A field experiment was undertaken to study the response of interspecific Bt cotton hybrid to different nutrientlevels and time of application of nitrogen and potassium in medium deep black soils at the Agriculture Research Station,Dharwad, Karnataka during the kharif season of 2014-15 under assured rainfall condition. The results indicated thatapplication of 150% RDF ha-1 (150:75:75 kg NPK ha-1) recorded significantly higher seed cotton yield per ha-1, highernitrogen, phosphorous and potassium uptake in stem, leaf, root and reproductive parts i.e 47.27, 51.39,2.60, 114.3 kg ha-1nitrogen, 6.91, 6.38, 0.76, 23.65 kg ha-1 phosphorous and 37.48, 12.60,1.07, 109.8 kg ha-1 potassium respectively. Drymatter production, nutrient content were also significantly higher in the treatment applied with 150% RDF ha-1 (150:75:75kg NPK ha-1) as against the treatments of 125% RDF and 100% RDF ha-1. Application of nitrogen and potassium in foursplits i.e., 25% N and K2O + 100% P2O5 as basal and 25% N and K2O each at 30, 60 and 90 DAS recorded significantlyhigher total nutrient uptake at harvesting stage as compared to the present practice of application of 50% N + 100% P2O5and K2O.