【摘要】：鄰苯二甲酸二丁酯(DBP)作為一種增塑劑,被廣泛的應(yīng)用于塑料制品中。近年來(lái),我國(guó)農(nóng)膜的使用量不斷增加,由于塑料農(nóng)膜的穩(wěn)定性差,極易導(dǎo)致PAEs從塑料薄膜中釋放。因此,越來(lái)越多的DBP進(jìn)入到周邊土壤中,導(dǎo)致農(nóng)業(yè)用地污染。另外,由于化肥、污灌等的不當(dāng)使用,也會(huì)使土壤中DBP的含量有所增加。受DBP污染土壤的影響,導(dǎo)致種植的作物對(duì)DBP的積累,進(jìn)而通過(guò)食物鏈進(jìn)入到人體中,對(duì)人體內(nèi)分泌、神經(jīng)系統(tǒng)等產(chǎn)生危害,因此土壤中DBP污染的防治刻不容緩。目前相關(guān)的研究主要集中在DBP降解菌的篩選及特性機(jī)理研究,而DBP降解菌功能菌群制劑的研發(fā)等相關(guān)方面的研究較少。微生物修復(fù)技術(shù)具有高效、成本低、無(wú)二次污染、操作簡(jiǎn)便且處理效果好等優(yōu)點(diǎn)。本試驗(yàn)旨在研發(fā)一種高效且協(xié)同修復(fù)效果良好的功能菌劑,在修復(fù)DBP污染土壤的同時(shí),又能夠?qū)ξ廴就寥婪柿M(jìn)行改良。通過(guò)盆栽試驗(yàn),考察修復(fù)菌劑對(duì)小白菜生長(zhǎng)的影響,進(jìn)一步確定功能菌劑修復(fù)DBP污染土壤效果及其對(duì)土壤肥力改良效果,考察功能菌劑對(duì)土壤生態(tài)安全的影響,探討DBP微生物修復(fù)技術(shù)的實(shí)際應(yīng)用價(jià)值,為PAEs污染土壤的修復(fù)提供科學(xué)依據(jù)及技術(shù)支持,為土壤有機(jī)污染的生物修復(fù)提供參考。主要研究結(jié)果如下:從土壤中篩選分離得到3株植物促生菌(解磷菌P6、解鉀菌K5、固氮菌N1),經(jīng)鑒定解磷菌為鞘脂菌(Sphingobium yanoikuyae)、解鉀菌為膠質(zhì)芽孢桿菌(Paenibacillus mucilaginosus)、固氮菌為產(chǎn)酸克雷伯氏菌(Enterobacter aerogenes)。對(duì)其各自活力進(jìn)行測(cè)定,最終確定P6的解磷能力為28.44 mg/L,K5解鉀量為22.61μg/m L,N1菌株的固氮能力為14.56 mg/L。將3株促生菌與DBP降解菌(實(shí)驗(yàn)室前期篩選所得,為假單胞菌(Pseudomonas sp.),并命名DNB-S1)進(jìn)行平板劃線以及搖瓶試驗(yàn),結(jié)果表明各菌株之間不存在拮抗反應(yīng),并且不會(huì)影響各自的活力。利用發(fā)酵培養(yǎng)基混合菌群發(fā)酵48 h,平板計(jì)數(shù)法得到固氮菌N1個(gè)數(shù)為5.3×108 cfu/m L,解磷菌P6個(gè)數(shù)為4.0×109 cfu/m L,解鉀菌K5個(gè)數(shù)為5.0×109 cfu/m L,DBP降解菌DNB-S1個(gè)數(shù)為7.6×109 cfu/m L。經(jīng)發(fā)酵的各菌株數(shù)量均在108 cfu/m L以上,發(fā)酵效果良好。以凹凸棒土、高嶺土、腐殖酸作為載體,制備功能菌劑,常溫保存?zhèn)溆谩?duì)菌劑內(nèi)活菌數(shù)測(cè)定發(fā)現(xiàn),室內(nèi)常溫保35 d時(shí),四種細(xì)菌的活細(xì)胞數(shù)量還可以保持較高的活性,保存效果良好。功能菌劑中降解菌和促生菌配比優(yōu)化試驗(yàn)表明,通過(guò)綜合考察各處理組對(duì)于污染土壤中35 d的DBP降解率、土壤肥力改良情況(速效氮、速效磷、速效鉀)以及對(duì)土壤中微生物多樣性的影響情況,最終確定功能菌劑中降解菌與促生菌的配比為1:1(g:g)的時(shí)候,為最佳的配比方式。采用溫室盆栽試驗(yàn)檢驗(yàn)功能菌劑修復(fù)DBP污染土壤(20 mg/kg)效果,通過(guò)對(duì)經(jīng)濟(jì)作物小白菜生理指標(biāo)和品質(zhì)、土壤酶活性以及土壤肥力改良的效果表明:在小白菜生長(zhǎng)過(guò)程中,添加功能菌劑的處理組葉綠素的含量顯著高于污染處理組,MDA含量也較污染處理低,菌劑的加入,明顯緩解了DBP對(duì)小白菜的毒害作用;收獲期時(shí)小白菜體內(nèi)干物質(zhì)積累量明顯高于對(duì)照,可溶性蛋白含量以及維生素C含量較污染處理組分別提升了24.93%、13.67%,提高了小白菜的品質(zhì)。在修復(fù)過(guò)程中功能菌劑對(duì)土壤脲酶、蔗糖酶活性起到了激活作用,而對(duì)過(guò)氧化氫酶活性不會(huì)產(chǎn)生很大的影響;土壤中速效氮、速效磷及速效鉀含量較對(duì)照分別提高了11.52%、11.01%、9.19%。
[Abstract]:Dibutyl phthalate (DBP) is widely used in plastic products as a plasticizer. In recent years, the use of agricultural film in our country has been increasing. Because of poor stability of plastic agricultural film, PAEs can easily be released from plastic films. Therefore, more and more DBP enters into the surrounding soil, which leads to the pollution of agricultural land. In addition, due to improper use of chemical fertilizer, sewage irrigation and the like, the content of DBP in soil can be increased. Due to the influence of DBP polluted soil, the accumulation of DBP in planted crops is caused, and the damage to endocrine and nervous system of human body is caused by the food chain entering into human body, so the prevention and cure of DBP pollution in soil is urgent. At present, the research on the screening and characterization mechanism of DBP-degrading bacteria is mainly focused on the research of the related aspects such as the research and development of DBP-degrading bacteria group preparation. The microbial remediation technology has the advantages of high efficiency, low cost, no secondary pollution, simple and convenient operation, good treatment effect and the like. The aim of this experiment is to develop a kind of functional microbial agent with high efficiency and good synergistic effect, which can improve the soil fertility while repairing DBP polluted soil. The effects of remediation agent on the growth of Chinese cabbage were studied by pot experiment, and the effect of functional microbial inoculum on soil fertility was further determined. The