微生物分子生态学研究方法及其存在的缺陷

The research methods for molecular microbial ecology and their defects

微生物群落多样性是微生物生态学和环境学研究的重点之一。分子生物学方法应用于微生物群落结构分析使得对环境样品中占大多数的不可培养微生物的研究成为了可能。然而,PCR过程中的偏差(bias)会引起结果并不能如实地再现原始的群落结构,随着多个宏基因组项目的研究,发现所谓的"通用引物"并不能覆盖全部的微生物类型,即使可以添加简并碱基,也无法与通过宏基因组得到的rRNA序列完全匹配,这导致了在诸多研究中会忽略环境中的微生物群落。即使是不经过PCR扩增的元基因组和元转录组学研究方法,对于分子微生物群落也存在着一定的问题。

Microbial community diversity is an essential issue in both microbial ecology and environmental microbiology.The features and characteristics of uncultured microorganisms,also known as the majority population in environments,could be accomplished because of the introduction of molecular methods.However,the PCR bias caused by PCR amplification especially by primer mismatches usually makes quantitative analysis of microbial communities inaccessible and hampered the analysis of biosphere even with massive sequencing.Even the methods without PCR amplification methods of metagenomic and metatranscriptomic strategies also have shortcomings when analyzing the microbial communities.