Phytochemical-induced defense activation in chickpea by Calotropis procera L. extract to control Fusarium wilt
Abstract
Fungicide application offers a solution for crop protection, but synthetic fungicides pose greater harm to the environment. This study explores the potential of Calotropis procera L. leaf extracts as a nature-friendly alternative to combat Fusarium wilt in chickpea. Methanol (MeOH) extracts were tested at various concentrations (0-6.5%) against Fusarium oxysporum, with complete growth suppression observed at 5% and 6.5%. Bioassay-guided fractionation of the crude extract was performed using n-hexane, dichloromethane (DCM), ethyl acetate (EtOAc), acetone (Ace), and methanol (MeOH). The results indicated that EtOAc and Ace fractions at 0.6% completely inhibited fungal growth. Crude extracts (3.5%, 5%, and 6%) along with EtOAc (0.2% and 0.6%) and Ace (0.2% and 0.6%) fractions were further subjected to phytotoxicity analysis on chickpea seedlings. Among these, the EtOAc-0.6% fraction exhibited the least phytotoxicity. In a pot experiment, the group inoculated with F. oxysporum and treated with the botanical extract (T6) showed a lower disease incidence of 41.7%, compared to 50% in the F. oxysporum-inoculated group treated with carbendazim (T5). T6 also exhibited improved root elongation (124%), seed germination (88.89%), catalase activity (52 U/mg protein), and guaiacol peroxidase activity (26.63 U/mg protein). The highest carotenoid content (1.6 µg/mL) was observed in T5, while the uninoculated group treated with the botanical extract (T4) recorded the highest SOD activity (78.24 U/mg protein). Overall, these findings suggest that the EtOAc-0.6% fraction of C. procera leaf extract could serve as a safer and effective candidate for managing Fusarium wilt in chickpea.
Keywords
Full Text:
PDFReferences
Abo-Elyousr, K.A.M., Ali, E.F., Sallam, N.M.A., 2022. Alternative control of tomato wilt using the aqueous extract of Calotropis procera. Horticulturae 8, 197.
Aizaz, M., Ahmad, W., Asaf, S., Khan, I., Saad Jan, S., Salim Alamri, S., Bilal, S., Jan, R., Kim, K.-M., Al-Harrasi, A., 2023. Characterization of the seed biopriming, plant growth-promoting and salinity-ameliorating potential of halophilic fungi isolated from hypersaline habitats. International Journal of Molecular Sciences 24, 4904.
Akram, W., Waqar, S., Hanif, S., Anjum, T., Aftab, Z.-H., Li, G., Ali, B., Rizwana, H., Hassan, A., Rehman, A., Munir, B., Umer, M., 2024. Comparative effect of seed coating and biopriming of Bacillus aryabhattai Z-48 on seedling growth, growth promotion, and suppression of Fusarium wilt disease of tomato plants. Microorganisms 12, 792.
Ali, F., Saddiqe, Z., Shahzad, M., Rafi, A., Javed, M., Haq, F., Saleem, S., Kusar, S., 2023. Crotalaria medicaginea Lamk.: An unexplored source of anticancer, antimicrobial and antioxidant agents. European Journal of Integrative Medicine 58, 102226.
Ali, Q., Ashraf, M., 2011. Induction of drought tolerance in maize (Zea mays L.) due to exogenous application of trehalose: growth, photosynthesis, water relations and oxidative defence mechanism. Journal of Agronomy and Crop Science 197, 258-271.
Azeem, W., Mukhtar, T., Haq, M.I., Khan, M.A., Ibrahim, M.S., Hassan, A., Regmi, H., Duncan, L.W., 2025. The nematicidal potential of Moringa oleifera extracts and rhizobacteria against Meloidogyne incognita in tomato. Frontiers in Plant Science 16, 1562074. https://doi.org/10.3389/fpls.2025.1562074
Bhattacharya, S., Saha, T., Debnath, S., Krishna, S.A., 2022. Allelopathic potential of weed extracts on seed germination and seedling growth of Vigna radiata L. Wilzek and Vigna mungo L. International Journal of Life Sciences Research 10, 13-20.
Bonaterra, A., Badosa, E., Cabrefiga, J., Francés, J., Montesinos, E., 2012. Prospects and limitations of microbial pesticides for control of bacterial and fungal pomefruit tree diseases. Trees 26, 215-226.
