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The application relates to the field of flexible functional materials and human-computer interaction technology, and discloses a preparation method of a trehalose-enhanced anti-freezing conductive hydrogel fiber and application of the trehalose-enhanced anti-freezing conductive hydrogel fiber in intelligent gloves. The hydrogel is an acrylamide / polyvinyl alcohol double-network structure, and trehalose and lithium chloride are introduced. The trehalose forms a reversible hydrogen bond network with polymer chains through multiple hydroxyl groups, and the lithium chloride exists in the form of a hydrated ion in network pores to form an ion conductive path. The hydrogel forms a continuous linear fiber structure, the cross-sectional diameter is 0.3-2.5 mm, the flexibility is maintained at an environment of-20 DEG C, the conductivity retention rate is greater than or equal to 80%, the elongation at break is greater than or equal to 1000%, and the mechanical hysteresis is less than or equal to 5%. The application solves the problems of low-temperature failure and difficulty in fiberization of existing conductive hydrogels, and can be applied to flexible strain sensors, intelligent gloves and remote mechanical control systems.