The sugarcane genotype YZ 07-86 is a distinct natural mutant with dual-axillary buds. Exploitation of this valuable trait in sugarcane breeding requires an understanding of the physiological mechanism of dual-axillary bud formation. This experiment determined the contents of indole-3-acetic acid (IAA), abscisic acid (ABA), gibberellins (GA3 and GA4), and three cytokinins (CKs), namely isopentenyl adenosine (iPA), zeatin riboside (ZR), and dihydrozeatin riboside (DHZR), in the shoot apical meristem tissues of YZ 07-86 and its sister line YZ 07-87, which served as the control genotype. The field experiment followed a completely randomized design with three replications for each genotype. Sampling was done at 5-day intervals, starting 106 days after planting. The ELISA technique was used to measure the contents of endogenous hormones. The study revealed that the mean contents of IAA, GA3, GA4, DHZR, and iPA were not significantly different (P > 0.05) among the genotypes. Conversely, in YZ 07-86, the mean ZR content was significantly higher (P < 0.05), and the mean ABA content was significantly lower than in the control genotype. Moreover, the mean content ratios of ZR/IAA, ZR/ABA, ZR/GA3, and ZR/GA4 were also significantly higher in YZ 07-86. The study concludes that elevated ZR content and an increased ZR/ABA ratio directly contributes to dual-axillary bud formation in YZ 07-86, offering valuable insights for the advancement of sugarcane genotypes.