滇池沉积物中反硝化细菌及产甲烷菌的分子鉴别研究  

Molecular Identification of Denitrifying and Methanogens Bacteria in Sediments of Dianchi Lake

作　　者：尹利方 陈泽斌[1] 夏体渊[1] 王定康[1] 姚丽媛[1] 华金珠[1] 徐胜光 王家银[3] 

机构地区：[1]昆明学院农学院,云南昆明650214 [2]云南省高校生物炭工程研究中心,云南昆明650214 [3]云南省农业科学院农业经济与信息研究所,云南昆明650205

出　　处：《西南农业学报》2018年第1期154-159,共6页Southwest China Journal of Agricultural Sciences

基　　金：昆明学院科学研究项目(XJZD1606);昆明学院"人才引进项目"(YJL14005);云南省应用基础研究青年项目(2013FD040);云南省高校优势特色重点学科(生态学)建设项目(05000511311);云南省特色生物资源开发与利用重点实验室开放基金项目(GXKJ201631)

摘　　要：【目的】为了解滇池沉积物中反硝化细菌及产甲烷菌的种类组成。【方法】采用引物NirS1F/NirS6R和Me1/Me2分别对沉积物总DNA进行PCR扩增,得到1000 bp的反硝化细菌和760 bp的产甲烷菌特异性片段。【结果】证实了滇池沉积物中存在反硝化细菌和产甲烷菌,且推测产甲烷菌的丰度高于反硝化细菌。2个克隆文库分别获得阳性克隆子55和99个,利用Mothur软件将序列分别划分为10和5个OUT,说明尽管反硝化细菌的丰度低于产甲烷菌,但物种多样性高于产甲烷菌。运用BLASTN程序在GenBank数据库中进行序列比对,发现55个反硝化细菌阳性克隆子可分为10个种类,OTU10类序列与细菌Dechloromonas aromatica的相似性达86%,是反硝化细菌中的优势类群;99个产甲烷菌阳性克隆子可分为5个种类,OTU5类序列与甲酸甲烷规则菌(Methanoregula formicicum)相似性达86%,该类群是反硝化细菌中的优势类群。【结论】滇池沉积物中的反硝化细菌与产甲烷菌多为潜在的未被培养的新种。【Objective】The study aims to understand the species composition of denitrifying bacteria and methanogens in Dianchi sediments.【Method】The total DNA of sediments was amplified by PCR using primers NirS1F/NirS6R and Me1/Me2 respectively,A 1000 bp denitrifying bacterium and a 760 bp methanogenic bacterial specific fragment were obtained. 【Result】The experiments confirmed that there were denitrifying bacteria and methanogens in the sediments of Dianchi,the abundance of methanogens bacteria was higher than that of denitrifying bacteria. 55 clones and 99 clones were obtained of 2 clone libraries. Although the abundance of denitrifying bacteria was lower than that of methanogens,the species diversity was higher than that of methanogens bacteria. Using BLASTN sequence alignment in GenBank database,we found 55 positive clones of denitrifying bacteria could be divided into 10 species. The similarity of OTU10 sequence to Dechloromonas aromatica was 86 %,which was the dominant group in denitrifying bacteria. 99 methanogens positive clones could be divided into 5 species. 【Conclusion】The similarity between OTU5 sequence and Methanoregula formicicum was 86 %,which was the dominant group in denitrifying bacteria. The denitrifying bacteria and methanogenic bacteria in sediments of Dianchi were mostly new species that were not cultured.

关 键 词：滇池 沉积物 反硝化细菌 产甲烷菌 分子鉴别 

分 类 号：Q948[生物学—植物学]