Abstract
Chemical fungicides are mainly used to control Colletotrichum orbiculare, but this fungal pathogen usually develops resistance to the pesticides. Natural compounds are therefore desirable for controlling C. orbiculare. Here, a culture filtrate (CF) from ME202, a fungal strain isolated from Trifolium incarnatum in Shimane Prefecture in 2020, was tested in vitro and found to inhibit conidial germination of C. orbiculare. The inhibitory compound in ME202-CF was soluble in ethyl acetate. The ethyl acetate extract of ME202-CF (ME202-ECF) significantly inhibited in vitro conidial germination of C. orbiculare in a dose-dependent manner and lesion formation on cucumber leaves. The inhibitory compounds were heat stable at 40–121 °C and molecular mass ≥ 500 and ≥ 1000 Da. Thin layer chromatography–bioautography of ME202-ECF showed that the compounds inhibiting C. orbiculare growth had an Rf of 0.81 and 0.91. Furthermore, ME202 was found to be a member of the genus Cercospora through sequence analysis of the internal transcribed spacer and 5.8 S rDNA. These results suggest that secondary metabolites of Cercospora spp., such as ME202, can be used to develop new fungicides against anthracnose.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bakhshi M, Arzanlou M, Babai-Ahari A, Groenewald JZ, Braun U, Crous PW (2015) Application of the consolidated species concept to Cercospora spp. from Iran. Persoonia 34:65–86. https://doi.org/10.3767/003158515X685698
Daub ME, Chung K-R (2007) Cercosporin: a photoactivated toxin in plant disease. Internet Resource. In: APSnet Features. Available at https://www.apsnet.org/edcenter/apsnetfeatures/Pages/Cercosporin.aspx. Cited 16 Sep 2021
Dean R, Van Kan JAL, Pretorius ZA, Hammond-Kosack KE, Di Pietro A, Spanu PD, Rudd JJ, Dickman M, Kahmann R, Ellis J, Foster GD (2012) The top 10 fungal pathogens in molecular plant pathology. Mol Plant Pathol 13:414–430. https://doi.org/10.1111/j.1364-3703.2011.00783.x
Duan Y, Xin W, Lu F, Li T, Li M, Wu J, Wang J, Zhou M (2019) Benzimidazole- and QoI-resistance in Corynespora cassiicola populations from greenhouse-cultivated cucumber: an emerging problem in China. Pestic Biochem Physiol 153:95–105. https://doi.org/10.1016/j.pestbp.2018.11.006
Ehrenshaft M, Upchurch RG (1991) Isolation of light-enhanced cDNAs of Cercospora kikuchii.. Appl Environ Microbiol 57:2671–2676. https://doi.org/10.1128/aem.57.9.2671-2676.1991
Farr DF, Rossman AY (2021) Fungal databases, systematic mycology and microbiology laboratory. Accessed Apr 16, 2021. http://nt.ars-grin.gov/fungaldatabases/. Anaesthetics Research Society, United States Department of Agriculture
Groenewald JZ, Nakashima C, Nishikawa J, Shin H-D, Park J-H, Jama AN, Groenewald M, Braun U, Crous PW (2013) Species concepts in Cercospora: spotting the weeds among the roses. Stud Mycol 75:115–170. https://doi.org/10.3114/sim0012
Guo Y, Fan Z, Yi X, Zhang Y, Khan RAA, Zhou Z (2021) Sustainable management of soil-borne bacterium Ralstonia solanacearum in vitro and in vivo through fungal metabolites of different Trichoderma spp. Sustainability 13:1491. https://doi.org/10.3390/su13031491
Holmes GJ, Ojiambo PS, Hausbeck MK, Quesada-Ocampo L, Keinath AP (2015) Resurgence of cucurbit downy mildew in the United States: a watershed event for research and extension. Plant Dis 99:428–441. https://doi.org/10.1094/PDIS-09-14-0990-FE
Ino M, Kihara J, Ueno M (2021) Inhibitory potential of fungi isolated from several weeds in Matsue city against Colletotrichum orbiculare, the causal agent of anthracnose disease in cucurbit crops. Bull Fac Life Environ Sci Shimane Univ 26:3–7
Ishii H, Yano K, Date H, Furuta A, Sagehashi Y, Yamaguchi T, Sugiyama T, Nishimura K, Hasama W (2007) Molecular characterization and diagnosis of QoI resistance in cucumber and eggplant fungal pathogens. Phytopathology 97:1458–1466. https://doi.org/10.1094/PHYTO-97-11-1458
Ishii H, Bryson PK, Kayamori M, Miyamoto T, Yamaoka Y, Schnabel G (2021) Cross-resistance to the new fungicide mefentrifluconazole in DMI-resistant fungal pathogens. Pestic Biochem Physiol 171:104737. https://doi.org/10.1016/j.pestbp.2020.104737
Khan RAA, Najeeb S, Mao Z, Ling J, Yang Y, Li Y, Xie B (2020) Bioactive secondary metabolites from Trichoderma spp. against phytopathogenic bacteria and root-knot nematode. Microorganisms 8:401. https://doi.org/10.3390/microorganisms8030401
