Bitter melon, a bitter tropical plant, was used as a medicinal plant from the past. Due to the observation of toxic effects of bitter melon extract against pathogenic bacteria and fungi, this study was performed to investigate ripe fruit extracts effects on reproduction and growth phytopathogenic bacteria and fungi. For this purpose, the ripe fruits of bitter melon were collected and dried and then was extracted by ethanol and ethylacetate solvents. To study of antimicrobial activity of extract, Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and Minimum Fungicidal Concentration (MFC) and inhibition zone of the sample were determined using microdilution broth method and disc diffusion method, respectively. For comparison of the extract effects against fungi and bacteria, a sample without plant extract was prepared as the control. Also, gas chromatography coupled to a mass spectrometer (GC- MS) analysis was performed to identify and determine the percentage of extracts metabolites. The results indicated different antimicrobial activities of the extracts against the studied phytopathogenic bacteria and fungi and it was more affected on fungi than bacteria. In addition, the phytochemical results showed that among the detected compounds in bitter melon, Tetradecanoic acid (49.62%) and n-Hexadecanoic acid (19.53%) in the ethyl acetate extract and Thiirane (72.95%) in the ethanol extract were the main compounds that their antimicrobial effects have already been reported. According to the results of this study, could be observed that these identified metabolites of bitter melon have the antimicrobial activity against studied phytopathogenic bacteria and fungi.