Endophytic fungal isolation from Blumea axillaris: Identification and biological activity of secondary metabolites

  • Nehru LAVANYA Bharathiar University, Department of Microbial Biotechnology, Coimbatore, Tamil Nadu (IN)
  • Vellingiri MANON MANI RVS Arts and Science College, Department of Biotechnology, Coimbatore, Tamil Nadu (IN)
  • Nachimuthu SARANYA Bharathiar University, Department of Microbial Biotechnology, Coimbatore, Tamil Nadu (IN)
  • Rajendran DEEPAKKUMAR Bharathiar University, Department of Microbial Biotechnology, Coimbatore, Tamil Nadu (IN)
  • Kathirvel PREETHI Bharathiar University, Department of Microbial Biotechnology, Coimbatore, Tamil Nadu (IN)
Keywords: antioxidant, antimicrobial activity, Blumea axillaris, endophytic fungi, metabolites, molecular analysis

Abstract

Medicinal plants are a wealthy source of natural medicinal properties and remain as base for new drug discoveries. Endophyte from the specific medicinal plants produce the analogous metabolites as that of the host plant. The metabolites from the endophytes comprise maximum therapeutic properties and have been extensively applied in treating various diseases and disorders. This study was focused on identification of the endophytic fungi from the medicinal plant Blumea axillaris and investigates the diversity of endophytic fungi from various explants of the same plant. The explants were cultured on potato dextrose agar and 6 endophytic fungi were successfully isolated from Blumea axillaris. They were identified morphologically and confirmed with molecular analysis as Xylaria arbuscula, Paraphoma radicina, Phomopsis phaseoli, Sordaria fimicola, Aspergillus amstelodami, Diaporthe eucalyptorum. The DNA sequences were analyzed by BLAST and the phylogenetic tree was constructed with neighbor joining method. The six isolates were subjected to antagonistic activity for the selection of potential strain and the bioactive strain Xylaria arbuscula was selected for the production of secondary metabolites by optimization. The parameters like pH, temperature, incubation period, carbon and nitrogen (organic and inorganic source) were optimized for secondary metabolite production. The fungal metabolite was extracted by solvent extraction method using polar and non-polar solvents like propanol, methanol, chloroform, acetone and ethyl acetate. To investigate the bioactivities of the fungal crude extract was subjected first for its antioxidant activity using DPPH radical scavenging method, followed by antimicrobial activity of methanolic (MeOH) extract of Xylaria arbuscula, that were also analyzed by the agar well-diffusion method against the clinical pathogens Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pnuemoniae, Proteus mirabilis, Aspergillus niger and Candida albicans.

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References

Barry AL, Garcia F, Thrupp LD (1970). An improved single-disk method for testing the antibiotic susceptibility of rapidly-growing pathogens. American Journal of Clinical Pathology 53:149-158. https://doi.org/10.1093/ajcp/53.2.149

Bauer AW, Kirby WM, Sherris JC, Turck M (1966). Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology 45:493-496.

Bérdy J (2005). Bioactive microbial metabolites – A personal view. Review. The Journal of Antibiotics 58(1):1-26. https://doi.org/10.1038/ja.2005.1

Berdy J (2012). Thoughts and facts about antibiotics: where we are now and we are heading. The Journal of Antibiotics 65:385-395. https://doi.org/10.1038/ja.2012.27

Boonsong S, Klaypradit W, Wilaipun P (2016). Antioxidant activities of extracts from five edible mushrooms using different extractants. Agriculture and Natural Resources 50:89-97. http://dx.doi.org/10.1016/j.anres.2015.07.002

Brunner F, Petrini O (1992). Taxonomy of some Xylaria species and xylariaceous endophytes by isozyme electrophoresis. Mycological Research 96:723-733. https://doi.org/10.1016/s0953-7562(09)80440-1

Bukelskienė V, Baltriukienė D, Repečkienė J (2006). Study of health risks associated with Aspergillus amstelodami and its mycotoxic effects. Ekologija 3:42-47.

