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Background:  Endophytic fungi are largely underexplored in the discovery of natural bioactive products though being rich sources of novel compounds with promising pharmaceutical potential. In this study,  Taxus wallichiana,  which has huge medicinal value, was investigated for its endophytic diversity and capability to produce bioactive secondary metabolites by analyzing antioxidant, antimicrobial and cytotoxic properties.    Methods:  The endophytes were identified by ITS-PCR using genomic DNA samples. The secondary metabolites were extracted by solvent extraction method using ethyl acetate. The antioxidant activity was analyzed by Thin Layer Chromatography, Total Phenol Content (TPC), Total Flavonoid Content (TFC) and DPPH assay, and the antimicrobial activity was analyzed by agar-well diffusion method. Brine shrimp lethality assay was used to analyze the cytotoxicity of the fungal extracts.    Results:  Out of 16 different  Taxus  trees sampled from different locations of Dhorpatan, 13 distinctive endophytic fungi were isolated and grouped into 9 different genera:  Bjerkandera, Trichoderma, Preussia, Botrytis, Arthrinium, Alternaria, Cladosporium, Sporormiella  and  Daldinia . The ethyl acetate extracts isolated from three endophytic fungi:  Alternaria alternata ,  Cladosporium cladosporioides  and  Alternaria brassicae  showed significant TPC values of 204±6.144, 312.3±2.147 and 152.7±4.958µg GAE/mg of dry extract, respectively, and TFC values of 177.9±2.911, 644.1±4.202 and 96.38±3.851µg RE/mg of dry extract, respectively. Furthermore, these three extracts showed a dose dependent radical scavenging activity with IC  50  concentration of 22.85, 22.15 and 23.001 µg/ml, respectively. The extracts of  C. cladosporioides  and  A. brassicae  also showed promising antimicrobial activity against  Escherichia coli ,  Staphylococcus aureus  and  Bacillus subtilis  with a minimum inhibitory concentration of 250μg/ml for all bacteria. Both the samples showed cytotoxic property against shrimp nauplii with LC  50  of 104.2 and 125.9µg/ml, respectively.    Conclusions:  The crude fungal extracts obtained from endophytes:  A. alternata ,  C. cladosporioides  and  A. brassicae  upon purification and further identification of the bioactive compounds can be a fascinating source for novel pharmaceutical agents.

f1000research

Evaluation of antimicrobial, antioxidant and cytotoxic properties of bioactive compounds produced from endophytic fungi of Himalayan yew (Taxus wallichiana) in Nepal