Mechanistic elucidation of a terpenoid nano-bionematicide for the management of root-knot nematodes, Meloidogyne incognita infecting tomato

mechanistic-elucidation-of-a-terpenoid-nano-bionematicide-for-the-management-of-root-knot-nematodes,-meloidogyne-incognita-infecting-tomato
Mechanistic elucidation of a terpenoid nano-bionematicide for the management of root-knot nematodes, Meloidogyne incognita infecting tomato

Scientific Reports , Article number:  (2026) Cite this article

We are providing an unedited version of this manuscript to give early access to its findings. Before final publication, the manuscript will undergo further editing. Please note there may be errors present which affect the content, and all legal disclaimers apply.

Subjects

Abstract

Plant-parasitic nematodes, particularly Meloidogyne incognita, represent a major constraint to global vegetable production and cause substantial yield losses. Although azadirachtin exhibits strong nematicidal potential, its practical application is limited by instability under field conditions. In this study, a terpenoid-based azadirachtin nano-formulation (Terpaz®, TNF) was developed and evaluated through an integrated approach encompassing physicochemical characterization, in vitro bioassays, molecular docking and molecular dynamics simulations, enzyme inhibition assays, root invasion studies, field evaluation, and biosafety assessment. The characterisation of TNF exhibited a nanoscale droplet size (79.69 nm) with narrow PDI (0.17). In vitro assays revealed substantial inhibition of egg hatching (up to 91.3%) and juvenile mortality (up to 88%), outperforming the synthetic nematicide fluopyram at equivalent doses. Molecular docking demonstrated a strong binding affinity of azadirachtin to acetylcholinesterase (AChE, − 5.37 kcal/mol) and ATPase (–4.8 kcal/mol), which was further supported by stable molecular dynamics trajectories. Enzyme assays confirmed dose-dependent inhibition of AChE (65.7%) and ATPase (73.4%) activities, validating the dual-target mechanism. Root penetration studies showed that TNF at 5 mL/L prevented juvenile invasion beyond the epidermis. Field trials demonstrated that TNF significantly suppressed nematode population densities (Pf = 1.5 J2/g soil), reduced galling, and enhanced tomato yield (25,400 kg/acre), outperforming the synthetic nematicide. Importantly, TNF showed no inhibitory effects on beneficial soil microbes. Collectively, this study establishes TNF as an effective, environmentally safe, and multi-targeted bionematicide, offering a promising sustainable alternative for M. incognita management in horticultural crops.

Data availability

All datasets generated during this study are included in this published article and its Supplementary files (Supplementary_file.xlsx). Other specific data will be available made on the reasonable request.

References

  1. Kyndt, T., Fernandez, D. & Gheysen, G. Plant-parasitic nematode infections in rice: molecular and cellular insights. Annu. Rev. Phytopathol. 52 (1), 135–153. https://doi.org/10.1146/annurev-phyto-102313-050111 (2014).

    Google Scholar 

  2. Jones, J. T. et al. Top 10 plant-parasitic nematodes in molecular plant pathology. Molecular Plant Pathology 14 (9), 946–961. https://doi.org/10.1111/mpp.12057 (2013).

  3. Nicol, J. M., Turner, S. J., Coyne, D. L., den Nijs, L. & Hockland, S. and Z. Tahna Maafi. Current nematode threats to world agriculture. In Genomics and Molecular Genetics of Plant-Nematode Interactions 21–43. https://doi.org/10.1007/978-94-007-0434-3_2 (Springer Netherlands, 2011).

  4. Li, J. et al. Molecular mechanisms of nematode-nematophagous microbe interactions: basis for biological control of plant-parasitic nematodes. Annu. Rev. Phytopathol. 53 (1), 67–95 (2015).

    Google Scholar 

  5. Kim, J. H. et al. Assessment of nematicidal and plant growth-promoting effects of Burkholderia sp. JB-2 in root-knot nematode-infested soil. Front. Plant Sci. 14, 1216031. https://doi.org/10.3389/fpls.2023.1216031 (2023).

    Google Scholar 

  6. Vos, C. et al. Bart Panis, and Vivienne Gianinazzi-Pearson. Mycorrhiza-induced resistance against the root–knot nematode Meloidogyne incognita involves priming of defense gene responses in tomato. Soil Biol. Biochem. 60, 45–54 (2013).

