采用PCR-SSCP技术分析蚯蚓粪便的微生物群落结构

通过SDS/蛋白酶的方法抽提蚯蚓粪便微生物总DNA,以总DNA为模板,采用通用引物分别成功扩增细菌16S rRNA基因的V4-V5可变区,真菌18S rRNA基因的V8-V9可变区,应用单链构象多态性技术（SSCP）分析了蚯蚓粪便中不同空间层次的微生物种群的多样性。结果表明,在粪便的不同空间层次上呈现出明显的空间分布多样性,并且与纯蚯蚓粪微生物种群的多样性有很大的差别。为进一步的蚯蚓粪生态学功能研究提供了有益的指导。

Study on Microbial Community Structure in Earthworms Faecal by Single Strand Conformation Polymorphism(SSCP) Technique

Total DNA of microbe in earthworms faecal was extracted by SDS/protease method.Taking total DNA as template,the V4-V5 variable region of bacterial 16S rRNA genes and the V8-V9 variable region of fungal 18S rRNA genes were successfully amplified by universal primers.The microbial diversity of different spatial levels was analyzed by SSCP technique.The results indicated that microbial diversity was obvious on the different spacial level of earthworms faecal and the microbial diversity between earthworms faecal and pure earthworms faecal was much more different.It also provided useful guidance and theoretical basis for futher research of ecological function of earthworms faecal.