污染土壤菌群结构研究.doc

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摘要:微生物对土壤中物质转化起到重要作用,研究污染土壤菌群结构有利于保护土壤环境,改良土壤,对农业生产有着重要的意义。本次实验通过对受到油泥污染的土壤分组进行4种前处理,并采用酚-氯仿法提取基因组DNA [1],从油泥污染土壤中提取出DNA。以此提出的DNA作为PCR反应模板,进行16S rRNA基因V3区有效扩增,并将扩增产物进行琼脂糖电泳测定,随后采用变性梯度凝胶电泳(DGGE)。根据DGGE电泳结果,采用Quantity One软件对DGGE图谱进行相关性分析。DGGE分子指纹图谱结果表明,单独种植高羊茅的油泥土壤与未种植植物、未接菌、定期浇水保持土壤湿润的油泥土壤最高相似系数为0.66;单独种植高羊茅、接菌D5A的油泥土壤与单独种植高羊茅的油泥土壤的最高相似系数为0.72;单独种植高羊茅、接种S-B细菌的油泥土壤与独种植高羊茅的油泥土壤的最高相似系数为0.53。即在植被、接菌等不同处理条件下的油泥土壤菌群结构存在明显差异。

关键词:油泥污染土壤; 提取DNA; PCR; DGGE; 菌群结构 

 

Abstract: Micro-organisms play an important role in the material transformation in the soil, contaminated soil flora structure conducive to the protection of the soil environment, soil improvement on agricultural production is of great significance. This experiment by the sludge-contaminated soil grouping the four kinds of pre-treatment with phenol - chloroform extraction of genomic DNA[1], DNA extracted from the sludge contaminated soil. This proposed DNA template for PCR reaction, the 16S rRNA gene V3 region of effective amplification, and PCR products were agarose gel electrophoresis and determination, followed by denaturing gradient gel electrophoresis (DGGE). Correlation analysis of DGGE profiles based on DGGE electrophoresis, using Quantity One software.DGGE molecular fingerprinting results show that separate planting of tall fescue clay soil and growing plants, not inoculated, and regular watering to keep the soil moist clay soil the highest similarity coefficient of 0.66; planting tall fescue alone, then the bacteria the D5A the sludgesoil and separate planting of tall fescue clay soil of the highest similarity coefficient of 0.72; separate planting of tall fescue, clay soil of the clay soil with the sole planting of tall fescue inoculated with SB bacteria the highest similarity coefficient of 0.53. There are obvious differences in the vegetation, then monocytogenes under different processing conditions, the sludge soil microflora.

Keywords: Sludge pollution of soil; DNA was extracted; PCR; DGGE; Microflora

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