Excessive or insufficient supplementation of trace elements (TEs) limits the progression of anaerobic digestion. The main reason for this is the lack of sufficient understanding of digestion substrate characteristics, which significantly affects the demand for TEs. In this review, the relationship between TEs requirements and substrate characteristics is discussed. We mainly focus on three aspects. 1) The basis for TE optimization and existing problems: The optimization of TEs often based on the total solids (TS) or volatile solids (VS) of substrates, does not fully consider substrate characteristics. 2) TE deficiency mechanisms for different types of substrates: nitrogen-rich, sulfur-rich, TE-poor, and easily hydrolyzed substrates are the four main types of substrates. The mechanisms underlying TEs deficiency in the different substrates are investigated. 3) Regulation of TE bioavailability: characteristics of substrates affect digestion parameters, which disturb the bioavailability TE. Therefore, methods for regulating bioavailability of TEs are discussed.