity and the pollutan
10、t removal ability of the plant, and came to it can effectively removed the nutrient and organic pollutants in the water. Through Pistia and Microcystis aeruginosa coculture experiments, the paper analyzed the influence of algal cell density and the concentration of chlorophyll a, as well as that of
11、the concentration of extracellular protein and polysaccharides by the aquatic plants, found that the plants has significant inhibitory effect on the algal cell proliferation, produces damage to the photosynthesis of algae cells, and damaged to the integrity of the cell membrane . In addition, this p
12、aper also studied the allelopathy of Pistia on Microcystis aeruginosa. cultivation of M aeruginosa by the liquor cultured with P stratiotes, the result was the liquor effect on algal growth appeared to promotive on low concentration and suppression on high. By algal and aquatic plants isolation cult
13、ivation experiments, it showed the growth of the plant suppressed effectively on the algae growth, and when there was algae stimulation, inhibition would strengthen. At the same time ,this paper also studied the effect of the aquatic plants initial biomass on its algal inhibiting ability. by curve f
14、itting ,we calculated out the required initial biomass of the plant when half inhibition rate. By analyzing change situation of TN and TP in the coculture water of plant and algae, it indicated the aquatic plants inhibitory effect on algae was not due to nutritional competition, but allelopathy. Fin
15、ally, using the direct method and indirect method, we extracted alkaline extraction,having obvious inhibitory effect on algae growth,from Pistia stratiotes. Using GC-MS,we found the effective components of inhibition was trans-(2,3- Diphenylcyclopropyl) methyl benzene sulfoxide and 9-(2,2-Dimethylph
16、enyl hydrazono)-3, 6-dichloro-2,7-bis-2-(diethylamine) ethoxy fluorene. In addition to 哈尔滨工业大学工学硕士学位论文 - III - P stratiotes, we selected Polygonum hydropipe , Trapa japonica , Acorus calamus and Phragmites australis ,4 species of aquatic plants, and worked inhibiting experiments of their alkaline extract on algal. By GC-MS analysis of their extracts, it was found activ
