Skip to main content

Advertisement

SpringerLink
Log in

A Secondary Metabolite of Cercospora sp., Associated with Rosa damascena Mill., Inhibits Proliferation, Biofilm Production, Ergosterol Synthesis and Other Virulence Factors in Candida albicans

  • Fungal Microbiology
  • Published:
Microbial Ecology Aims and scope Submit manuscript

Abstract

Here we describe the antimicrobial potential of secondary metabolites, fulvic acid (F.A.) and anhydrofulvic acid (AFA), produced by RDE147, an endophyte of Rosa damascena Mill. The endophyte was identified as Cercospora piaropi by ITS and β-tubulin–based phylogenetic analyses, while chemoprofiling of the endophyte by column chromatography and spectroscopy yielded two pure compounds, F.A. and AFA. The compounds demonstrated different antimicrobial profiles, with AFA suppressing the growth of C. albicans at 7.3 µg ml−1 IC50. Further studies revealed that AFA strongly restricted the biofilm production and hyphae formation in C. albicans by down-regulating several biofilm and morphogenesis-related genes. The time-kill assays confirmed the fungicidal activity of AFA against C. albicans, killing 83.6% of the pathogen cells in 24 h at the MIC concentration, and the post-antibiotic effect (PAE) experiments established the suppression of C. albicans growth for extended time periods. The compound acted synergistically with amphotericin B and nystatin and reduced ergosterol biosynthesis by the pathogen, confirmed by ergosterol estimation and comparative expression profiling of selected genes and molecular docking of AFA with C. albicans squalene epoxidase. AFA also suppressed the expression of several other virulence genes of the fungal pathogen. The study determines the anti-C. albicans potential of AFA and its impact on the biology of the pathogen. It also indicates that Cercospora species may yield potential bioactive molecules, especially fulvic acid derivatives. However, it is imperative to conduct in vivo studies to explore this molecule’s therapeutic potential further.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Data Availability

The pure culture (Voucher no. MRCJ–1029) was deposited in the institute’s in-house microbial repository (WDCM 1117). The ITS and β-tubulin gene sequences were deposited with GenBank accession numbers OK335281 and OL862418, respectively.

Code Availability

Not applicable.

References

  1. Brown GD, Denning DW, Gow NA, Levitz NA, Netea MG, White TC (2012) Hidden killers human fungal infections. Sci Transl Med 4:165113–165113. https://doi.org/10.1086/376906

    Article  Google Scholar 

  2. Pappas PG, Rex JH, Lee J, Hamill RJ, Larsen RA, Powderly W, Kauffman CA, Hyslop N, Mangino JE, Chapman S (2003) A prospective observational study of candidemia: epidemiology, therapy, and influences on mortality in hospitalized adult and pediatric patients. Clin Infect Dis 37:634–643. https://doi.org/10.1086/376906

    Article  PubMed  Google Scholar 

  3. Kachroo AH, Laurent JM, Yellman CM, Meyer AG, Wilke CO, Marcotte EM (2015) Systematic humanization of yeast genes reveals conserved functions and genetic modularity. Science 348:921–925. https://doi.org/10.1126/science.aaa0769

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Cragg GM, Newman D (1830) J (2013) Natural products: a continuing source of novel drug leads. Biochim Biophys Acta 6:3670–3695. https://doi.org/10.1016/j.bbagen.2013.02.008

    Article  CAS  Google Scholar 

  5. Arora P, Ahmad T, Farooq S, Riyaz-Ul-Hassan S (2019) Endophytes: a hidden treasure of novel antimicrobial metabolites antibacterial drug discovery to combat MDR. Springer, 165–192 https://doi.org/10.1007/978-981-13-9871-1_8

  6. Mileva M, Ilieva Y, Jovtchev G, Gateva S, Zaharieva MM, Georgieva A, Dimitrova L, Dobreva A, Angelova T, Vilhelmova-Ilieva N (2021) Rose flowers—a delicate perfume or a natural healer? Biomolecules 11:127. https://doi.org/10.3390/biom11010127

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Rangel LI, Spanner RE, Ebert MK, Pethybridge SJ, Stukenbrock EH, de Jonge R, Secor GA, Bolton MD (2020) Cercospora beticola: the intoxicating lifestyle of the leaf spot pathogen of sugar beet. Mol Plant Pathol 21:1020–1041. https://doi.org/10.1111/mpp.12962

