Abstract: The study aims to screen out good cold-resistance varieties through a comprehensive evaluation of cold resistance. 9 sugarcane varieties, which had excellent agronomic traits and were bred by Yunnan Agricultural University, were taken as the test materials, low temperature stress processing was conducted at sugarcane seedling stage, the change of physiological and biochemical indexes were observed, and the comprehensive evaluation of the varieties (or lines) was carried out with pole sorting method. Results showed that: (1) low temperature (3℃) processing increased the conductivity of the sugarcane leaf cell membrane, soluble sugar content, MDA content, proline content, soluble protein content, chlorophyll content, peroxidase (POD) activity by an average of 21.21%, 134.1%, 83.60%, 35.47%, 47.72%, 9.07%, 565.2%, respectively; (2) low temperature (3℃) processing reduced sugarcane leaf superoxide dismutase (SOD) activity by 19.67% on average. Therefore, the screening results indicating that the best cold resistance sugar cane variety was ‘Yunnan sugarcane 03-91’, the worst cold resistance sugarcane variety was ‘Yunnan sugarcane 02-39’, while varieties (or lines) ‘Yunnan sugarcane 08-5’, ‘Yunnan sugarcane 05-103’, ‘Yunnan sugarcane 01-58’ had stronger cold resistance than ‘Yunnan sugarcane 04-14’, ‘Yunnan sugarcane 04-429’, ‘Yunnan sugarcane 05-522’ and ‘Yunnan sugarcane 02-227’ .