| Summary: | Endophytic fungi are widely present in internal plant tissues and provide different benefits to their host. Medicinal plants have unexplored diversity of functional fungal association; therefore, this study aimed to isolate endophytic fungi associated with leaves of medicinal plants <i>Ephedra pachyclada</i> and evaluate their plant growth-promoting properties. Fifteen isolated fungal endophytes belonging to Ascomycota, with three different genera, <i>Penicillium, Alternaria,</i> and <i>Aspergillus</i>, were obtained from healthy leaves of <i>E. pachyclada</i>. These fungal endophytes have varied antimicrobial activity against human pathogenic microbes and produce ammonia and indole acetic acid (IAA), in addition to their enzymatic activity. The results showed that <i>Penicillium commune</i> EP-5 had a maximum IAA productivity of 192.1 ± 4.04 µg mL<sup>−1</sup> in the presence of 5 µg mL<sup>−1</sup> tryptophan. The fungal isolates of <i>Penicillium crustosum</i> EP-2<i>, Penicillium chrysogenum</i> EP-3, and <i>Aspergillus flavus</i> EP-14 exhibited variable efficiency for solubilizing phosphate salts. Five representative fungal endophytes of <i>Penicillium crustosum</i> EP-2, <i>Penicillium commune</i> EP-5, <i>Penicillium caseifulvum</i> EP-11, <i>Alternaria tenuissima</i> EP-13, and <i>Aspergillus flavus</i> EP-14 and their consortium were selected and applied as bioinoculant to maize plants. The results showed that <i>Penicillium commune</i> EP-5 increased root lengths from 15.8 ± 0.8 to 22.1 ± 0.6. Moreover, the vegetative growth features of inoculated maize plants improved more than the uninoculated ones.
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