Bioactive Components and Health Potential of Endophytic Micro-Fungal Diversity in Medicinal Plants
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sample Collection
2.2. Surface Sterilization
2.3. Sample Inoculation
2.4. Morphological Identification of Endophytic Fungi
2.5. Lactophenol Cotton Blue Mounting
2.6. Diversity Analysis
2.7. Production of Fermentation Medium
2.8. Extraction of Metabolites
2.9. FTIR Analysis
2.10. GC-MS Analysis
2.11. Antibacterial Assay
2.12. Phytochemical Analysis
2.13. Flavonoids Test
2.13.1. Alkaline
2.13.2. Lead Acetate
2.13.3. Ferric Chloride
2.13.4. Sulphuric Acid
2.13.5. Saponins
2.14. Tannins Test
2.14.1. Ferric Chloride
2.14.2. Gelatin
2.14.3. Lead Acetate
2.15. Proteins Test
2.15.1. Millon
2.15.2. Biuret
2.15.3. Ninhydrin
2.15.4. Aminoacids
2.16. Carbohydrate Test
2.16.1. Benedicts
2.16.2. Iodine
2.16.3. Fehlings
2.16.4. Terpenoids
2.16.5. Steroids
2.16.6. Glycosides
2.16.7. Alkaloids
Mayer Test
2.16.8. Phenols
Ferric Chloride Test
Litmus Paper Test
2.17. Antioxidant Analysis
2.17.1. DPPH Radial Scavenging Activity
2.17.2. H2O2 Radical Scavenging Activity
2.17.3. Nitric Oxide Scavenging Activity
2.18. Molecular Docking
2.19. FT-IR Spectrophotometer Analysis
2.20. GC–MS Analysis
2.21. Statistical Analysis
3. Results and Discussion
3.1. Endophytic Microfungal Isolation
3.2. FTIR Analysis
3.3. GC-MS Analysis for Bioactive Compound Screening
3.4. Antibacterial Potential of the Microfungal Extract
3.5. Phytochemical Analysis of the Endophytic Microfungal Extract
3.6. Antioxidant Activity of Endophytic Microfungal Extracts
3.6.1. DPPH Antioxidant Scavenging Activity of Fungal Extracts
3.6.2. H2O2 and Nitric Oxide Antioxidant Scavenging Activity of Fungal Extracts
3.6.3. Molecular Docking Study
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Extracellular (Ethyl Acetate) Extract | Intracellular (Methanol Mat) Extract | ||||||
---|---|---|---|---|---|---|---|
Gaminicolous helminthosporium | Bipolaris australiensis | Cladosporium cladosporioides | Gaminicolous helminthosporium | Bipolaris australiensis | Cladosporium cladosporioides | ||
Peak Range | Peak Range | Peak Range | Peak Range | Peak Range | Peak Range | Group | Compound |
517.85 | 517.85 | 456.13 | 516.89 | 516.89 | 516.89 | C-I stretching | halo compound |
580.53 | 547.75 | 517.85 | 596.93 | 595.96 | 593.07 | C-I stretching | halo compound |
635.50 | 592.11 | 592.11 | 640.48 | 648.04 | 648.04 | C-I stretching | halo compound |
692.40 | 636.47 | 635.50 | 678.90 | 679.89 | 678.90 | C-H bending | Monosubstituted |
806.19 | - | - | - | - | 874.66 | C=C bending | Alkene |
947.95 | 814.87 | - | - | 1007.74 | 940.23 | N-O stretching | Oxidized Nitrogen |
1056.92 | 1055.95 | 1057.88 | 1081.03 | 1056.92 | 1008.70 | CO-O-CO stretching | Anhydride |
1171.68 | 1101.28 | - | 1148.53 | 1147.57 | 1055.95 | P=O stretching | phosphine oxide |
1243.04 | 1243.04 | 1243.04 | 1384.79 | 1384.79 | 1147.57 | C-N stretching | Amine |
- | 1315.36 | 1312.26 | 1454.23 | 1315.36 | 1213.14 | S=O stretching | Sulfonamide |
1371.29 | 1338.51 | 1394.44 | 1338.51 | 1488.94 | 1398.30 | N-O stretching | nitro compound |
1644.20 | 1510.16 | 1510.16 | 1511.12 | 1589.23 | 1511.12 | C=C stretching | α,β-unsaturated ketone |
1679.88 | 1643.24 | - | - | - | 1605.63 | C=O stretching | tertiary amide |
1712.67 | 1712.67 | 1712.67 | 1717.49 | - | - | C=O stretching | carboxylic acid |
1767.64 | 1767.64 | 1759.92 | - | 1800.43 | - | N=C=S stretching | Isothiocyanate |
2314.42 | 2312.49 | 2360.71 | 2309.60 | 2309.60 | 2309.60 | O=C=O stretching | carbon dioxide |
