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Multigene Phylogeny, Beauvericin Production and Bioactive Potential of Fusarium Strains Isolated in India
Shiwali Rana1,2, Sanjay Kumar Singh1,2, Laurent Dufossé3
1National Fungal Culture Collection of India, Biodiversity and Palaeobiology Group, MACS' Agharkar Research Institute, G.G. Agarkar Road, Pune 411004, India.
Abstract:
The taxonomy of the genus Fusarium has been in a flux because of ambiguous circumscription of species-level identification based on morphotaxonomic criteria. In this study, multigene phylogeny was conducted to resolve the evolutionary relationships of 88 Indian Fusarium isolates based on the internal transcribed spacer region, 28S large subunit, translation elongation factor 1-alpha, RNA polymerase second largest subunit, beta-tubulin and calmodulin gene regions. Fusarium species are well known to produce metabolites such as beauvericin (BEA) and enniatins. These identified isolates were subjected to fermentation in Fusarium-defined media for BEA production and tested using TLC, HPLC and HRMS. Among 88 isolates studied, 50 were capable of producing BEA, which varied from 0.01 to 15.82 mg/g of biomass. Fusarium tardicrescens NFCCI 5201 showed maximum BEA production (15.82 mg/g of biomass). The extract of F. tardicrescens NFCCI 5201 showed promising antibacterial activity against Staphylococcus aureus MLS16 MTCC 2940 and Micrococcus luteus MTCC 2470 with MIC of 62.5 and 15.63 µg/mL, respectively. Similarly, the F. tardicrescens NFCCI 5201 extract in potato dextrose agar (40 µg/mL) exhibited antifungal activity in the food poison technique against plant pathogenic and other fungi, Rhizoctonia solani NFCCI 4327, Sclerotium rolfsii NFCCI 4263, Geotrichum candidum NFCCI 3744 and Pythium sp. NFCCI 3482, showing % inhibition of 84.31, 49.76, 38.22 and 35.13, respectively. The antibiotic effect was found to synergize when Fusarium extract and amphotericin B (20 µg/mL each in potato dextrose agar) were used in combination against Rhizopus sp. NFCCI 2108, Sclerotium rolfsii NFCCI 4263, Bipolaris sorokiniana NFCCI 4690 and Absidia sp. NFCCI 2716, showing % inhibition of 50.35, 79.37, 48.07 and 76.72, respectively. The extract also showed satisfactory dose-dependent DPPH radical scavenging activity with an IC50 value of 0.675 mg/mL. This study reveals the correct identity of the Indian Fusarium isolates based on multigene phylogeny and also throws light on BEA production potential, suggesting their possible applicability in the medicine, agriculture and industry.
Insights
This study clarifies the identity of Indian Fusarium species using multigene analysis. It identifies Fusarium tardicrescens as a high producer of beauvericin (BEA), a compound with significant antibacterial, antifungal, and antioxidant properties.
Area of Science:
- Mycology
- Molecular Biology
- Natural Products Chemistry
Background:
- The taxonomic classification of Fusarium species is challenging due to ambiguous species identification based on traditional morphotaxonomic methods.
- Fusarium species are known producers of valuable secondary metabolites, including beauvericin (BEA) and enniatins.
Purpose of the Study:
- To resolve the evolutionary relationships of 88 Indian Fusarium isolates using multigene phylogenetic analysis.
- To assess the beauvericin (BEA) production capabilities of these isolates.
- To evaluate the bioactivity of BEA-producing isolates, including antibacterial, antifungal, and antioxidant properties.
Main Methods:
- Multigene phylogenetic analysis using ITS, 28S LSU, TEF1-α, RPB2, β-tubulin, and calmodulin gene regions.
- Fermentation of Fusarium isolates in Fusarium-defined media for BEA production.
- Quantification of BEA using Thin Layer Chromatography (TLC), High-Performance Liquid Chromatography (HPLC), and High-Resolution Mass Spectrometry (HRMS).
- Bioactivity assays including antibacterial (Minimum Inhibitory Concentration - MIC), antifungal (food poison technique), and antioxidant (DPPH radical scavenging) assays.
Main Results:
- Multigene phylogeny successfully resolved the evolutionary relationships of the 88 Indian Fusarium isolates.
- Fifty out of 88 isolates produced BEA, with concentrations ranging from 0.01 to 15.82 mg/g of biomass.
- Fusarium tardicrescens NFCCI 5201 exhibited the highest BEA production (15.82 mg/g).
- Extracts of F. tardicrescens NFCCI 5201 demonstrated potent antibacterial activity against Staphylococcus aureus and Micrococcus luteus.
- The extract showed significant antifungal activity against plant pathogenic fungi like Rhizoctonia solani and Sclerotium rolfsii, with synergistic effects observed when combined with amphotericin B.
- The extract displayed dose-dependent DPPH radical scavenging activity with an IC50 value of 0.675 mg/mL.
Conclusions:
- Multigene phylogenetic analysis provides a robust method for accurate Fusarium species identification.
- Fusarium tardicrescens NFCCI 5201 is a promising source of BEA with significant potential for medicinal, agricultural, and industrial applications.
- The identified Fusarium isolates and their metabolites warrant further investigation for their therapeutic and practical uses.
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