Abstract: High soil calcium content poses a serious threat to the normal growth and development of plants. This is an important limiting factor for agricultural production and environmental protection in the karst region of southwest China. Understanding the response mechanism of plants to high calcium stress can promote sustainable agricultural development and ecological environment protection in karst areas. In order to reveal the corresponding molecular mechanism of high calcium stress response in plants, this study made full use of the abundant biological information resources of Arabidopsis thaliana, and systematically compares and analyzes the gene expression of A.thaliana roots under control and high calcium stress treatment. 40 mmol/L CaCl2 treatment significantly inhibited A. thaliana root growth. Transcriptome analysis showed that under high calcium treatment, the expression of ethylene, abscisic acid and jasmonic acid in A. thaliana roots were up-regulated, and the expression of jasmonic acid synthesis, response and metabolic pathway related genes were up-regulated, indicating that these three hormone-related pathways plants had an important role in dealing with high calcium stress, especially the jasmonic acid-related pathways.