不同16SrDNA靶序列PCR-DGGE分析油页岩细菌多样性

为全面了解油页岩细菌群落组成结构，同时筛选出适于油页岩PCR-DGGE的最佳引物，采用改进SDS高盐提取法，提取抚顺西露天矿油页岩微生物总DNA，并用968F/1401R，338F/518R，341F/907R和1055F/1406R，对16SrDNAV6-V8，V3，V8和V9区进行PCR-DGGE分析.结果表明，4组引物均能较好扩增出目的基因，但不同靶序列对多样性检出有显著影响(P<0001)；与其他引物相比，968F/1401R和338F/518R获得的指纹图谱物种丰富度更高，检出细菌类群更丰富，较适合油页岩样品.油页岩细菌群落组成较简单，优势细菌包括不动杆菌属、假单胞菌属和埃希氏菌属，同时还有假单胞菌目和肠杆菌目尚未被鉴定的一些种类.

Analysis of Bacterial Diversity in Oil Shale by PCR DGGE with Different 16S rDNA Target Sequences

For a comprehensive understanding of bacterial community structure of the oil shale from Fushun west open pit mine in China and screening out best prime for PCR DGGE analysis of oil shale， an improved SDS high salt extraction method was used to extract microbial total DNA from the oil shale. Four set primers (968F/1401R， 338F/518R， 341F/907R and 1055F/1406R) of 16S rDNA high variable target regions， V6 V8， V3， V8， V9， were compared to obtain the optimal target sequences suitable for PCR DGGE. The results from PCR DGGE patterns showed that four set primers can amplify the target sequences， but different target sequences of primers have a significant effect on the detection of the bacterial diversity (P<0001). Compared with the other primers， 968F/1401R and 338F/518R are more suitable for the bacterial diversity analysis of oil shale samples due to higher finger print species richness and abundant bacteria group obtained from their PCR DGGE. The community structure of bacteria in oil shale is not much rich. Acinetobacter， Pseudomonas and Escherichia are the dominant bacterial communities， as well as some unidentified bacterium which belong to Pseudomonadales and Enterobacteriales.