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A mixture of Azotobacter, Azospirillum, and Klebsiella strains improves root-rot disease complex management and promotes growth in sunflowers in calcareous soil

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Abstract

Biological farming using a mixture of bio-agents that are compatible with each other and adapted to the plant rhizosphere is a strategic approach to manage crop disease. This study is a unique approach to manage the root-rot disease complex caused by Macrophomina phaseolina, Rhizoctonia solani, and Fusarium solani in sunflower plants grown in calcareous soil using a mixture of rhizobacteria strains, namely Azotobacter chroococcum ZCR, Azospirillum brasilense SBR, and Klebsiella pneumoniae KPR. These strains were screened in vitro for N2 fixation and phosphate solubilization, as well as the production of indoleacetic acid, siderophore, and hydrogen cyanide. Interestingly, the rhizobacteria strains exhibited promising biofertilizer and biocontrol properties. Furthermore, they significantly inhibited the growth of root-rot pathogens in a dual culture assay. Notably, the rhizospheric bacteria exhibited successful colonization of the rhizoplane of sunflower plants and persisted at high levels for up to 60 days (8 × 104–12 × 104 cfu g−1 fresh root). In field experiments under naturally infested soil conditions, application of a bio-formulation containing a plant growth-promoting rhizobacteria mixture (HALEX bio-formulation) as a seed treatment as well as by soil drenching was found to be effective in controlling sunflower root-rot disease and significantly decreased the disease severity rating by 44–55% of the non-treated control in 2 years. In addition, treating with rhizobacteria significantly improved plant growth, yield, and oil content and decreased weight loss due to pathogen-induced stress. Consequently, it is suggested that farmers use this rhizobacteria mixture as a promising eco-friendly approach for effective management of root-rot disease complex in sunflowers in semi-arid regions.

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References

  1. Abdallah, D. B., Frikha-Gargouri, O., & Tounsi, S. (2018). Rizhospheric competence, plant growth promotion and biocontrol efficacy of Bacillus amyloliquefaciens subsp. plantarum strain 32a. Biological Control, 124, 61–67.

  2. Adriansen, H. K. (2009). Land reclamation in Egypt: a study of life in the new lands. Geoforum, 40(4), 664–674.

  3. Ahemad, M., & Khan, M. S. (2011). Effects of insecticides on plant-growth-promoting activities of phosphate solubilizing rhizobacterium Klebsiella sp. strain PS19. Pesticide Biochemistry and Physiology, 100(1), 51–56.

  4. Ahemad, M., & Kibret, M. (2014). Mechanisms and applications of plant growth promoting rhizobacteria: current perspective. Journal of King Saud University –Science, 26(1), 1–20.

  5. Ahmad, F., Ahmad, I., & Khan, M. S. (2005). Indole acetic acid production by the indigenous isolates of Azotobacter and fluorescent Pseudomonas in the presence and absence of tryptophan. Turkish Journal of Biology, 29, 29–34.

  6. Ali Siddiqui, I., Ehetshamul-Haque, S., & Shahid Shaukat, S. (2001). Use of rhizobacteria in the control of root rot-root knot disease complex of mungbean. Journal of Phytopathology, 149(6), 337–346.

  7. Ata, A., Salem, T. N., & Hassan, R. (2018). Geotechnical characterization of the calcareous sand in northern coast of Egypt. Ain Shams Engineering Journal, 9(4), 3381–3390.

  8. Bakker, A. W., & Schippers, B. (1987). Microbial cyanide production in the rhizosphere in relation to potato yield reduction and Pseudomonas spp.-mediated plant growth-stimulation. Soil Biology and Biochemistry, 19(4), 451–457.

  9. Baldani, V. L. D., & Döbereiner, J. (1980). Host-plant specificity in the infection of cereals with Azospirillum spp. Soil Biology and Biochemistry, 12(4), 433–439.

  10. Bouizgarne, B. (2013). Bacteria for plant growth promotion and disease management. Bacteria in agrobiology: Disease management (pp. 15–47). Berlin, Heidelberg: Springer.

  11. Chauhan, S., Wadhwa, K., Vasudeva, M., & Narula, N. (2012). Potential of Azotobacter spp. as biocontrol agents against Rhizoctonia solani and Fusarium oxysporum in cotton (Gossypium hirsutum), guar (Cyamopsis tetragonoloba) and tomato (Lycopersicum esculentum). Archives of Agronomy and Soil Science, 58(12), 1365–1385.

