Exploration of Rhizosphere Bacteria for Cyclic Lipopeptide-Mediated Control against Tomato Pathogens

latha sukumaran

doi:10.56093/ijas.v96i7.175802

Authors

latha sukumaran Dr

https://doi.org/10.56093/ijas.v96i7.175802

Keywords:

Agricultural biotechnology; Biocontrol agents; Cyclic lipopeptides; Pseudomonas spp; Sustainable crop protection; Tomato (Solanum lycopersicum)

Abstract

Tomato (Solanum lycopersicum L.) is an economically important crop widely consumed in fresh and processed forms, yet its productivity is severely limited by bacterial and fungal diseases. Sustainable disease management strategies are therefore required to reduce reliance on chemical pesticides that negatively impact environmental health and beneficial soil microbiota. This study evaluated rhizosphere-derived bacteria for cyclic lipopeptide production and their biocontrol potential against major tomato phytopathogens. Five bacterial isolates (T1–T5) were obtained from diseased tomato rhizosphere soils and characterized using standard biochemical methods. Lipopeptide production was optimized and assessed through physicochemical and biological analyses. Among the isolates, T1, identified as Pseudomonas spp., exhibited the highest lipopeptide yield (18.5 mg per 50 mL) and protein content (780 µg/mL). Thin-layer chromatography revealed a prominent band with an Rf value of 0.76, while SDS–PAGE analysis confirmed the presence of low-molecular-weight cyclic lipopeptides in the range of 3-10 kDa. The extracted lipopeptides displayed strong antimicrobial activity against Xanthomonas campestris, Clavibacter michiganensis, Fusarium oxysporum, and Alternaria solani, along with significant inhibition of biofilm formation. In contrast, isolates T4 and T5 produced lower levels of metabolites and exhibited weaker antimicrobial activity. Overall, the results demonstrate that rhizosphere-derived cyclic lipopeptides, particularly those produced by Pseudomonas spp. (T1), have strong potential as eco-friendly biocontrol agents for sustainable tomato disease management.

Downloads

Download data is not yet available.

References

1. Collins, E.J., Bowyer, C., Tsouza, A. and Chopra, M. (2022) Tomatoes: an extensive review of the associated health impacts of tomatoes and factors that can affect their cultivation. 1st edn. Basel, Switzerland: MDPI.

2. Dorais, M., Ehret, D.L. and Papadopoulos, A.P. (2008) ‘Tomato (Solanum lycopersicum) health components: from the seed to the consumer’, Phytochemistry Reviews, 7(2), pp. 231–250. https://doi.org/10.1007/s11101-007-9085-x

3. Shanmugam, V., Atri, K., Gupta, S., Kanoujia, N. and Naruka, D.S. (2011) ‘Selection and differentiation of Bacillus spp. antagonistic to Fusarium oxysporum f. sp. lycopersici and Alternaria solani infecting tomato’, Folia Microbiologica, 56(2), pp. 170–177.

4. Mansfield, J., Genin, S., Magori, S. et al. (2012) ‘Top 10 plant pathogenic bacteria in molecular plant pathology’, Molecular Plant Pathology, 13(6), pp. 614–629. https://doi.org/10.1111/j.1364-3703.2012.00804.x

5. McManus, P.S., Stockwell, V.O., Sundin, G.W. and Jones, A.L. (2002) ‘Antibiotic use in plant agriculture’, Annual Review of Phytopathology, 40, pp. 443–465. https://doi.org/10.1146/annurev.phyto.40.120301.093927

6. Popowska, M., Rzeczycka, M., Miernik, A., Krawczyk-Balska, A., Walsh, F. and Duffy, B. (2012) ‘Influence of soil use on prevalence of tetracycline, streptomycin, and erythromycin resistance and associated resistance genes’, Antimicrobial Agents and Chemotherapy, 56(3), pp. 1434–1443.

