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Hemp (Cannabis sativa L.) is an important crop, yet little is known about how herbivory and soil microbial communities interact to influence plant performance. In this study, two hemp cultivars, BaOx and Cherry Citrus, were grown under identical greenhouse conditions and exposed to naturally occurring background populations of the two-spotted spider mite (Tetranychus urticae). Plant traits were measured, and rhizosphere soil was sampled for 16S rRNA gene sequencing to compare bacterial community composition and diversity between cultivars. Spider mite injury was assessed using a standardized 0–5 visual damage scale commonly applied in integrated pest management studies. Although the cultivars did not differ significantly in growth traits, Cherry Citrus experienced significantly less spider mite damage than BaOx, suggesting greater tolerance or resistance to herbivory under shared conditions. Rhizosphere bacterial communities differed markedly between cultivars despite identical soil and environmental conditions. BaOx rhizospheres were enriched in Actinobacteria, including taxa associated with decomposition and antimicrobial compound production, whereas Cherry Citrus rhizospheres were enriched in Alphaproteobacteria, particularly nitrogen-cycling and root-associated taxa such as Rhizobium and Reyranella. Alpha diversity metrics did not differ between cultivars; however, beta diversity analyses revealed significant cultivar-level separation, particularly in phylogenetic community structure. Because herbivore pressure and microbial communities were not experimentally manipulated, this observational study identifies ecological associations rather than direct causal relationships. Nevertheless, the results demonstrate that hemp cultivar identity is associated with distinct rhizosphere microbiomes and differential susceptibility to spider mite damage. These findings highlight the potential for integrating cultivar selection and microbiome-informed strategies into sustainable pest management programs for hemp.