山仔水库两种水华蓝藻对不同形态磷吸收及响应机制研究

发布时间：2018-05-20 23:21

本文选题：水华蓝藻 + 吸收动力学　；参考：《福建师范大学》2015年硕士论文

【摘要】：福建省山仔水库自1992年建库以来,承担着福州地区农田灌溉、防洪、发电的功能,更是福州地区居民的第二大饮用水源地,水库发挥的作用不可估量。然而,由于人类的生产活动和生活方式的改变,山仔水库自1997年首次发现蓝藻以来,富营养化引起的蓝藻水华现象便屡屡发生。水华产生的微囊藻毒素可危及饮用水的安全和人类的健康。因此,山仔水库水华蓝藻种类的分类属性,是否产毒,以及对主要营养盐的吸收与生理响应机制研究,对于进一步揭示山仔水库蓝藻水华成因以及有毒蓝藻水华的预警监测,具有现实意义。本论文的主要内容有：(1)对山仔水库主要水华蓝藻进行分离纯化,在形态学鉴定的基础上,进一步以16SrRNA基因为分子标记对分离获得的2株蓝藻进行分子鉴定。鉴定结果为铜绿微囊藻(Microcystis aeruginosa B1-14)和浮游鱼腥藻(Anabaena planctonica B1-15);(2)利用全细胞PCR法和高效液相法对这两株蓝藻进行微囊藻毒素测定。发现分离的藻株Aicrocystis aeruginosa B1-14(缩写“M.aeruginosa B1-14")有微囊藻毒素基因mcyA和mcyB并且能产生MC-LR,藻株Anabaena planctonica B1-15缩写“A.planctonica B1-15")不含这些基因和毒素,故铜绿微囊藻Microcystis aeruginosa B1-14株能够产生微囊藻毒素,而浮游鱼腥藻Anabaena planctonica B1-15株不产毒；(3)研究两株水华蓝藻在不同形态磷4种无机磷和2种有机磷)条件下的生长效应。通过测定比生长速率、胞外总磷含量及碱性磷酸酶活性,比较两者对不同形态磷的摄取能力并探讨其响应机制。结果显示,两种水华蓝藻对正磷酸盐(磷酸氢二钾,磷酸二氢钾)的吸收速率和碱性磷酸酶酶活性均大于多聚磷酸盐(焦磷酸钠,聚磷酸钠)和有机磷盐(ATP,甘油磷酸钠),表明正磷酸盐是两种水华蓝藻生长的最适磷盐,但在碱性磷酸酶的作用下,能够吸收部分聚磷和有机磷。M.aeruginosaB1-14对磷的吸收速率大于A.planctonica,B1-15,但A.planctonica B1-15的碱性磷酸酶活性高于M.aeruginosa B1-14,证明了M.aeruginosa B1-14具有过量吸收积累磷的特性以及藻细胞产生碱性磷酸酶的能力与其表面积和体积之比正相关；(4)在上述研究基础上,分别选择正磷酸盐(磷酸氢二钾)和ATP为磷源,研究两株水华蓝藻的磷吸收动力学特征,并通过动力学参数比较,探讨两株蓝藻的竞争能力和可能的优势演替顺序。结果表明,在不同磷质量浓度下,两种藻对磷有明显反应的低限浓度均为0.54 mg·L1,在初始磷浓度为054-5.4mg·-1范围内,M.aeruginosa B1-14的生长速率随磷浓度的提高而明显增加,相同条件下,A.planctonic B1-15比生长速率的增加不明显且低于M.aeruginosa B1-14的生长速率。大于5.4 mg·L-1时,两种藻的生长速率不再增加。无机磷条件下,M.aeruginosaB1-14的最大比生长率和半饱和常数分别为0.18418d-1和0.079785mg.L-1,大于A.planctonica B1-15的最大比生长速率(0.12825d-1)和半饱和常数(0.032230mg·L-1)据此推断,当无机磷浓度大于054mg·L-1时,M.aeruginosa B1-14易成为优势藻,反之,A.planctonica B1-15成优势藻。有机磷条件下,两种藻的半饱和常数和最大比生长率均较小,且差异不明显。此也证实两株蓝藻主要吸收可溶性无机磷,但可以利用有机磷；(5)研究两种水华蓝藻问的竞争作用和相互化感作用。结果为竞争条件下M.aeruginosa B1-14 比A.planctonica B1-15更具生长优势,A.plantonica B1-15滤液对M.aeruginosa B1-14的的生长的促进作用大于M.aeruginosa B1-14滤液对A.planctonica B1-15的作用；(6)研究常见的有机磷农药对两种水华蓝藻生理生化的影响。在EC50下,M.aeruginosa B1-14对有机磷农药的敏感强度大小依次为：乐果辛硫磷草甘膦；A.planctonica B1-15对有机磷农药的敏感强度大小依次为：乐果草甘膦辛硫磷,且M.aeruginosa B1-14 比A.planctonica B1.15敏感。
[Abstract]:Since the establishment of the reservoir in Fujian Province in 1992, Fujian province has undertaken the functions of farmland irrigation, flood control and power generation in Fuzhou area. It is also the second major drinking water source of residents in the region of Fuzhou. The function of the reservoir is immeasurable. However, the reservoir has been rich since the first discovery of blue algae since the first discovery of blue algae in 1997 because of the changes in human production activities and life style. The cyanobacteria blooms caused by the cultivation are frequently occurring. The microcystins produced by the water bloom can endanger the safety of drinking water and human health. Therefore, the classification of the species of cyanobacteria in the mountain Zai reservoir, whether it is poisonous, and the mechanism of the absorption and physiological response to the main nutrients are studied to further reveal the cyanobacteria bloom in Sam reservoir. The causes and early warning monitoring of toxic cyanobacteria bloom are of practical significance. The main contents of this paper are as follows: (1) to separate and purify the main cyanobacteria of szai reservoir, and on the basis of morphological identification, the molecular identification of 2 cyanobacteria separated by 16SrRNA gene is further identified. The results are Microcystis aeruginosa (Microcystis aeruginosa). Microcystis aeruginosa B1-14) and planktonic Anabaena (Anabaena planctonica B1-15); (2) the microcystins were measured by the whole cell PCR method and high performance liquid phase method. It was found that the isolated algal strain Aicrocystis aeruginosa B1-14 (abbreviated "M.aeruginosa B1-14") had the microcystin gene and could produce the toxin. The Anabaena planctonica B1-15 abbreviation of the algae plant "A.planctonica B1-15") does not contain these genes and toxins, so Microcystis