Influence of silver nanoparticles on a common contaminant isolated during the establishment of Stevia rebaudiana Bertoni
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ORIGINAL ARTICLE
Influence of silver nanoparticles on a common contaminant isolated during the establishment of Stevia rebaudiana Bertoni culture Marco A. Ramírez‑Mosqueda1 · Lino Sánchez‑Segura2 · Sandra L. Hernández‑Valladolid3 · ·Elohim Bello‑Bello4 · Jericó J. Bello‑Bello5 Received: 2 July 2020 / Accepted: 5 October 2020 © Springer Nature B.V. 2020
Abstract Contamination by fungi and bacteria during the in vitro propagation of plants leads to considerable losses of biological material and precludes phytosanitary certification. The anti-microbial effect of silver nanoparticles (AgNPs) may be an alternative for the eradication of in vitro contaminants. This study evaluated the microbicidal activity of AgNPs on a recurrent fungus during the micropropagation of stevia (Stevia rebaudiana Bertoni). First, the fungus was isolated and identified at a molecular level by the sequencing and analysis of the ITS4/ITS5 rDNA region. The results of the phylogenetic analysis of various fungal species showed that the strain under study (16-166-H) belongs to the genus Sordaria and is 99.64% similar to S. tomento-alba (strain CBS 260.78). Subsequently, the inhibition of the growth of S. tomento-alba was tested under different concentrations of AgNPs (0, 25, 50, 100, and 200 mg L−1), observing that 50 and 100 mg L−1 achieve ca. 50% growth inhibition (IC50), while 200 mg L−1 produces a drastic inhibition. On the other hand, the shape and size of AgNPs was examined using transmission electron microscopy (TEM), and the transport and accumulation of AgNPs in S. tomento-alba cells were monitored through multiphoton microscopy. The morphological and fluorescence analyses showed that AgNPs display different sizes, with larger nanoparticles retained in fungal cell walls while smaller AgNPs penetrate into fungal cells. Probably, apoplastic and symplastic mechanisms involved in the accumulation and transport of AgNPs affect the metabolic processes of the fungus, thus inhibiting its growth. These results suggest that AgNPs possess antifungal activity and can be used in the eradication of contaminants during the in vitro culture of plant species. Key message The microbicidal potential of AgNPs in the micropropagation of different plant species. Keywords Nanobiotechnology · Silver nanoparticles · Transmission electron microscopy · Antifungal activity
Communicated by Sergio J. Ochatt. * Jericó J. Bello‑Bello [email protected] 1
Colegio de Postgraduados Campus Córdoba, Km. 348 de la Carretera Federal Córdoba‑Veracruz, Congregación Manuel León, Amatlán de Los Reyes, Veracruz, Mexico
2
Departamento de Ingeniería Genética, Unidad Irapuato, CINVESTAV-Irapuato, Libramiento Norte Carr. Irapuato‑León Km 9.6. Irapuato, Guanajuato, Mexico
3
Agricultura Sustentable y Protegida, Universidad Tecnológica del Centro de Veracruz, Av. Universidad 350, Dos caminos, 94910 Cuitláhuac, Veracruz, Mexico
4
Laboratorio Nacional de Genómica Para La Biodiversidad/Unidad de Genómica Avanzada, Centro de Investigación Y Estudios Avanzados, I
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