  12. Chen, Y., & Barak, P. (1982). Iron nutrition of plants in calcareous soils. Advances in Agronomy, 35, 217–240.

  13. de Weert, S., & Bloemberg, G. V. (2006). Rhizosphere competence and the role of root colonization in biocontrol. In S. S. Gnanamanickam (Ed.), Plant-associated bacteria (pp. 317–333). Berlin: Springer.

  14. Enebe, M. C., & Babalola, O. O. (2018). The influence of plant growth-promoting rhizobacteria in plant tolerance to abiotic stress: a survival strategy. Applied Microbiology and Biotechnology, 102(18), 7821–7835.

  15. Figueroa-López, A. M., Cordero-Ramírez, J. D., Martínez-Álvarez, J. C., López-Meyer, M., Lizárraga-Sánchez, G. J., Félix-Gastélum, R., & Maldonado-Mendoza, I. E. (2016). Rhizospheric bacteria of maize with potential for biocontrol of Fusarium verticillioides. SpringerPlus, 5, 330.

  16. Galloway, J. N., Townsend, A. R., Erisman, J. W., Bekunda, M., Cai, Z. C., Freney, J. R., Martinelli, L. A., Seitzinger, S. P., & Sutton, M. A. (2008). Transformation of the nitrogen cycle: recent trends, questions, and potential solutions. Science, 320(5878), 889–892.

  17. Gomez, K. A., & Gomez, A. A. (1984). Statistical procedures for agricultural research. Wiley.

  18. Harveson, R. M., Markell, S. G., Block, C. C., & Gulya, T. J. (2016). Compendium of sunflower diseases and pests. APS press, St Paul, MN, USA. https://doi.org/10.1094/9780890545096.

  19. Hassouna, M. G. (1973). Biological curing of nitrogen deficiency in Gramineae. I. Inoculation of barley grown in pots with non-symbiotic N2-fixers. Proceedings of the First Congress of the Egyptian Phytopathological Society, Nov. 12–15, 1973, Cairo, Egypt. Academy of Scientific Research and Technology.

  20. Hassouna, M. G., El-Saedy, M. A. M., & Saleh, H. M. (1998). Biocontrol of soil-borne plant pathogens attacking cucumber (Cucumis sativus) by Rhizobacteria in a semiarid environment. Arid Land Research and Management, 12(4), 345–357.

  21. Hoben, H. J., & Somasegaran, P. (1982). Comparison of the pour, spread, and drop plate methods for enumeration of Rhizobium spp. in inoculants made from presterilized peat. Applied and Environmental Microbiology, 44, 1246–1247.

  22. Hollomon, D. W. (2015). Fungicide resistance: facing the challenge-a review. Plant Protection Science, 51(4), 170–176.

  23. Husen, E. (2016). Screening of soil bacteria for plant growth promotion activities in vitro. Indonesian Journal of Agricultural Science, 4(1), 27–31.

  24. Hymowitz, T., Collins, F. I., Panczner, J., & Walker, W. M. (1972). Relationship between the content of oil, protein, and sugar in soybean seed 1. Agronomy Journal, 64(5), 613–616.

  25. Kalantari, S., Marefat, A., Naseri, B., & Hemmati, R. (2018). Improvement of bean yield and Fusarium root rot biocontrol using mixtures of Bacillus, Pseudomonas and Rhizobium. Tropical Plant Pathology, 43(6), 499–505.

  26. Khan, M. R., Haque, Z., Rasool, F., Salati, K., Khan, U., Mohiddin, F. A., & Zuhaib, M. (2019). Management of root-rot disease complex of mungbean caused by Macrophomina phaseolina and Rhizoctonia solani through soil application of Trichoderma spp. Crop Protection, 119, 24–29.

  27. Muleta, D., Assefa, F., & Granhall, U. (2007). In vitro antagonism of rhizobacteria isolated from Coffea arabica L. against emerging fungal coffee pathogens. Engineering in Life Sciences, 7(6), 577–586.

  28. Nandakumar, R., Babu, S., Viswanathan, R., Sheela, J., Raguchander, T., & Samiyappan, R. (2001). A new bio-formulation containing plant growth promoting rhizobacterial mixture for the management of sheath blight and enhanced grain yield in rice. Biocontrol, 46(4), 493–510.

  29. Ouda, S. (2015). Major crops and water scarcity in Egypt: Irrigation water management under changing climate. Springer.

  30. Parewa, H. P., Meena, V. S., Jain, L. K., & Choudhary, A. (2018). Sustainable crop production and soil health management through plant growth-promoting Rhizobacteria. In Role of rhizospheric microbes in soil (pp. 299–329). Singapore: Springer.

  31. Pikovskaya, R. I. (1948). Mobilization of phosphorus in soil in connection with vital activity of some microbial species. Microbiology, 17, 362–370.

