本文選題：生物炭 + 微生物固定化　； 參考：《上海交通大學(xué)》2015年博士論文

【摘要】：論文以水稻秸稈、稻殼和牛糞為原料,在350和500℃條件下限氧熱解制備生物炭的基礎(chǔ)上,研究了生物炭對(duì)土著微生物生態(tài)效應(yīng)影響,并采用末端限制性多態(tài)分析(T-RFLP)和熒光定量PCR(q PCR)技術(shù),對(duì)土壤微生物多樣性及功能基因進(jìn)行定量分析。在此基礎(chǔ)上,以持久性有機(jī)污染物多環(huán)芳烴(PAHs)為研究對(duì)象,采用微生物固定化技術(shù),將自主篩選的PAHs降解菌固定于生物炭上,開展了生物炭固定化微生物對(duì)復(fù)合污染土壤中16種PAHs生物修復(fù)及其作用機(jī)理研究。為生物炭強(qiáng)化和刺激土著微生物修復(fù)PAHs污染土壤提供科學(xué)依據(jù),論文對(duì)于發(fā)展污染土壤原位生物修復(fù)技術(shù),闡明生物炭固定化降解菌強(qiáng)化修復(fù)PAHs污染土壤機(jī)理及其生態(tài)效應(yīng)具有重要的理論和現(xiàn)實(shí)意義,主要研究結(jié)果如下:1.對(duì)生物炭主要元素和理化性質(zhì)研究表明,牛糞、稻殼和水稻秸稈生物炭表面存在羰基(C=O)、羧基(-COOH)、羥基(-OH)、醚(-O-)、芳香族C=N鍵和N-H3鍵等主要官能基團(tuán)。生物炭除了含有大量C元素以外,還含有N、P、Ca、Na、Mg、K及Mn、Zn、Cu等微量元素;供試6種生物炭均呈堿性,水稻秸稈生物炭RS500的p H值最高,為10.50;500℃熱解溫度制備的生物炭中的C元素和灰分含量高于350℃制備的生物炭;相同熱解溫度條件下,稻殼和水稻秸稈生物炭中的C/N大于牛糞生物炭。2.生物炭對(duì)土壤蔗糖酶活性具有顯著的刺激作用,對(duì)放線菌具有抑制作用,對(duì)土壤磷酸酶、脲酶活性、細(xì)菌和真菌數(shù)量、CO2和N2O的釋放量的影響與施炭量和生物炭制備溫度密切相關(guān),其中,在添加4%生物炭的土壤中,生物炭對(duì)土壤細(xì)菌生長(zhǎng)具有顯著的刺激作用;500℃熱解制備生物炭對(duì)土壤中細(xì)菌的刺激作用要大于350℃熱解制備的生物炭。生物炭對(duì)土壤N2O釋放的影響具有典型hormesis效應(yīng)特征。3.生物炭處理土壤中細(xì)菌群落發(fā)生了顯著變化,主要形成5個(gè)類群;所有供試生物炭處理土壤中nir S基因拷貝數(shù)增加了67.22~70.18%;生物炭處理對(duì)土壤中微生物群落的多樣性和amo A基因拷貝數(shù)的影響與生物炭制備溫度、原料和施炭量密切相關(guān)。其中,施炭量為4%時(shí),350℃低溫?zé)峤庵苽涞纳锾匡@著提高土壤微生物的多樣性。除DM350和RH350兩個(gè)處理外,土壤中amo A基因拷貝數(shù)比對(duì)照減少了93~98.10%。生物炭對(duì)土壤細(xì)菌16S r DNA和PAH-RHDαGP基因拷貝數(shù)量增加具有刺激作用。其中,低溫生物炭處理的刺激作用大于高溫?zé)峤庵苽涞纳锾�。處理土壤中PAH-RHDαGP基因拷貝數(shù)量增加最多的是水稻秸稈RS350,其拷貝數(shù)達(dá)到1.73×107 copies g-1 dry soil。4.生物炭對(duì)PAHs復(fù)合污染土壤中amo A基因的拷貝數(shù)具有抑制作用,抑制率最大的是水稻秸稈生物炭RS500;然而,PAH-RHDαGP和nir S基因的數(shù)量則顯著增加。其中,生物炭對(duì)污染土壤amo A基因的拷貝數(shù)的抑制率達(dá)到23.53~85.13%,PAH-RHDαGP基因拷貝數(shù)量增加最多的是水稻秸稈RS350生物炭,拷貝數(shù)達(dá)到1.73×107 copies g-1 dry soil;相同處理?xiàng)l件下,在500℃高溫?zé)峤馍锾刻幚硗寥乐衝ir S基因的拷貝數(shù)大于350℃低溫?zé)峤庵苽涞纳锾?增加最多是水稻秸稈生物炭RS350處理,為對(duì)照土壤的2.9倍,拷貝數(shù)達(dá)到6.45×107 copies g-1 dry soil。5.采用富集培養(yǎng)方法,篩選和分離到1株能在重金屬存在條件下對(duì)PAHs具有高效降解作用的降解菌,經(jīng)形態(tài)學(xué)觀察和16S r DNA分子鑒定,屬于貪銅菌屬,將其命名為B35。6.低溫生物炭固定化B35菌劑對(duì)培養(yǎng)基中PAHs的吸附與降解作用大于高溫生物炭處理。相同熱解制備溫度條件下,植物類的稻殼和水稻秸稈生物炭固定化降解菌劑對(duì)PAHs的吸附作用大于牛糞類。固定化菌劑對(duì)PAHs吸附作用的大小依次為:稻殼生物炭RH350水稻秸稈生物炭RS350牛糞生物炭RM350稻殼生物炭RH500牛糞生物炭DM500水稻秸稈生物炭RS500。對(duì)PAHs的降解效率最高的是稻殼生物炭RH350菌劑,降解率達(dá)到31.34%。7.不同生物炭固定化菌劑對(duì)不同分子量PAHs均具有吸附與降解作用,不同環(huán)PAHs之間存在顯著差異。除水稻秸稈生物炭RS500外,所有生物炭固定化菌劑對(duì)4環(huán)PAHs吸附率均高于2~3環(huán)PAHs,平均吸附率分別為44.98%和24.50%,熱解溫度對(duì)不同生物炭固定化菌劑吸附5~6環(huán)PAHs的影響差異比2~4環(huán)的差異大。350℃熱解制備的生物炭固定化菌劑對(duì)2~4環(huán)PAHs的降解率高于500℃熱解制備的生物炭固定化菌劑。其中,對(duì)2~3環(huán)、4環(huán)和5~6環(huán)PAHs的上海交通大學(xué)博士學(xué)位論文平均降解率分別為:15.03%、25.08%和10.08%。降解率最高分別達(dá)到23.33%、36.50%和22.75%。8.與培養(yǎng)基中生物炭固定化菌劑PAHs吸附作用相同,350℃低溫制備的生物炭固定化菌劑對(duì)污染土壤PAHs的吸附和降解率高于高溫500℃制備的生物炭處理,并且對(duì)2~4環(huán)PAHs的吸附作用大于5~6環(huán)。其中,吸附作用最強(qiáng)的是水稻秸稈生物炭RS350處理,吸附率達(dá)到22.80%。9.生物強(qiáng)化和生物刺激協(xié)同作用是生物炭固定化B35降解菌劑修復(fù)PAHs污染土壤的重要作用機(jī)理。不同生物炭固定化菌劑對(duì)污染土壤中PAHs的修復(fù)作用的大小依次為:水稻秸稈生物炭RS350稻殼生物炭RH350牛糞生物炭RM350稻殼生物炭RH500牛糞生物炭DM500水稻秸稈生物炭RS500。
[Abstract]:In this paper, rice straw, rice husk and cow dung were used as raw materials. On the basis of bio carbon preparation by limiting oxygen pyrolysis at 350 and 500 degrees, the effects of biological charcoal on the ecological effects of indigenous microorganisms were studied. The microbial diversity and functional genes of soil microbial diversity and functional genes were quantified by terminal restricted polymorphism analysis (T-RFLP) and fluorescence quantitative PCR (Q PCR) technique. On this basis, based on the persistent organic pollutants polycyclic aromatic hydrocarbons (PAHs) as the research object, the microbial immobilization technology was used to immobilize the independent PAHs degrading bacteria on biochar, and the mechanism of the biological charcoal immobilized microorganism to repair 16 kinds of PAHs species in the contaminated soil was carried out and the mechanism of biological charcoal enhancement and stimulation was studied. The aboriginal microorganism provides scientific basis for the remediation of PAHs contaminated soil. This paper is of great theoretical and practical significance to the development of in situ bioremediation technology of contaminated soil and clarifying the mechanism and ecological effects of Biocharcoal immobilized degradation bacteria to strengthen the remediation of PAHs contaminated soil. The main fruits are as follows: 1. main elements and physicochemical properties of biological charcoal Studies have shown that there are carbonyl (C=O), carboxyl (-COOH), hydroxyl (-OH), ether (-O-), aromatic C=N and N-H3 key on the surface of biological charcoal of cow dung, rice hull and rice straw. Biological charcoal also contains N, P, Ca, Na, Mg, Mg, Ca, Na, and other trace elements in addition to a large number of C elements; the 6 kinds of biological charcoal are alkaline and rice straw organisms. The P H value of carbon RS500 is the highest, which is 10.50. The C element and ash content in the Biocharcoal prepared at 500 C is higher than that of the biological carbon prepared at 350 C. Under the same pyrolysis temperature, the C/N in the rice husk and rice straw bio carbon is greater than that of the cow dung biological charcoal.2. Biocharcoal, which has a significant stimulating effect on the activity of the soil sucrase, and the actinomycetes have a significant effect on the activity of the actinomycetes. The effects of inhibition on soil phosphatase, urease activity, bacteria and fungi, the release of CO2 and N2O are closely related to the amount of carbon and the preparation temperature of biological carbon. In the soil adding 4% biological charcoal, biological carbon has significant stimulation for the growth of soil bacteria, and the pyrolysis of biochar at 500 degrees centigrade to stimulate the bacteria in the soil The effect of bio carbon on the release of soil N2O has a typical hormesis effect characteristic of the soil N2O release. The bacterial community in the soil treated by.3. biological charcoal has changed significantly, mainly formed 5 groups, and the copy number of NIR S gene in the soil of all the tested biochar treated soil increased by 67.22~70.18%; Biocharcoal treatment of soil The diversity of microbial communities in the soil and the effect of AMO A gene copy number are closely related to the preparation temperature of biological carbon and the amount of carbon applied in the soil. Among them, the Biocharcoal prepared by low temperature pyrolysis at 350 centigrade can significantly improve the diversity of soil microorganism. The amo A gene copy number in soil is less than that of the control, except DM350 and RH350. 