Weeds significantly reduce sugarcane (Saccharum officinarum L.) production by 30% to 50% and cause complete crop loss in severe cases. Guangxi, a central sugarcane-growing region in southern China, faces significant challenges due to the proliferation of weeds severely impacting crop tillering, yield, and quality. In this study, we surveyed and identified 35 weed species belonging to 16 families in Longzhou, Nongqin, and Qufeng, with significant threats posed by purple nutsedge (Cyperus rotundus L.), bermudagrass [Cynodon dactylon (L.) Pers.], hairy crabgrass [Digitaria sanguinalis (L.) Scop.], black nightshade (Solanum nigrum L.), white-edge morningglory [Ipomoea nil (L.) Roth], and ivy woodrose [Merremia hederacea (Burm. f.) Hallier f.]. The application of 81% MCPA-ametryn-diuron achieved greater than 90% control within 15 d. Although herbicides are effective, they can unintentionally harm sugarcane, indicating a need for tolerant genotypes. Therefore, we comprehensively evaluated herbicide-induced phytotoxic responses and identified tolerant sugarcane genotypes over 3 yr of trials conducted on 222 genotypes across Guangxi. We quantified phytotoxicity by counting the number and severity of affected leaves. The ANOVA revealed statistically significant main and interaction effects among genotype, crop cycle, and location. Cluster and discriminant analyses classified the genotypes into five groups: 21 highly tolerant (HT), 68 tolerant, 75 moderately tolerant, 18 susceptible, and 40 highly susceptible. The 21 HT genotypes demonstrated strong potential to be used as parental lines for breeding herbicide-tolerant varieties, to inform precision breeding strategies, and to increase tolerance to herbicide stress in sugarcane.