Bioactivity of Volatile Compounds Present in Crinum zeylanicum Leaf Extracts Identified by GC-MS for the Control of Cercospora malayensis Isolates of Okra (Abelmoschus esculentus L.) In Vitro

  • Marie Elvia Akong Laboratory of Biotechnologies and Environment, Phytopathology and Plant Protection Research Unit, Department of Plant Biology, Faculty of Science, University of Yaounde I, PO. Box: 812 Yaounde, Cameroon https://orcid.org/0009-0007-2624-0255
  • Patrice Zemko Ngatsi Laboratory of Biotechnologies and Environment, Phytopathology and Plant Protection Research Unit, Department of Plant Biology, Faculty of Science, University of Yaounde I, PO. Box: 812 Yaounde, Cameroon https://orcid.org/0000-0001-6416-4918
  • Nobert William Tueguem Kuate Laboratory of Biotechnologies and Environment, Phytopathology and Plant Protection Research Unit, Department of Plant Biology, Faculty of Science, University of Yaounde I, PO. Box: 812 Yaounde, Cameroon https://orcid.org/0000-0002-3591-0085
  • Thierry Songwe Atindo Laboratory of Biotechnologies and Environment, Phytopathology and Plant Protection Research Unit, Department of Plant Biology, Faculty of Science, University of Yaounde I, PO. Box: 812 Yaounde, Cameroon https://orcid.org/0000-0001-5618-0192
  • Hubert Boli Laboratory of Biotechnologies and Environment, Phytopathology and Plant Protection Research Unit, Department of Plant Biology, Faculty of Science, University of Yaounde I, PO. Box: 812 Yaounde, Cameroon
  • Claire Stéphane Tsogo Laboratory of Biotechnologies and Environment, Phytopathology and Plant Protection Research Unit, Department of Plant Biology, Faculty of Science, University of Yaounde I, PO. Box: 812 Yaounde, Cameroon
  • Bekolo Ndongo Laboratory of Biotechnologies and Environment, Phytopathology and Plant Protection Research Unit, Department of Plant Biology, Faculty of Science, University of Yaounde I, PO. Box: 812 Yaounde, Cameroon https://orcid.org/0000-0002-2479-8382
Keywords: Antifungal properties, Biochemical compounds, C. malayensis, Crinum zeylanicum, Extracts, GC–MS analysis

Abstract

Cercospora malayensis induced leaf spot is a dangerous okra disease that reduces crop yields. The aim of this study is to use leaf extract from Crinum zeylanicum to regulate the growth of Cercospora malayensis in vitro. The pathogen was isolated from okra leaves from the Meyomessala and Akonolinga localities that had typical disease symptoms. Phytochemical screening and GC-MS analysis of C. zeylanicum leaf extracts were carried out. The mycelial growth and germination of C. malayensis isolates 1 and 2 were evaluated with concentrations of 15, 30, 60, and 120 μL mL-1, fungicide (3.33 g L-1), and control. Minimum inhibition concentrations (MIC50 and MIC90) were evaluated. The results showed the presence of alkaloids, phenols, terpenoids and sterols in the leaf extracts. In acetone (AcE), methanol (ME) and aqueous (AqE) extracts, 32; 39 and 10 chemical compounds, respectively were found by GC-MS analysis. The most prevalent biochemically active compounds were n-Hexadecanoic acid (35.04 %), Cis-Vaccenic acid (31.76 %), Quinoline-7-carboxylic acid, 2-phenyl-, methyl ester (26.63 %), 9,12-Octadecadienoic acid (Z,Z)- methyl ester (16.89 %) and 9,12-Octadecadienoic acid (Z,Z) (10.83 %). AqE, AcE and ME extracts at 120 μL mL-1 inhibited 100 % of mycelial growth and conidial germination of isolates 1 and 2 compared to the control. The lowest MICs (MIC50 and MIC90) were 6.79 and 10.98 μL mL-1 for isolate 1 and 7.48 and 11.22 μL mL-1 for isolate 2, respectively. C. zeylanicum it is possible to use extracts for their volatile biochemical substances in a C. malayensis control program.

