Infestations of glyphosate-resistant (GR) horseweed have become widespread in the eastern United States. This biotype is problematic in no-tillage production that relies extensively on glyphosate for weed control. Because horseweed is treated at various stages of growth, a greenhouse study explored rate response of glyphosate-resistant and -susceptible horseweed at three growth stages. GR horseweed was more responsive to glyphosate at the seedling stage than at the large rosette or bolting stages. A field study evaluated GR horseweed response when treated with glyphosate at soybean planting time, POST in-crop (about 45 d after planting), or both at planting and POST in-crop. There was a cumulative effect of the at-planting followed by POST in-crop glyphosate applications. When evaluating single glyphosate applications, the at-planting application was more effective at suppressing GR horseweed than a POST in-crop application. Because glyphosate cannot control GR horseweed, this biotype should be controlled with an herbicide with an alternate mode of action and applied at the most effective timing.