海岸带生物学与生物资源保护实验室知识库

王中华, 宁波大学生命科学与生物工程学院, 宁波, 浙江 315211, 中国.; 李太武, 宁波大学生命科学与生物工程学院, 宁波, 浙江 315211, 中国.; 苏秀榕, 宁波大学生命科学与生物工程学院, 宁波, 浙江 315211, 中国.; 秦松, 中国科学院烟台海岸带可持续发展研究所, 烟台, 山东 264003, 中国.; 李佳霖, 中国科学院烟台海岸带可持续发展研究所, 烟台, 山东 264003, 中国.

为对原油降解过程中微生物群落结构的变化,本文采用直接向原油中加入营养物刺激的方法,获得原油降解菌体系。提取不同培养时间条件下降解菌体系的总DNA,并以之为模板采用PCR-DGGE技术研究了在原油降解过程中微生物群落的组成,结果表明,随着培养时间的不同,微生物的群落结构表现出差异,随着时间的变化其微生物群落也会发生改变。回收了DGGE图谱中的18个条带,测序结果经过Blast比对表明其中10个条带代表的细菌是不可培养的,显示了DGGE技术的优越性。同时采用气相色谱技术对降解过程中的原油烃的变化进行了分析,结果显示原油烃类组分会随着降解菌多样性的变化而发生变化,降解菌体系表现出了其对烷烃类物质的降解能力。  

The biodegradation bacterium system was made with the method of adding nutrition to oil.The techniques of molecular biology, including the extraction of bacterial DNA, PCR amplification of bacterial 16S rDNA by universal primers, and DGGE analysis were used in the experiment.The change of bacterial community structure and diversity were investigated and analyzed by the method of denaturing gradient gel electrophoresis(DGGE).The results showed that there were different type amount of strips among samples.Eighteen DGGE bands recovered were re-amplified, sequenced and aligned with Blast.The results showed that ten of the fragments belong to uncultivated bacteria implying DGGE technique having priority in analyzing uncultivated bacteria in the biodegraded crude oil.GC analysis showed that the low-carbon chain alkanes and high-carbon-chain alkanes as well as aromatic hydrocarbons can be degraded.