Kimura M (1980) A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16:111–120. https://doi.org/10.1007/BF01731581
Kong W-L, Rui L, Ni H, Wu X-Q (2020) Antifungal effects of volatile organic compounds produced by Rahnella aquatilis JZ-GX1 against Colletotrichum gloeosporioides in Liriodendron chinense × tulipifera. Front Microbiol 11:1114. https://doi.org/10.3389/fmicb.2020.01114
Kumarihamy M, Khan SI, Jacob M, Tekwani BL, Duke SO, Ferreira D, Nanayakkara NPD (2012) Antiprotozoal and antimicrobial compounds from the plant pathogen Septoria pistaciarum. J Nat Prod 75:883–889. https://doi.org/10.1021/np200940b
Liu S, Ruan W, Li J, Xu H, Wang J, Gao Y, Wang J (2008) Biological control of phytopathogenic fungi by fatty acids. Mycopathologia 166:93–102. https://doi.org/10.1007/s11046-008-9124-1
McGrath MT, Wyenandt CA (2017) First detection of Boscalid resistance in Podosphaera xanthii in the United States associated with failure to control cucurbit powdery mildew in New York and New Jersey in 2009. Plant Health Prog 18:93. https://doi.org/10.1094/PHP-03-17-0014-BR
Mitchell TK, Chilton WS, Daub ME (2002) Biodegradation of the polyketide toxin cercosporin. Appl Environ Microbiol 68:4173–4181. https://doi.org/10.1128/AEM.68.9.4173-4181.2002
Monggoot S, Pichaitam T, Tanapichatsakul C, Pripdeevech P (2018) Antibacterial potential of secondary metabolites produced by Aspergillus sp., an endophyte of Mitrephora wangii. Arch Microbiol 200:951–959. https://doi.org/10.1007/s00203-018-1511-5
Mookherjee A, Mitra M, Kutty NN, Mitra A, Maiti MK (2020) Characterization of endo-metabolome exhibiting antimicrobial and antioxidant activities from endophytic fungus Cercospora sp. PM018. S. Afr J Bot 134:264–272. https://doi.org/10.1016/j.sajb.2020.01.040
Moutassem D, Belabid L, Bellik Y (2020) Efficiency of secondary metabolites produced by Trichoderma spp. in the biological control of Fusarium wilt in chickpea. J Crop Prot 9:217–231
Pohl CH, Kock JLF, Thibane VS (2011) Antifungal free fatty acids. In: Méndez-Vilas A (ed) Science against microbial pathogens: communicating current research and technological advances. Formatex Research Center, Badajoz, Spain, pp 61–71
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
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425. https://doi.org/10.1093/oxfordjournals.molbev.a040454
Santos MS, Orlandelli RC, Polonio JC, dos Santos Ribeiro MA, Sarragiotto MH, Azevedo JL, Pamphile JA (2017) Endophytes isolated from passion fruit plants: molecular identification, chemical characterization, and antibacterial activity of secondary metabolites. J Appl Pharm Sci 7:38–43
Souza AGC, Rodrigues F, Maffia LA, Mizubuti ESG (2011) Infection process of Cercospora coffeicola on coffee leaf. J Phytopathol 159:6–11. https://doi.org/10.1111/j.1439-0434.2010.01710.x
Tessmann DJ, Charudattan R, Preston JF (2008) Variability in aggressiveness, cultural characteristics, cercosporin production and fatty acid profile of Cercospora piaropi, a biocontrol agent of water hyacinth. Plant Pathol 57:957–966. https://doi.org/10.1111/j.1365-3059.2008.01867.x
Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position- specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680. https://doi.org/10.1093/nar/22.22.4673
White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic Press, San Diego, CA, USA, pp 315–322
Acknowledgements
The authors thank the Faculty of Life and Environmental Science at Shimane University for providing the financial support to publish this report.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflicts of interest.
Ethical approval
Not applicable.
Additional information
Publisher’s Note
Springer Nature and the Phytopathological Society of Japan remain neutral with regard to jurisdictional claims in geographical names, published maps and institutional affiliations.
The nucleotide sequence data presented in this paper are available in the DDBJ/EMBL/GenBank database under accession number LC651161.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Ino, M., Kihara, J. & Ueno, M. Inhibitory potential of metabolites produced by Cercospora sp. strain ME202 isolated from Trifolium incarnatum against anthracnose caused by Colletotrichum orbicular. J Gen Plant Pathol 88, 309–317 (2022). https://doi.org/10.1007/s10327-022-01080-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10327-022-01080-7