Abstract: We applied GC/MS detection method to different metabolites of Dioscorea bulbifera L. microtubers conserved in vitro at low temperature. Compared with D.bulbifera L. microtubers conserved in vitro at 25℃, different metabolites of D.bulbifera L. microtubers conserved in vitro at 4℃ had alanine, catechin, N,N-Di-(2-Hydroxyethyl)-methanamine, salicylic acid, citric acid and sorbose. In the conservation in vitro of microtuber at 4℃, alanine mainly involved in cyanoamino acid metabolism; Catechin mainly involved in biosynthesis of secondary metabolites, flavonoid biosynthesis and biosynthesis of phenylpropanoids; Salicylic acid mainly involved in polycyclic aromatic hydrocarbon degradation, microbial metabolism in diverse environments, plant hormone signal transduction, biosynthesis of secondary metabolites, dioxin degradation, phenylalanine metabolism, degradation of aromatic compounds, biosynthesis of plant hormones, biosynthesis of siderophore group non-ribosomal peptides and biosynthesis of phenylpropanoids; Citric acid mainly involved in biosynthesis of alkaloids derived from ornithine, lysine and nicotinic acid, biosynthesis of alkaloids derived from histidine and purine, microbial metabolism in diverse environments, biosynthesis of plant secondary metabolites, 2-Oxocarboxylic acid metabolism, biosynthesis of terpenoids and steroids, carbon fixation pathways in prokaryotes, biosynthesis of secondary metabolites, biosynthesis of alkaloids derived from shikimate pathway, biosynthesis of alkaloids derived from terpenoid and polyketide, citrate cycle(TCA cycle), biosynthesis of plant hormones, glyoxylate and dicarboxylate metabolism, two-component system, biosynthesis of phenylpropanoids and biosynthesis of alkaloids derived from ornithine, lysine and nicotinic acid. The preliminary findings of different metabolites of D.bulbifera L. microtubers conserved in vitro at low temperature would lay the foundation for further understanding the molecular mechanism of conservation in vitro at low temperature and provide a theoretical basis for low temperature conservation breaking dormancy and its subsequent germination D.bulbifera L. microtubers.