2816.84 | 2882.42 | 2882.42 | 2884.35 | 2867.95 | 2868.92 | C-H stretching | Aldehyde |
2882.42 | 2980.78 | 2991.39 | 2981.74 | 2977.89 | 2942.21 | C-H stretching | Alkane |
2993.32 | 3290.33 | - | - | 3284.55 | 3336.62 | C-H stretching | Alkane |
3645.21 | 3588.32 | 3525.63 | - | - | - | N-H stretching | primary amine |
3746.47 | 3738.75 | 3739.72 | - | 3786.97 | 3723.32 | O-H stretching | Alcohol |
Compounds | Extracellular (Ethyl Acetate) | Intracellular (Methanol Mat) | Application |
---|---|---|---|
Benzo [h] quinoline | Bipolaris australiensis, Cladosporium cladosporioides | Cladosporium cladosporioides | Wound healing, antibacterial, antioxidant, antimalarial, antiviral, anticonvulsant, anti-HIV |
2-Ethylacridine | Graminicolous helminthosporium, Bipolarisaustraliensis, Cladosporium cladosporioides | Bipolaris australiensis, Cladosporium cladosporioides | Antitumor Antioxidant |
Cyclotrisiloxane | Graminicolous helminthosporium, Bipolaris australiensis, Cladosporium cladosporioides | Graminicolou shelminthosporium, Bipolaris australiensis, Cladosporium cladosporioides | Antibacterial, antiinflammatory Anticancer |
Anthracene | Bipolaris australiensis, Cladosporium cladosporioides | - | Insecticidal, wood preservative, used as a preparation of cool-tar, fossil fuel, cosmetic, anticancer, antibacterial, anti-inflammatory |
5-methyl-2-phenylindolizine | - | Graminicolous helminthosporium, Bipolaris australiensis | Inhibitory act against TB |
Indole-2-one | Graminicolous helminthosporium, Bipolaris australiensis | - | Enrich metabolism Anticancer Antiviral |
Tris(tert-butyldimethylsilyloxy)arsane | Graminicolous helminthosporium, Cladosporium cladosporioides | Graminicoloushelminthosporium, Bipolarisaustraliensis, Cladosporium cladosporioides | Antioxidant Antibacterial Antifungal |
3-quinolinecarboxylic acid | Graminicolous helminthosporium | - | Antibacterial Antifungal |
Benzene | Graminicolous helminthosporium, Bipolaris australiensis | - | Industrial manufacturing dyes, lubricants, drugs, fuel |
n-Hexadeconic acid | Bipolaris australiensis | - | Antiinflammatory antioxidant, anticancer, Antinociceptive, cytotoxic compound |
Oleic acid, Oxacyclotetradecan-2-one, Cis-Vaccenic acid | Bipolaris australiensis | - | Emulsifying agent, anticancer effect, reduces blood pressure, Reduces coronary heart disease (CHD), Antioxidant and antipolymerization agents, Used in asthma inhalers. |
Oxacyclotetradecan-2-one | Bipolaris australiensis | - | Antibacterial activity |
Cis-Vaccenic acid | Bipolaris australiensis | - | Antibacterial reduce sickle cell, reduction in the risk of coronary heart disease (CHD) |
Tetrasiloxane | Bipolaris australiensis | Bipolaris australiensis, Cladosporium cladosporioides | Anticancer |
Extracellular | Intracellular | |||||
---|---|---|---|---|---|---|
Cladosporium cladosporioides | Bipolaris australiensis | Graminicolous helminthosporium | Cladosporium cladosporioides | Bipolaris australiensis | Graminicolous helminthosporium | |
Flavonoid | +++ | +++ | +++ | +++ | +++ | +++ |
Saponin | ++ | - - | + | ++ | ++ | + |
Tannin | ++ | - - | - - | +++ | +++ | +++ |
Carbohydrate | - - | - - | - - | ++ | +++ | ++ |
Phenol | +++ | +++ | +++ | + | + | +++ |
Terpenoid | - - | +++ | +++ | + | ++ | ++ |
Concentration of Samples (µL) | C. cladosporioides | G. helminthosporium | B. australiensis | |||
---|---|---|---|---|---|---|
Absorption Value (OD) | % of Scavenging | Absorption Value (OD) | % of Scavenging | Absorption Value (OD) | % of Scavenging | |
10 | 0.2 | 68.99 c | 0.209 | 67.6 c | 0.195 | 69.76 c |
