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污染控制技术及原理

刘有胜,杨朝晖,曾光明,肖勇,杨恋,徐峥勇.PCR-DGGE技术对城市餐厨垃圾堆肥中细菌种群结构分析[J].环境科学学报,2007,27(7):1151-1156

Application of PCR-DGGE to analyzing bacterial communities in culinary waste compost

基金项目：国家重点基础研究(973)发展计划项目(No.2005CB724203);国家自然科学基金资助项目(No.50478053);湖南省自然科学基金资助项目(No.04JJ3004,05JJ2004);湖南省科技计划项目(No.05FJ3001);长沙市科技计划重点项目(No.K051132-72)

摘要：使用基于16SrDNA的PCR-DGGE(变性梯度凝胶电泳)技术对城市餐厨垃圾堆肥过程中细菌种群结构随时间的变化进行了研究.在堆肥不同时间取样,进行了堆肥温度、pH、含水率、有机质、C/N的变化分析和DGGE分析.结果显示,堆肥温度高于50℃的天数为7d,最高达65℃,可有效杀灭致病菌;最终pH值接近7.5;C/N为20.04.PCR-DGGE图谱显示,不同时间堆肥样中细菌DGGE图谱有着明显的差异性;堆肥升温期细菌种群丰富,优势种群不明显;高温期细菌种群减少,优势种群明显;降温期细菌种群结构基本保持稳定.温度对堆肥过程中细菌种群具有明显的筛选作用.堆肥各阶段DGGE图谱相似性Cs值比较低,堆肥处理后细菌种群结构与堆肥原料之间存在明显差异.

Abstract：Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis was used to study the dynamic succession of bacterial communities during composting of culinary waste. Composts were sampled during different composting periods, and the changes of temperature, pH, water content, organic substance content and the ratio of carbon to nitrogen were measured. The results showed that the pile was insulated and temperatures above 50℃ were maintained for 7 days. The highest temperature in the compost was 65℃, which was high enough to kill pathogens. The final pH was 7.5 and the C/N was 20.04,and thus the final product could be used as organic fertilizer. After extraction and purification of genomic DNA from the sample compost, the 16SrRNAgenes (V3 region) were amplified by using the bacterial primer pair GC-341F/907R. Then, the amplified 16SrDNAfragments were separated by DGGE. Different DGGEbands appeared during different composting periods. The Cs values of pairwise similarity coefficient were analyzed for samples from composting days 0, 4, 10, 14, and 18. The Cs value of micro-ecological structure was the highest on the 4th day,Temperature appears to play a key role in determining the composition of bacteria present at the respective stages of the composting process.