Glyphosate-resistant (GR) Palmer amaranth has become a serious pest in parts of the Cotton Belt. Some GR cotton cultivars also contain the WideStrike™ insect resistance trait, which confers tolerance to glufosinate. Use of glufosinate-based management systems in such cultivars could be an option for managing GR Palmer amaranth. The objective of this study was to evaluate crop tolerance and weed control with glyphosate-based and glufosinate-based systems in PHY 485 WRF cotton. The North Carolina field experiment compared glyphosate and glufosinate alone and in mixtures applied twice before four- to six-leaf cotton. Additional treatments included glyphosate and glufosinate mixed with S-metolachlor or pyrithiobac applied to one- to two-leaf cotton followed by glyphosate or glufosinate alone on four- to six-leaf cotton. All treatments received a residual lay-by application. Excellent weed control was observed from all treatments on most weed species. Glyphosate was more effective than glufosinate on glyphosate-susceptible (GS) Palmer amaranth and annual grasses, while glufosinate was more effective on GR Palmer amaranth. Annual grass and GS Palmer amaranth control by glyphosate plus glufosinate was often less than control by glyphosate alone but similar to or greater than control by glufosinate alone, while mixtures were more effective than either herbicide alone on GR Palmer amaranth. Glufosinate caused minor and transient injury to the crop, but no differences in cotton yield or fiber quality were noted. This research demonstrates glufosinate can be applied early in the season to PHY 485 WRF cotton without concern for significant adverse effects on the crop. Although glufosinate is often less effective than glyphosate on GS Palmer amaranth, GR Palmer amaranth can be controlled with well-timed applications of glufosinate. Use of glufosinate in cultivars with the WideStrike trait could fill a significant void in current weed management programs for GR Palmer amaranth in cotton.