湖底表层沉积物中蓝藻定量检测方法研究

发布时间：2018-05-29 05:38

本文选题：越冬蓝藻 + 冷冻干燥法　；参考：《合肥工业大学》2017年硕士论文

【摘要】：本文以室内培养藻与野外水华蓝藻为研究对象,以提取藻蓝蛋白的浓度为指标,以荧光检测法为手段,通过对破碎方法和提取剂的选择,分别探究了水样中蓝藻生物量与藻蓝蛋白浓度的关系以及沉积物样品中蓝藻生物量与藻蓝蛋白浓度的关系。实验结果表明:水样中通过对藻屑的显微镜检及控制相同实验条件下藻蓝蛋白的提取效率可知,冻干法对蓝藻细胞的破碎效率高于反复冻融法。控制破碎方法及其他实验条件不变时,对比去离子水、磷酸盐溶液和Tris试剂对藻蓝蛋白的提取效率发现,去离子水对藻蓝蛋白的提取效率最高。在反复冻融实验中,冷冻温度与提取剂添加顺序,对蓝藻细胞破碎率无显著影响。在冻干实验中,蓝藻真空抽滤在滤膜上能提高藻蓝蛋白的破碎率。采用荧光检测法测定藻蓝蛋白时,高效的前处理技术不仅为在湖泊环境监测中能实现快速准确的定量蓝藻的生物量提供可能,同时对蓝藻水华的预防和控制也起到积极作用。对沉积物样品检测发现,相同实验条件下,对沉积物中蓝藻细胞进行冻干破碎和反复冻融破碎,所提取的藻蓝蛋白浓度没有明显差别,两种破碎方法均可选用。其中沉积物含水率对实验结果影响很大,由于沉积物颗粒对藻蓝蛋白的吸附与掩蔽作用,沉积物的含水率越高,藻蓝蛋白提取效率就越高。提高沉积物含水率是提高实验准确定的重要措施。沉积物中蓝藻生物量与藻蓝蛋白的浓度具有良好的线性相关性,可以通过对沉积物中藻蓝蛋白的测定计算蓝藻生物量。对比显微镜计数法、流式细胞仪计数法和藻蓝蛋白间接定量方法可知,显微镜和流式细胞仪只能对水样中蓝藻生物量进行准确计数,对于沉积物中的蓝藻却无计可施。而通过藻蓝蛋白间接定量蓝藻细胞不仅可以准确计算水样中蓝藻的生物量,还能准确计算出沉积物中蓝藻的生物量。并且实验结果稳定性好,准确率高。
[Abstract]:In this paper, indoor culture algae and field Shui Hua cyanobacteria were used as research objects, the concentration of phycocyanin was taken as the index, fluorescence detection method was used as the method, and the method of crushing and the selection of extractant were used. The relationship between cyanobacteria biomass and phycocyanin concentration in water samples and the relationship between cyanobacteria biomass and phycocyanin concentration in sediment samples were studied. The results showed that the efficiency of phycocyanin extraction in water samples was higher than that by repeated freeze-thawing method through microscopical examination and control of phycocyanin extraction under the same experimental conditions. Compared with deionized water, phosphate solution and Tris reagent, the extraction efficiency of phycocyanin was the highest in deionized water and other experimental conditions. In the repeated freeze-thaw experiment, the freezing temperature and the adding order of extractant had no significant effect on the cell breakage rate of cyanobacteria. In freeze-drying experiment, cyanobacteria vacuum filtration on the membrane can improve the breaking rate of phycocyanin. In the determination of phycocyanin by fluorescence detection, the efficient pretreatment technique not only makes it possible to realize rapid and accurate quantitative biomass of cyanobacteria in lake environment monitoring, but also plays an active role in the prevention and control of cyanobacteria Shui Hua. It was found that under the same experimental conditions, the concentration of phycocyanin extracted by freeze-drying and repeated freezing and thawing of cyanobacteria in sediment was not significantly different, and the two methods could be used. The sediment moisture content has a great influence on the experimental results. Because of the adsorption and masking of phycocyanin by sediment particles, the higher the water content of sediment is, the higher the extraction efficiency of phycocyanin is. Increasing the moisture content of sediment is an important measure to improve the accuracy of the experiment. There is a good linear correlation between cyanobacteria biomass and phycocyanin concentration in sediments, and the biomass of cyanobacteria can be calculated by the determination of phycocyanin in sediments. Compared with microscope counting method flow cytometry method and indirect quantitative method of phycocyanin we can see that microscope and flow cytometry can only accurately count the biomass of cyanobacteria in water samples but there is no way to do anything about cyanobacteria in sediment. The indirect quantification of cyanobacteria by phycocyanin can not only accurately calculate the biomass of cyanobacteria in water samples, but also accurately calculate the biomass of cyanobacteria in sediments. The experimental results are stable and accurate.
【学位授予单位】：合肥工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：X524;X832

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