【摘要】：納米材料的特殊結(jié)構(gòu)使得它們具有許多特異的優(yōu)良性質(zhì),其中納米銀的廣譜抗菌性給納米銀的廣泛應(yīng)用開辟了廣闊的前景,同樣也給土壤環(huán)境帶來了脅迫風(fēng)險。本文通過在室內(nèi)培養(yǎng)條件下,以硝酸銀與石墨烯為對照,測定納米銀對土壤呼吸、土壤酶以及土壤微生物群落功能多樣性的影響,分析它們對土壤微生物的影響,預(yù)估納米銀給土壤生態(tài)環(huán)境帶來的風(fēng)險;探討土傳真菌病害腐皮鐮刀菌(Fusarium solani)生長、細(xì)胞結(jié)構(gòu)及轉(zhuǎn)錄組對納米銀的響應(yīng)。得到的主要結(jié)果如下:1.納米銀對土壤呼吸強(qiáng)度存在抑制作用,較同濃度硝酸銀顯著。石墨烯對土壤呼吸無影響。2.納米銀與硝酸銀均抑制土壤可培養(yǎng)細(xì)菌、真菌、放線菌的數(shù)量;100 mg·kg-1和150 mg·kg-1的納米銀與硝酸銀處理顯著降低了三種土壤微生物的數(shù)量。低于100 mg·kg-1的石墨烯處理土壤細(xì)菌、真菌、放線菌數(shù)量均無顯著變化;但1000 mg·kg-1的石墨烯顯著增加土壤細(xì)菌與真菌的數(shù)量。3.納米銀顯著抑制土壤脲酶與土壤脫氫酶活性,且抑制作用大于同濃度硝酸銀處理,其中納米銀≥10 mg·kg-1時,土壤脲酶、脫氫酶活性均顯著降低,但硝酸銀達(dá)到100mg·kg-1以上這兩種酶活性才顯著下降;納米銀與硝酸銀對土壤過氧化氫酶和堿性磷酸酶活性無影響;石墨烯存在顯著抑制了土壤堿性磷酸酶活性。4.納米銀對土壤微生物群落的總體活性、Shannon、Simpson、Mc Intosh指數(shù)的負(fù)面影響大于硝酸銀,兩者在150 mg·kg-1處理下均最為顯著,石墨烯存在降低了土壤微生物群落功能多樣性。5.納米銀對F.solani生長的抑制存在明顯的劑量效應(yīng),EC50值為3.6 mg·L-1,10mg·L-1的納米銀可完全抑制F.solani的生長。納米銀可破壞F.solani的細(xì)胞壁、細(xì)胞膜、細(xì)胞器等,且在細(xì)胞膜上觀察到疑似納米銀顆粒。6.基于對納米銀處理后F.solani轉(zhuǎn)錄組數(shù)據(jù)的分析,共得到23791條Unigenes,分別在6 h、12 h、24 h時發(fā)現(xiàn)862、867、774個差異表達(dá)基因(P0.05)。GO功能富集分析表明納米銀處理后F.solani在蛋白質(zhì)加工、氧化還原酶活性、分解代謝、細(xì)胞成分作用于碳-氮的水解酶活性、結(jié)構(gòu)分子活性、翻譯、外部包裝結(jié)構(gòu)、RNA結(jié)合和前體代謝物與能量的產(chǎn)生等類別表達(dá)差異較大。KEGG代謝途徑分析表明,納米銀處理后F.solani在轉(zhuǎn)錄、翻譯、折疊,分選和降解、碳水化合物代謝、能量代謝、信號轉(zhuǎn)導(dǎo)與信號分子和相互作用等途徑表達(dá)差異較大。7.差異表達(dá)基因在能量和物質(zhì)代謝、信號轉(zhuǎn)導(dǎo)和遺傳信息途徑的比較集中,主要包括TCA循環(huán)、硫代謝、抗氧化酶活性、熱激蛋白、蛋白質(zhì)合成與分解、DNA修復(fù)等過程。納米銀可通過擾亂F.solani的能量物質(zhì)代謝、產(chǎn)生ROS、造成蛋白質(zhì)甚至DNA損傷殺死F.solani細(xì)胞。
[Abstract]:The special structure of nanomaterials makes them have many special and excellent properties. The broad-spectrum antimicrobial properties of nanocrystalline silver have opened up a broad prospect for the wide application of nano-silver, and have also brought about stress risks to the soil environment. In this paper, the effects of silver nitrate and graphene on soil respiration, soil enzymes and the functional diversity of soil microbial communities were determined under indoor culture conditions, and their effects on soil microbes were analyzed. To estimate the risk of nano-silver to soil ecological environment and to study the growth, cell structure and transcriptional response of soil-borne fungal disease Fusarium vulgaris (Fusarium solani) to nano-silver. The main results are as follows: 1. Nano silver inhibited soil respiration, which was more significant than that of silver nitrate at the same concentration. Graphene has no effect on soil respiration. 2. Both silver nanoparticles and silver nitrate inhibited the number of culturable bacteria, fungi and actinomycetes in the soil, and the treatment of 100 mg kg-1 and 150 mg kg-1 of silver nanoparticles and silver nitrate significantly reduced the number of three kinds of soil microbes. The amount of soil bacteria, fungi and actinomycetes treated with graphene less than 100 mg kg-1 had no significant change, but 1000 mg kg-1 of graphene significantly increased the number of soil bacteria and fungi. The activity of soil urease and soil dehydrogenase was significantly inhibited by nano-silver, and the inhibitory effect was greater than that of the same concentration of silver nitrate. When nano-silver was more than 10 mg kg-1, the activity of soil urease and dehydrogenase decreased significantly. However, silver nitrate and silver nitrate had no effect on the activities of catalase and alkaline phosphatase, and graphene significantly inhibited the activity of alkaline phosphatase. The negative effect of silver nanoparticles on the total activity of soil microbial communities was greater than that of silver nitrate, and the existence of graphene decreased the functional diversity of soil microbial communities. The growth of F.solani was inhibited by nano-silver in a dose-dependent manner, and the growth of F.solani was completely inhibited by silver nanoparticles with a EC50 value of 3.6 mg L ~ (-1) ~ (-1) ~ (10 mg 路L ~ (-1). Silver nanoparticles can destroy the cell wall, cell membrane and organelle of F.solani, and the suspected silver nanoparticles can be observed on the cell membrane. 6. 6. Based on the analysis of F.solani transcriptome data after silver nanoparticles treatment, a total of 23791 Unigenes, differentially expressed genes were found at 6 h or 12 h or 24 h. (P0.05). GO functional enrichment analysis showed that F.solani was involved in protein processing and redox enzyme activity after treatment with silver nanoparticles. Catabolism, cellular components acting on carbon-nitrogen hydrolase activity, structural molecular activity, translation, external packaging structure, RNA binding and the production of precursor metabolites and energy were significantly different. KEGG metabolic pathway analysis showed that, There were significant differences in the expression of F.solani in transcription, translation, folding, sorting and degradation, carbohydrate metabolism, energy metabolism, signal transduction and signal molecules and interactions. The differentially expressed genes are concentrated in energy and substance metabolism, signal transduction and genetic information pathway, including TCA cycle, sulfur metabolism, antioxidant enzyme activity, heat shock protein, protein synthesis and decomposition, DNA repair and so on. Silver nanoparticles can damage F.solani cells by disrupting the energy metabolism of F.solani and producing ROS, causing protein and even DNA damage.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S154.3;S432.44

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