Fluorescence Detection of Pyrene-Stained Bacillus amyloliquefaciens MHR24 in Tomato (Solanum lycopersicum L.) Stem Tissues

Hernández-Rodríguez, Mónica; Turlakov, Gleb; Lozano, Celín; Arias, Eduardo; Flores-Olivas, Alberto; Moggio, Ivana; Valenzuela-Soto, José Humberto

doi:10.3390/ijms27094013

Open AccessArticle

Fluorescence Detection of Pyrene-Stained Bacillus amyloliquefaciens MHR24 in Tomato (Solanum lycopersicum L.) Stem Tissues

by

Mónica Hernández-Rodríguez

¹,

Gleb Turlakov

¹,

Celín Lozano

¹,

Eduardo Arias

¹

,

Alberto Flores-Olivas

²,

Ivana Moggio

^1,*

and

José Humberto Valenzuela-Soto

^3,*

¹

Centro de Investigación en Química Aplicada, Departamento de Materiales Avanzados, Boulevard Enrique Reyna Hermosillo 140, Saltillo 25294, Mexico

²

Departamento de Parasitología, Calz. Antonio Narro, Buenavista, Universidad Autónoma Agraria Antonio Narro, Saltillo 25315, Mexico

³

SECIHTI-Centro de Investigación en Química Aplicada, Departamento de Biociencias y Agrotecnología, Boulevard Enrique Reyna Hermosillo 140, Saltillo 25294, Mexico

^*

Authors to whom correspondence should be addressed.

Int. J. Mol. Sci. 2026, 27(9), 4013; https://doi.org/10.3390/ijms27094013

Submission received: 28 March 2026 / Revised: 24 April 2026 / Accepted: 27 April 2026 / Published: 30 April 2026

(This article belongs to the Special Issue New Advances in Plant–Microbe Interaction)

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Abstract

The PGPR strain of Bacillus amyloliquefaciens MHR24 (MHR24) was recently reported as a strong biocontrol strain. In this study, MHR24 was used to investigate phyllosphere effects during inoculations of tomato leaves (Solanum lycopersicum L.). When MHR24 was inoculated on foliar tissue, it caused apical chlorosis symptoms at 3–6 days after infiltration or submersion, which suggests that the bacterium may adopt a potentially pathogenic lifestyle in the phyllosphere. In order to detect the MHR24 interaction with the plant, it was stained with the commercial fluorophore 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt, selected from a pyrene series bearing diverse functional groups, based on several in vitro staining assays. Fluorescence used as a detection signal was observed by LSCM mainly in the vascular bundles, suggesting that rhizobacteria may preferentially colonize these tissue regions. Molecular docking, performed by analyzing the possible interactions between the outer membrane protein assembly factor BamB of the family protein B. amyloliquefaciens and the fluorophore, indicates that hydrogen bonds with serine 126 (SER126), serine 182 (SER182), isoleucine 180 (ILE180), and tryptophan 66 (TRP66), charges attraction and π-stacking with TRP66, and non-bonded attractions with leucine 224 (LEU224) can occur, which likely gives rise to a stable complex. These results are important in view of the application of MHR24 as part of a sustainable approach for increasing tomato crop production.

Keywords: pyrene markers; rhizobacteria; membrane proteins; pH

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MDPI and ACS Style

Hernández-Rodríguez, M.; Turlakov, G.; Lozano, C.; Arias, E.; Flores-Olivas, A.; Moggio, I.; Valenzuela-Soto, J.H. Fluorescence Detection of Pyrene-Stained Bacillus amyloliquefaciens MHR24 in Tomato (Solanum lycopersicum L.) Stem Tissues. Int. J. Mol. Sci. 2026, 27, 4013. https://doi.org/10.3390/ijms27094013

AMA Style

Hernández-Rodríguez M, Turlakov G, Lozano C, Arias E, Flores-Olivas A, Moggio I, Valenzuela-Soto JH. Fluorescence Detection of Pyrene-Stained Bacillus amyloliquefaciens MHR24 in Tomato (Solanum lycopersicum L.) Stem Tissues. International Journal of Molecular Sciences. 2026; 27(9):4013. https://doi.org/10.3390/ijms27094013

Chicago/Turabian Style

Hernández-Rodríguez, Mónica, Gleb Turlakov, Celín Lozano, Eduardo Arias, Alberto Flores-Olivas, Ivana Moggio, and José Humberto Valenzuela-Soto. 2026. "Fluorescence Detection of Pyrene-Stained Bacillus amyloliquefaciens MHR24 in Tomato (Solanum lycopersicum L.) Stem Tissues" International Journal of Molecular Sciences 27, no. 9: 4013. https://doi.org/10.3390/ijms27094013

APA Style

Hernández-Rodríguez, M., Turlakov, G., Lozano, C., Arias, E., Flores-Olivas, A., Moggio, I., & Valenzuela-Soto, J. H. (2026). Fluorescence Detection of Pyrene-Stained Bacillus amyloliquefaciens MHR24 in Tomato (Solanum lycopersicum L.) Stem Tissues. International Journal of Molecular Sciences, 27(9), 4013. https://doi.org/10.3390/ijms27094013

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Fluorescence Detection of Pyrene-Stained Bacillus amyloliquefaciens MHR24 in Tomato (Solanum lycopersicum L.) Stem Tissues

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