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Feng Y, Ren F, Niu S, Wang L, Li L, Liu X, Che Y (2014) Guanacastane diterpenoids from the plant endophytic fungus Cercospora sp. J Nat Prod 77:873–881. https://doi.org/10.1021/np4009688

    Article  CAS  PubMed  Google Scholar 

  9. Daub ME (1982) Peroxidation of tobacco membrane lipids by the photosensitizing toxin, cercosporin. Plant Physiol 69:1361–1364. https://doi.org/10.1104/pp.69.6.1361

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Trigos A, Espinoza C, Martínez M, Márquez O, León LG, Padron JM, Norte M, Fernandez JJ (2013) Antiproliferative activity of epi-cercosporin in human solid tumor cell lines. Nat Prod Commun 8: https://doi.org/10.1177/1934578X1300800214

  11. Fujita K-I, Nagamine Y, Ping X, Taniguchi M (1999) Mode of action of anhydrofulvic acid against Candida utilis ATCC 42402 under acidic condition. The J Antibiot 52:628–634. https://doi.org/10.7164/antibiotics.52.628

    Article  CAS  Google Scholar 

  12. Kumla D, Pereira JA, Dethoup T, Gales L, Freitas-Silva J, Costa PM, Lee M, Silva A, Sekeroglu N, Pinto MM (2018) Chromone derivatives and other constituents from cultures of the marine sponge-associated fungus Penicillium erubescens KUFA0220 and their antibacterial activity. Mar Drugs 16:289. https://doi.org/10.3390/md16080289

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Farooq S, Qayum A, Nalli Y, Lauro G, Chini MG, Bifulco G, Chaubey A, Singh SK, Riyaz-Ul-Hassan S, Ali A (2020) Discovery of a secalonic acid derivative from Aspergillus aculeatus, an endophyte of Rosa damascena Mill., triggers apoptosis in MDA-MB-231 triple negative breast cancer cells. ACS Omega 5:24296–24310. https://doi.org/10.1021/acsomega.0c02505

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR protocols: a guide to methods and applications 18:315–322

    Google Scholar 

  15. Samson RA, Visagie CM, Houbraken J, Hong S-B, Hubka V, Klaassen CH, Perrone G, Seifert KA, Susca A, Tanney JB (2005) Phylogeny, identification and nomenclature of the genus Aspergillus. Stud Mycol 53:1–27. https://doi.org/10.1016/j.simyco.2014.07.004

    Article  Google Scholar 

  16. Tamura K, Nei M (1993) Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Mol Biol Evol 10:512–526. https://doi.org/10.1093/oxfordjournals.molbev.a040023

    Article  CAS  PubMed  Google Scholar 

  17. Eloff JN (1998) A sensitive and quick microplate method to determine the minimal inhibitory concentration of plant extracts for bacteria. Planta Med 64:711–713. https://doi.org/10.1055/s-2006-957563

    Article  CAS  PubMed  Google Scholar 

  18. Arora P, Wani ZA, Nalli Y, Ali A, Riyaz-Ul-Hassan S (2016) Antimicrobial potential of Thiodiketopiperazine derivatives produced by Phoma sp., an endophyte of Glycyrrhiza glabra Linn. Microb Ecol 72:802–812. https://doi.org/10.1007/s00248-016-0805-x

    Article  CAS  PubMed  Google Scholar 

  19. Clinical and Laboratory Standards Institute (2015) Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. Approved Standard M07–A10. Wayne, PA: CLSI, USA

  20. O’Toole GA, Pratt LA, Watnick PI, Newman DK, Weaver VB, Kolter R (1999) [6] Genetic approaches to study of biofilms. Methods Enzymol 310:91–109. https://doi.org/10.1016/S0076-6879(99)10008-9

    Article  CAS  PubMed  Google Scholar 

  21. Sun L, Liao K, Wang D (2015) Effects of magnolol and honokiol on adhesion, yeast-hyphal transition, and formation of biofilm by Candida albicans. PLoS ONE 10:e0117695. https://doi.org/10.1371/journal.pone.0117695

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Sopirala MM, Mangino JE, Gebreyes WA, Biller B, Bannerman T, Balada-Llasat J-M, Pancholi P (2010) Synergy testing by Etest, microdilution checkerboard, and time-kill methods for pan-drug-resistant Acinetobacter baumannii. Antimicrob Agents Chemother 54:4678–4683. https://doi.org/10.1128/AAC.00497-10

