Laticifers are hypothesized to mediate both plant-herbivore and plant-microbe interactions. However, there is little evidence for the dual function of these secretory structures. We investigated whether the major constituent of natural rubber, cis-1,4-polyisoprene, a phylogenetically widespread and economically important latex polymer, alters plant resistance and the root microbiome of the Russian dandelion (Taraxacum koksaghyz) under attack of a root herbivore, the larva of the May cockchafer (Melolontha melolontha). Rubber-depleted transgenic plants lost more shoot and root biomass upon herbivory than normal rubber content near-isogenic lines. M. melolontha preferred to feed on artificial diet supplemented with rubber-depleted latex rather than normal content latex. Likewise, adding purified cis-1,4-polyisoprene in ecologically relevant concentrations to diet deterred larval feeding and reduced larval weight gain. Metagenomics and metabarcoding revealed that abolishing biosynthesis of natural rubber alters the structure but not the diversity of the rhizosphere and root microbiota in a herbivore-dependent manner. Roots from rubber-depleted plants, however, did not exhibit a higher pathogen load compared to normal rubber content roots. Taken together, our data demonstrate that natural rubber biosynthesis reduces herbivory and alters the plant microbiota in a herbivore-dependent manner, which highlights the role of plant specialized metabolites and secretory structures in shaping multitrophic interactions.