Due to widespread adoption of no-till management and use of glyphosate-resistant transgenic crops, glyphosate is the most widely used herbicide worldwide. However, its effect on soil microbial communities is inconsistent. We studied the effects of glyphosate, tillage, and crop rotation on the diversity and composition of soil bacterial communities in wheat (Triticum turgidum var. durum Desf.) rhizosphere after 6 and 7 yr of glyphosate applications. In a 2 × 2 × 2 factorial design, there were two crop rotation treatments: continuous wheat (W–W) and wheat in rotation with field pea (Pisum sativum L.) (P–W); two tillage treatments: minimum tillage (MT) and no-till (NT); and two glyphosate treatments: no application or pre-seeding application at the recommended rate. None of the treatments affected wheat rhizobacterial α-diversity or the relative abundances of most bacterial groups. The most abundant phyla were Proteobacteria (25.1% relative abundance), Actinobacteria (21.7%), Acidobacteria (8.7%), Bacteroidetes (5.9%), Firmicutes (1.4%), Armatimonadetes (1.3%), and Verrucomicrobia (1.2%). Glyphosate reduced the relative abundance of Alphaproteobacteria in W–W rotation but increased it in P–W rotation, and it reduced the relative abundance of Opitutus spp. The W–W rotation had greater relative abundances of the classes Bacilli (Firmicutes) and Gammaproteobacteria, and genera Bacillus and Opitutus (Verrucomicrobia), than the P–W rotation. Compared with MT, NT increased the relative abundance of the phylum candidate division WPS-1, but it reduced that of Phenylobacterium spp. in W–W rotation. These treatment effects probably had implications for soil functioning, including nutrient cycling and biological disease/pest control.