安太堡露天煤礦復(fù)墾土壤微生物群落結(jié)構(gòu)研究
發(fā)布時(shí)間：2019-06-08 15:42
【摘要】：露天煤礦開采造成嚴(yán)重的環(huán)境污染和生態(tài)破壞。礦區(qū)廢棄地的生態(tài)修復(fù)已成為維持人類生存環(huán)境穩(wěn)定、煤炭工業(yè)可持續(xù)發(fā)展的必要措施。本研究以安太堡露天煤礦復(fù)墾區(qū)16個(gè)不同復(fù)墾模式樣地(包括不同的復(fù)墾植被和復(fù)墾年限)為研究對(duì)象,首先,用PCR-DGGE技術(shù)分析不同復(fù)墾模式對(duì)土壤細(xì)菌和真菌的影響。結(jié)果表明,土壤細(xì)菌和真菌多樣性分析,細(xì)菌和真菌多樣性指數(shù)均因不同復(fù)墾模式而顯著變化;從細(xì)菌多樣性的角度來看,楊樹和油松比較有優(yōu)勢;從真菌多樣性的角度來看,榆樹比較有優(yōu)勢;隨復(fù)墾年限的延長,真菌多樣性指數(shù)有增大的趨勢。聚類分析和PCA均顯示,相近復(fù)墾年限的土樣細(xì)菌和真菌群落結(jié)構(gòu)相似性較高。相關(guān)性分析表明,細(xì)菌和真菌多樣性指數(shù)與土壤pH、OC(有機(jī)碳)、TN(總氮)、C/N(碳氮比)和WC(絕對(duì)含水量)之間都不存在顯著相關(guān)性。RDA結(jié)果表明,土壤pH、OC、TN、C/N和WC對(duì)土壤細(xì)菌和真菌的群落演替分布均沒有顯著影響。細(xì)菌優(yōu)勢和差異條帶測序鑒定出Nitrospira、Sphingomonas、Arthrobacter、Brachybacterium、Streptomyces、Rhizobium、Devosia、Microvirga以及Mesorhizobium等或參與氮循環(huán)、或與植物共生固氮、或降解多環(huán)芳烴類有機(jī)物等有利于污染土壤的生態(tài)修復(fù)和肥力恢復(fù)的功能細(xì)菌屬。真菌則鑒定出Phoma、Fusarium、Myrothecium、Pycnoporus以及Cladosporium等或吸附重金屬、或產(chǎn)漆酶等降解多環(huán)芳烴類有機(jī)物、或與植物共生等有利于礦區(qū)生態(tài)修復(fù)的功能真菌屬;其中也存在一些致病菌,危害動(dòng)植物甚至人類的健康。其次,采用熒光定量PCR技術(shù)定量分析不同復(fù)墾模式土壤與氮循環(huán)相關(guān)功能微生物(包括與硝化作用相關(guān)的氨氧化細(xì)菌(AOB)、氨氧化古細(xì)菌(AOA)的amoA基因、固氮微生物nifH基因和與反硝化作用相關(guān)的反硝化細(xì)菌的nirS、nirK基因)的豐度差異。并用PCR-DGGE技術(shù)分析不同復(fù)墾模式對(duì)AOB-amoA、AOA-amoA和固氮微生物nif H基因的群落結(jié)構(gòu)及多樣性的影響。結(jié)果表明,AOB-amoA、AOA-amoA、nifH、nirS和nirK基因的豐度均因不同復(fù)墾模式而顯著變化,其豐度最高的植被樣地分別是復(fù)墾20年的榆樹土樣、復(fù)墾22年的楊樹土樣、復(fù)墾5年的油松土樣、復(fù)墾8年的沙棘土樣和復(fù)墾20年的杏樹土樣;土樣AOB豐度均低于AOA,nirS和nirK豐度差異不大;隨著復(fù)墾年限的延長,各功能基因的相對(duì)豐度變化不大。AOB-amoA和AOA-amoA的多樣性指數(shù)(H)均因不同復(fù)墾模式而變化,但差異不顯著,固氮微生物nifH的H因不同復(fù)墾模式而顯著變化;AOB-amoA和AOA-amoA的H均是復(fù)墾20年的云杉土樣最高,固氮微生物的H則是復(fù)墾5年的油松土樣顯著最高。相關(guān)性分析表明,AOA-amoA、nifH和nirS的豐度均與WC有相關(guān)性,前一者是極顯著負(fù)相關(guān),后兩者是顯著正相關(guān);AOB-amoA的H只是與TN有極顯著負(fù)相關(guān)性;而AOA-amoA的H恰恰只與TN沒有相關(guān)性,與pH呈顯著正相關(guān)、與OC、C/N和WC呈顯著負(fù)相關(guān);固氮微生物nifH的H與C/N呈顯著負(fù)相關(guān)。RDA結(jié)果顯示,土壤pH、OC、TN、C/N和WC對(duì)功能微生物豐度及H有不同程度的影響,并且土壤OC和TN達(dá)到極顯著水平,WC達(dá)到顯著水平�？梢�,土壤pH、WC、OC、TN和C/N可能是影響功能微生物群落的重要環(huán)境因子。最后,通過對(duì)土樣理化因子和微生物的H及豐度結(jié)果進(jìn)行PCA分析,計(jì)算獲得土壤綜合肥力指標(biāo)(IFI),結(jié)果表明,復(fù)墾年限較長的土樣其IFI較高,刺槐、油松、楊樹和榆樹樣地的IFI都比較高,可作為礦區(qū)復(fù)墾土壤肥力恢復(fù)的優(yōu)選植被。
[Abstract]:The mining of open-pit coal mine has serious environmental pollution and ecological damage. The ecological restoration of the mining area in the mining area has become the necessary measure to maintain the stability of the human living environment and the sustainable development of the coal industry. The effects of different reclamation modes on soil bacteria and fungi were analyzed by PCR-DGGE technique. The results showed that the diversity of bacterial and fungal diversity of soil and the index of bacterial and fungal diversity were significantly changed due to different reclamation modes; from the perspective of the diversity of the bacteria, the poplar and Pinus tabulaeformis had the advantages; from the perspective of the diversity of the fungi, the elm trees were more advantageous; With the extension of the number of reclamation, the index of the fungus diversity has a tendency to increase. The cluster analysis and PCA show that the similarity of the soil sample bacteria and the fungal community with similar reclamation years is high. The correlation analysis showed that there was no significant correlation between the bacterial and fungal diversity index and the soil pH, OC (organic carbon), TN (total nitrogen), C/ N (carbon-nitrogen ratio) and WC (absolute water content). The results showed that the soil pH, OC, TN, C/ N and WC had no significant effect on the community succession distribution of soil bacteria and fungi. The bacterial and differential band sequencing identified Nitzira, Sphingomonas, Arthorobacter, Braachyarteria, Streptomyces, Rhizobium, Devosia, Microvira