Dereeper A, Guignon V, Blanc G, Audic S, Buffet S, Chevenet F, … Gascuel O (2008). Phylogeny. fr: robust phylogenetic analysis for the non-specialist. Nucleic Acids Research 1:36. https://doi.org/10.1093/nar/gkn180

Doss RP, Welty RE (1995). A polymerase chain reaction-based procedure for detection of Acremonium coenophialum in tall fescue. Phytopathology 85:913-917. https://doi.org/10.1094/Phyto-85-913

Edgar RC (2004). MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research 32(5):1792-1797. https://doi.org/10.1093/nar/gkh340

Fathima BS, Balakrishnan RM (2014). Biosynthesis and optimization of silver nanoparticles by endophytic fungus Fusarium solani. Materials Letters 132:428-431. https://doi.org/10.1016/j.matlet.2014.06.143

Ferreira I, Baptista P, Vilas-Boas M, Barros L (2007). Free-radical scavenging capacity and reducing power of wild edible mushrooms from northeast Portugal: Individual cap and stipe activity. Food Chemistry 1511-1516. http://dx.doi.org/10.1016/j.foodchem.2005.11.043

Fokkema NJ (1978). Fungal antagonism in the phylosphere. Annals of Applied Biology 89:115-117. http://dx.doi.org/10.2298/ABS1003611Z

Gade AK, Bonde P, Ingle AP, Marcato PD, Duran N, Rai MK (2008). Exploitation of Aspergillus niger for synthesis of silver nanoparticles. Journal of Biobased Materials and Bioenergy 2(3):243-247. http://dx.doi.org/10.1166/jbmb.2008.401

Gerber GK (2014). The dynamic microbiome. FEBS Letters 588:4131-4139. https://doi.org/10.1016/j.febslet.2014.02.037

Gond SK, Mishra A, Sharma VK, Verma SK, Kumar J, Kharwar RN, Kumar A (2012). Diversity and antimicrobial activity of endophytic fungi isolated from Nyctanthes arbor-tristis, a well-known medicinal plant of India. Mycoscience 53(2):113-121.

Gond SK, Verma VC, Kumar A, Kumar V, Kharwar RN (2007). Study of endophytic fungal communities from different parts of Aeglemarmelos correae (Rutaceae) from Varanasi (India). World Journal of Microbiology and Biotechnology 23:1371-1375.

Han WB, Zhai YJ, Gao Y, Zhou HY, Xiao J, Pescitelli G, Gao JM (2019). Cytochalasins and an abietane-type diterpenoid with allelopathic activities from the endophytic fungus Xylaria species. Journal of Agricultural and Food Chemistry 67:3643-3650. https://doi.org/10.1021/acs.jafc.9b00273

Hung PV, Nhi NNY (2012). Nutritional composition and antioxidant capacity of several edible mushrooms grown in the Southern Vietnam. International Food Research Journal 19(2):611-615. https://www.researchgate.net/publication/267036593

Ivanová H (2015). Sordaria fimicola (Ascomycota, Sordariales) on Acer palmatum. Folia Oecologica 42:67-71.

Johan C, Janse van Rensburg Sandra C, Lamprecht Johannes Z, Groenewald Lisa A, Castlebury, Pedro W Crous (2006). Characterisation of Phomopsis spp. associated with die-back of rooibos (Aspalathus linearis) in South Africa. Studies in Mycology 55:65-74. https://doi.org/10.3114/sim.55.1.65

Joseph B, Priya RM (2011). Bioactive compounds from endophytes and their potential in pharmaceutical effect: a review. American Journal of Biochemistry and Molecular Biology 1:291-309. https://dx.doi.org/10.3923/ajbmb.2011.291.309

Keles A, Koca I, Genccelep H (2011). Antioxidant properties of wild edible mushrooms. Journal of Food Processing Technology 2(6):1-6. https://doi.org/10.4172/2157-7110.1000130