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Ahmad T, Arora P, Nalli Y, Ali A, Riyaz-Ul-Hassan S (2020) Antibacterial potential of Juglomycin A isolated from Streptomyces achromogenes, an endophyte of Crocus sativus Linn. J Appl Microbiol 128:1366–1377. https://doi.org/10.1111/jam.14568

    Article  CAS  PubMed  Google Scholar 

  24. Khan MSA, Ahmad I, Cameotra SS (2013) Phenyl aldehyde and propanoids exert multiple sites of action towards cell membrane and cell wall targeting ergosterol in Candida albicans. AMB Express 3:1–16. https://doi.org/10.1186/2191-0855-3-54

    Article  CAS  Google Scholar 

  25. Yang J, Zhang Y (2015) I-TASSER server: new development for protein structure and function predictions. Nucleic Acids Res 43:W174–W181. https://doi.org/10.1093/nar/gkv342

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Wass MN, Kelley LA, Sternberg MJ (2010) 3DLigandSite: predicting ligand-binding sites using similar structures. Nucleic Acids Res 38:W469–W473. https://doi.org/10.1093/nar/gkq406

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Biovia Dassault Systèmes (2017) Discovery studio modeling environment. Release, San Diego, Dassault Systèmes. https://www.3ds.com/products-services/biovia/products/molecular-modeling-simulation/biovia-discovery-studio/

  28. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001

    Article  CAS  PubMed  Google Scholar 

  29. Yamauchi M, Katayama S, Todoroki T, Watanabe T (1987) Studies on the syntheses of heterocyclic compounds containing benzopyrone. Part 5. Total synthesis of fulvic acid. J Chem Soc Perkin Transactions 1:389–394. https://doi.org/10.1039/P19870000389

    Article  Google Scholar 

  30. Groenewald J, Nakashima C, Nishikawa J, Shin H-D, Park J-H, Jama A, Groenewald M, Braun U, Crous P (2006) Species concepts in Cercospora: spotting the weeds among the roses. Stud Mycol 55:ii–ii. https://doi.org/10.3114/sim0012

    Article  Google Scholar 

  31. Winkler J, Ghosh S (2018) Therapeutic potential of fulvic acid in chronic inflammatory diseases and diabetes. J Diabetes Res. https://doi.org/10.1155/2018/5391014

    Article  PubMed  PubMed Central  Google Scholar 

  32. Georgousaki K, Tsafantakis N, Gumeni S, González-Menéndez V, de Pedro N, Tormo JR, Almeida C, Lambert C, Genilloud O, Trougakos IP (2019) Cercospora sp. as a source of anti-aging polyketides targeting 26S proteasome and scale-up production in submerged bioreactor. J Biotechnol 301:88–96. https://doi.org/10.1016/j.jbiotec.2019.05.015

    Article  CAS  PubMed  Google Scholar 

  33. Lee D-S, Jang J-H, Ko W, Kim K-S, Sohn JH, Kang M-S, Ahn JS, Kim Y-C, Oh H (2013) PTP1B inhibitory and anti-inflammatory effects of secondary metabolites isolated from the marine-derived fungus Penicillium sp. JF-55. Mar Drugs 11:1409–1426. https://doi.org/10.3390/md11041409

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Cavalheiro M, Teixeira MC (2018) Candida biofilms: threats, challenges, and promising strategies. Front Med 5:28. https://doi.org/10.3389/fmed.2018.00028

    Article  Google Scholar 

  35. Finkel JS, Mitchell AP (2011) Genetic control of Candida albicans biofilm development. Nat Rev Microbiol 9:109–118. https://doi.org/10.1038/nrmicro2475

    Article  CAS  PubMed  Google Scholar 

  36. Engku Nasrullah Satiman EAF, Ahmad H, Ramzi AB, Abdul Wahab R, Kaderi MA, Wan Harun WHA, Dashper S, McCullough M, Arzmi MH (2020) The role of Candida albicans candidalysin ECE1 gene in oral carcinogenesis. J Oral Pathol Med 49:835–841. https://doi.org/10.1111/jop.13014

    Article  CAS  PubMed  Google Scholar 

  37. Ramage G, VandeWalle K, López-Ribot JL, Wickes BL (2002) The filamentation pathway controlled by the Efg1 regulator protein is required for normal biofilm formation and development in Candida albicans. FEMS Microbiol Lett 214:95–100. https://doi.org/10.1111/j.1574-6968.2002.tb11330.x

