The objective of this study was to determine the association of physiological responses and root distribution patterns on yield of the second ratoon cane and the relationships among these traits. Seventeen sugarcane genotypes were planted in a randomized complete block design with four replications. The second ratoon crop was evaluated for germination percentage, cane yield, SPAD chlorophyll meter reading (SCMR), chlorophyll fluorescence, relative water content (RWC), specific leaf area (SLA) and stomatal conductance. Root length density (RLD) was evaluated by auger method. The root samples were divided into upper soil layer and lowers soil layers to study root distribution patterns. Sugarcane genotypes were significantly different for RLD, germination percentage and cane yield. Root distribution patterns were classified into three groups based on the RLD. High RLD between plants in the upper soil layers at 90 DAH was positively correlated with high germination, whereas high RLD between rows in the lower soil layers at 90 and 270 DAH was associated with high cane yield. RWC at 90 DAH and stomatal conductance at 180 DAH were closely related to germination percentage, whereas chlorophyll fluorescence and stomatal conductance at 180 DAH were closely related to cane yield.