References

Abayhne, M.A. & Chauhan, N.M., 2016. Antifungal activity of various medicinal plants against late blight of potato from Ethiopia. Journal of Scientific Research and Reports, 12(5), pp.1-9. doi: 10.9734/JSRR/2016/30217

Alawode, TT. et al., 2021. Stigmasterol and β‑Sitosterol: Antimicrobial Compounds in the Leaves of Icacina trichantha identified by GC–MS. Beni-Suef University Journal of Basic and Applied Sciences, 10, 80. doi: 10.1186/s43088-021-00170-3.

Ali, M. & Houssain, M.Z., 2000. Inheritence of yellow vein Mosaic Virus (YVMV) tolerance in cultivar of okra (Abelmoschus esculentus (L.) Moench). Euphytica, 111, pp.205-209. doi: 10.1023/A:1003855406250.

Ambang, Z. et al., 2011. Control of leaf spot disease caused by Cercospora sp. on groundnut (Arachis hypogaea) using methanolic extracts of yellow oleander (Thevetia peruviana) seeds. Australian Journal of Crops Science, 5(3), pp.227-232.

Atindo, S.T. et al., 2020. Effectiveness of Moringa oleifera (Lam) Extracts against Sclerotinia sclerotorium (Lib) De Bary, the Causative Agent of White Mold of Common Bean (Phaseolus vulgaris L.). International Journal of Current Microbiology and Applied Sciences, 9(11), pp.812-820. doi: 10.20546/ijcmas.2020.911.098.

Baltussen, T.J.H. et al., 2020. Molecular mechanisms of conidial germination in Aspergillus spp. Microbiology and Molecular Biology Reviews, 84(1), e00049-19. doi: 10.1128/mmbr.00049-19.

Banu, K.S. & Cathrine L., 2015. General Techniques Involved in Phytochemical Analysis. International Journal of Advanced Research in Chemical science, 2(4), pp.25-32.

Beilharz, V., Pascoe, I. & Parbery, D., 2002. Three new Pseudocercospora species, one with a Mycosphaerella teleomorph, from Kennedia in Australia. Mycotaxon 82, pp.397-408.

Berkov, S. et al., 2011. Antiproliferative Alkaloids from Crinum zeylanicum. Phytotherapy Research 25(11), pp.1686-1692. doi: doi.org/10.1002/ptr.3468.

Bolie, H. et al., 2021. Antifungal Activity of Annona muricata Seed Extracts against Cercospora malayensis, Causal Agent of Cercospora Leaf Spot Disease of Okra (Abelmoschus esculentus L.). International Journal of Pathogen Research, 6(4), pp.12-24. doi: 10.9734/IJPR/2021/v6i430167.

Braun, U. & Freire F., 2004. Some cercosporoid hyphomycetes from Brazil-III. Cryptogamie Mycologie 25, pp.221–244.

Braun, U., Crous, P.W. & Nakashima, C., 2014. Cercosporoid fungi (Mycosphaerellaceae) 2. Species on monocots (Acoraceae to Xyridaceae, excluding Poaceae). IMA Fungus, 5, pp.203–390. doi: 10.5598/imafungus.2014.05.02.04.

Chai, A. et al., 2021. First Report of Cercospora Leaf Spot Caused by Cercospora cf. flagellaris on Okra in China. Plant disease, 105(7). doi: 10.1094/PDIS-10-20-2155-PDN.

Crous, P.W. & Braun, U. 2003. Mycosphaerella and Its Anamorphs: 1. Names Published in Cercospora and Passalora, Netherlands: Centraalbureau voor Schimmelcultures (CBS).

Da, X. et al., 2019. Antifungal activity and mechanism of action of Ou-gon (Scutellaria root extract) components against pathogenic fungi. Scientific Reports, 9, 1683. doi: 10.1038/s41598-019-38916-w.

Daub, M.E., 1982. Peroxidation of tobacco membrane lipids by the photosensitizing toxin, cercosporin. Plant Physiology, 69(6), pp.1361-1364. doi: 10.1104/pp.69.6.1361.

Dida, S.L.L. et al., 2024. First report of different mating types of Phytophthora infestans isolates from potatoes in Cameroon and their control using Thevetia peruviana (Pers.) K. Schum. seeds extracts analyzed by GC–MS. Indian Phytopathology, 77, pp.363–375. doi: 10.1007/s42360-024-00720-6.

