Rachmi Masnilah, Ilfa I. Dewi, Ankardiansyah P. Pradana


The study aimed to investigate the inhibitory potential of betel and neem leaf extracts against the growth of the Sclerotium rolfsii, which causes damping-off disease in groundnut plants. The research employed both in vitro and in vivo methods to determine the antifungal activity of the extracts. The isolation of the pathogen inoculum was done from the base stem of the infected groundnut plants by S. rolfsii in the field. The betel and neem leaves were dried at a controlled temperature of 40°C for 7 days, ground, and then immersed in 96% ethanol for 48 hours. The resulting extracts were tested for their inhibitory potential against S. rolfsii in vitro by admixing them with PDA media, inoculating with S. rolfsii, and measuring colony diameters. The study found that the colony diameter of S. rolfsii was significantly influenced by the use of betel leaf extract, neem leaf extract, and a mixture of both extracts at a concentration of 1.5% (v/v). The widest colony diameter was observed in the control treatment on day 5, measuring 7.47 cm. In contrast, the colony diameter of S. rolfsii in the treatment with betel leaf extract, neem leaf extract, and a mixture of both extracts on day 5 was 5.58 cm, 4.35 cm, and 1.41 cm, respectively. The addition of each extract to the parameters of colony diameter and inhibition percentage was capable of inhibiting the growth of S. rolfsii, with the betel-neem extract treatment exhibiting the highest inhibition percentage of 81.09%. In the in vivo suppression test, the study found that treating groundnut seeds with a 1.5% concentration of betel leaf and neem leaf extracts, as well as a combination of both extracts, significantly reduced the incidence of damping-off disease. The betel-neem extract treatment produced the lowest disease incidence and exhibited the highest percentage of pre-emergence in groundnut seeds. Overall, the study provides evidence that betel and neem leaf extracts have antifungal activity against S. rolfsii and could be used as a potential alternative to synthetic fungicides for controlling damping-off disease in groundnut.


Antifungal, Fungicide, In vitro, In vivo, Sclerotium.

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Ahmad, A. 2012. Potential applications of neem based products as biopesticides. The Health Journal, 3: 116-120.

Akgul, D. S., H. Ozgonen and A. Erkilic. 2011. The effects of seed treatments with fungicides on stem rot caused by Sclerotium rolfsii Sacc., in peanut. Pakistan Journal of Botany, 43: 2991-2996.

Alcazar-Fuoli, L. and E. Mellado. 2013. Ergosterol biosynthesis in Aspergillus fumigatus: its relevance as an antifungal target and role in antifungal drug resistance. Frontiers in Microbiology, 3: 439-445.

Al-Obaidi, H., R. M. Kowalczyk, R. Kalgudi and M. G. Zariwala. 2019. Griseofulvin solvate solid dispersions with synergistic effect against fungal biofilms. Colloids and Surfaces B: Biointerfaces, 184: 110540.

Altine, S., M. Sabo, N. Muhammad, A. Abubakar and L. Saulawa. 2016. Basic nutrient requirements of the domestic quails under tropical conditions: A review. World Scientific News, 2: 223-235.

Arif, T., J. Bhosale, N. Kumar, T. Mandal, R. Bendre, G. Lavekar and R. Dabur. 2009. Natural products–antifungal agents derived from plants. Journal of Asian Natural Products Research, 11: 621-638.

Arockianathan, P. M., M. Mishra and R. Niranjan. 2019. Recent status and advancements in the development of antifungal agents: highlights on plant and marine based antifungals. Current Topics in Medicinal Chemistry, 19: 812-830.

Bera, S., S. Kasundra, J. Kamdar, A. B. C, C. Lal, P. P. Thirumalasmy, P. Dash and A. K. Maurya. 2014. Variable response of interspecific breeding lines of groundnut to Sclerotium rolfsii infection under field and laboratory conditions. Electronic Journal of Plant Breeding, 5: 22-29.

Bhatia, S., R. C. Dubey and D. K. Maheshwari. 2005. Enhancement of plant growth and suppression of collar rot of sunflower caused by Sclerotium rolfsii through fluorescent Pseudomonas. Indian Phytopathology, 58: 17-24.

Bhuiyan, M. A. H, B., M. T. Rahman and K. A. Bhuiyan. 2012. In vitro screening of fungicides and antagonists against Sclerotium rolfsii. African Journal of Biotechnology, 11: 14822-14827.

Billah, K. M. M., M. B. Hossain, M. H. Prince and M. M. P. Sumon. 2017. Pathogenicity of Sclerotium rolfsii on different host, and its over wintering survival; A mini review. International Journal of Advances in Agriculture Sciences, 2: 1-6.

