安太堡露天煤矿复垦土壤微生物群落结构研究

发布时间：2019-06-08 15:42

【摘要】：露天煤矿开采造成严重的环境污染和生态破坏。矿区废弃地的生态修复已成为维持人类生存环境稳定、煤炭工业可持续发展的必要措施。本研究以安太堡露天煤矿复垦区16个不同复垦模式样地(包括不同的复垦植被和复垦年限)为研究对象,首先,用PCR-DGGE技术分析不同复垦模式对土壤细菌和真菌的影响。结果表明,土壤细菌和真菌多样性分析,细菌和真菌多样性指数均因不同复垦模式而显著变化;从细菌多样性的角度来看,杨树和油松比较有优势;从真菌多样性的角度来看,榆树比较有优势;随复垦年限的延长,真菌多样性指数有增大的趋势。聚类分析和PCA均显示,相近复垦年限的土样细菌和真菌群落结构相似性较高。相关性分析表明,细菌和真菌多样性指数与土壤pH、OC(有机碳)、TN(总氮)、C/N(碳氮比)和WC(绝对含水量)之间都不存在显著相关性。RDA结果表明,土壤pH、OC、TN、C/N和WC对土壤细菌和真菌的群落演替分布均没有显著影响。细菌优势和差异条带测序鉴定出Nitrospira、Sphingomonas、Arthrobacter、Brachybacterium、Streptomyces、Rhizobium、Devosia、Microvirga以及Mesorhizobium等或参与氮循环、或与植物共生固氮、或降解多环芳烃类有机物等有利于污染土壤的生态修复和肥力恢复的功能细菌属。真菌则鉴定出Phoma、Fusarium、Myrothecium、Pycnoporus以及Cladosporium等或吸附重金属、或产漆酶等降解多环芳烃类有机物、或与植物共生等有利于矿区生态修复的功能真菌属;其中也存在一些致病菌,危害动植物甚至人类的健康。其次,采用荧光定量PCR技术定量分析不同复垦模式土壤与氮循环相关功能微生物(包括与硝化作用相关的氨氧化细菌(AOB)、氨氧化古细菌(AOA)的amoA基因、固氮微生物nifH基因和与反硝化作用相关的反硝化细菌的nirS、nirK基因)的丰度差异。并用PCR-DGGE技术分析不同复垦模式对AOB-amoA、AOA-amoA和固氮微生物nif H基因的群落结构及多样性的影响。结果表明,AOB-amoA、AOA-amoA、nifH、nirS和nirK基因的丰度均因不同复垦模式而显著变化,其丰度最高的植被样地分别是复垦20年的榆树土样、复垦22年的杨树土样、复垦5年的油松土样、复垦8年的沙棘土样和复垦20年的杏树土样;土样AOB丰度均低于AOA,nirS和nirK丰度差异不大;随着复垦年限的延长,各功能基因的相对丰度变化不大。AOB-amoA和AOA-amoA的多样性指数(H)均因不同复垦模式而变化,但差异不显著,固氮微生物nifH的H因不同复垦模式而显著变化;AOB-amoA和AOA-amoA的H均是复垦20年的云杉土样最高,固氮微生物的H则是复垦5年的油松土样显著最高。相关性分析表明,AOA-amoA、nifH和nirS的丰度均与WC有相关性,前一者是极显著负相关,后两者是显著正相关;AOB-amoA的H只是与TN有极显著负相关性;而AOA-amoA的H恰恰只与TN没有相关性,与pH呈显著正相关、与OC、C/N和WC呈显著负相关;固氮微生物nifH的H与C/N呈显著负相关。RDA结果显示,土壤pH、OC、TN、C/N和WC对功能微生物丰度及H有不同程度的影响,并且土壤OC和TN达到极显著水平,WC达到显著水平。可见,土壤pH、WC、OC、TN和C/N可能是影响功能微生物群落的重要环境因子。最后,通过对土样理化因子和微生物的H及丰度结果进行PCA分析,计算获得土壤综合肥力指标(IFI),结果表明,复垦年限较长的土样其IFI较高,刺槐、油松、杨树和榆树样地的IFI都比较高,可作为矿区复垦土壤肥力恢复的优选植被。
[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.
【学位授予单位】：山西大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TD88;TD824

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