    Google Scholar 

  7. Riga, E. The effects of Brassica green manures on plant parasitic and free living nematodes used in combination with reduced rates of synthetic nematicides. J. Nematology. 43 (2), 119 (2011).

    Google Scholar 

  8. Subhash, S., Raghavendra, K. V., Balodi, R. & Dubey, N. K. Deepika, Use of green chemicals in pest and disease management. In Sustainable Management of Potato Pests and Diseases 495–524. (Springer Singapore, 2022).

  9. Adusei, S. & Azupio, S. Neem: A novel biocide for pest and disease control of plants. J. Chem. 2022 (1), 6778554. https://doi.org/10.1155/2022/6778554 (2022).

    Google Scholar 

  10. Ahmad, F., Rather, M. A. & Siddiqui, M. A. Nematicidal activity of leaf extracts from Lantana camara L. against Meloidogyne incognita (kofoid and white) chitwood and its use to manage roots infection of Solanum melongena L. Brazilian archives biology Technol. 53, 543–548 (2010).

    Google Scholar 

  11. Dang, Q. L. et al. Nematicidal and antifungal activities of annonaceous acetogenins from Annona squamosa against various plant pathogens. J. Agric. Food Chem. 59 (20), 11160–11167. https://doi.org/10.1021/jf203017f (2011).

    Google Scholar 

  12. Zhang, W. P., Ruan, W. B., Deng, Y. Y. & Gao, Y. B. Potential antagonistic effects of nine natural fatty acids against Meloidogyne incognita. J. Agric. Food Chem. 60 (46), 11631–11637. https://doi.org/10.1021/jf3036885 (2012).

    Google Scholar 

  13. Boeke, S. J. et al. Safety evaluation of neem (Azadirachta indica) derived pesticides. J. Ethnopharmacol. 94 (1), 25–41. https://doi.org/10.1016/j.jep.2004.05.011 (2004).

    Google Scholar 

  14. Mojumder, V., Kamra, A. & Dureja, P. Effect of neem extracts on activity and mortality of second-stage juveniles of Meloidogyne incognita (Nematologia mediterranea, 2002).

  15. d’Errico, G., Sasanelli, N., Guastamacchia, F., Stillittano, V. & D’Addabbo, T. Efficacy of azadirachtin in the integrated management of the root knot nematode Meloidogyne incognita on short-and long-cycle crops. Plants 12 (6), 1362. https://doi.org/10.3390/plants12061362 (2023).

    Google Scholar 

  16. Cromwell, W. A., Yang, J., Starr, J. L. & Jo, Y. K. Nematicidal effects of silver nanoparticles on root-knot nematode in bermudagrass. J. nematology. 46 (3), 261 (2014).

    Google Scholar 

  17. Priyadharshini, V. et al. Smart delivery of nanofiber matrix encapsulated botanical nematicide α-terthienyl against root knot nematode on tomato. Physiol. Mol. Plant Pathol. 136, 102508. https://doi.org/10.1016/j.pmpp.2024.102508 (2025).

    Google Scholar 

  18. Kundu, A. et al. A comprehensive in vitro and in silico analysis of nematicidal action of essential oils. Front. Plant Sci. 11, 614143. https://doi.org/10.3389/fpls.2020.614143 (2021).

    Google Scholar 

  19. Arun, A. et al. An insight into occurrence, biology, and pathogenesis of rice root-knot nematode Meloidogyne graminicola. Biology 12 (7), 987. https://doi.org/10.3390/biology12070987 (2023).

    Google Scholar 

  20. Shelley, M. J. The dynamics of microtubule/motor-protein assemblies in biology and physics. Annu. Rev. Fluid Mech. 48 (1), 487–506 (2016).

    Google Scholar 

  21. Friesner, R. A. Combined quantum and molecular mechanics (QM/MM). Drug Discov. Today Technol. 1 (3), 253–260. (2004).

  22. Sharmila, D., Jeya Sundara & Jino Blessy, J. Molecular dynamics of sialic acid analogues complex with cholera toxin and DFT optimization of ethylene glycol-mediated zinc nanocluster conjugation. J. Biomol. Struct. Dynamics. 35 (1), 182–206. https://doi.org/10.1080/07391102.2015.1136689 (2017).

