Broomrape parasitism on faba bean (Vicia faba L.) is the most destructive factor for this crop in Egypt. Pot experiments were conducted during the two successive seasons 2017/2018 and 2018/2019 to study the mitigation of broomrape stress on faba bean using a ten-fold dilution of 10% (w/v) spent mushroom substrate extract (SMSE) of Pleurotus ostreatus and the same dilution of culture filtrate of mushroom (MCF) grown in potato dextrose broth (PDB) at a rate of 48 l hectare−1 compared with the commercial herbicide Roundup (Glyphosate 48% emulsifiable concentrate) at a rate of 144 cm3 ha−1 on the two varieties (Misr3 and Sakha3) cultivated in broomrape-infested soil. The treatments include the use of mushroom products as foliar spray and/or soil amendment in addition to Roundup spraying as a recommended treatment. Using Gas Chromatography-Mass Spectrometry (GC-MS) spectroscopy, our results indicate that the major components of the two mushroom products were bioactive compounds such as polyphenol and high molecular weight aliphatic and aromatic hydrocarbons that may interfere with parasite and host metabolism. These results indicated that SMSE of P. ostreatus and MCF of the same mushroom grown in potato dextrose broth (PDB) gave the best control of broomrape, and increased plant height, root length, leaf area, chlorophyll concentration, relative water content and seed yield (g plant−1), as well as anatomical characters of leaves in the two faba bean varieties (Misr3 and Sakha3), such as upper and lower epidermis, palisade tissue, spongy tissue and vascular bundles. Additionally, electrolyte leakage was decreased in the treated plants compared to control plants and the plants treated with Roundup (glyphosate) because of the important role of SMSE and MCF in the improvement of faba bean water status.