aeruginosa Microcystis aeruginosa B1-14 strain can produce microcystin, and the Anabaena planctonica B1-15 strain of Anabaena Anabaena is not produced. (3) two species of cyanobacteria in different forms of phosphorus and 2 kinds of organic phosphorus are studied. The uptake of different forms of phosphorus and its response mechanism were compared by measuring the ratio of growth rate, total phosphorus content and alkaline phosphatase activity. The results showed that the absorption rate and alkaline phosphatase activity of two species of cyanobacteria (two potassium hydrogen phosphate and potassium two hydrogen phosphate) were both large. In polyphosphate (sodium pyrophosphate, sodium polyphosphate) and organophosphorus salts (ATP, sodium glycerophosphate), Biao Mingzheng phosphate is the most suitable phosphorus salt for the growth of two kinds of cyanobacteria. But under the action of alkaline phosphatase, the absorption rate of partial phosphorus and organophosphorus.M.aeruginosaB1-14 is greater than A.planctonica, B1-15, but A.planctonica B1-1 5 alkaline phosphatase activity was higher than M.aeruginosa B1-14, which proved that M.aeruginosa B1-14 had the characteristics of excessive absorption and accumulation of phosphorus, and the ability of algal cells to produce alkaline phosphatase was positively related to the ratio of surface area and volume. (4) on the basis of the above study, we selected two strains of orthophosphate (two potassium hydrogen phosphate) and ATP as the phosphorus source respectively. The kinetic characteristics of phosphorus absorption of cyanobacteria were studied, and the competitive ability of two cyanobacteria and the possible dominant succession sequence were discussed by comparison of kinetic parameters. The results showed that under different phosphorus concentrations, the low limiting concentration of two kinds of algae to phosphorus was 0.54 mg. L1, and the initial phosphorus concentration was 054-5.4mg -1, M.aeruginosa B1- The growth rate of 14 increased obviously with the increase of phosphorus concentration. Under the same condition, the growth rate of A.planctonic B1-15 was not obvious and lower than the growth rate of M.aeruginosa B1-14. The growth rate of two kinds of algae was no longer increased when it was greater than 5.4 mg. L-1. The maximum specific growth rate and semi saturation constant of M.aeruginosaB1-14 under the inorganic phosphorus condition. The maximum specific growth rate (0.12825d-1) and semi saturation constant (0.032230mg L-1) of A.planctonica B1-15, respectively 0.18418d-1 and 0.079785mg.L-1, are deduced. When the concentration of inorganic phosphorus is greater than 054mg. L-1, M.aeruginosa B1-14 becomes the dominant algae. On the contrary, A.planctonica becomes the dominant algae. Under the condition of organophosphorus, the half full of the two kinds of algae The two strains of cyanobacteria mainly absorb soluble inorganic phosphorus, but they can use organophosphorus, and (5) study the competition and Allelopathy of the two species of cyanobacteria. The result is that M.aeruginosa B1-14 has a better growth advantage than A.planctonica B1-15 under the competitive condition, A.plant The effect of onica B1-15 filtrate on the growth of M.aeruginosa B1-14 is greater than the effect of M.aeruginosa B1-14 filtrate on A.planctonica B1-15; (6) the study of the physiological and biochemical effects of common organophosphorus pesticides on two species of cyanobacteria. At EC50, the sensitivity of M.aeruginosa B1-14 to organophosphorus pesticides is in sequence: dioxin sulphur. The sensitivity of A.planctonica B1-15 to organophosphorus pesticides is: glyphosate parathion in dimethoate, and M.aeruginosa B1-14 is more sensitive than A.planctonica B1.15.
【学位授予单位】：福建师范大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：X524

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