The invention discloses a wheat low temperature-resistant conditioning agent and an application method thereof. The wheat low temperature-resistant conditioning agent comprises the following components in parts by weight: 18-27 parts of melatonin, 13-19 parts of potassium humate, 12-17 parts of a-naphthylacetic acid, 8-14 parts of jasmonic acid, 6-14 parts of phytohormone brassinolide, 3-7 parts of cytokinin, 2-8 parts of an antifreeze, 0.3-0.7 parts of antibacterial peptide, 1-5 parts of a dispersant, and 20-30 parts of water. The wheat low temperature-resistant conditioning agent has the beneficial effects that the conditioning agent takes water as a medium, has obvious environment benefit and social benefit, can increase the security; protects a cell membrane, stabilizes the structure of biomacromolecules such as antioxidant enzyme, enhances capability for effectively removing ROS by cells, reduces intracellular water potential and ice point, and enhances the anti-pathogen capability of the wheat; the active ingredient source is abundant, the active ingredient is regenerable, is easily degraded in natural environment, has the characteristics of low cost and convenient usage, and conditioning agent has the advantages of easy popularization and application.