Fig 1.
Photomicrograph showing the morphology of fungal colony on PDA plate after 10 days at 27±2 °C, corresponding light microscope image of fungal endophytes (a) C. lunata, (b) D. eschscholtzii, (c) Colletotrichum sp. WF134, (d) D. tulliensis, (e) C. fusiforme, (f) C. gloeosporioides, (g) E. multirostrata and (h) D. tectonendophytica. Scale bar = 100 μm.
Fig 2.
Phylogenetic tree of fungal endophytes constructed by maximum likelihood bootstrap (MLBS) method.
The sequences were aligned through MUSCLE alignment program and the evolutionary history was inferred by using the Maximum Likelihood bootstrap (MLBS) method and General Time Reversible model.
Table 1.
Total phenolic and flavonoid content present in EA extract of identified fungal endophytes associated with the leaf of O. indicum.
Fig 3.
HPTLC fingerprint profiling of EA extract of fungal endophytes (a) TLC plate scanned at 254 nm, (b) TLC plate scanned at 366 nm, (c) TLC plate under white light after derivatization with anisaldehyde sulphuric acid reagent and (d) HPTLC fingerprint profiling of EA extract of fungal endophyte C. gloeosporioides showing unique bioactive component with respective value of retention factor (Rf).
Fig 4.
Free radical scavenging potential of EA extract of fungal endophytes (a) C. lunata, (b) D. eschscholtzii, (c) Colletotrichum sp. WF134, (d) D. tulliensis, (e) C. fusiforme, (f) C. gloeosporioides, (g) E. multirostrata and (h) D. tectonendophytica. All the experiments were performed in triplicate. P-value was calculated by comparing means ± SD of the free radical scavenging potential (%), using one-way ANOVA followed by Tukey to determine statistical significance. Statistical significance are as follows; ***, P≤0.001; **, P ≤0.002; *, P ≤0.033.
Table 2.
Antioxidant activity of EA extract of identified fungal endophytes associated with the leaf of O. indicum.
Fig 5.
Superoxide anion scavenging potential of EA extract of fungal endophytes (a) C. lunata, (b) D. eschscholtzii, (c) Colletotrichum sp. WF134, (d) D. tulliensis, (e) C. fusiforme, (f) C. gloeosporioides, (g) E. multirostrata and (h) D. tectonendophytica. All experiments were performed in triplicate. P-value was calculated by comparing means ± SD of the superoxide radical scavenging potential (%), using one-way ANOVA followed by Tukey to determine statistical significance. Statistical significance are as follows; ***, P≤0.001; **, P ≤0.002; *, P ≤0.033.
Fig 6.
Nitric oxide scavenging potential of EA extract of fungal endophytes (a) C. lunata, (b) D. eschscholtzii, (c) Colletotrichum sp. WF134, (d) D. tulliensis, (e) C. fusiforme, (f) C. gloeosporioides, (g) E. multirostrata and (h) D. tectonendophytica. All the experiments were performed in triplicate. P-value was calculated by comparing means ± SD of the nitric oxide radical scavenging potential (%), using one-way ANOVA followed by Tukey to determine statistical significance. Statistical significance are as follows; ***, P≤0.001; **, P ≤0.002; *, P ≤0.033.
Fig 7.
Hydroxyl radical scavenging potential of EA extract of fungal endophytes (a) C. lunata, (b) D. eschscholtzii, (c) Colletotrichum sp. WF134, (d) D. tulliensis, (e) C. fusiforme, (f) C. gloeosporioides, (g) E. multirostrata and (h) D. tectonendophytica. All experiments were performed in triplicate. P-value was calculated by comparing means ± SD of the hydroxyl anion radical scavenging potential (%), using one-way ANOVA followed by Tukey to determine statistical significance. Statistical significance are as follows; ***, P≤0.001; **, P ≤0.002; *, P ≤0.033.
Fig 8.
Cytotoxic activity of EA extract of fungal isolates C. gloeosporioides (OI-L6) against (a) HEK 293T cell line, (b) HCT 116 cell line, (c) HeLa cell line and (d) HepG2 cell line. All experiments were performed in triplicate. P-value was calculated by comparing means ± SD of percentage of cell viability of non-cancer and cancer cell, using one-way ANOVA followed by Tukey to determine statistical significance. Statistical significance are as follows; ***, P≤0.001; **, P ≤0.002; *, P ≤0.033.
Table 3.
Cytotoxic activity of EA extracts of eight fungal endophytes associated with the leaf of O. indicum against non-cancer and cancer cell lines.
Fig 9.
Comparative antioxidant activity and HPTLC fingerprint profiling of EA extract of C. gloeosporioides.
(a, b) Free radical scavenging potential of OI-L6 and MCC 9008, (c, d) Superoxide anion scavenging potential of OI-L6 and MCC 9008, (e, f) Nitric oxide scavenging potential of OI-L6 and MCC 9008, (g, h) Hydroxyl radical scavenging potential of OI-L6 and MCC 9008, (i) TLC plate scanned at 254 nm, (j) TLC plate scanned at 366 nm, (k) TLC plate under white light after derivatization with anisaldehyde sulphuric acid reagent. MCC 9008: C. gloeosporioides, associated with dead twigs. Experiments of antioxidant assays were performed in triplicate. P-value was calculated by comparing the means ± SD of scavenging potential (%) of EA extract of OI-L6 and MCC 9008, using one-way ANOVA followed by Tukey to determine statistical significance. Statistical significance are as follows; ***, P≤0.001; **, P ≤0.002; *, P ≤0.033.
Table 4.
Comparative antioxidant activity of EA extract of fungal strains C. gloeosporioides (OI-L6) and MCC 9008.