东北大学学报（自然科学版）

东北大学学报:自然科学版 ›› 2014, Vol. 35 ›› Issue (7): 1033-1037.DOI: 10.12068/j.issn.1005-3026.2014.07.027

• 资源与土木工程 • 上一篇    下一篇

不同16SrDNA靶序列PCR-DGGE分析油页岩细菌多样性

蒋绍妍1，王文星1,2，薛向欣1，杨寿浩2   

  1. (1. 东北大学 材料与冶金学院， 辽宁 沈阳110819； 2. 东北大学 生命科学与健康学院， 辽宁 沈阳110819)
  • 收稿日期:2013-09-01 修回日期:2013-09-01 出版日期:2014-07-15 发布日期:2014-04-11
  • 通讯作者: 蒋绍妍
  • 作者简介:蒋绍妍(1985-)，女，辽宁沈阳人，东北大学博士研究生；薛向欣(1954-)，男，辽宁沈阳人，东北大学教授，博士生导师.
  • 基金资助:
    国家自然科学基金资助项目(51204055)；中国博士后科学基金资助项目(20100481205).

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

JIANG Shaoyan1， WANG Wenxing1,2， XUE Xiangxin1， YANG Shouhao2   

  1. 1. School of Materials & Metallurgy， Northeastern University， Shenyang 110819， China; 2. School of Life & Health Sciences， Northeastern University， Shenyang 110819， China.
  • Received:2013-09-01 Revised:2013-09-01 Online:2014-07-15 Published:2014-04-11
  • Contact: XUE Xiangxin
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摘要： 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.

关键词: 油页岩, 16SrDNA, PCR-DGGE, 细菌多样性, 靶序列

Abstract: 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.

Key words: oil shale, 16S rDNA, PCRDGGE, bacterial diversity, target sequences

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