Dubey, V.K. & Bhagat, K.P., 1998. Effect of insecticides and plant products against shoot and fruit borer of okra, Earias vittella (Fab.). Agricultural Science Digest, 18(2), pp.120–122.

Edeoga, H.O., Okwu, D.E. & Mbaebie, B.O., 2005. Phytochemical constituents of some Nigerian medicinal plants. African Journal of Biotechnology, 4(7), pp.685-688. doi: 10.5897/AJB2005.000-3127.

España, M.D. et al., 2017. Eucalyptus leaf byproduct inhibits the anthracnose-causing fungus Colletotrichum gloeosporioides. Industrial Crops and Products, 108, pp.793-797. doi: 10.1016/j.indcrop.2017.08.002.

Essomé, C.E. et al., 2022. Antifungal Potential of Acetone and Ethyl Acetate Extracts of Thevetia peruviana on Development of Phytophthora colocasiae, Causal Agent of Late Blight of Taro (Colocasia esculenta (L.) Schott) from Three Agro-Ecological Zones of Cameroon. International Journal of Pathogen Research, 11(2), pp.20-31. doi: 10.9734/ijpr/2022/v11i2208

FAOSTAT, 2024, ‘Statistics databases; annual production of tuber crops in Africa’, in Food and Agriculture Organization of the United Nations, viewed June 2024 from https://www.fao.org/faostat/

Farrag, E.S.H., 2011. First record of Cercospora leaf spot disease on okra plants and its control in Egypt. Plant Pathology, 10, pp.175-180. doi: 10.3923/ppj.2011.175.180

Fulano, A.M. et al., 2016. Antifungal Activity of Local Microbial Isolates against Snap Bean Pathogens. International Journal of Current Microbiology and Applied Science, 5(12), pp.112-122. doi: 10.20546/ijcmas.2016.512.013

Griffin, S.G, Markham, J.L. & Leach, D.N., 2000. An agar dilution method for the determination of the minimum inhibitory concentration of essential oils. Journal of Essential Oil Research, 12, pp.249-255. doi: 10.1080/10412905.2000.9699509.

Gurjar, M.S. et al., 2012. Efficacy of plant extracts in plant disease management. Agricultural Sciences, 3(3), pp.425-433. doi: 10.4236/as.2012.33050.

Hasanin, M.S. & Hashem, A.H., 2020. Eco-friendly, economic fungal universal medium from watermelon peel waste. Journal of Microbiological Methods, 168, 105802. doi: 105802. 10.1016/j.mimet.2019.105802

Hashem, M. & Farrag E.S.H., 2005. Biological control of Cercospora beticola leaf spot of sugar beet and its associated invaders. Egyptian Journal of Biotechnology, 20, pp.312-327.

Hernández-Gutiérrez, A. & Dianese, J.C., 2009. New cercosporoid fungi from the Brazilian Cerrado 2. Species on hosts of the subfamilies Caesalpinioideae, Faboideae and Mimosoideae (Leguminosae S. Lat.). Mycotaxon, 107, pp.1-24. doi: 10.5248/107.1

Jafri, S.A.A. et al., 2022. Evaluation of phytochemical and antioxidant potential of various extracts from traditionally used medicinal plants of Pakistan. Open Chemistry, 20(1), pp.1337-1356. doi: 10.1515/chem-2022-0242

Jasso de Rodríguez, D., Angulo-Sánchez, J.L. & Hernández-Castillo, F.D., 2006. An Overview of the Antimicrobial Properties of Mexican Medicinal Plants. In Naturally Occurring Bioactive Compounds, Elsevier, Amsterdam, pp.325-377. doi: 10.9734/ARRB/2014/5777.

Ju, H.J. et al., 2020. First Report of Cercospora malayensis Causing Leaf Spot on Okra in Korea. Plant disease, 104(6), pp.1858-1858. doi: 10.1094/PDIS-11-19-2468-PDN.

Jukte, S.R. et al., 2016. Symptomatology, isolation, identification and pathogenicity test of damping off disease in okra. International Journal of Plant Protection, 9, pp.358-361.

Kahlon, T.S., Chapman M.H. & Smith G.E., 2007. In vitro binding of bile acids by okra beets asparagus eggplant turnips green beans carrots and cauliflower. Food Chemistry, 103: 676-680. doi: 10.1016/j.foodchem.2006.07.056.