93~98.10%. bio carbon has a stimulating effect on the increase of the copy number of 16S R DNA and PAH-RHD alpha GP in soil bacteria. Among them, the stimulation effect of low temperature biochar treatment is greater than that of high temperature pyrolysis. The maximum number of PAH-RHD alpha GP gene copies in the soil is increased by the rice straw RS350, and the copy number is 1.73 * 107 copies G. -1 dry soil.4. biochar could inhibit the copy number of AMO A gene in the PAHs compound contaminated soil, and the maximum inhibition rate was the rice straw bio carbon RS500; however, the number of PAH-RHD a GP and NIR S genes increased significantly. The copy number increased most because of the rice straw RS350 biological carbon, the copy number reached 1.73 * 107 copies g-1 dry soil. Under the same treatment conditions, the copy number of NIR S gene in the soil treated at 500 C high temperature pyrolytic biochar was greater than the low temperature pyrolytic carbon from 350 C, and the most increased is the rice straw bio carbon RS350 treatment, which was the control soil. 2.9 times of the soil, the copy number reached 6.45 * 107 copies g-1 dry soil.5., and the enrichment culture method was used to screen and isolate 1 degrading bacteria which could effectively degrade PAHs under the presence of heavy metals. By morphological observation and 16S R DNA molecular identification, it belonged to the genus cupric, and named it as a B35.6. low temperature biological charcoal immobilized B35 microbial agent. The adsorption and degradation of PAHs in the medium is greater than that of high temperature biochar treatment. Under the same pyrolysis temperature conditions, the adsorption of rice husk and rice straw bio carbon immobilized microbial agent on PAHs is greater than that of cow dung. The size of the adsorption of PAHs by immobilized bacteria is the following: Rice hull bio carbon RH350 rice straw biology Carbon RS350 cow dung biological charcoal RM350 rice husk biological charcoal RH500 cow dung biological carbon DM500 rice straw bio carbon RS500. degradation efficiency of PAHs is the highest in rice hull Biocharcoal RH350 bacteria, degradation rate to 31.34%.7. different biological charcoal immobilized bacteria have adsorption and degradation of different molecular weight PAHs, there is significant difference between different loop PAHs. Except for rice straw biological carbon RS500, all Biocharcoal immobilized bacteria have higher adsorption rates for 4 ring PAHs than 2~3 ring PAHs, and the average adsorption rate is 44.98% and 24.50% respectively. The difference of the pyrolysis temperature on the adsorption of 5~6 ring PAHs with different Biocharcoal immobilized bacteria is greater than that of 2~4 ring. The degradation rate of PAHs was higher than that of Biocharcoal immobilized bacteria prepared by pyrolysis at 500 C. The average degradation rate of Shanghai Jiao Tong University doctorate in the Shanghai Jiao Tong University was 15.03%, 25.08% and 10.08%. respectively reached 23.33%, 36.50% and 22.75%.8., respectively, and PAHs adsorption of Biocharcoal immobilized bacteria in the medium, respectively. The adsorption and degradation rate of Biocharcoal immobilized bacteria on contaminated soil PAHs was higher than that prepared at 500 C at 350 C, and the adsorption of 2~4 ring PAHs was greater than that of 5~6 ring. Among them, the strongest adsorption was RS350 treatment of rice straw biological carbon, the adsorption rate was 22.80%.9. biofortified and biostimulation Co The same effect is the important mechanism of the biological charcoal immobilized B35 degradation bacteria for the remediation of PAHs contaminated soil. The size of the remediation of PAHs in contaminated soil by different Biocharcoal immobilized bacteria is in turn: rice straw biological carbon, RS350 rice hull, RH350 cattle manure, RM350 rice husk, RH500 cattle dung, Biocharcoal, Biocharcoal and rice straw biology Carbon RS500.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：X53;S154.3

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本文編號(hào)：1902589