文章摘要
夏耘,郁二蒙,谢骏,余德光,王广军,李志斐,王海英,龚望宝.基于PCR-DGGE技术分析生物絮团的细菌群落结构[J].水产学报,2012,36(10):1563~1571
基于PCR-DGGE技术分析生物絮团的细菌群落结构
Analysis of bacterial community structure of Bio-Floc by PCR-DGGE
投稿时间:2012-02-23  修订日期:2012-07-10
DOI:10.3724/SP.J.1231.2012.27998
中文关键词: 生物絮团  细菌群落结构  α-变形菌属  PCR-DGGE
英文关键词: Bio-Floc  bacterial communities  α-proteobacteria  PCR-DGGE
基金项目:现代农业产业技术体系建设专项(CARS-46-17);公益性行业科研专项(201203083);国家科技支撑计划课题(2012BAD25B04);广东省海洋渔业科技推广项目(A200901D04)
作者单位
夏耘 中国水产科学研究院珠江水产研究所,农业部热带亚热带水产资源利用与养殖重点实验室 
郁二蒙 中国水产科学研究院珠江水产研究所,农业部热带亚热带水产资源利用与养殖重点实验室 
谢骏 中国水产科学研究院珠江水产研究所,农业部热带亚热带水产资源利用与养殖重点实验室 
余德光 中国水产科学研究院珠江水产研究所,农业部热带亚热带水产资源利用与养殖重点实验室 
王广军 中国水产科学研究院珠江水产研究所,农业部热带亚热带水产资源利用与养殖重点实验室 
李志斐 中国水产科学研究院珠江水产研究所,农业部热带亚热带水产资源利用与养殖重点实验室 
王海英 中国水产科学研究院珠江水产研究所,农业部热带亚热带水产资源利用与养殖重点实验室 
龚望宝 中国水产科学研究院珠江水产研究所,农业部热带亚热带水产资源利用与养殖重点实验室 
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中文摘要:
      在草鱼养殖过程中添加碳源(葡萄糖)维持水体C∶N为20∶1以培养生物絮团,通过对生物絮团细菌群落构成进行种类鉴定来评价生物絮团中功能微生物的组成。采用PCR-DGGE(变性梯度凝胶电泳)技术分析生物絮团形成第5、10和15天的细菌群落结构。DGGE指纹图谱结果分析表明:第5天和第10天的相似性最高,达67.4%;第5天和第15天相似性系数最低仅为40.5%。第10天时微生物多样性最高,第15天时多样性最低。对DGGE指纹图谱特征条带进行回收、克隆测序,结果表明,生物絮团培养过程主要微生物类群隶属于以下6个纲:α-变形菌纲(Alphaproteobacteria)、γ-变形菌纲(Gammaproteobacteria)、β-变形菌纲(Betaproteobacteria)、放线菌纲(Actinobacteria)、芽孢杆菌纲(Bacilli)、拟杆菌纲(Bacteroidetes),其中α-、β-及γ-变形菌占据主要位置。α-proteobacteria为3个阶段的共有优势菌,第5天时特异菌包括食酸菌属(Acidovorax)、气单胞菌属(Aeromonas)、土壤杆菌属(Agrobacterium),第10天和15天分别为芽孢杆菌属(Bacillus)与红球菌属(Rhodococcus)。研究首次发现,生物絮团应用于淡水养殖系统时细菌的组成和多样性都极其丰富,通过结合分析这些微生物的功能特点,为生物絮团技术在实际养殖生产中的进一步应用奠定基础。
英文摘要:
      Bio-Floc consists of phytoplankton,bacteria,aggregates of living and dead particulate organic matter,and grazers of the bacteria.It not only can regulate water quality,but also can supply the food protein for breeding animals.For the study of bacterial community structure of Bio Floc in freshwater culture systems,the key technologies of Israeli Avnimelech team about Bio-Floc in the field of aquaculture were used in this paper.Glucose was added as the carbon source which makes a C∶N ratio of 20∶1 be maintained during the experiment.The bacterial communities of Bio Floc were analyzed by using the PCR-DGGE(PCR-denaturing gradient gel electrophoresis)on the 5th,10th,and 15th day,respectively.The results indicated that the similarity of bacterial communities was highest on the 5th day and 10th day which was 67.4%,and was lowest on the 5th day and 15th day which was 40.5%.Diversity of bacterial species in the Bio-Floc was maximal on the 10th day and minimal on the 15 day.The acquired sequences of 24 bands in DGGE gel were performed by BLAST searches against NCBI database,and the sequences including the closely related sequences were aligned with Clustal W program in BioEdit software for phylogenetic analysis.The main microbes represented by 24 main bands in DGGE gel were Alphaproteobacteria,Gammaproteobacteria,Betaproteobacteria,Actinobacteria,Bacilli and Bacteroidetes.Among these bacteria,the α-,β-,and γ-proteobacteria are the major bacterial groups,but Actinobacteria bacteria exist on the 10th day and the 15th day.α-proteobacteria was the dominant bacteria during the entire process.The specific bacteria at different stages were usually the flocculant-producing bacteria:Acidovorax,Aeromonas,Agrobacterium only for 5th day;Bacillus,Rhodococcus for 10th day and 15th day,respectively.This study first found that the composition and diversity of bacteria of the Bio-Floc used in freshwater aquaculture systems are extremely rich.The study of functional microbial composition of the Bio-Floc may help to lay the foundation for the further application of the bio-floc technology in actual aquaculture production.
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