本文關(guān)鍵詞： 融合菌株F14 多環(huán)芳烴 細(xì)胞膜 微生物降解 土壤修復(fù)　出處：《中北大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：多環(huán)芳烴(PAHs)具有較大的固水分配系數(shù)且生物可利用性偏低,容易對(duì)土壤造成大面積污染,使人類活動(dòng)及生態(tài)健康遭受不良影響可能性增大。土壤PAHs污染已成為主要生態(tài)問題,PAHs的修復(fù)越來越引起人們的重視。融合菌株F14是一株高效降解PAHs的菌株,由假單胞菌和鞘氨醇單胞菌作為親本,利用原生質(zhì)體融合技術(shù)融合而成。為進(jìn)一步了解降解菌F14對(duì)PAHs代表物菲的降解機(jī)制,本文從微生物細(xì)胞膜的角度考察了菌株降解菲過程中細(xì)胞膜上表面物質(zhì)的變化情況。利用室內(nèi)模擬的方法,探討了融合菌株F14對(duì)土壤環(huán)境中PAHs代表物菲的降解特性及其影響因子并分析了土壤酶活性以及降解菌的生長(zhǎng)情況。分別以校園內(nèi)清潔土壤和焦化廠廠區(qū)內(nèi)土壤為研究對(duì)象,考察菌株F14在PAHs污染土壤中的降解性能,為更好地修復(fù)土壤環(huán)境中的PAHs污染提供理論依據(jù)。主要研究結(jié)果如下:(1)細(xì)胞膜表面脂多糖、磷脂及脂質(zhì)過氧化物與降解菲的濃度具有密切的關(guān)系,隨著菲的濃度增大,磷脂及脂多糖含量也隨之增大,當(dāng)濃度大于150 mg/L時(shí),磷脂含量減小,脂多糖含量繼續(xù)增大,丙二醛的含量隨著菲濃度的增大而增大。融合菌株F14具有較高的表面疏水性,處于對(duì)數(shù)期以及穩(wěn)定期的F14表面疏水率達(dá)到69.7%,65.2%。這對(duì)污染物的去除有很強(qiáng)的促進(jìn)作用。(2)單因素實(shí)驗(yàn)研究了含水率、接種量及添加營(yíng)養(yǎng)元素對(duì)菌株F14降解土壤中的污染物菲的影響情況。結(jié)果表明:菌株F14對(duì)土壤中菲的降解效果較好,且在接種量為2 m L(濃度0.5 g/mL)、含水率約為60%的條件下最適合菌種生長(zhǎng),C:N:P比例對(duì)菌株F14的降解作用影響并不明顯。在菲的污染水平為100 mg/kg時(shí),仍有大約30%的菲被降解,說明菌株F14至少能耐受環(huán)境中100 mg/kg的污染水平。(3)為探究F14對(duì)菲降解過程中生物反應(yīng)活性,測(cè)定土壤中過氧化氫酶、多酚氧化酶(polyphenol oxidase,PPO)和脫氫酶(Dehydrogenase,DHA)的活性,結(jié)果表明,土壤中菲的添加刺激了DHA和PPO,對(duì)過氧化氫酶活性無刺激作用。此外,融合菌株F14可以在土壤中存活,菌數(shù)量隨著菲的降解有不同變化。在菲污染土壤中檢測(cè)到的F14的數(shù)量大于無菲土壤中檢測(cè)到的菌數(shù)量。(4)為了探討菌株F14對(duì)焦化土壤中PAHs的微生物修復(fù)可行性,應(yīng)用菌株F14對(duì)PAHs工業(yè)污染土壤進(jìn)行修復(fù)實(shí)驗(yàn)。結(jié)果表明,接種F14對(duì)PAHs的修復(fù)有促進(jìn)作用,降解60d后,PAHs總降解率平均達(dá)到42%;接種F14對(duì)2-3環(huán)PAHs降解率的提高更加明顯,低環(huán)PAHs的平均降解率大于空白對(duì)照組12個(gè)百分點(diǎn)。
[Abstract]:Polycyclic aromatic hydrocarbons (PAHs) have a large solid water partition coefficient and low bioavailability, which can easily pollute the soil in a large area. The possibility of adverse effects on human activities and ecological health is increased. Soil PAHs pollution has become the main ecological problem. More and more attention has been paid to the repair of PAHs. The fusion strain F14 is a highly efficient PAHs degrading strain with Pseudomonas sphingomonas and sphingomonas as parents. In order to further understand the degradation mechanism of PAHs phenanthrene by using protoplast fusion technology. In this paper, the changes of surface matter on the membrane of phenanthrene in the process of degrading phenanthrene were investigated from the point of view of microbial cell membrane. The degradation characteristics and influencing factors of phenanthrene (phenanthrene), a representative of PAHs in soil environment, were studied by the fusion strain F14. The enzyme activity of soil and the growth of degrading bacteria were analyzed. The clean soil and coking plant were used to clean the campus. The soil in this area is the object of study. The degradation of strain F14 in PAHs contaminated soil was investigated to provide theoretical basis for better remediation of PAHs pollution in soil environment. The main results were as follows: 1) lipopolysaccharide on cell membrane. The concentration of phospholipid and lipid peroxide was closely related to the concentration of degrading phenanthrene. The content of phospholipid and lipopolysaccharide also increased with the increase of the concentration of phenanthrene. When the concentration was more than 150 mg/L, the content of phospholipid decreased. The content of lipopolysaccharide continued to increase, and the content of malondialdehyde increased with the increase of phenanthrene concentration. The fusion strain F14 had higher surface hydrophobicity. The surface hydrophobic rate of F14 in logarithmic period and stable period reached 69.7% and 65.2%, which has a strong promoting effect on the removal of pollutants. The results showed that strain F14 had a better effect on the degradation of phenanthrene in soil. Under the condition of inoculation amount of 2 mL (concentration 0.5 g / mL) and moisture content of about 60%, it is most suitable for strain growth. The effect of C: n: P ratio on the degradation of strain F14 was not obvious. When the pollution level of phenanthrene was 100 mg/kg, about 30% of phenanthrene was still degraded. The results showed that strain F14 could at least tolerate the pollution level of 100 mg/kg in the environment. In order to study the bioactivity of F14 in the degradation of phenanthrene, the catalase in soil was determined. The activities of polyphenol oxidase (PPO) and dehydrogenase (DHA) were studied. The addition of phenanthrene in soil stimulated DHA and PPO, but did not stimulate the activity of catalase. In addition, the fusion strain F14 could survive in soil. The amount of F14 detected in phenanthrene contaminated soil was higher than that in non-phenanthrene soil. In order to investigate the feasibility of microbial remediation of PAHs in coking soil by strain F14. Strain F14 was used to repair PAHs contaminated soil. The results showed that inoculation of F14 could promote the restoration of PAHs and degrade it for 60 days. The average degradation rate of PAHs was 42%. The degradation rate of 2-3 ring PAHs was significantly increased by inoculation with F14, and the average degradation rate of low ring PAHs was 12 percentage points higher than that of control group.
【學(xué)位授予單位】：中北大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：X53;X172

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