7. Hancock, R.E., Haney, E.F. and Gill, E.E. (2016) ‘The immunology of host defence peptides: beyond antimicrobial activity’, Nature Reviews Immunology, 16(5), pp. 321–334. https://doi.org/10.1038/nri.2016.29

8. Jenssen, H., Hamill, P. and Hancock, R.E.W. (2006) ‘Peptide antimicrobial agents’, Clinical Microbiology Reviews, 19(3), pp. 491–511. https://doi.org/10.1128/CMR.00056-05

9. Berendsen, R.L., Pieterse, C.M.J. and Bakker, P.A.H.M. (2012) ‘The rhizosphere microbiome and plant health’, Trends in Plant Science, 17(8), pp. 478–486. https://doi.org/10.1016/j.tplants.2012.04.001

10. Huang, J., Wei, Z., Tan, S., Mei, X., Yin, S., Shen, Q. and Xu, Y. (2013) ‘The rhizosphere soil of diseased tomato plants as a source for novel microorganisms to control bacterial wilt’, Applied Soil Ecology, 72, pp. 79–84.

11. Ongena, M. and Jacques, P. (2008) ‘Bacillus lipopeptides: versatile weapons for plant disease biocontrol’, Trends in Microbiology, 16(3), pp. 115–125. https://doi.org/10.1016/j.tim.2007.12.009

12. Mendes, R., Kruijt, M., de Bruijn, I. et al. (2011) Deciphering the rhizosphere microbiome for disease-suppressive bacteria. 1st edn. Washington, DC: American Association for the Advancement of Science.

13. Bouassida, M., Mnif, I., Hammami, I., Triki, M.A. and Ghribi, D. (2023) ‘Bacillus subtilis SPB1 lipopeptide biosurfactant: antibacterial efficiency against the phytopathogenic bacteria Agrobacterium tumefaciens’, Food Science and Biotechnology, 32(11), pp. 1595–1609.

14. Dimkić, I., Stanković, S., Nišavić, M., Petković, M., Ristivojević, P., Fira, D. and Berić, T. (2017) ‘The profile and antimicrobial activity of Bacillus lipopeptide extracts of five potential biocontrol strains’, Frontiers in Microbiology, 8, p. 925.

15. Grover, M., Nain, L., Singh, S.B. and Saxena, A.K. (2009) ‘Molecular and biochemical approaches for characterization of antifungal trait of a potent biocontrol agent Bacillus subtilis RP24’, Current Microbiology, 60(2), pp. 99–106. https://doi.org/10.1007/s00284-009-9508-6

16. Laemmli, U.K. (1970) ‘Cleavage of structural proteins during the assembly of the head of bacteriophage T4’, Nature, 227(5259), pp. 680–685.

17. Zhao, X., Zhao, F., Wang, J. et al. (2023) ‘Antimicrobial peptides inhibit biofilm formation of multidrug-resistant Pseudomonas aeruginosa by targeting extracellular DNA’, Frontiers in Microbiology, 14, p. 1122768. https://doi.org/10.3389/fmicb.2023.1122768

18. O’Toole, G.A. (2011) ‘Microtiter dish biofilm formation assay’, Journal of Visualized Experiments, (47), p. 2437.

19. Compant, S., Clément, C. and Sessitsch, A. (2010) ‘Plant growth-promoting bacteria in the rhizo- and endosphere of plants’, Soil Biology and Biochemistry, 42(5), pp. 669–678. https://doi.org/10.1016/j.soilbio.2009.11.024

20. Raaijmakers, J.M., Vlami, M. and de Souza, J.T. (2002) ‘Antibiotic production by bacterial biocontrol agents of plant diseases’, Phytopathology, 92(11), pp. 1272–1282. https://doi.org/10.1094/PHYTO.2002.92.11.1272

21. Kloepper, J.W., Ryu, C.-M. and Zhang, S. (2004) ‘Induced systemic resistance and promotion of plant growth by Bacillus spp.’, Phytopathology, 94(11), pp. 1259–1266. https://doi.org/10.1094/PHYTO.2004.94.11.1259

22. Glick, B.R. (2012) ‘Plant growth-promoting bacteria: mechanisms and applications’, Scientifica, 2012, p. 963401. https://doi.org/10.6064/2012/963401

23. Rosenblueth, M. and Martínez-Romero, E. (2006) ‘Bacterial endophytes and their interactions with hosts’, Molecular Plant–Microbe Interactions, 19(8), pp. 827–837. https://doi.org/10.1094/MPMI-19-0827

24. Hamed, H.A., Moustafa, Y.A. and Abdel-Aziz, S.M. (2011) ‘In vivo efficacy of lactic acid bacteria in biological control against Fusarium oxysporum for protection of tomato plant’, Life Science Journal, 8(4), pp. 462–468.

