The use of mepiquat chloride (MC) in cotton (Gossypium hirsutum L.) cultivation has increased significantly in recent years. The use of MC, a growth regulator, results in higher precocity and lower height and consequently increases yield and reduces crop costs. The objective of this study was to evaluate the effects of nitrogen (N) and MC doses on fiber quality and cotton yield. This study was conducted during the 2012/13 and 2013/14 harvests in the experimental area of the Fundação de Apoio à Pesquisa Agropecuária de Chapadão, located in the municipality of Chapadão do Sul, MS, Brazil, at 18° 48’ 45.9” S, 52° 36’ 3” W, having an altitude of 820 m. The experimental design was a randomized blocks in a 5 × 4 factorial scheme comprised of five N doses (0, 40, 80, 120, 160 kg ha-1) and four MC doses (0, 50, 100, 150 mL ha-1) with four repetitions. Analyses of fiber quality included: length, uniformity, elongation, strength, micronaire index, reflectance, yellowing degrees and short fiber content. Regarding cotton yield, green color index (GCI), plant height at harvest time, and cotton seed yield were determined. The best results for fiber quality and productivity occurr with the application of 76.8 mL ha-1 of MC to the cotton crops when the N dose is greater than 80 kg ha-1. Increasing N doses causes an increase in cotton yield and micronaire index, but there is a decrease in fiber uniformity.