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Due to the emergence of bacterial resistance in phytopathogenic microorganisms, it is necessary to search for new treatment alternatives for these pathogens. Natural extracts are a potential source of bioactive compounds that can act against such bacterial strains. The antibacterial activity of <i>Larrea tridentata</i> against bacteria with public health importance has been documented; however, few reports cover pathogens associated with the agricultural sector. The aim of the present study was to evaluate the antibacterial activity of <i>Larrea tridentata</i> hydroalcoholic extract (LTHE) and fractions against phytopathogenic bacteria. LTHE was obtained by the maceration technique and then subjected to bipartition using solvents of different polarities. <i>Clavibacter michiganensis</i> sbsp. <i>michiganensis</i>, <i>Pseudomonas syringae</i>, and <i>Xanthomonas campestris</i> strains were used, and their antibiotic sensitivity was determined. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of LTHE and its fractions (aqueous: LTAq-F; organic: LTEtOAc-F) were determined. LTHE and its organic fraction showed bactericidal activity against the three bacteria, showing better activity against <i>X. campestris</i>, exhibiting an MIC of 0.39 mg/mL and an MBC of 0.78 mg/mL. The results show that LTHE and its organic fraction have bactericidal activity in vitro against <i>Clavibacter michiganensis</i> sbsp. <i>michiganensis</i>, <i>Pseudomonas syringae</i>, and <i>Xanthomonas campestris</i>.