30 | 0.195 | 69.76 b | 0.192 | 70.23 b | 0.192 | 70.23 b |
50 | 0.192 | 70.23 a | 0.187 | 71 a | 0.187 | 71 a |
Concentration of Samples (µL) | C. cladosporioides | G. helminthosporium | B. australiensis | |||
---|---|---|---|---|---|---|
Absorption Value (OD) | % of Scavenging | Absorption Value (OD) | % of Scavenging | Absorption Value (OD) | % of Scavenging | |
10 | 0.187 | 70.01 c | 0.196 | 69.61 c | 0.132 | 79.53 c |
30 | 0.167 | 74.11 b | 0.189 | 70.7 b | 0.105 | 83.72 b |
50 | 0.147 | 77.21 a | 0.182 | 71.78 a | 0.026 | 95.97 a |
Concentration of Samples (µL) | C. cladosporioides | G. helminthosporium | B. australiensis | |||
---|---|---|---|---|---|---|
Absorption Value (OD) | % of Scavenging | Absorption Value (OD) | % of Scavenging | Absorption Value (OD) | % of Scavenging | |
10 | 0.123 | 93.35 b | 0.17 | 90.82 c | 0.167 | 90.98 c |
30 | 0.092 | 95.01 a | 0.118 | 93.63 b | 0.091 | 95.03 b |
50 | 0.087 | 95.31 a | 0.06 | 96.76 a | 0.058 | 96.87 a |
Concentration of Samples (µL) | C. cladosporioides | G. helminthosporium | B. australiensis | |||
---|---|---|---|---|---|---|
Absorption Value (OD) | % of Scavenging | Absorption Value (OD) | % of Scavenging | Absorption Value (OD) | % of Scavenging | |
10 | 0.266 | 85.63 c | 0.255 | 86.23 b | 0.273 | 85.25 c |
30 | 0.213 | 88.49 b | 0.203 | 29.03 c | 0.194 | 89.52 b |
50 | 0.107 | 94.23 a | 0.161 | 91.3 a | 0.15 | 91.9 a |
Concentration of Samples (µL) | C. cladosporioides | G. helminthosporium | B. australiensis | |||
---|---|---|---|---|---|---|
Absorption Value (OD) | % of Scavenging | Absorption Value (OD) | % of Scavenging | Absorption Value (OD) | % of Scavenging | |
10 | 0.065 | 86.82 c | 0.071 | 85.6 c | 0.068 | 86.21 b |
30 | 0.061 | 87.63 b | 0.065 | 86.82 b | 0.064 | 87.01 a |
50 | 0.059 | 88.03 a | 0.063 | 87.22 a | 0.061 | 87.63 a |
Concentration of Samples (µL) | C. cladosporioides | G. helminthosporium | B. australiensis | |||
---|---|---|---|---|---|---|
Absorption Value (OD) | % of Scavenging | Absorption Value (OD) | % of Scavenging | Absorption Value (OD) | % of Scavenging | |
10 | 0.273 | 44.62 c | 0.193 | 60.85 c | 0.32 | 35.1 c |
30 | 0.239 | 51.52 b | 0.073 | 85.19 a | 0.141 | 71.4 b |
50 | 0.065 | 86.82 a | 0.08 | 83.77 b | 0.092 | 81.34 a |
Ligand | E min |
---|---|
Benzo [h] quinolone | −213.20 |
Cyclotrisiloxane | −146.53 |
Ethylacridine | −213.24 |
Anthracene | −199.45 |
5 methyl 2 phenylindolizine | −224.85 |
Brallobarbital | −227.89 |
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Muthukrishnan, S.; Prakathi, P.; Sivakumar, T.; Thiruvengadam, M.; Jayaprakash, B.; Baskar, V.; Rebezov, M.; Derkho, M.; Zengin, G.; Shariati, M.A. Bioactive Components and Health Potential of Endophytic Micro-Fungal Diversity in Medicinal Plants. Antibiotics 2022, 11, 1533. https://doi.org/10.3390/antibiotics11111533
Muthukrishnan S, Prakathi P, Sivakumar T, Thiruvengadam M, Jayaprakash B, Baskar V, Rebezov M, Derkho M, Zengin G, Shariati MA. Bioactive Components and Health Potential of Endophytic Micro-Fungal Diversity in Medicinal Plants. Antibiotics. 2022; 11(11):1533. https://doi.org/10.3390/antibiotics11111533
Chicago/Turabian StyleMuthukrishnan, Sundaram, Paranivasakam Prakathi, Thangavel Sivakumar, Muthu Thiruvengadam, Bindhu Jayaprakash, Venkidasamy Baskar, Maksim Rebezov, Marina Derkho, Gokhan Zengin, and Mohammad Ali Shariati. 2022. "Bioactive Components and Health Potential of Endophytic Micro-Fungal Diversity in Medicinal Plants" Antibiotics 11, no. 11: 1533. https://doi.org/10.3390/antibiotics11111533