Cucumber is regrettably greatly impacted by a variety of diseases in the field, which results in significant yield losses. Cucumber root rot disease, which is brought on by the pathogenic soil-borne fungus Fusarium oxysporum, Rhizoctonia solani, Fusarium solani, and negatively impacts cucumber production in Egypt. The Experimental Farm of the Faculty of Agriculture at Benha University served as the site for this experiment. Five biological agents were examined for their antagonistic effects on the growth of R. solani (RS1) in vitro, including the fungi Trichoderma harzianum, T. viride, and T. album, and the bacteria Pseudomonas fluorescens and Bacillus subtilis. For preventing Rhizoctonia root rot disease, root dipping treatments including salicylic acid (SA), ascorbic acid (As), calcium chloride (CaCl2), and dipotassium hydrogen phosphate (K2HPO4) at an 8 mM concentration were tried. When compared with the control, R. solani (RS1)'s growth was inhibited by all the biocontrol agents in use. In terms of growth reduction percentage, Trichoderma viride was the most effective antagonistic fungus (50.33%), followed by Trichoderma album (40.77%). Every tested fungicide increased total, conjugated, and free phenols as well as peroxidase, polyphenoloxidase and chitinase. This study determines the effectiveness of bioagents, chemical inducers, and fungicides in controlling R. solani caused cucumber root rot disease