Kumarihamy M, Ferreira D, Croom EM Jr, Sahu R, Tekwani BL, Duke SO, … Nanayakkara NP (2019). Antiplasmodial and cytotoxic cytochalasins from an endophytic fungus, Nemania sp. UM10M, isolated froma diseased Torreya taxifolia leaf. Molecules 24:777. https://doi.org/10.3390/molecules24040777

Lacap DC, Hyde KD, Liew EC (2003). An evaluation of the fungal 'morphotype' concept based on ribosomal DNA sequences. Fungal Diversity 12:53-66. http://dx.doi.org/10.3897/mycokeys.32.23670

Majumdar MK, Majumdar SK (1965). Effects of minerals on neomycin production by Streptomyces fradiae. Journal of Applied Microbiology 13:190-193. https://dx.doi.org/10.1128%2Faac.8.4.391

Mani VM, Priya MS, Dhayalini S, Preethi K (2015b). Antioxidant and antimicrobial evaluation of bioactive pigment from Fusarium sp isolated from stressed environment. International Journal of Current Microbiology and Applied Sciences 4(6): 1147-1158.

Mani VM, Soundari AJPG, Balasubramanian B, Park S, Issara U, Preethi K, Liu W-C (2021). Evaluation of dimer of epicatechin from an endophytic fungus Curvularia australiensis FC2AP on Acute toxicity levels, anti-inflammatory and anti-cervical cancer activity in animal models. Molecules 26(3):654. https://doi.org/10.3390/molecules26030654

Mani VM, Soundari APG, Ashokraj S, Priyadarisini VB (2017). Molecular and functional characterization of metabolites from terrestrial Streptomyces hygroscopicus AVS7 isolated from Western Ghats, India. Journal of Pure and Applied Microbiology 11(4):1823-1831.

Mani VM, Soundari APG, Karthiyaini D, Preethi K (2015a). Bioprospecting for endophytic fungi and their metabolites from medicinal tree Aegle marmelos in Western Ghats, India. Mycobiology 43(3):303-310. https://dx.doi.org/10.5941%2FMYCO.2015.43.3.303

Mani VM, Soundari APG, Tamilarasi S (2018). Determination of in vitro cytotoxicity and anti- angiogenesis for a bioactive compound from Aspergillus terreus FC36AY1 isolated from Aeglemarmelosaround Western Ghats, India. In Tech Open- Book:Medicinal Chemistry, pp 13- 28.

Molina G, Pimentel MR, Bertucci TCP, Pastore GM (2012). Application of fungal endophytes in biotechnological processes. Chemical Engineering Transactions 27:289-294. https://doi.org/10.7324/JAPS.2012.2807

Moslemi A, Ades PK, Groom T, Crous PW, Nicolas ME, Taylor PWJ (2016). Pharaphoma crown rot of pyrethrum (Tanacetum cinerariifolium). Plant Disease Journal (12):2363-2369. https://doi.org/10.1094/PDIS-05-16-0628-RE

Naik MK, Hiremath PC, Hegde RK (1988). Physiological and nutritional studies on C. gloeosporioides, a causal agent of anthracnose of beetlevine. Mysore Journal of Agricultural Science 22:471-474.

Newman DJ, Cragg GM (2014). Marine-sourced anti-cancer and cancer pain control agents in clinical and late pre-clinical development. Marine Drugs 12:255-278. https://doi.org/10.3390/md12010255

Nicoletti R, and Fiorentino A (2015). Plant bioactive metabolites and drugs produced by endophytic fungi of Spermatophyta. Agriculture 5:918-970. https://doi.org/10.3390/agriculture5040918

Petrini O, Petrini LE, Rodrigues K (1995). Xylariaceaous endo-phytes: an exercise in biodiversity. Fitopatologica Brasiliensis 20:531-539.

