Invasive weeds cause substantial ecological, economical, and social problems, and are currently being controlled by herbicide applications. However, how herbicides affect other ecological interactions of invasive weeds, including their symbiosis with arbuscular mycorrhizal fungi (AMF), remains poorly understood. In this study, we therefore conducted field investigation to understand how the herbicide glyphosate affects the AMF diversity in the rhizosphere of the invasive weed Solidago canadensis. We also performed a greenhouse experiment to study if AMF can contribute to herbicide resistance. The results showed that the AMF colonization rate was significantly higher in S. canadensis when exposed to glyphosate in the field or in greenhouse settings. AMF diversity was also found to be higher in the rhizosphere soil after glyphosate application in the field. AMF colonization in greenhouse experiments also positively correlated with plant growth and reduced amounts of damaged leaves and the plant's content of the stress markers flavonol and anthocyanin. Chlorophyll content was significantly enhanced by AMF colonization, regardless of glyphosate application. These results indicate that herbicide can promote AMF colonization and diversity, and that AMF can enhance the herbicide resistance of S. canadensis. These findings suggest that herbicide application may promote the spread of S. canadensis through enhanced microbial interactions, posing new eco-environmental risks.