Recommended rates of glyphosate for noncultivated areas destroy theaboveground shoots of the perennial plant leafy spurge. However, suchapplications cause little or no damage to underground adventitious buds(UABs), and thus the plant readily regenerates vegetatively. Highconcentrations of glyphosate, applied under controlled environmentalconditions, have been shown to cause sublethal effects in UABs of leafyspurge that produce stunted and bushy phenotypes in subsequent generationsof shoots. We treated leafy spurge plants in the field with glyphosate (0,1.1, 3.4, or 6.7 kg ai ha−1) to determine its effects onvegetative growth from UABs and on molecular processes. The number of shootsderived from UABs of glyphosate-treated plants was significantly increasedcompared to controls in subsequent years after application, and new shootsdisplayed various phenotypical changes, such as stunted and bushyphenotypes. Quantifying the abundance of a selected set of transcripts inUABs of nontreated vs. treated plants (0 vs. 6.7 kg ha−1)indicated that glyphosate impacted molecular processes involved inbiosynthesis or signaling of tryptophan or auxin (ARF4, CYP79B2, PIN3, TAA1, TRP6, YUC4), gibberellic acid (GA1/CPS1, GA2/KS), ethylene (ACO1, ACS10), cytokinins (AHP1, AK2, CKX1), andthe cell cycle (CDC2A, CDC2B, CYCD3;1). Glyphosate-induced effects on vegetativegrowth and transcript abundance were persistent for at least 2 yr aftertreatment. Determining the molecular mechanisms associated with vegetativereproduction in leafy spurge following foliar glyphosate-treatment couldidentify limiting factors or new targets for manipulation of plant growthand development in perennial weeds.