Kumar, S.V. et al., 2010. Phytochemical investigation and chromatographic evaluation of the ethanolic extract of whole plant extract of Dendrophthoe falcata (L.F) Ettingsh. International Journal of Pharmaceutical sciences and Research, 1(1), pp.39-45. doi: 10.13040/IJPSR.0975-8232.1(1).39-45.

Manga, A.D. et al., 2021. Effectiveness of Anonidium mannii seed extracts against Colletotrichum lindemuthianum and Fusarium solani, agents responsible for anthracnose and fusariose of common beans (Phaseolus vulgaris L.) in the Centre region Cameroon. International Journal of Current Research in Biosciences and Plant Biology, 8(2), pp.23-33. doi: 10.20546/ijcrbp.2021.802.003.

Mboussi, S.B. et al., 2018. Control of cocoa mirids using aqueous extracts of Thevetia puriviana and Azadirachta indica. Cogent Food and Agriculture, 4, pp.430-470. doi: 10.1080/23311932.2018.1430470.

Ngo, T.V. et al., 2017. Impact of different extraction solvents on bioactive compounds and antioxidant capacity from the root of Salacia chinensis L. Journal of Food Quality, 1, pp.1–8. doi: 10.1155/2017/9305047.

Park, S.H. et al., 2017. Identification and Characterization of Cercospora malayensis Causing Leaf Spot on Kenaf. Mycobiology. 45(2), pp.114-118. doi: 10.5941/myco.2017.45.2.114.

Perera, F.P. et al., 2005. A Summary of Recent Findings on Birth Outcomes and Developmental Effects of Prenatal ETS, PAH, and Pesticide Exposures. NeuroToxicology, 26, pp.573-587. doi: 10.1016/j.neuro.2004.07.007

QuEChERS, 2004. A Mini-Multi-Residue Method for the Analysis of Pesticide Residues in Low-Fat Products. Baden-Württemberg: CVUA Stuttgart.

Raid, R. & Palmateer A., 2006. Florida plant diseases management guide: Okra. IFAS Extension, PDMG-V3-41, University of Florida, USA.

Ramírez-Gómez, X.S. et al., 2019. Plant metabolites in plant defense against pathogens. In Plant Diseases - Current Threats and Management Trends. IntechOpen. doi: 10.5772/intechopen.87958.

Rashid, A. et al., 2004. Efficiency of different neem (Azadirachta indica A. Juss) products against various life stages of Phytophthora infestans (Mont.) Debary. Pakistan Journal of Botany, 36(4), pp.881-886.

Refaat, J. et al., 2012. Crinum; an endless source of bioactive principles: a review, Part 1. Crinum alkaloids: lycorine-type alkaloids. International Journal of Pharmaceutical Sciences and Research, 3, pp.3091-3100.

Rongai, D. et al., 2017. Antifungal activity of pomegranate peel extract against fusarium wilt of tomato. European Journal of Plant Pathology, 147, pp.229–238 doi: 10.1007/s10658-016-0994-7.

Rosen, T. & Stein Gold, L.F. 2016. Antifungal drugs for onychomycosis: Efficacy, safety, and mechanisms of action. Seminars in Cutaneous Medicine and Surgery, 35 (Supplement 3), S51-5, doi: 10.12788/j.sder.2016.009.

Saifullah, M. & Rabbani, M.G., 2009. Evaluation and characterization of okra (Abelmoscus esculentus L. Moench.) genotypes. SAARC Journal of Agriculture, 7, pp.92-99.

Singh, G. et al., 1993. Chimical and fongistatic investigation out the essential oil citrus. Pers. Z. dentshe zeeits Halft fur Pflanzenfrankenen und flanzenschutz 100, pp.69-74

Souza, A.G.C. et al., 2015. A time series analysis of brown eye spot progress in conventional and organic coffee production systems. Plant Pathology, 64, pp.157–166. doi: 10.1111/ppa.12250.

Steinkamp, M.P., et al., 1979. Ultrastructure of lesions produced in leaves of Beta vulgaris. Physiol. Plant Pathology, 15, pp.13–16. doi: 10.1016/0048-4059(79)90035-3.