Preethi K, Mani VM, Lavanya N (2021). Endophytic fungi: a potential source of bioactive compounds for commercial and therapeutic applications. In: Patil RH, Maheshwari VL (Eds). Book: Endophytes. Springer, Singapore. https://doi.org/10.1007/978-981-15-9371-0_12

Ramesh V, Santosh K, Anand TD., Shanmugaiah V, Kotamraju S, Karunakaran C, Rajendran A (2015). Novel bioactive wild medicinal mushroom−Xylaria sp. R006 (Ascomycetes) against multidrug resistant human bacterial pathogens and human cancer cell lines. International Journal of Medicinal Mushrooms 17:1005-1017. https://doi.org/10.1615/IntJMedMushrooms.v17.i10.100

Ripa FA, Nikkon F, Zaman S, Khondkar P (2009). Optimal conditions for antimicrobial metabolites production from a new Streptomyces sp. RUPA-08PR isolated from Bangladeshi soil. Mycobiology 37(3):211-214. https://doi.org/10.4489/MYCO.2009.37.3.211

Rodriguez RJ, Woodward CJ Redman RS (2012). Fungal influence on plant tolerance to stress. In: Blackwell W (Ed). Biocomplexity of Plant-Fungal Interactions. Oxford, UK pp 155-163.

Rogers JD (1984). Xylaria acuta, Xylaria cornu-damae, and Xylaria mali in continental United States. Mycologia 76:22-33 https://doi.org/10.2307/3792832

Rogers JD, Samuels GS (1986). Ascomycetes of New Zealand. 8. Xylaria. New Zealand Journal of Botany 24:615-650. https://doi.org/10.1080/0028825X.1986.10409947

Saikkonen K, Saari S, Helander M (2010). Defensive mutualism between plants and endophyticfungi? Fungal Diversity 41:101-113. http://dx.doi.org/10.1007/s13225-010-0023-7

Schulz B, Boyle C (2005). The endophytic continuum. Mycological Research 109:661-686. https://doi.org/10.1017/S095375620500273X

Selim KA, El-Beih AA, AbdEl RTM, Diwany AI (2012). Biology of endophytic fungi. Current Research in Environmental & Applied Mycology 2:31-82. https://doi.org/10.5943/cream/2/1/3

Senthilkumar A, Kannathasan K, Venkatesalu V (2008). Chemical constituents and larvicidal properties of the essential oil of Blumea mollis (D. Don) Merr. against Culex quinquefasciatus. Parasitology Research 103(4):959-962. https://doi.org/10.1007/s00436-008-1085-2

Singh D, Rathod V, Singh AK, Joshi R, Nagaratna H, Azmathunnisa, Avinash B (2015). Antibacterial activity and phytochemical analysis of the crude extracts of endophytic fungus, Alternaria sp. from the medicinal plant Euphorbia hirta (L). International Journal of Green Chemistry and Bioprocess 5:14-20.

Singh LS, Mazumder S, Bora TC (2009). Optimisation of process parameters for growth and bioactive metabolite produced by a salt-tolerant and alkaliphilic actinomycete, Streptomyces tanashiensis strain A2D. Journal of Medical Mycology 19:225-233.

Sivanandhan S, Ganesan P, Jackson A, Darvin S, Paulraj MG, Ignacimuthu S (2018). Activity of some medicinal plants against phytopathogenic fungi. International Journal of Scientific Research in Biological Sciences 5(5):124-137. https://doi.org/10.26438/ijsrbs/v5i5.124137

Sreelekha KP, Ajeesh Krishna TP, Adarsh Krishna TP, Deepa PE, Udayan D, Juliet S, ... Ravindran R (2017). Pharmaco-chemical characterization of leaves of Blumea mollis (D. Don) merr. from Western Ghats of Wayanad region of Kerala, India. Journal of Pharmacognosy and Phytochemistry 6(4):319-323.

