Abstract:Microorganisms are ubiquitous on the earth, with high diversity and a variety of functions. They play an irreplaceable role in human health, ecological stability, and species evolution. Despite the endeavor in the isolation, research, and utilization of microorganisms since the 17th century, most microorganisms cannot be obtained by the pure culture method, which has seriously hindered the research on microbial life activities and the development of microbial resources. The acquisition of the “dark matter” has always been a serious challenge. However, it is also an opportunity to break this shackle and obtain more microbial resources. This review introduces the influencing factors that restrict the isolation and incubation of microorganisms in the environments, describes the new culture techniques such as optimization of traditional culture strategy, co-occurrence network-based culture, membrane diffusion-based culture, microfluidics, and cell sorting, and puts forward the strategy of combining multiple techniques, with a view to providing reference for the development and utilization of microbial resources.
