The invention discloses a rapid positioning method for industrial hemp trait related genes, and belongs to the technical field of industrial hemp research. The method comprises the following steps: variety selection, extreme group construction, specific site amplified fragment library construction and high-throughput sequencing, marker development, correlation analysis, gene annotation and real-time fluorescent quantitative PCR. The method comprises the following steps: constructing a high-low mixing pool for F1-generation groups obtained by hybridizing parents with relatively large target character differences, re-sequencing the parents, sequencing the high-low mixing pool by adopting a simplified genome deep sequencing technology, developing an SLAF tag, carrying out single nucleotide polymorphism detection, and carrying out marker correlation analysis on genotype frequency differences among the mixing pools. According to the method, a target character related candidate region is obtained, most plant populations at least need to be propagated to F2 generations to perform gene localization, and due to the high heterozygosity characteristic of industrial hemp, gene localization isrealized through F1 generations, so that rapid localization of industrial hemp genes can be realized in a high-efficiency and low-cost manner.