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为了解北京山区油松(Pinus tabuliformis)林地点柄乳牛肝菌(Suillus granulatus)出菇期间菌塘内的细菌群落变化规律,收集出菇期前后(5-11月)的菌塘土样,通过聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)方法进行评估与分析.利用土壤总DNA提取试剂盒提取不同时期总DNA,以细菌通用引物338F和518R进行PCR扩增和DGGE,进一步对细菌群落的DGGE指纹图谱分析后进行切胶回收和测序.结果显示在点柄乳牛肝菌出菇期间,菌塘中细菌群落较为丰富,群落多样性指数维持在较高水准,并且菌塘中的细菌群落呈现出季节性变化.菌塘细菌分属于5个门(酸杆菌门、放线菌门、拟杆菌门、厚壁菌门和变形菌门)22个属(酸杆菌属、氨基酸杆菌属等).菌塘中变形菌门和酸杆菌门菌群的相对丰度较高,为优势菌群,其中α-变形菌纲为菌塘中最丰富的菌群.γ-变形菌纲菌群的相对丰度变化与点柄乳牛肝菌子实体发生周期一致.酸杆菌门菌群的相对丰度在点柄乳牛肝菌出菇前期(7月)明显增大.这些发现为点柄乳牛肝菌野生资源保育与人工驯化等后续研究提供了理论和实验依据.
In order to understand the variation of bacterial community in ponds during the fruiting period of Pinus tabuliformis forest in Beijing mountainous area, soil samples from ponds collected before and after mushroom stage (May-November) PCR-DGGE method was used to evaluate and analyze the total DNA extracted from soil total DNA at different periods, PCR amplification and DGGE were carried out with the common bacterial primers 338F and 518R, Bacterial community DGGE fingerprinting analysis was carried out gel recovery and sequencing.The results showed that during the fruiting of fructus coleus, the bacterial community in the ponds was more abundant and the community diversity index maintained at a higher level, The bacterial communities showed seasonal changes, and the bacteria belong to 22 genera (Acidobacteria, Amino acidum, Bacillus subtilis, Etc.) The relative abundance of Proteus mirabilis and acidobacteria in the ponds is relatively high, which is the dominant microflora, of which the α-proteobacteria are the most abundant in the ponds, and the γ-proteobacteria The relative abundance changes with the point-handled Suillus fruiting bodies The relative abundance of Acidobacter spp. Population increased significantly in the early fruiting stage (July) of the point of emergence of Dairy Boletus.These findings provide theoretical basis for the follow-up study on the conservation and artificial domestication of Dairy Boletus And experimental basis.