Carrillo-Munoz, A. J., G. Giusiano, P. A. Ezkurra and G. Quindós. 2006. Antifungal agents: mode of action in yeast cells. Revista Espanola de Quimioterapia, 19: 130-139.

Dalavayi H., M. F. Ahmed, S. Bala and D. Choudhury. 2021. Eco-friendly plant based on botanical pesticides. Plant Archives, 21: 2197-2204.

Datta, A., S. Ghoshdastidar and M. Singh. 2011. Antimicrobial property of Piper betel leaf against clinical isolates of bacteria. International Journal of Pharma Sciences and Research, 2: 104-109.

Dwivedi, S. and G. Prasad. 2016. Integrated management of Sclerotium rolfsii: an overview. European Journal of Biomedical and Pharmaceutical Sciences, 3: 137-146.

Ezeonu, C. S., C. Imo, D. I. Agwaranze, A. Iruka and A. Joseph. 2018. Antifungal effect of aqueous and ethanolic extracts of neem leaves, stem bark and seeds on fungal rot diseases of yam and cocoyam. Chemical and Biological Technologies in Agriculture, 5: 18.

Fariña, J., F. Sineriz, O. Molina and N. Perotti. 2001. Isolation and physicochemical characterization of soluble scleroglucan from Sclerotium rolfsii. Carbohydrate Polymers, 44: 41-50.

Zore, G. B., D. T. Archana, J. Sitara and S. M. Karuppayil. 2011. Terpenoids inhibit Candida albicans growth by affecting membrane integrity and arrest of cell cycle. Phytomedicine, 18: 1181-1190.

Guclu, V., M. Aydogdu, M. Basak, S. Kizil, B. Uzun and E. Yol. 2020. Characterization of a groundnut collection to stem rot disease caused by Sclerotium rolfsii. Australasian Plant Pathology, 49: 691-700.

He, Y., P. Du, T. Zhao, F. Gao, M. Wang, J. Zhang, L. He, K. Cui and L. Zhou. 2022. Baseline sensitivity and bioactivity of tetramycin against Sclerotium rolfsii isolates in Huanghuai peanut-growing region of China. Ecotoxicology and Environmental Safety, 238: 113580.

Hidayati, N. 2018. Identification of pathogen causes of damping off diseases on kaliandra. Jurnal Pemuliaan Tanaman Hutan, 12: 137-144.

Hoesain, M., S. Prastowo, S. Suharto, A. P. Pradana, I. N. Asyiah, F. K. Alfarizy and M. Adiwena. 2021. Combination of plant growth-promoting bacteria and botanical pesticide increases organic red rice yield and reduces the Leptocorisa acuta population. Biodiversitas Journal of Biological Diversity, 22: 1686-1694.

Hussainy, S. A. H. and R. Vaidyanathan. 2019. Relative performance of groundnut (Arachis hypogaea) based intercropping systems under different irrigation levels. International Journal of Agriculture and Biology, 22: 841-848.

Kiprono, P. C., F. Kaberia, J. M. Keriko and J. N. Karanja. 2000. The in vitro anti-fungal and anti-bacterial activities of β-sitosterol from Senecio lyratus (Asteraceae). Zeitschrift für Naturforschung C, 55: 485-488.

Koike, S. 2004. Southern blight of Jerusalem artichoke caused by Sclerotium rolfsii in California. Plant Disease, 88: 769-769.

Kumar, P., S. Bharty and K. Kumar. 2017. Management of collar rot of elephant foot yam caused by Sclerotium rolfsii Sacc.-A Review. Journal of Pharmacognosy and Phytochemistry, 6: 723-728.

Kwon, J. H., J. W. Kim, Y. H. Lee and H. S. Shim. 2012. Sclerotium rot of sponge gourd caused by Sclerotium rolfsii. Research in Plant Disease, 18: 54-56.

Lachke, A. H. and M. V. Deshpande. 1988. Sclerotium rolfsii: status in cellulase research. FEMS Microbiology Reviews, 4: 177-193.

Costa, L. B. C., J. E. B. P. Pinto, S. K. V. Bertolucci, J. C. B. Costa, P. B. Alves and E. S. Niculau. 2015. In vitro antifungal activity of Ocimum selloi essential oil and methylchavicol against phytopathogenic fungi. Revista Ciência Agronômica, 46: 428-435.

Madhumita, M., P. Guha and A. Nag. 2020. Bio‐actives of betel leaf (Piper betle L.): A comprehensive review on extraction, isolation, characterization, and biological activity. Phytotherapy Research, 34: 2609-2627.

Mahadevakumar, S., C. Chandana, Y. Deepika, K. Sumashri, V. Yadav and G. Janardhana. 2018. Pathological studies on the southern blight of China aster (Callistephus chinensis) caused by Sclerotium rolfsii. European Journal of Plant Pathology, 151: 1081-1087.