    Article  CAS  PubMed  Google Scholar 

  38. Ghannoum MA, Rice LB (1999) Antifungal agents: mode of action, mechanisms of resistance, and correlation of these mechanisms with bacterial resistance. Clin Microbiol Rev 12:501–517. https://doi.org/10.1128/CMR.12.4.501

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Zhou Y, Liao M, Zhu C, Hu Y, Tong T, Peng X, Li M, Feng M, Cheng L, Ren B (2018) ERG3 and ERG11 genes are critical for the pathogenesis of Candida albicans during the oral mucosal infection. Int J Oral Sci 10:1–8. https://doi.org/10.1038/s41368-018-0013-2

    Article  PubMed  PubMed Central  Google Scholar 

  40. Zhanel GG, Hoban DJ, Harding GK (1991) The postantibiotic effect: a review of in vitro and in vivo data. DICP 25:153–163. https://doi.org/10.1177/106002809102500210

    Article  CAS  PubMed  Google Scholar 

  41. Mukaremera L, Lee KK, Mora-Montes HM, Gow NA (2017) Candida albicans yeast, pseudohyphal, and hyphal morphogenesis differentially affects immune recognition. Front Immunol 8:629. https://doi.org/10.3389/fimmu.2017.00629

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Naglik J, Albrecht A, Bader O, Hube B (2004) Candida albicans proteinases and host/pathogen interactions. Cell Microbiol 6:915–926. https://doi.org/10.1111/j.1462-5822.2004.00439.x

    Article  CAS  PubMed  Google Scholar 

  43. Monroy-Pérez E, Paniagua-Contreras GL, Rodríguez-Purata P, Vaca-Paniagua F, Vázquez-Villaseñor M, Díaz-Velásquez C, ... & Vaca S (2016) High virulence and antifungal resistance in clinical strains of Candida albicans. Can J Infect Dis Med Microbiol https://doi.org/10.1155/2016/5930489

  44. Wang HX, Douglas LM, Vesela P, Rachel R, Malinsky J, Konopka JB (2016) Eisosomes promote the ability of Sur7 to regulate plasma membrane organization in Candida albicans. Mol Biol Cell 27:1663–1675. https://doi.org/10.1091/mbc.E16-01-0065

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

AB is supported by a Senior Research Fellowship of the University Grant Commission, New Delhi. T.A. and S.F. obtain fellowships from the Department of Science and Technology, New Delhi. Approval of the institutional IPR committee was obtained with the manuscript no. CSIR-IIIM/IPR/00386.

Funding

The project MLP1008 is supported by the Council of Scientific and Industrial Research (CSIR), New Delhi.

Author information

Authors and Affiliations

  1. Fermentation and Microbial Biotechnology Division, CSIR-Indian Institute of Integrative Medicine, Sanat Nagar, Srinagar, 190005, India

    Abid Bashir, Tanveer Ahmad, Sadaqat Farooq, Malik M. Manzoor, Phalisteen Sultan & Syed Riyaz-Ul-Hassan

  2. Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India

    Abid Bashir, Tanveer Ahmad, Sadaqat Farooq, Waseem I. Lone, Asha Chaubey, Asif Ali & Syed Riyaz-Ul-Hassan

  3. Natural Products Chemistry Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu, 180001, India

    Waseem I. Lone, Yedukondalu Nalli & Asif Ali

  4. Fermentation and Microbial Biotechnology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu, 180001, India

    Asha Chaubey

Authors
  1. Abid Bashir
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tanveer Ahmad
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sadaqat Farooq
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Waseem I. Lone
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Malik M. Manzoor
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yedukondalu Nalli
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Phalisteen Sultan
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Asha Chaubey
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Asif Ali
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Syed Riyaz-Ul-Hassan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Syed Riyaz-Ul-Hassan.

Ethics declarations

Ethics Approval

Not applicable.

Conflict of Interest

The authors declare no competing interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 1258 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bashir, A., Ahmad, T., Farooq, S. et al. A Secondary Metabolite of Cercospora sp., Associated with Rosa damascena Mill., Inhibits Proliferation, Biofilm Production, Ergosterol Synthesis and Other Virulence Factors in Candida albicans. Microb Ecol 85, 1276–1287 (2023). https://doi.org/10.1007/s00248-022-02003-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00248-022-02003-x

Keywords

Search

Navigation