Johnsongrass is one of the most troublesome weeds of the world and is listed as a noxious weed in Arkansas. Reduced johnsongrass control with the recommended application rate of glyphosate (840 g ae ha1) was reported in a continuous soybean field near West Memphis, AR, in the fall of 2007. A greenhouse study was conducted (1) to confirm and characterize glyphosate resistance in the johnsongrass biotype from West Memphis and (2) to determine whether resistant and susceptible biotypes have differential glyphosate absorption or translocation. Doseresponse studies revealed that the resistant biotype was five- to seven-fold less sensitive to glyphosate than the susceptible biotype. Glyphosate absorption was similar in resistant and susceptible biotypes at 72 h after treatment (HAT). However, the treated leaf of the resistant biotype retained 28 percentage points more absorbed14C glyphosate compared to the susceptible biotype at 72 HAT. Additionally, the resistant biotype had less14C glyphosate translocated to the aboveground tissue below the treated leaf and to roots compared to the susceptible biotype at 24 and 72 HAT. Reduced translocation and increased retention of glyphosate in treated leaves is a probable mechanism of resistance in this glyphosate-resistant johnsongrass biotype.