Abstract: Analysis of linkage inheritance of sub-red plant, a natural mutant from upland cotton, was conducted by two-point test using parents Gossypium barbadense Hai7124 and G. hirsutum B026 both with petal spots and green leaves, and G. hirsutum E083 with normal white flowers and, sub-red plants. The average percentage of recombination was 1.35% when calculating test-cross combining populations derived from E083/B026//Su9701 and B026/E083//Su9701. The average percentage of recombination was 2.94% when calculating test-cross combining populations derived from E083/Hai7124// Su9701 and Hai7124/E083//Su9701. Meanwhile, the genetic stocks having genotype Rs, Lc₁, r2, and rs, lc₁, and R₂ were used in the three-point test, which demonstrated that the mutant gene Rs was located between genes Lc and R2. The genetic distance between Rs and Lc₁ was 42.21 cM, and 1.68 cM between Rs and R2. Coefficient of interference among R₂, Lc₁, and Rs was 0.79, indicating that the single cross between Rs and R2 has less impact on the other single cross between Rs and Lc₁. Based on known genetic distance around gene Rs, the genetic linkage map was drawn. Rs is located between genes Lc₁ and R₂, molecular marker NAU2863, NAU3735 and NAU3735, NAU1048. The genetic distances between Rs and two markers NAU3735, NAU1048 are 0.1 and 0.2 cM, respectively.