粘虫肠道细菌群落多样性的PCR-DGGE指纹图谱分析

Analysis of the Gut Bacterial Diversity of Mythimna separata by PCR-DGGE Fingerprint

研究东方粘虫[Mythimna separata(Walker)]肠道细菌的多样性。采用常规分离培养与分子鉴定相结合的方法,并以16SrDNA作为分子标记的变性梯度凝胶电泳(DGGE)对肠道细菌进一步分离,DGGE分离样品是研磨提取粘虫中肠组织DNA、提取培养混合菌DNA及直接以培养混合菌为模板进行PCR扩增的产物。结果表明,共得到DGGE条带19条,其中肠组织研磨提取DNA的DGGE条带最多,为17条;直接以培养混合菌液为模板进行PCR扩增次之,为13条;培养混合菌液提取DNA的条带最少,为12条。传统方法和分子生物学方法相结合研究肠道微生物多样性时能获得更全面的微生物信息,可采用培养混合菌提取DNA再结合其他方法进行后续研究。

The diversity of intestinal bacteria of Mythimna separata （Walker） were investigated in this experiment. The intestinal bacteria profile was analyzed by denaturing gradient gel electrophoresis （PCR-DGGE） based on the sequence of 16S rDNA. The traditional culture and molecular method were combined to isolated template DNA for the PCR. PCR products, which were amplified from DNA of grinding intestinal tissue, mixed bacteria culture and DNA from cultured mixed bacteria, re- spectively, were isolated by DGGE. 19 DGGE bands were detected in this study. 17 DGGE bands were gained from the DNA of grinding intestinal tissue sample and 12 bands were amplified from DNA of cultured mixed bacteria. The combination of molecular and traditional culturing methods can be used to analyze and monitor the diversity of intestinal bacteria effectively, and that will give us more information of microorganism diversity. Traditional culturing methods could use DNA of cultured mixed bacteria and combine other methods to do further experiments.