摘要
利用限制性片段长度多态性和16s rRNA PCR扩增及其序列分析来研究中国云南东川黄铜矿酸性浸矿废水(AMD)中微生物群落结构。基于建立16s rRNA克隆文库的分子方法常常被用来研究AMD中微生物生态。一小部分的rRNA被PCR扩增,克隆,然后用限制性片段长度多态性来筛选出其不同种类。在三个样地中,共挑选了357个克隆子,经限制性酶切分析后聚为100个可操作分类单元(OTU)。一个OTU代表各个样地中出现的同一种酶切图谱。有58个OTU被测序分析,结果在基因数据库中比对分析,其系统发育分析表明它们聚类为六大类:γ-proteobacteria,Acidobacteria,Clostridia,Actinobacteria,Nitrospira,α-pro- teobacteria。其中,γ-proteobacteria为最大的群落,在三个样地DCK-Ⅰ,DCN-Ⅰand DC-Ⅱ中分别占36%,57%,62%的比例。但与浸矿相关的微生物如At.ferrooxidans;L ferrooxidans等却发现的很少,仅在两个样地中各发现四个相关的OTUs。在这个AMD环境中浸矿相关的微生物都不是优势种群。
The acid mine drainage, commonly referred to as acid mine drainage （AMD）, has become an economic and environmental burden. The major producer of such effluents is the mining industry. Microorganisms accelerate iron oxidation rates at low pH. Physiological experiments have shown that AMD microorganisms can impact rates of sulfur oxidation during dissolution of pyrite, arsenopyrite, chalcopyrite, marcasite, and sphalerite. The presence of various bacterial species included common prokaryotic chemolithotrophs, such as Aciditiobacillus. Ferrooxidans, Leptospirillum ferrooxidans, A. thiooxidans, and A. caldus in mining environments. PCR-RFLP analysis was used to study the microbial community of three acid mine drainage from Dongchuan copper pyrite, China,which locate in the eastern part of Yunnan Province and has a mining history of more than 400 years and now is still one of the three major copper producers in China. 16s rRNA genes were amplified, subsequently cloned and detected by a restriction fragment length polymorphism （RFLP） analysis. A total of 357 clones were grouped into 100 operational taxonomic units （OTUs） by the RFLP analysis and 58 OTUs were sequenced. The phylogenetic analysis revealed that all sequences were clustered to six groups （γ-Proteobacteria, Acidobacteria, Clostridia, Actinobacteria, Nitrospira, α-Proteobacteria families）. γ-Proteobacteria family was the dominant groups in three sites. However, only few acidophilic microorganisms were detected and they were respectively affiliated with At.ferrooxidans and L. ferrooxidans. This study researched the geochemistry character of three sites of Dongchuang copper mine, evaluated the community composition of acid mine drainage by 16s rRNA gene clone libraries and debated the influence of the geochemistry character, temperature, pH value and the ion concentration for microorganism community structure and we considered that perennial low temperature （even less 20℃ in summer） may be the primary causation that At. Ferrooxidans and L. ferrooxidans was both not the dominated species.
出处
《现代生物医学进展》
CAS
2007年第11期1608-1611,共4页
Progress in Modern Biomedicine
基金
国家自然科学基金项目(No.50321402)
国家重点基础研究项目(973项目
No.2004CB619201)
