核糖体RNA测序技术在海洋微生物多样性研究中的应用

Advances in ribosomal RNA sequencing technologies for studying marine microbial diversity

摘要:

海洋微生物总数高达10³⁰个，在海洋生态系统中发挥着驱动能量物质循环和维持生态平衡的重要作用。深入研究海洋微生物多样性有助于理解海洋生态系统运行机理、应对全球海洋生态危机，以及开发海洋微生物资源。由于目前可培养分离的海洋微生物种类极少，极大限制了对海洋微生物群落、丰度和生理生态特征的研究。核糖体RNA测序技术以较低的成本，根据遗传信息差异对微生物快速准确地进行分类鉴定，揭示群落结构、评估进化和生态学关系。近年来基于核糖体RNA序列测序分析的海洋微生物学研究随着测序技术本身的快速优化，在发现海洋微生物新类群，揭示海洋微生物生态规律，分析海洋微生物进化关系，以及与海洋微生物相关的代谢产物开发和海洋生态治理等研究中取得显著进展。本综述详细介绍了核糖体RNA序列测序技术的原理，各代测序技术在海洋微生物多样性研究中的应用，以及多种建库和测序技术的有机结合。最后对研究不同海洋微生物多样性问题提供了测序方案的建议和展望，以期为核糖体RNA测序技术在海洋微生物多样性研究中的应用提供参考。

Abstract:

The total number of marine microorganisms is estimated to be 10³⁰. They are the key components driving marine energy and material recycling and maintaining ecological balance. Research advances in marine microbial diversity contribute to our understanding of the operative mechanisms of marine ecosystems. These knowledges can guide our respondence to ecological crisis of global oceans and our exploration of marine microbial resources. As the number of culturable marine microorganisms is currently low, our knowledges to the abundance, physiological characteristics and ecological functions of marine microbial communities are greatly limited. Ribosomal RNA sequencing technologies can rapidly and accurately identify or classify microorganisms based on nucleic acid sequences at low-cost. These technologies have been used in studying community structure and evolutionary and ecological relationships between microorganisms. In recent years, with the benefit of rapid development in sequencing technologies, marine microbial studies based ribosomal RNA sequencing analyses have achieved remarkable progresses in discovering novel marine microbial lineages, revealing ecological mechanisms and evolutionary relationships between marine microorganisms, discovering novel metabolites and applying in marine bioremediation. In this review, we introduce the technical principles of ribosomal RNA sequencing, the application of three generations of sequencing technologies in studying marine microbial diversity, and variable combinations of library construction and sequencing technologies. Finally, we address prospectives in utilizing these sequencing strategies in studies with different purposes.