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[不同能源富集的嗜热微生物菌群的细菌多样性分析]

[Bacterial diversity analysis of moderately thermophilic microflora enriched by different energy sources].

作者信息

Liu Fei-fei, Zhou Hong-bo, Fu Bo, Qiu Guan-zhou

机构信息

School of Minerals Processing and Bioengineering, Central South University, Changsha 410083, China.

出版信息

Wei Sheng Wu Xue Bao. 2007 Jun;47(3):381-6.

Abstract

Bacterial biodiversities of three moderately thermophilic bioleaching microfloras grown at 50 degrees C on media with pyrite, chalcopyrite, and pure ferrous iron supplemented with sulfur as energy sources were investigated respectively. The 16S rRNA genes of the microorganisms in the cultures flasks were PCR amplified and cloned to identify the bacterial species by comparative sequence analysis, the structural differences of microfloras enriched by different energy sources were compared. A total of 303 clones were recovered and evaluated by restriction fragment length polymorphism (RFLP) analysis. Cluster analysis identified 29 unique RFLP patterns, and the inserted 16S rRNA genes sequences were determined and for phylogenetic analysis. Most of sequences obtained were similar (89.1%-99.7%) to the 16S rRNA gene sequences of the reported bioleaching microorganisms. The species identified from the flasks during bioleaching of pyrite, pure ferrous iron supplemented with sulfur, and chalcopyrite were closely related to Acidithiobacillus caldus, Sulfobacillus thermotolerans, Sulfobacillus thermosulfidooxidans, Leptospirillum ferriphilum, two uncultured forest soil bacterium clones and one uncultured proteobacterium clone. Among these bacteria, Acidithiobacillus caldus, Sulfobacillus thermotolerans and Leptospirillum ferriphilum were the dominant bacterial species. L. ferriphilum was the most dominant species in microfloras enriched in media with pyrite and ferrous iron supplemented with sulfur as energy sources, the abundance were 53.8% and 45.9% respectively. In the culture with chalcopyrite as energy sources, S. thermotolerans had the highest abundance of 70.1%.

摘要

分别研究了三种嗜温性生物浸出微生物菌群在50摄氏度下,以含有黄铁矿、黄铜矿的培养基以及添加硫作为能源的纯亚铁为底物生长时的细菌多样性。对培养瓶中微生物的16S rRNA基因进行PCR扩增和克隆,通过比较序列分析鉴定细菌种类,比较不同能源富集的微生物菌群的结构差异。通过限制性片段长度多态性(RFLP)分析共回收并评估了303个克隆。聚类分析确定了29种独特的RFLP模式,并对插入的16S rRNA基因序列进行了测定以进行系统发育分析。获得的大多数序列(89.1%-99.7%)与已报道的生物浸出微生物的16S rRNA基因序列相似。在黄铁矿、添加硫的纯亚铁和黄铜矿生物浸出过程中,从培养瓶中鉴定出的物种与嗜热嗜酸硫杆菌、耐热硫化芽孢杆菌、嗜热硫化氧化芽孢杆菌、嗜铁钩端螺旋菌、两个未培养的森林土壤细菌克隆和一个未培养的变形菌克隆密切相关。在这些细菌中,嗜热嗜酸硫杆菌、耐热硫化芽孢杆菌和嗜铁钩端螺旋菌是优势细菌种类。嗜铁钩端螺旋菌是在以黄铁矿和添加硫的亚铁为能源的培养基中富集的微生物菌群中最主要的物种,丰度分别为53.8%和45.9%。在以黄铜矿为能源的培养物中,耐热硫化芽孢杆菌的丰度最高,为70.1%。

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