Carril, P., Ghorbani, M., Azarnejad, N., Anselmi, S., Renzi, M., Loppi, S., 2025. Promoting early growth in tomato (Solanum lycopersicum L.) by co-application of biochar and beneficial bacteria. Journal of Soil Science and Plant Nutrition 25, 1493-1503.
Deresa, E.M., Diriba, T.F., 2023. Phytochemicals as alternative fungicides for controlling plant diseases: A comprehensive review of their efficacy, commercial representatives, advantages, challenges for adoption, and possible solutions. Heliyon 9, e13810.
Doshi, H., Satodiya, H., Thakur, M.C., Parabia, F., Khan, A., 2011. Phytochemical screening and biological activity of Calotropis procera (Ait). R. Br. (Asclepiadaceae) against selected bacteria and Anopheles stephansi larvae. International Journal of Plant Research 1, 29-33.
El-Esawi, M.A., Al-Ghamdi, A.A., Ali, H.M., Alayafi, A.A., 2019. Azospirillum lipoferum FK1 confers improved salt tolerance in chickpea (Cicer arietinum L.) by modulating osmolytes, antioxidant machinery and stress-related genes expression. Environmental and Experimental Botany 159, 55-65.
Ghasemi, S., Ghasemi, M., Moradi, N., 2011. Effect of Calotropis procera leaf extract on seed germination of some plants. Journal of Ornamental and Horticultural Plants 2(1), 27-32.
Haida, Z., Hakiman, M., 2019. A comprehensive review on the determination of enzymatic assay and nonenzymatic antioxidant activities. Food Science and Nutrition 7, 1555-1563.
Hiscox, J.D., Israelstam, G.F., 1979. A method for the extraction of chlorophyll from leaf tissue without maceration. Canadian journal of botany 57(12), 1332-1334.
Huang, H., Ullah, F., Zhou, D.X., Yi, M., Zhao, Y., 2019. Mechanisms of ROS regulation of plant development and stress responses. Frontiers in plant science 10, 800.
Hussain, M.A., Mukhtar, T., Kayani, M.Z., 2011. Efficacy evaluation of Azadirachta indica, Calotropis procera, Datura stramonium and Tagetes erecta against root-knot nematodes Meloidogyne incognita. Pakistan Journal of Botany 43 (Special Issue), 197-204.
Jabbar, A., Tariq, M., Gulzar, A., Mukhtar, T., Zainab, T., 2022. Lethal and sub-lethal effects of plant extracts and green silver nanoparticles against Culex pipiens mosquitoes. Pakistan Journal of Zoology 54(3), 1259-1267.
Jiménez-Díaz, R.M., Castillo, P., Jiménez-Gasco, M.M., Landa, B.B., Navas-Cortés, J.A., 2015. Fusarium wilt of chickpeas: Biology, ecology and management. Crop Protection 73, 16-27.
Kayani, M.Z., Mukhtar, T., Hussain, M.A., 2012. Evaluation of nematicidal effects of Cannabis sativa L. and Zanthoxylum alatum Roxb. against root-knot nematodes, Meloidogyne incognita. Crop Protection 39, 52-56.
Khan, H.S., Tariq, M., Mukhtar, T., Gulzar, A., 2021. Insecticidal toxicity of plant extracts and green silver nanoparticles against Aedes albopictus. Pakistan Journal of Zoology 53(6), 2123-2128.
Kumar, S., Li, G., Yang, J., Huang, X., Ji, Q., Liu, Z., Ke, W., Hou, H., 2021. Effect of salt stress on growth, physiological parameters, and ionic concentration of water dropwort (Oenanthe javanica) cultivars. Frontiers in plant science, 12, 660409.
Kumar, S., Singh, A.J.J.B.B., 2014. Biopesticides for integrated crop management: environmental and regulatory aspects. Journal of Biofertilizers and Biopesticides 5, e121.
Leong, W.-H., Teh, S.-Y., Hossain, M.M., Nadarajaw, T., Zabidi-Hussin, Z., Chin, S.-Y., Lai, K.-S., Lim, S.-H.E., 2020. Application, monitoring and adverse effects in pesticide use: The importance of reinforcement of Good Agricultural Practices (GAPs). Journal of Environmental Management 260, 109987.
Li, X., Yang, Y.Q., Zhang, M., Wang, X.F., 2010. Effect of PEG priming on plasma membrane H+-ATPase activities and mitochondrium function in soybean seeds. Seed Science and Technology 38, 49-60.