Strobel G, Daisy B (2003). Bioprospecting for microbial endophytes and their natural products. Microbiological and Molecular Biology Reviews 67:491-502. https://dx.doi.org/10.1128%2FMMBR.67.4.491-502.2003

Strobel GA, Dirkse E, Sears J, Markworth C (2001). Volatile antimicrobials from Muscodor albus, a novel endophytic fungus. Microbiology 147:2943-2950. https://doi.org/10.1099/00221287-147-11-2943

Talavera G, Castresana J (2007). Improvement of phylogenies after removing divergent and ambiguously aligned blocks from protein sequence alignments. Systematic Biology 56:564-577. https://doi.org/10.1080/10635150701472164

Uecker FA (1988). A world list of Phomopsis names with notes on nomenclature, morphology and biology. Mycologia Memoir 13:1-231.

Vellingiri MM, Arockiam Jeyasundar PGS, Venkatesan BP, Balakrishnan B, Liu W (2021). Statistical optimization of parameters for enhanced bioactive metabolites produced by Streptomyces hygroscopicus AVS7. Arabian Journal of Science and Engineering 46:5345-5360. https://doi.org/10.1007/s13369-020-05116-y

Vigneshwari A, Rakk D, Németh A, Kocsubé S, Kiss N, Csupor D, Papp T, … Szekeres A (2019). Host metabolite producing endophytic fungi isolated from Hypericum perforatum. Plos One 14(5):e0217060. https://doi.org/10.1371/journal.pone.0217060

Walsh CT, Fischbach MA (2010). Natural products version 2.0: connecting genes to molecules. Journal of American Chemical Society 132:2469-2493. https://doi.org/10.1021/ja909118a

Wang WX, Lei X, Ai HL, Bai X, Li J, He J, … Liu JK (2019). Cytochalasans from the endophytic fungus Xylaria cf. curta with resistance reversal activity against fluconazole-resistant Candida albicans. Organic Letters 21:1108-1111. https://doi.org/10.1021/acs.orglett.9b00015

Wang WX, Lei X, Yang YL, Li ZH, Ai HL, Li J, ... Liu JK (2019). Xylarichalasin A, a halogenated hexacyclic cytochalasan from the fungus Xylaria cf. curta. Organic Letters 21:6957-6960. https://doi.org/10.1021/acs.orglett.9b02552

Wang WX, Li ZH, Feng T, Li J, Sun H, Huang R, … Liu JK (2018). Curtachalasins A and B, two cytochalasans with a tetracyclic skeleton from the endophytic fungus Xylaria curta E10. Organic Letters 20:7758-7761. https://doi.org/10.1021/acs.orglett.8b03110

Wittstein K, Cordsmeier A, Lambert C, Wendt L, Sir EB, Weber J, … Stadler M (2020). Identification of Rosellinia species as producers of cyclodepsipeptide PF1022 A and resurrection of the genus Dematophora as inferred from polythetic taxonomy. Study of Mycology 96:1-16. https://doi.org/10.1016/j.simyco.2020.01.001

Wu W, Davis RW, Tran-Gyamfi MB, Kuo A, LaButti K, Mihalcheva S, … Gladden JM (2017). Characterization of four endophytic fungi as potential consolidated biosprocessing hosts for conversion of lignocellulose into advanced biofuels. Bioenergy and Biofuels 101:2603-2618. https://doi.org/10.1007/s00253-017-8091-1

Xu L, Zhou L, Zhao J, Jiang W (2008). Recent studies on the antimicrobial compounds produced by plant endophytic fungi. Natural Product Research and Development 20:731-740.

Yildirim A, Mavi A, Kara AA (2001). Determination of antioxidant and antimicrobial activities of Rumex crispus L. extracts. Journal of Agriculture and Food Chemistry 9:4083-4089. https://doi.org/10.1021/jf0103572

Published
2021-06-23
How to Cite
LAVANYA, N., MANON MANI, V., SARANYA, N., DEEPAKKUMAR, R., & PREETHI, K. (2021). Endophytic fungal isolation from Blumea axillaris: Identification and biological activity of secondary metabolites. Notulae Scientia Biologicae, 13(2), 10953. https://doi.org/10.15835/nsb13210953
Section
Research articles
CITATION
DOI: 10.15835/nsb13210953