Sukdee, S., 2023. Antifungal activity of Plant Extracts against Colletotrichum capsici Causal Agent of Chili Anthracnose. Rattanakosin Journal of Science and Technology, 5(1), pp.1–8.

Świderska-Burek, U., 2015. Cercosporoid Fungi of Poland. Volume 105 Polish Botanical Society; Wrocław, Poland.

Tiwari, P. et al., 2011. Phytochemical Screening and Extraction: A Review. Internationale Pharmaceutica Sciencia. 1(1), pp.98-106.

Toka, A.N. et al., 2023. Phytochemical analysis and antifungal activity of Azadirachta indica and Balanites aegyptiaca seed extracts against Fusarium oxysporum isolate on tomatoes. Journal of Global Innovations in Agricultural Sciences, 11(3), pp.293-304. doi: 10.22194/JGIAS/23.1159

Tram, N.T.N. et al., 2002. Crinum L. (Amaryllidaceae). Fitoterapia 73(3), pp.183-208. doi: 10.1016/S0367-326X(02)00068-0

VasudhaUdupa, A. et al., 2021. The antimicrobial and antioxidant property, GC–MS analysis of non-edible oil-seed cakes of neem, madhuca, and simarouba. Bulletin of the National Research Centre, 45, 41. doi: 10.1186/s42269-021-00498-x.

Vongsak, B. et al., 2013. Maximizing total phenolics, total flavonoids contents and antioxidant activity of Moringa oleifera leaf extract by the appropriate extraction method. Industrial crops and products, 44, pp.566–571. doi: 10.1016/j.indcrop.2012.09.021

Vuerich, M.E. et al., 2023. Antifungal activity of chili pepper extract with potential for the control of some major pathogens in grapevine. Pest Management Science, 79(7), pp.2503-2516. doi: 10.1002/ps.7435.

Watanabe-Akanuma, M., Ohta, T. & Sasaki, Y.F., 2005. A novel genotoxic aspect of thiabendazole as a photomutagen in bacteria and cultured human cells. Toxicology Letters, 158, 3, pp.213-219. doi: 10.1016/j.toxlet.2005.03.013

Widmer, T.L. & Laurent, N., 2006. Plant extracts containing caffeic acid and rosmarinic acid inhibit zoospore germination of Phytophthora spp. pathogenic to Theobroma cacao. European Journal of Plant Pathology, 115, pp.377–388. doi: 10.1007/s10658-006-9024-5

Yoshimi, A.K. et al., 2022. Cell Wall Integrity and Its Industrial Applications in Filamentous Fungi. Journal of Fungi, 8,435.

You, J. et al., 2016. Multiple criteria-based screening of Trichoderma isolates for biological control of Botrytis cinerea on tomato. Biological Control, 101, pp.31–38. doi: 10.1016/j.biocontrol.2016.06.006

Yusoff, S.F. et al., 2020. Antifungal Activity and Phytochemical Screening of Vernonia amygdalina Extract against Botrytis cinerea Causing Gray Mold Disease on Tomato Fruits. Biology, 11(9), 286. doi: 10.3390/biology9090286.

Zibbu, G. & Batra, A., 2010. A Review on Chemistry and Pharmacological activity of Nerium oleander L. Journal of Chemical and Pharmaceutical Research, 6, pp.351-358.

Zulbayu, L.O.M.A., Lukitaningsih, E.R. & Rumiyatu, R., 2021. GC-MS analysis of bioactive compounds in ethanol and ethyl acetate fraction of grape fruit (Citrus maxima L.) Rind. Borneo Journal of Pharmacy, 4(1), pp.29-35. doi: 10.33084/bjop.v4i1.1665

Published
2025-05-19
How to Cite
Akong, M. E., Ngatsi, P. Z., Kuate, N. W. T., Atindo , T. S., Boli, H., Tsogo, C. S. and Ndongo, B. (2025) “Bioactivity of Volatile Compounds Present in Crinum zeylanicum Leaf Extracts Identified by GC-MS for the Control of Cercospora malayensis Isolates of Okra (Abelmoschus esculentus L.) In Vitro”, Journal of Tropical Biodiversity and Biotechnology, 10(2), p. jtbb14097. doi: 10.22146/jtbb.14097.
Section
Research Articles