and Mesorhizobium, or other functional bacteria, which were involved in the nitrogen cycle, or with plant symbiotic nitrogen fixation, or the degradation of polycyclic aromatic hydrocarbons, and the like, which are beneficial to the ecological restoration and fertility restoration of the contaminated soil. The fungi are identified as the genus Phoma, Fusaridium, Myrothecium, Pycpoorus and Cladosporium, or other functional fungi which are beneficial to the ecological restoration of the mining area, such as heavy metal, or laccase, or the like, and are beneficial to the ecological restoration of the mining area, such as plant symbiosis, etc., in which there are also some pathogenic bacteria, which are harmful to the health of animals and plants and even human. secondly, the ampoA gene of the soil and the nitrogen circulation related functional microorganism (including the ammonia oxidizing bacteria (AOB) and the ammonia oxidizing archaea (AOA) related to the nitrification is quantitatively analyzed by the fluorescence quantitative PCR technology, Abundance of the nifH gene of the nitrogen-fixing microorganism and the nirS, nirK gene of the denitrifying bacteria associated with the denitrification. The effects of different reclamation modes on the community structure and diversity of AOB-amoA, AOA-amoA and nf H genes of nitrogen-fixing microorganisms were analyzed by PCR-DGGE technique. The results showed that the abundance of AOB-amoA, AOA-amoA, nfH, nirS and nirK was significantly changed due to different reclamation modes. The highest abundance of the vegetation was the 20-year-old elm-tree soil sample, the 22-year-old poplar soil sample was reclaimed, and the soil samples of Pinus tabulaeformis were reclaimed for 5 years. 8-year-old sea-buckthorn soil samples and 20-year-old apricot-tree soil samples were reclaimed; the abundance of AOB in soil samples was lower than that of AOA, nirS and nirk, and the relative abundance of each functional gene was not large with the extension of the number of reclamation. The diversity index (H) of AOB-amoA and AOA-amoA was changed due to different reclamation modes, but the difference was not significant, and the H of the nitrogen-fixing microorganism nfH was significantly changed due to the different reclamation modes; both the AOB-amoA and the AOA-amoA were the highest in the 20-year-old spruce soil sample, The H of the nitrogen-fixing microorganism is the highest in the 5-year-old Chinese pine. The correlation analysis showed that the abundance of AOA-amoA, nifH and nirS was positively correlated with WC, and the first one was significantly negative and the latter were significantly positively correlated; the H of AOB-amoA was only positively correlated with TN; while the H of AOA-amoA was directly related to TN, which was significantly positively correlated with the pH, and with OC, There was a significant negative correlation between C/ N and WC, and there was a negative correlation between H and C/ N of nfH. The results of RDA show that the soil pH, OC, TN, C/ N and WC have different effects on the functional microbial abundance and H, and the soil OC and TN reach a very significant level, and the WC reaches a significant level. It can be seen that the soil pH, WC, OC, TN and C/ N may be an important environmental factor affecting the functional microbial community. In the end, the soil comprehensive fertility index is obtained by PCA analysis of the soil and chemical factors and the H and abundance results of the micro-organisms. The results show that the soil samples with longer reclamation years are higher than that of the high-yield, Robinia pseudoacacia, Pinus tabulaeformis, poplar and elm. And can be used as the preferred vegetation for recovering the soil fertility of the mining area.
