利用PCR-DGGE方法研究添加复合菌剂对堆肥微生物群落的影响

Effects of Adding to Complex Bacteria on Microbial Community of Compost Assessed by PCR-DGGE Techniques

为研究接种复合菌剂对堆肥微生物群落变化的影响,以奶牛粪便和稻草为原料进行堆肥试验,设添加复合菌剂BLD(由3株木质纤维素降解细菌组成,经测序鉴定分别为枯草芽孢杆菌、地衣芽孢杆菌、假单胞菌属中未鉴定种)和不加菌剂两个处理,利用传统平板培养与聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)方法相结合研究两种堆肥处理对微生物群落演替的影响。结果表明,基于传统培养方式,微生物数量呈升高-降低-升高-降低的趋势,且细菌数量明显大于真菌数量;DGGE图谱显示,两种堆肥处理的条带数呈现与传统培养相似的变化趋势。接种菌株在堆肥初期成功定殖,高温期成为优势菌株,降温期优势逐渐减弱。接种菌剂增加了堆肥中细菌数量,提高了堆肥微生物群落多样性,从而促进堆肥微生物群落演替,缩短堆肥腐熟时间。

In order to study the influence of inoculated strains on composting microbial community strcture,composting experiment was conducted,which used the cow dung and straw as raw materials.Two groups were designed,bacterial strains were added to one group（code name as BLD,which were composed of three kinds of strains,and sequencing showed that they were B.subtilis,B.licheniformis and Pseudomonas）,the other was the control.Variation of microbial communities in inoculation and natural composts was studied with the traditional plate-count method and the PCR-DGGE technology.Results indicated that：based on the traditional method,the population of microbes showed wave-like fluctuation：up-down-up-down,and the total bacterial count was significantly more than fungi.The DGGE graphs showed similar trend.And the inoculated strains colonized successfully in the early stage of compost,they became the superior strains in the high temperature phase,the strength of bands reduced in the cooling and maturation phases.Inoculatation could increase the total count of bacterial,enhance microbial diversity and accelerate microbial succession,and then shorten the maturation phase.