effect of functional agent on soil ecological safety was studied, and the practical application value of DBP microbial repair technology was discussed. provides scientific basis and technical support for remediation of PAEs polluted soil, and provides a reference for bioremediation of contaminated soil. The main results are as follows: 3 plant growth-promoting bacteria (P. P. P. 6, K _ 5, N1N1) were screened and isolated from the soil, and the Bacillus mucilaginosus was identified as Bacillus mucilaginosus and Enterobacter aerogenes. The nitrogen fixation capacity of NP1 was 14.56 mg/ L, and the nitrogen fixation capacity of N1 strain was 14.56 mg/ L. 3 strains of growth promoting bacteria and DBP degrading bacteria were screened for Pseudomonas sp. in the early stage of the laboratory. (2) and named DNB-S1 for plate marking and shaking flask test, and the results showed that there was no antagonistic reaction between the strains, and the activity was not affected. The fermentation medium was used to mix the bacteria group to ferment for 48h, and the number of NN1 in the plate counting method was 5. 3, 108 cfu/ m L, the number of P-solubilizing bacteria P6 was 4. 0-109 cfu/ m L, and the number of KK5 was 5. 0-109 cfu/ m L. The number of DBP-degrading bacteria DNB-S1 was 7. 6-109 cfu/ m L. The number of strains tested was above 108cfu/ m L and the fermentation effect was good. taking attapulgite, kaolin and humic acid as a carrier, preparing a functional microbial inoculum, and keeping standby at normal temperature. It was found that the living cells of four kinds of bacteria could keep higher activity at normal temperature for 35 days, and the preservation effect was good. The results showed that the degradation rate of DBP and soil fertility (quick-acting nitrogen, quick-acting phosphorus, available potassium) and the influence on microbial diversity in soil were investigated by comprehensive investigation of the degradation rate of DBP and soil fertility (available nitrogen, available phosphorus, available potassium) in the contaminated soil. finally, when the ratio of the degrading bacteria and the growth promoting bacteria in the functional microbial inoculum is 1 to 1 (g: g), the optimal matching mode is provided. The effects of functional microbial inoculum on the recovery of DBP contaminated soil (20 mg/ kg) were tested by pot pot experiment. The effects of physiological index and quality, soil enzyme activity and soil fertility on the physiological indexes and quality, soil enzyme activity and soil fertility of the young Chinese cabbage showed that during the growth of Chinese cabbage, the content of chlorophyll in the treatment group added with the functional microbial inoculum is obviously higher than that of the pollution treatment group, the content of MDA is lower than that of the pollution treatment group, the addition of the microbial inoculum obviously relieves the toxic effect of the DBP on the Chinese cabbage, the accumulation amount of dry matter in the Chinese cabbage is obviously higher than that of the control during the period of the treatment period, The soluble protein content and vitamin C content were increased by 24.93% and 13.67% respectively, and the quality of Chinese cabbage was improved. in that soil, the activity of the functional microbial agent on the soil, the activity of the sucrose enzyme play an active role, and the activity of the catalase doe not greatly affect the activity of the catalase; the quick-acting nitrogen, the quick-acting phosphorus and the quick-acting potassium content in the soil are respectively increased by 11.52 percent, 11.01 percent and 9.19 percent respectively.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：X53

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