  32. Prasad, M., Srinivasan, R., Chaudhary, M., Choudhary, M., & Jat, L. K. (2019). Chapter seven - plant growth promoting Rhizobacteria (PGPR) for sustainable agriculture: Perspectives and challenges. In: Singh, A.K., Kumar, A., Singh, P. K. (Eds.), PGPR Amelioration in Sustainable Agriculture. Woodhead Publishing, pp. 129–157.

  33. Richardson, A. E., & Simpson, R. J. (2011). Soil microorganisms mediating phosphorus availability update on microbial phosphorus. Plant physiology, 156(3), 989–996.

  34. Ruano-Rosa, D., Cazorla, F. M., Bonilla, N., Martín-Pérez, R., De Vicente, A., & López-Herrera, C. J. (2014). Biological control of avocado white root rot with combined applications of Trichoderma spp. and rhizobacteria. European Journal of Plant Pathology, 138(4), 751–762.

  35. Saleh, A. A., Sharafaddin, A. H., El_Komy, M. H., Ibrahim, Y. E., Hamad, Y. K., & Molan, Y. Y. (2017). Fusarium species associated with date palm in Saudi Arabia. European Journal of Plant Pathology, 148(2), 367–377.

  36. SAS Institute Inc. (2003). SAS/STATA guide for personal computers version 9.1 edition. Carry: SAS Institute.

  37. Shen, H., He, X., Liu, Y., Chen, Y., Tang, J., & Guo, T. (2016). A complex inoculant of N2-fixing, P-and K-solubilizing bacteria from a purple soil improves the growth of kiwifruit (Actinidia chinensis) plantlets. Frontiers in Microbiology, 7, 841.

  38. Shirinbayan, S., Khosravi, H., & Malakouti, M. J. (2019). Alleviation of drought stress in maize (Zea mays) by inoculation with Azotobacter strains isolated from semi-arid regions. Applied Soil Ecology, 133, 138–145.

  39. Singh, R., Tiwari, S., Patel, R. P., Soni, S. K., & Kalra, A. (2018). Bioinoculants and AM fungus colonized nursery improved management of complex root disease of Coleus forskohlii Briq. under field conditions. Biological Control, 122, 11–17.

  40. Sundaramoorthy, S., Raguchander, T., Ragupathi, N., & Samiyappan, R. (2012). Combinatorial effect of endophytic and plant growth promoting rhizobacteria against wilt disease of Capsicum annum L. caused by Fusarium solani. Biological Control, 60(1), 59–67.

  41. Tarkka, M., Schrey, S., & Hampp, R. (2008). Plant associated soil micro-organisms. In Molecular mechanisms of plant and microbe coexistence (pp. 3–51). Berlin, Heidelberg: Springer.

  42. Tortora, M. L., Díaz-Ricci, J. C., & Pedraza, R. O. (2011). Azospirillum brasilense siderophores with antifungal activity against Colletotrichum acutatum. Archives of microbiology, 193(4), 275–286.

  43. Vatchev, T., & Maneva, S. (2012). Chemical control of root rot complex and stem rot of greenhouse cucumber in straw-bale culture. Crop Protection, 42, 16–23.

  44. Yu, X., Ai, C., Xin, L., & Zhou, G. (2011). The siderophore-producing bacterium, Bacillus subtilis CAS15, has a biocontrol effect on Fusarium wilt and promotes the growth of pepper. European Journal of Soil Biology, 47(2), 138–145.

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Acknowledgments

The authors extend their appreciation to the Deanship of Scientific Research at King Saud University for funding this work through research group No. RG-1440-028.

Author information

MGH and EMA designed research experiments. MHK conducted the experiments, collected the data, and wrote the manuscript. YA and ASA analyzed the experimental data and helped to write the manuscript. All authors critically reviewed the manuscript.

Correspondence to Mahmoud H. El_Komy.

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Authors declared that this manuscript has not published elsewhere. All authors read and approved the final version of this manuscript. The authors declare that the present work was developed without any potential conflict of interest, with no human or animal participants.

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The authors declare that they have no conflict of interest.

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El_Komy, M.H., Hassouna, M.G., Abou-Taleb, E.M. et al. A mixture of Azotobacter, Azospirillum, and Klebsiella strains improves root-rot disease complex management and promotes growth in sunflowers in calcareous soil. Eur J Plant Pathol (2020). https://doi.org/10.1007/s10658-019-01921-w

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Keywords

  • Rhizobacteria inoculate
  • Root-rot index
  • Sunflower
  • Plant growth-promotion
  • Biocontrol
  • Calcareous soil stress