Lichtenthaler, H.K., 1987. Chlorophylls and carotenoids: Pigments of photosynthetic biomembranes, in: Methods in Enzymology, Plant Cell Membranes. Academic Press, pp. 350-382.
Morsy, N., 2014. Phytochemical analysis of biologically active constituents of medicinal plants. Main Group Chemistry 13, 7-21.
Mukhtar, T., Kayani, M.Z., Hussain, M.A., 2013. Nematicidal activities of Cannabis sativa L. and Zanthoxylum alatum Roxb. against Meloidogyne incognita. Industrial Crops and Products 42, 447-453.
Nagpal, S., Sharma, P., Sirari, A., 2021. Induction of antioxidant response with compatible combination of Mesorhizobium sp. and Pseudomonas sp. against Fusarium oxysporum sp. ciceris in chickpea. Bangladesh Journal of Botany 50, 359-364.
Peng, Y., Li, S.J., Yan, J., Tang, Y., Cheng, J.P., Gao, A.J., Yao, X., Ruan, J.J., Xu, B.L., 2021. Research progress on phytopathogenic fungi and their role as biocontrol agents. Frontiers in Microbiology 12, 670135.
Radwan, A.M., Alghamdi, H.A., Kenawy, S.K.M., 2019. Effect of Calotropis procera L. plant extract on seeds germination and the growth of microorganisms. Annals of Agricultural Sciences 64, 183-187.
Raja, R., 2023. Pulses outlook (Agricultural Situation, Market Development Reports No. PK2023-0015). United States Department of Agriculture Foreign Agricultural Service, Pakistan.
Saeed, M., Mukhtar, T., Haq, M.I., Khan, M.A., 2021. Assessment of nematicidal potential of Cannabis sativa and Azadirachta indica in the management of root-knot nematode (Meloidogyne javanica) on peach. Pakistan Journal of Agricultural Sciences 58, 1555-1561.
Siraj, Khan, N., 2021. Quantitative ethnobotany of Calotropis procera and associated vegetation: a step forward for conservation and management practice in northern areas of Pakistan. Ethnobotany Research and Applications 22, 1-17.
Tembo, Y., Mkindi, A.G., Mkenda, P.A., Mpumi, N., Mwanauta, R., Stevenson, P.C., Ndakidemi, P.A., Belmain, S.R., 2018. Pesticidal plant extracts improve yield and reduce insect pests on legume crops without harming beneficial arthropods. Frontiers in Plant Science 9, 1425.
Thite, S., Aparadh, V., Kore, B., 2013. Effect of powdery mildew infection on DPPH radical scavenging activity and ferric-reducing antioxidant power of plants. World Journal of Pharmaceutical Research 2, 1-6.
Wang, Q., Pokhrel, A., Coleman, J.J., 2021. The extracellular superoxide dismutase Sod5 from Fusarium oxysporum is localized in response to external stimuli and contributes to fungal pathogenicity. Frontiers in Plant Science 12, 608861.
Wojtyla, L., Lechowska, K., Kubala, S., Garnczarska, M., 2016. Different modes of hydrogen peroxide action during seed germination. Frontiers in Plant Science 7, 66.
Xu, J., Xiong, H., Zhang, X., Muhayimana, S., Liu, X., Xue, Y., Huang, Q., 2020. Comparative cytotoxic effects of five commonly used triazole alcohol fungicides on human cells of different tissue types. Journal of Environmental Science and Health, Part B 55, 438-446.
Yago, J., Roh, J., Bae, S., Yoon, Y., Kim, H., Nam, M., 2011. The effect of seed-borne mycoflora from sorghum and foxtail millet seeds on germination and disease transmission. Mycobiology 39, 206-218.
Zhang, J., Wang, J., Zhu, C., Singh, R.P., Chen, W., 2024. Chickpea: its origin, distribution, nutrition, benefits, breeding, and symbiotic relationship with Mesorhizobium species. Plants 13, 429.
Zubairi, T., Jabeen, K., Khalid, S., Iqbal, S., 2021. Isolation and molecular characterization of causal agent of blue mold on Allium cepa L. and its control by Pennistum flaccidum Griseb. Saudi Journal of Biological Sciences 28, 6774-6781.
Zucconi, F., Pera, A., Forte, M., DeBertolli, M., 1981. Evaluating toxicity of immature compost. Biocycle, 22, 54-57.
DOI: https://doi.org/10.33804/pp.009.02.5603
Refbacks
- There are currently no refbacks.





