本文關(guān)鍵詞： 石油污染 根際菌群 豆科植物 根瘤菌 盆栽試驗(yàn)　出處：《西北農(nóng)林科技大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：石油污染嚴(yán)重地危害著土壤環(huán)境、農(nóng)作物的生長和人類的健康。建立高效、環(huán)保的石油污染修復(fù)技術(shù)是目前研究的迫切需求。本論文通過盆栽實(shí)驗(yàn),在農(nóng)田土壤和石油污染土壤中分別種植12種豆科植物,利用植物生長自然馴化根際菌群和建立共生,測定根際菌群對菲的降解效率,獲得高效降解的菲的豆科植物-根際菌群體系;然后通過16S rDNA高通量測序技術(shù),解析根際降解菌群的結(jié)構(gòu);再通過16S rDNA序列對分離的根瘤菌進(jìn)行系統(tǒng)發(fā)育分析,探究石油污染對豆科植物-根瘤菌共生關(guān)系的影響。結(jié)果如下:通過種植豆科植物獲得24個根際菌群(包括污染土和農(nóng)田土),測定和比較菌群對初始濃度為250 mg/L的菲的5天后降解效率,結(jié)果顯示從石油污染土壤中馴化的小冠花、豇豆、百脈根和紅三葉4個根際降解菌群對菲降解效率均在65%以上,與農(nóng)田土壤獲得的根際菌群相比,降解效率均提高30%以上,其中從石油污染土壤中馴化的小冠花根際菌群降解效率高達(dá)80%,與從農(nóng)田土壤獲得的根際菌群相比,降解效率提高了67%。通過對根際土DNA進(jìn)行16S rDNA V4-V5區(qū)高通量測序分析,發(fā)現(xiàn)在石油污染土中富集的根際菌群與在農(nóng)田土中富集的根際菌群在群落結(jié)構(gòu)上具有顯著差異,其中,小冠花、豇豆、百脈根和紅三葉在石油污染土富集的根際菌群差異顯著的物種為α-變形菌綱(Alphaproteobacteria)和γ-變形菌綱(Gammaproteobacteria),而在農(nóng)田土中則為β-變形菌綱(Betaproteobacteria),可見,在石油污染土中,差異顯著的物種α-變形菌(Alphaproteobacteria)和γ-變形菌綱(Gammaproteobacteria)與菲降解效率有關(guān)。通過對根瘤菌的分離純化,發(fā)現(xiàn)在石油污染土壤與農(nóng)田土壤中,僅從豇豆和紅三葉分離到了根瘤菌,其中從豇豆中分離的根瘤菌均歸屬于Bradyrhizobium,從紅三葉中分離的根瘤菌均歸屬于Rhizobium,說明石油污染對豇豆、紅三葉與根瘤菌的匹配類型沒有影響,只是影響了根瘤數(shù)量。在石油污染土壤中,百脈根、檸條、刺槐等植物沒有結(jié)瘤,可見,石油污染抑制了百脈根、檸條、刺槐等植物與根瘤菌共生關(guān)系的建立。
[Abstract]:Oil pollution seriously endangers the soil environment, crop growth and human health. It is an urgent need to establish efficient and environmentally friendly oil pollution remediation technology. Twelve species of legumes were planted in farmland soil and petroleum contaminated soil. The natural domestication and symbiosis of rhizosphere microflora were used to determine the degradation efficiency of phenanthrene. The biodegradable phenanthrene legume rhizosphere flora system was obtained. Then the structure of the rhizosphere degrading bacteria was analyzed by 16s rDNA high-throughput sequencing technique. The phylogeny of isolated rhizobia was analyzed by 16s rDNA sequence. To explore the effect of petroleum pollution on the symbiotic relationship between legumes and rhizobia. The results are as follows: 24 rhizosphere flora (including contaminated soil and farmland soil) were obtained by planting legumes. The degradation efficiency of phenanthrene at initial concentration of 250 mg/L was determined and compared after 5 days. The results showed that cowpea was domesticated from petroleum contaminated soil. The degradation efficiency of phenanthrene was more than 65% in the four rhizosphere biodegradable colonies of Baimai root and red clover, and the degradation efficiency was 30% higher than that of the rhizosphere flora obtained from farmland soil. Among them, the degradation efficiency of rhizosphere microflora domesticated from petroleum contaminated soil was as high as 80%, which was compared with that obtained from farmland soil. The degradation efficiency was increased by 67.The high throughput sequencing analysis of 16s rDNA V4-V5 region was carried out on DNA in rhizosphere soil. It was found that there were significant differences in community structure between rhizosphere microflora enriched in petroleum contaminated soil and rhizosphere bacteria enriched in farmland soil, among which, small crown flower, cowpea. The species of Alphaproteobacteria and 緯 -Proteobacteriae that are enriched in petroleum contaminated soil are the species of Alphaproteobacteria. Gamma proteobacteria. But in the farmland soil is 尾 -Proteobacter teriae, we can see, in the oil-contaminated soil. Alphaproteobacteria) and Gamma proteobacteria (Gamma proteobacteria). Related to the degradation efficiency of phenanthrene. Through the isolation and purification of rhizobia. It was found that rhizobia were isolated only from cowpea and red clover in oil-contaminated soil and farmland soil, and rhizobia isolated from cowpea belonged to Bradyrhizobium. All rhizobia isolated from red clover belong to Rhizobium, indicating that oil pollution has no effect on matching types of cowpea, red clover and rhizobia. In the soil polluted by oil, there were no nodules in the soil, such as Caragana korshinskii, Robinia pseudoacacia and so on. It can be seen that oil pollution has inhibited the establishment of symbiotic relationship between rhizobia, Caragana, Robinia and other plants.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：X17;X53

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