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Quantifying and visualizing Nitrospirillum amazonense strain CBAmC in sugarcane after using different inoculation methods

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Abstract

Background and Aims

Nitrospirillum amazonense strain CBAmC is a nitrogen-fixing bacterium that has been used for sugarcane inoculation. Despite its beneficial effects, however, little is known about the establishment and colonization of this microorganism in its inoculated host plant. Furthermore, alternative inoculation methods need to be tested in the sugarcane crop. The objective of the present work was to quantify and visualize CBAmC in sugarcane cultivar RB867515 after using three inoculation methods.

Methods

Utilizing one node as propagation material, three bacterial inoculation methods were tested under greenhouse conditions. For up to 60 days after inoculation (DAI), qPCR was used to quantify the bacterial loads in the plant or substrate while FISH-CLSM was used to confirm the establishment and colonization profile of CBAmC in different plant tissues.

Results

FISH-CLSM indicated that the CBAmC strain could establish itself on different tissues of sugarcane, especially roots. qPCR revealed variable bacterial loads in the plants inoculated according to the three methods tested. In addition, little or no bacterial loads were detected in the substrate a few days after inoculation, regardless of the method used.

Conclusions

N. amazonense CBAmC proves to be a bacterium that preferentially associates with the surface of the root tissues of sugarcane propagated by one node, instead of colonizing internal plant tissues or remaining in the soil. The inoculation method consisting of immersion of the mini-setts in bacterial suspension proved to be more efficient than the inoculation methods of applying the suspension onto the post-sprouting substrate or to the lowest leaf.

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Data availability

Key data supporting the findings of this study are available in the paper and in supplemental material published online. Cited data not shown in the paper or in the Online Resource material, unnecessary for understanding and reproducing the experiments, are available from the corresponding author, SS, upon request.

Notes

  1. Corn steep solids (CSS): 100 g in sufficient amount of H2O for 1000 mL.

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Funding

This work was supported by the authors’ institutions: Basf S.A. and Embrapa Agrobiologia.

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Author notes

  1. J. P. Ferreira

    Present address: Solubio, Rodovia GO-184, km 09, Jataí, Goiás State, CEP 75809-899, Brazil

Authors and Affiliations

  1. Embrapa Agrobiologia, Rodovia BR-465, km 7, Seropédica, Rio de Janeiro State, CEP 23891-000, Brazil

    S. Schwab, J. C. A. Amaral, C. G. N. da Silva, J. P. Ferreira, L. V. da Silva, J. R. C. Rouws, L. F. M. Rouws, J. I. Baldani & V. M. Reis

  2. Agricultural Experimental Station – BASF S.A, Rodovia SP-340, km 144, Santo Antônio de Posse, São Paulo State, CEP 13830- 000, Brazil

    E. S. Hirata

  3. Instituto de Ciências Biológicas e da Saúde, Universidade Federal Rural do Rio de Janeiro, Rodovia BR-465, km 7, Seropédica, Rio de Janeiro State, CEP 23897-000, Brazil

    J. C. A. Amaral

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Contributions

SS: conceptualization, formal analysis, investigation, project administration, supervision, writing – original draft, review & editing; ESH: conceptualization, project administration, resources, supervision; JRCR: formal analysis, writing – original draft; JCAA, CGNS, JPF, LVS: investigation; LFMR, JIB, VMR: conceptualization, writing – review & editing.

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Correspondence to S. Schwab.

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Schwab, S., Hirata, E.S., Amaral, J.C.A. et al. Quantifying and visualizing Nitrospirillum amazonense strain CBAmC in sugarcane after using different inoculation methods. Plant Soil 488, 197–216 (2023). https://doi.org/10.1007/s11104-023-05940-9

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