Persisters are a group of special subpopulation of bacteria, only occupying < 0.1% of the whole population but having the characteristics different from the ordinary bacteria and resistant mutants. They have complex formation mechanism, and are difficult to isolate and culture. The persisters can adapt to the adverse environment via "dormancy-growth-proliferation" to maintain their survival and cell structure stability, and play a vital role in the multi-drug and multi-metal tolerance of microbial biofilm, being of great significance in maintaining the stability of microbial community structure. This paper reviewed the research progress on the characteristics of persisters, their gene regulation mechanisms, and their effects on the multi-drug and multi-metal tolerance of microbial biofilm. The related research directions in the future were also prospected.