Alfalfa is a leguminous plant with strong autotoxicity, which seriously affects regeneration and stability. In order to clarify the relationship between the key autotoxic substances and autotoxicity of alfalfa, this experiment determined the content of phenolic autotoxic substances in different varieties of alfalfa and the effect of different concentrations of alfalfa extracts on seed germination, seedling growth and physiology. The results showed that the content of single autotoxic substances in the eight alfalfa varieties was highest for total coumarin. The variety with the highest total coumarin content was "LZ", and the lowest content was "656". Principal component analysis of the autotoxicity of eight alfalfa varieties revealed that the variety with the strongest autotoxicity was "LZ" and the weakest was "656". After treatment with extracts, the germination potential, germination rate, germination index and vigor index of 656 were higher than those of LZ, and the seeds of LZ and 656 did not germinate when the concentration was higher than C0.025 and C0.05, respectively. Compared with LZ, 656 had stronger osmotic regulation and antioxidant capacity, while the degree of membrane lipid peroxidation and ROS accumulation were lower. Further correlation analysis between the autotoxic substance content and autotoxicity observed that the content of total coumarin and autotoxic substances showed a significant positive association with autotoxicity (p < 0.01), and the total coumarin content showed a significant positive correlation with the content of autotoxic substances (p < 0.05). The total coumarin content is the major contributor to autotoxicity, and the higher the coumarin content, the higher the autotoxic substance content and the stronger the autotoxicity. Eight alfalfa varieties were systematically clustered on the basis of total coumarin content and autotoxicity, and the high-autotoxic alfalfa variety "LZ" and low-autotoxic alfalfa variety "656" were screened.
Keywords: alfalfa; autotoxicity; content; coumarin; phenolic autotoxic substances.