Endophytic bacteria promote plant growth, reduce stress caused by biotic and abiotic factors, and can trigger active defense reactions in plants. This study aimed to evaluate enzyme activity of in vitro jojoba (Simmondsia chinensis) plants inoculated with endophytic bacteria. In vitro shoots of female and male plants were inoculated with strains of Azospirillum brasilense (Cd), Methylobacterium aminovorans (JRR11), Rhodococcus pyridinivorans (JRR22) or co-inoculated with a mixture of JRR11 + JRR22. A total of 10 treatments were performed to evaluate shoot and root length; changes in key enzymes involved in plant defense (superoxide dismutase, catalase, peroxidase, ascorbate peroxidase and phenylalanine ammonia lyase) after post-inoculation (45 days). All endophytic bacteria strains used promoted plant growth and rhizogenesis. Differences were found in enzyme activity between female and male plants. The plants inoculated with JRR22 strain, showed the highest enzyme activity suggesting an induced systemic response and a potential increase in plant resistance to pathogen attack.