    Google Scholar 

  23. Abdel-Rahman, A. A. et al. Ahmed. Sublethal concentrations of conventional nematicides alter the physiological activities of Meloidogyne incognita and suppress parasitism. Sci. Rep. 13 (1), 229. https://doi.org/10.1038/s41598-022-27270-z (2023).

    Google Scholar 

  24. Li, J. et al. Acetylcholinesterase inhibitory activity of sesquiterpenoids isolated from Laggera pterodonta. Front. Plant Sci. 14, 1074184. https://doi.org/10.3389/fpls.2023.1074184 (2023).

    Google Scholar 

  25. Wang, H. L. et al. A novel ATPase gene, Ab-atps, plays an important role in the interaction of rice and white tip nematode, Aphelenchoides besseyi. Sci. Rep. 11 (1), 18521. https://doi.org/10.1038/s41598-021-97981-2 (2021).

    Google Scholar 

  26. Shandeep, G. et al. and. Exploiting the nematicidal compounds from guava endo microbiome against root-knot nematodes, Meloidogyne enterolobii. Physiol. Mol. Plant Pathol. 131 102268. https://doi.org/10.1016/j.pmpp.2024.102268 (2024).

  27. Al-Rehiayani, S. M. Acetylcholinesterase in selected plant-parasitic nematodes: Inhibition, kinetic and comparative studies. Pestic. Biochem. Physiol. 90 (1), 19–25. https://doi.org/10.1016/j.pestbp.2007.07.004 (2008).

    Google Scholar 

  28. Allman, E., Johnson, D. & Keith Nehrke. Loss of the apical V-ATPase a-subunit VHA-6 prevents acidification of the intestinal lumen during a rhythmic behavior in C. elegans. Am. J. Physiology-Cell Physiol. 297 (5), C1071–C1081. https://doi.org/10.1152/ajpcell.00284.2009 (2009).

    Google Scholar 

  29. Heinick, A. et al. Eva Liebau, and Kai Lüersen. Caenorhabditis elegans P5B-type ATPase CATP-5 operates in polyamine transport and is crucial for norspermidine-mediated suppression of RNA interference. FASEB J. 24 (1), 206. https://doi.org/10.1096/fj.09-135889 (2010).

    Google Scholar 

  30. Ntalli, N. G. & Caboni, P. Botanical nematicides in the mediterranean basin. Phytochem. Rev. 11 (4), 351–359 (2012).

    Google Scholar 

Download references

Acknowledgements

The authors extend their sincere appreciation to the Indian Farmers Fertiliser Cooperative Limited (IFFCO) and IFFCO Nanoventions for their generous provision of materials and financial support that made this research possible.

Funding

This research was funded by IFFCO Nanoventions Private Limited, Coimbatore, Tamil Nadu, India (Project ID -202501).

Author information

Authors and Affiliations

  1. IFFCO Nanoventions Private Limited, Coimbatore, Tamil Nadu, India

    Lakshmanan Arunachalam, Sivashankari Lakshmanan & Shandeep Ganeshan

Authors

  1. Lakshmanan Arunachalam
  2. Sivashankari Lakshmanan
  3. Shandeep Ganeshan

Contributions

Ganeshan Shandeep: Writing—original draft, Software, Methodology, Investigation, Formal analysis, Data curation. Sivashankarai Lakshmanan: Writing—review & editing, Supervision, Lakshmanan Arunachalam: Conceptualization, Validation, Supervision, Resources, Methodology, review & editing.

Corresponding author

Correspondence to Shandeep Ganeshan.

Ethics declarations

Competing interests

The authors declare no competing interests.

Permission/ Ethical compliance statement

The field studies did not involve any endangered or protected species; no specific permits were required for the described field studies, and the landowner/farmer permitted these field studies to be carried out. The methods were carried out in accordance with the relevant guidelines and regulations, and all data in this paper would be available based on the request to authors.

Additional information

Publisher’s note

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

Supplementary Information

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Arunachalam, L., Lakshmanan, S. & Ganeshan, S. Mechanistic elucidation of a terpenoid nano-bionematicide for the management of root-knot nematodes, Meloidogyne incognita infecting tomato. Sci Rep (2026). https://doi.org/10.1038/s41598-026-41775-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1038/s41598-026-41775-x

Keywords