Abstract:Many kinds of microorganisms with several robust functions are widely distributed and abundant in the natural environment, playing critical roles in human health and safety, ecological stability and species evolution. Although microbial cultivation technologies have been developed for more than one hundred years, by far, succeeded isolated microorganisms only account for 0.1%?1.0% of the total microorganisms in the planet due to plenty of limitation factors. Abundant microbial resources in the natural environments remain to be explored and applied. Therefore, it is particularly necessary to understand the restricting elements for cultivation of microbes and elucidate mechanisms, develop novel and effective methods of microbial isolation and culture. This paper analyzes the constraint factors for the growth of typical environmental microorganisms, then shows findings on the improvements of culture media and conditions. Subsequently, applications of novel techniques in microbial isolation such as in situ cultivation, co-culture, microfluidics cultivation and cell-sorting are reviewed, finally, the present bottleneck problems constraining uncultured microbes are discussed. The integration of multiple technologies, microbial interactions, metabolic pathway and mediation mechanisms, resuscitation mechanisms should be considered in the near future. Hopefully, it can provide a basic reference for the development and utilization of microbial resources.