Fungal endophytes associated with medicinal plants have potential role to promote plant growth through different mechanisms. However, the biological and ecological roles of fungal endophytes still totally unexplored. In this study, three different fungal endophytes were isolated from the medicinal plant of Asclepias sinaica and identified as Penicillium chrysogenum Pc_25, Alternaria alternata Aa_27 and the third fungal strain was described as sterile hyphae Sh_26. It was recorded that, these endophytes had various ability to produce several extracellular enzymes including amylase, pectinase, cellulase, gelatinase, xylanase and tyrosinase. Their antimicrobial activities against different specific test organisms were investigated as well. In addition, both endophyte isolates i.e. Sh_26 and Aa_27 were found to promote root growth higher than Pc_25 and control treatments. These fungal isolates had a considerable impact on plant growth parameters including root elongation as a result of ammonia and IAA production.