Mahmoud, D., N. Hassanein, K. Youssef and M. A. Zeid. 2011. Antifungal activity of different neem leaf extracts and the nimonol against some important human pathogens. Brazilian Journal of Microbiology, 42: 1007-1016.

Maimunah, M., A. Azwana and C. Pandala. 2019. The effectiveness of kenikir and betel leaves extract as bio fungicide to the causes of anthracnose disease (Colletotrichum capsici) on chili plants (Capsicum annum L.) with In vitro. Budapest International Research in Exact Sciences, 1: 29-36.

Didehar, M., Z. Chegini and A. Shariati. 2022. Eugenol: A novel therapeutic agent for the inhibition of Candida species infection. Frontiers in Pharmacology, 13: 872127.

Nawaz, M., J. I. Mabubu and H. Hua. 2016. Current status and advancement of biopesticides: microbial and botanical pesticides. Journal of Entomology and Zoology Studies, 4: 241-246.

Ngegba, P. M., G. Cui, M. Z. Khalid and G. Zhong. 2022. Use of botanical pesticides in agriculture as an alternative to synthetic pesticides. Agriculture, 12: 600-609.

Nugroho, C., E. Mirnia and C. J. R. Cumagun. 2019. Antifungal activities of sweet basil (Ocimum basilicum L.) aqueous extract against Sclerotium rolfsii, causal agent of damping-off on tomato seedling. Agrivita, 41: 149-157.

Odds, F. C., A. J. P. Brown and N. A. R. Gow. 2003. Antifungal agents: mechanisms of action. Trends in Microbiology, 11: 272-279.

Oliveira, V., E. Carraro, M. Auler and N. Khalil. 2016. Quercetin and rutin as potential agents antifungal against Cryptococcus spp. Brazilian Journal of Biology, 76: 1029-1034.

Panth, M., S. C. Hassler and F. Baysal-Gurel. 2020. Methods for management of soilborne diseases in crop production. Agriculture, 10: 16-24.

Pawar, S., V. Kalyankar, B. Dhamangaonkar, S. Dagade, S. Waghmode and A. Cukkemane. 2017. Biochemical profiling of antifungal activity of betel leaf (Piper betle L.) extract and its significance in traditional medicine. Journal of Advanced Research in Biotechnology, 2: 1-4.

Raghavendra, S. S. and K. D. Balsaraf. 2014. Antifungal efficacy of Azadirachta indica (neem)-An in vitro study. Brazilian Journal of Oral Sciences, 13: 242-245.

Rani, L., K. Thapa, N. Kanojia, N. Sharma, S. Singh, A. S. Grewal, A. L. Srivastav and J. Kaushal. 2021. An extensive review on the consequences of chemical pesticides on human health and environment. Journal of Cleaner Production, 283: 124657.

Ristaino, J. B., P. K. Anderson, D. P. Bebber, K. A. Brauman, N. J. Cunniffe, N. V. Fedoroff, C. Finegold, K. A. Garrett, C. A. Gilligan and C. M. Jones. 2021. The persistent threat of emerging plant disease pandemics to global food security. Proceedings of the National Academy of Sciences, 118: e2022239118.

Sadeghi-Ghadi, Z., A. Vaezi, F. Ahangarkani, M. Ilkit, P. Ebrahimnejad and H. Badali. 2020. Potent in vitro activity of curcumin and quercetin co-encapsulated in nanovesicles without hyaluronan against Aspergillus and Candida isolates. Journal de Mycologie Medicale, 30: 101014.

Sarma, B., U. Singh and K. Singh. 2002. Variability in Indian isolates of Sclerotium rolfsii. Mycologia, 94: 1051-1058.

Schilling, B., A. Henning and U. Rau. 2000. Repression of oxalic acid biosynthesis in the unsterile scleroglucan production process with Sclerotium rolfsii ATCC 15205. Bioprocess Engineering, 22: 51-55.

Semangun, H. 1991. Penyakit-penyakit tanaman pangan di Indonesia, Gadjah Mada University Press, Indonesia.

Singh, U., S. Maurya and D. Singh. 2005. Phenolic acids in neem (Azadirachta indica) a major pre-existing secondary metabolites. Journal of Herbal Pharmacotherapy, 5: 35-43.

Suleiman, M. 2011. Antifungal properties of leaf extract of neem and tobacco on three fungal pathogens of tomato (Lycopersicon esculentum Mill). Advances in Applied Science Research, 2: 217-220.

Xu, H., D. Lai and Z. Zhang. 2017. Research and application of botanical pesticide azadirachtin. Journal of South China Agricultural University, 38: 1-11.

Zhang, P., E. F. Sari, M. J. McCullough and N. Cirillo. 2022. Metabolomic profile of Indonesian betel quids. Biomolecules, 12: 1469.



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