Glyphosate-resistant (GR) horseweed has become an especially problematicweed in different crop production systems across the United States and theworld. In this field study, we used a nondestructive measurement system toanalyze the pollen production, deposition, and dispersion of a Tennesseeglyphosate resistant (TNR) horseweed biotype in Knoxville, TN during the2013 pollination season. We observed that the pollination season of TNRhorseweed lasted about 2 mo (54 d). About 78.93% of horseweed pollen wasreleased between 9:00 A.M. and 7:00 P.M. during each sampling day and therelease peak was at about 1:30 P.M. The seasonal release of pollen grainswas estimated to be 5.11 million grains plant−1. The release ratedata indicated that the integrated horizontal flux density and depositionflux density contributed to 78.17% and 21.83% of the release rate,respectively. We also found that pollen concentration decreased withdistance from the source field; the average pollen concentration decreasedto 50.69% at a distance of 16 m from the source plot. This is the firstresult of a systematic, direct examination of the release rate (emission anddeposition), release pattern (daily and seasonal), and dispersion pattern ofGR horseweed pollen.