【學(xué)位授予單位】：山西大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TD88;TD824
本文編號(hào)：2495404
[Abstract]:The mining of open-pit coal mine has serious environmental pollution and ecological damage. The ecological restoration of the mining area in the mining area has become the necessary measure to maintain the stability of the human living environment and the sustainable development of the coal industry. The effects of different reclamation modes on soil bacteria and fungi were analyzed by PCR-DGGE technique. The results showed that the diversity of bacterial and fungal diversity of soil and the index of bacterial and fungal diversity were significantly changed due to different reclamation modes; from the perspective of the diversity of the bacteria, the poplar and Pinus tabulaeformis had the advantages; from the perspective of the diversity of the fungi, the elm trees were more advantageous; With the extension of the number of reclamation, the index of the fungus diversity has a tendency to increase. The cluster analysis and PCA show that the similarity of the soil sample bacteria and the fungal community with similar reclamation years is high. The correlation analysis showed that there was no significant correlation between the bacterial and fungal diversity index and the soil pH, OC (organic carbon), TN (total nitrogen), C/ N (carbon-nitrogen ratio) and WC (absolute water content). The results showed that the soil pH, OC, TN, C/ N and WC had no significant effect on the community succession distribution of soil bacteria and fungi. The bacterial and differential band sequencing identified Nitzira, Sphingomonas, Arthorobacter, Braachyarteria, Streptomyces, Rhizobium, Devosia, Microvira and Mesorhizobium, or other functional bacteria, which were involved in the nitrogen cycle, or with plant symbiotic nitrogen fixation, or the degradation of polycyclic aromatic hydrocarbons, and the like, which are beneficial to the ecological restoration and fertility restoration of the contaminated soil. The fungi are identified as the genus Phoma, Fusaridium, Myrothecium, Pycpoorus and Cladosporium, or other functional fungi which are beneficial to the ecological restoration of the mining area, such as heavy metal, or laccase, or the like, and are beneficial to the ecological restoration of the mining area, such as plant symbiosis, etc., in which there are also some pathogenic bacteria, which are harmful to the health of animals and plants and even human. secondly, the ampoA gene of the soil and the nitrogen circulation related functional microorganism (including the ammonia oxidizing bacteria (AOB) and the ammonia oxidizing archaea (AOA) related to the nitrification is quantitatively analyzed by the fluorescence quantitative PCR technology, Abundance of the nifH gene of the nitrogen-fixing microorganism and the nirS, nirK gene of the denitrifying bacteria associated with the denitrification. The effects of different reclamation modes on the community structure and diversity of AOB-amoA, AOA-amoA and nf H genes of nitrogen-fixing microorganisms were analyzed by PCR-DGGE technique. The results showed that the abundance of AOB-amoA, AOA-amoA, nfH, nirS and nirK was significantly changed due to different reclamation modes. The highest abundance of the vegetation was the 20-year-old elm-tree soil sample, the 22-year-old poplar soil sample was reclaimed, and the soil samples of Pinus tabulaeformis were reclaimed for 5 years. 8-year-old sea-buckthorn soil samples and 20-year-old apricot-tree soil samples were reclaimed; the abundance of AOB in soil samples was lower than that of AOA, nirS and nirk, and the relative abundance of each functional gene was not large with the extension of the number of reclamation. The diversity index (H) of AOB-amoA and AOA-amoA was changed due to different reclamation modes, but the difference was not significant, and the H of the nitrogen-fixing microorganism nfH was significantly changed due to the different reclamation modes; both the AOB-amoA and the AOA-amoA were the highest in the 20-year-old spruce soil sample, The H of the nitrogen-fixing microorganism is the highest in the 5-year-old Chinese pine. The correlation analysis showed that the abundance of AOA-amoA, nifH and nirS was positively correlated with WC, and the first one was significantly negative and the latter were significantly positively correlated; the H of AOB-amoA was only positively correlated with TN; while the H of AOA-amoA was directly related to TN, which was significantly positively correlated with the pH, and with OC, There was a significant negative correlation between C/ N and WC, and there was a negative correlation between H and C/ N of nfH. The results of RDA show that the soil pH, OC, TN, C/ N and WC have different effects on the functional microbial abundance and H, and the soil OC and TN reach a very significant level, and the WC reaches a significant level. It can be seen that the soil pH, WC, OC, TN and C/ N may be an important environmental factor affecting the functional microbial community. In the end, the soil comprehensive fertility index is obtained by PCA analysis of the soil and chemical factors and the H and abundance results of the micro-organisms. The results show that the soil samples with longer reclamation years are higher than that of the high-yield, Robinia pseudoacacia, Pinus tabulaeformis, poplar and elm. And can be used as the preferred vegetation for recovering the soil fertility of the mining area.
【學(xué)位授予單位】：山西大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TD88;TD824
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