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Application of Two-Photon Microscopy to Study Sclerotium cepivorum Berk Sclerotia Isolated from Naturally Infested Soil and Produced In Vitro

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Abstract

The danger of Sclerotium cepivorum lies in the strength of its survival structure: sclerotia. Sclerotia comprising hardened mycelium contains food reserves that allow it to remain dormant for long period, which makes the sclerotia-infested soil useless to grow any crop of the Allium species, including onion and garlic. This paper would be the first report on the application of two-photon fluorescence microscopy to the analysis of the structure of sclerotia from S. cepivorum. For this study and, in order to test the method, two different types of sclerotia were used: (1) sclerotia isolated from naturally infested soil and (2) sclerotia produced in vitro (from 20-day-old cultures). Both types of sclerotia were processed by cryopreservation and eight µm histological cuts were used to obtain an autofluorescence image. For both sclerotia, the fluorescence spectrum has three peak signals at their wall. Sclerotia from infested soil presented fluorescence peaks at 400–436, 436–475, and 515–575 nm, while signals from sclerotia produced in vitro presented fluorescence peaks at 400–442, 500–600, and 655–700 nm. Peaks at the violet electromagnetic region (400–436 and 400–442) are like that of the signals reported by the melanin. This study showed that two-photon microscopy is a novel and valuable tool for the study of sclerotia structure and their fluorescence signal, and the possibility of using it as a specific marker to direct detection in the field should be explored.

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Acknowledgements

This work forms part of the Integral Program for Strengthening Academic Groups of the PRODEP program (PIFOCA 2017). Many thanks to Dr. Lino Sánchez-Segura for microscopic analysis and thanks to the SOLENA Company for collecting and providing soil samples. We are grateful to MSc María Karina Manzo-Valencia for their invaluable help in technical assistance. Ayabeth Ordaz-Arias received a scholarship granted by SICES through the agreement SICES/CONV/140/2018 ITESI during XI State Summer Research. Laura Valdés-Santiago and Rafael Vargas-Bernal (He reviewed the English translation of the document and realized the photonic analysis) are members of the National System of Researchers (SNI), Mexico.

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Author notes

  1. Laura Valdés-Santiago and Rafael Vargas-Bernal have contributed equally to this work.

Authors and Affiliations

  1. Ingeniería Bioquímica, Instituto Tecnológico Superior de Irapuato, Carretera Irapuato-Silao Km 12.5, 36821, Irapuato, Guanajuato, Mexico

    Laura Valdés-Santiago & Ayabeth Ordaz-Arias

  2. Ingeniería en Materiales, Instituto Tecnológico Superior de Irapuato, Carretera Irapuato-Silao Km 12.5, 36821, Irapuato, Guanajuato, Mexico

    Rafael Vargas-Bernal & Gabriel Herrera-Pérez

  3. Biología, Instituto Tecnológico Superior de Irapuato, Carretera Irapuato-Silao Km 12.5, 36821, Irapuato, Guanajuato, Mexico

    Juan G. Colli-Mull

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  1. Laura Valdés-Santiago
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Correspondence to Laura Valdés-Santiago.

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Valdés-Santiago, L., Vargas-Bernal, R., Herrera-Pérez, G. et al. Application of Two-Photon Microscopy to Study Sclerotium cepivorum Berk Sclerotia Isolated from Naturally Infested Soil and Produced In Vitro. Curr Microbiol 78, 749–755 (2021). https://doi.org/10.1007/s00284-020-02341-4

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