25. Goswami, M., Deka, S. and Bora, S.S. (2021) ‘Nutrient supplementation strategies to improve secondary metabolite production in microbial fermentation: a review’, World Journal of Microbiology and Biotechnology, 37(1), p. 20. https://doi.org/10.1007/s11274-020-02947-4

26. Shaligram, S. and Singhal, R.S. (2010) ‘Surfactin: a review on biosynthesis, fermentation, purification and applications’, Food Technology and Biotechnology, 48(2), pp. 119–134.

27. Zhou, S., Zhuang, X., Wang, Y., Li, J. and Yang, X. (2019) ‘Metabolic pathways and regulation of microbial biosynthesis of lipopeptides: progress and prospects’, Applied Microbiology and Biotechnology, 103(6), pp. 2491–2502. https://doi.org/10.1007/s00253-019-09627-z

28. Geudens, N. and Martins, J.C. (2018) ‘Cyclic lipodepsipeptides from Pseudomonas spp.: diversity, activity and biogenesis’, Environmental Microbiology, 20(3), pp. 735–746.

29. Meena, K.R. and Kanwar, S.S. (2015) ‘Lipopeptides as antifungal and antibacterial agents: applications in food safety and therapeutics’, BioMed Research International, 2015, p. 473050. https://doi.org/10.1155/2015/473050

30. Meena, K.R., Kanwar, S.S., Sharma, S. and Yadav, S.K. (2018) ‘Enhanced production and characterization of antifungal lipopeptides by Bacillus subtilis strain K1’, Journal of Basic Microbiology, 58(6), pp. 538–549. https://doi.org/10.1002/jobm.201700451

31. Gong, A., Li, H., Yuan, Q. et al. (2015) ‘Antagonistic mechanism of iturin A and plipastatin A from Bacillus amyloliquefaciens S76-3 from wheat spikes against Fusarium graminearum’, PLOS ONE, 10(2), e0116871. https://doi.org/10.1371/journal.pone.0116871

32. Hazarika, D.J., Goswami, G., Gautom, T. et al. (2019) ‘Lipopeptide mediated biocontrol activity of endophytic Bacillus subtilis against fungal phytopathogens’, BMC Microbiology, 19, pp. 1–13.

33. Yuan, Q.S., Yang, P., Liu, Y.K. et al. (2025) ‘Iturin and fengycin lipopeptides inhibit pathogenic Fusarium by targeting multiple components of the cell membrane and their regulative effects in wheat’, Journal of Integrative Plant Biology, advance online publication. https://doi.org/10.1111/jipb.13933

34. Grover, M., Ali, S.Z., Sandhya, V., Rasul, A. and Venkateswarlu, B. (2010) ‘Role of microorganisms in adaptation of agriculture crops to abiotic stresses’, World Journal of Microbiology and Biotechnology, 27(5), pp. 1231–1240.

35. Raaijmakers, J.M., de Bruijn, I. and de Kock, M.J.D. (2006) ‘Cyclic lipopeptide production by plant-associated Pseudomonas spp.: diversity, activity, biosynthesis and regulation’, Molecular Plant–Microbe Interactions, 19(7), pp. 699–710. https://doi.org/10.1094/MPMI-19-0699

36. Gross, H. and Loper, J.E. (2009) ‘Genomics of secondary metabolite production by Pseudomonas spp.’, Natural Product Reports, 26(11), pp. 1408–1446.

37. Mnif, I. and Ghribi, D. (2015) ‘Lipopeptide surfactants: production, recovery and pore forming capacity’, Peptides, 71, pp. 100–112. https://doi.org/10.1016/j.peptides.2015.05.003

38. Bais, H.P., Fall, R. and Vivanco, J.M. (2004) ‘Biocontrol of Bacillus subtilis against infection of Arabidopsis roots by Pseudomonas syringae is facilitated by biofilm formation and surfactin production’, Plant Physiology, 134(1), pp. 307–319.